Systematic Studies in Abildgaardieae (Cyperaceae) (Thesis part 2)

Kerri Clarke

The tribe Abildgaardieae Lye is composed of 5-7 genera: Crosslandia, Fimbristylis, Abildgaardia (= Fimbristylis section Abildgaardia), Bulbostylis, Nemum, Nelmesia and Tylocarya (= Fimbristylis nelmesii). There has been little disagreement about the general boundaries of the tribe. However, limits of the main genera are disputed and unresolved. Some species and generic boundaries of Crosslandia, Abildgaardia and Australian Bulbostylis require assessment across their morphological and geographical range of distribution. The general aim of this thesis is to test monophyly of and within the tribe Abildgaardieae. To address the aim, the limits for Crosslandia, Abildgaardia and Australian Bulbostylis are assessed to determine the species and generic limits on a global level. Data from morphology, vegetative anatomy and embryo morphology were used in phenetic and cladistic analyses. Phenetic analyses of morphological data were used to test and set species limits. Additional characters from morphology, vegetative anatomy and embryo morphology were used in cladistic analyses to test monophyly of the tribe and previous classifications. Representative samples from Fimbristylis, Nemum, and the monotypic genera Nelmesia and Tylocarya (= Fimbristylis nelmesii) were added to those species defined in phenetic analyses. Data were polarised using the outgroup method; with outgroup taxa selected from the provisional Arthrostylideae, Schoenoplectus and Schoenoplectiella. Cladistic analysis revealed that the tribe Abildgaardieae is not monophyletic when Nemum, Nelmesia and Tylocarya are included. Members of ‘Arthrostylideae’ violated monophyly of Abildgaardieae. The limits of Crosslandia and Abildgaardia are revised: Crosslandia 4 spp., all endemic to Australia and Abildgaardia 11 spp., 9 in Australia, 8 endemic. The limits for Australian species of Bulbostylis remain unresolved, with further testing of a wider sample of overseas species needed. A new species of Bulbostylis, B. kakadu in ed., is recognised. Species of Nemum are not monophyletic and Nelmesia rendered Abildgaardia non-monophyletic in the full tribal analysis. Tylocarya formed a clade with Fimbristylis depauperata. Species of Fimbristylis did not form a monophyletic group. Suggestions for further work are given.

136 Chapter 5 Phenetic and cladistic analyses of Australian Bulbostylis Kuuth Introduction This chapter focuses on the limits of Australian taxa within the genus Bulbostylis Kunth nom. cons. Bulbostylis is a large and mostly pantropical genus comprising around c. 200 species, with the centre of diversity in the African continent (Haines and Lye 1983; Goetghebeur and Coudijzer 1985); the generic rank is now widely accepted (see Chapter 1 for historical perspective). Currently, there are six species of Bulbostylis recognised that occur in Australia; B. burbidgeae K.L. Wilson, B. turbinata S.T.Blake, and B. pyriformis S.T.Blake are endemic, while B. barbata (Rottb.) C.B.Clarke and B. densa (Wall.) Hand.-Mazz. have a widespread distribution. Bulbostylis humilis (Kunth) C.B.Clarke (sYn B. striatella C.B.Clarke; see World Checklist of Mononcotyledons) is a relatively recent introduction to Australia, and known in the Arrnidale - Glenn Innes area of New South Wales. Intermediate morphology due to clinal variation or hybridisation is COlnn10n for some species within the genus e.g. B. schoeinoides complex (Gordon-Gray 1988), B. hispidula complex (Haines and Lye 1983; Lye 1995), B. densa complex (Haines 137 and Lye 1983; Gordon-Gray 1995; Lye 1996), creating uncertainties in species lin1its and confusion for the identification of specimens. Bulbostylis densa (as Bulbostylis capillaris var. trifida (Kunth) C.B.Clarke) was separated from B. capillaris (L.) Kunth ex C.B.Clarke due to morphological differences between the American and Asian/Pacific specimens. Blake (1941), however, commented that the Australian material that Clarke assigned with the Indian B. capillaris var. trifida, was indistinguishable from the American B. capillaris var. capillaris. Since then, other species and subspecies have been separated from and within B. densa (Haines and Lye 1983), e.g. B. pusilla (Hochst. ex A.Rich) C.B.Clarke and B. densa subsp. afromontana (Lye) R.W.Haines were split to delimit the African variation. Collections from Kwazulu-Natal, South Africa, which have nuts at the plant base that are distinctly larger than the aerial counterparts (i.e. the plants are amphicarpic), do not fit the current description of B. densa or similar species (Haines and Lye 1983; Gordon-Gray 1995). In addition, two collections, one from China and the other from Queensland, Australia (E. sp. afC densa 1), have morphology similar to B. densa s.s., but with different nut characteristics; the Queensland collection does not fit descriptions of any accepted Australian species. It is necessary to compare Australian (including Paci fic), African, and Asian material of B. densa, \vith the American material of B. capillaris, to assess species limits of B. densa in Australia. Bulbostylis barbata is another cosmopolitan species where the morphological variation needs to be compared globally. Bulbostylis barbata subsp. pulchella (Thwaites) T.Koyama was separated to demarcate the variation in plants from southern India and Indo-China generally. A putative new species of Bulbostylis (B. sp. aff. barbata), with distinctly piliferous glume margins, and hairy sheaths and leaf blades, was collected fronl the Kakadu National Park in the Northern Territory. These plants grow interspersed with plants of B. barbata and have a similar growth habit and inflorescence-synflorescence structure. Although the nuts of the putative new species are similar to those seen in B. barbata there are apparent differences between the collections of both taxa. The species limits for B. sp. aff. barbata required testing before a new species can be defined. Blake (1941) described B. pyriformis S.T.Blake and commented on the style base that mayor may not persist on the nut. This feature of the style base has been a cause of great confusion in the placement of B. hispidula (Vahl) R.W.Haines, as is evident fron1 the many nomenclatural synonyms (World Checklist of Monocotyledons 2004); the persistence of the style base on the nut was a key character in assigning taxa to Bulbostylis, with non-persistence characteristic for Fimbristylis. Embryo morphology provided the evidence that united specimens of B. hispidula with Bulbostylis through sharing the Bulbostylis-type embryo (Van der Veken 1965). Some specimens of Bulbostylis turbinata also show variability in the persistence of the style base on the nut. The Australian B. pyriformis shares similar characteristics in general plant morphology and nut micromorphology with taxa of the B. hispidula complex. It was necessary to compare the Australian material with SOlue of the African B. hispidula subspecies to test the species limits. \Nilson (1980) described Bulbostylis burbidgeae as a new species endeluic to Australia. Two separate collections, P.K. Latz 11364 (NSW 452329) (B. sp. aff. puberula) and C.R. Dunlop (DNA 14302, NSW) (B. sp. aff. burbidgeae) superficially resemble B. burbidgeae, but have nuts that are quite distinct from each 139 other and from those typical of B. burbidgeae. Both collections are fron1 the Northern Territory and therefore outside the known Western Australian distribution range for B. burbidgeae. The collections of B. sp. aff. puberula and B. sp. aff. burbidgeae need to be compared to the other Australian species and to B. puberula (Poir.) C.B.Clarke, to assess the species boundaries. As a recent introduction to Australia (Wilson 1993), the African species Bulbostylis humilis, needs to be included to assess the extended range of distribution. The limits of all species and putative species of Australian Bulbostylis were tested using phenetic analysis. The relationships of those species and of the genus as sarrlpled here were then assessed for monophyly in the cladistic analysis. Materials and methods Taxa All Australian taxa currently recognised as Bulbostylis, i.e. B. barbata, B.densa, B.turbinata, B. pyriformis and B. burbidgeae (Wilson 1980, 1993), formed the basis of the phenetic study. Putative new species, i.e. B. sp. aff. barbata, B. sp. aff. burbidgeae, B. sp. aff. puberula and B. sp. aff. densa l, were included for species level assessment (Table 5.1). Overseas specimens for the widespread Bulbostylis barbata and B. densa were included with Australian material in the phenetic analyses to define the species on a global level. Representative specilnens of B. capillaris (TYPE species for the genus) B. humilis, B. puberula, and samples from the B. hispidula complex (B. hispidula (Yahl) R.W.Haines subsp. pyr{formis Table 5.1 Specimens sampled as the focus group in the phenetic assessment of Australian Bulbostylis. The 'OTU' corresponds to the label used in phenetic analyses. States are given for Australian collections and the Country of origin for all other samples collected overseas. N.T. = Northern Territory, W.A. = Western Australia, S.A. = South Australia, Qld = Queensland, N.S.W. = New South Wales, P.N.G = Papua New Guinea. See Appendix 1 for specimen details. Species Bulbostylis sp. aff. barbata Bulbostylis barbata Bulbostylis turbinata Bu/bostylis densa OTU State or Country baffba1 N.T. baffba2 N.T. baffba3 N.T. baffba4 N.T. baffba5 N.T. baffba6 N.T. baffba7 N.T. bba1 Qld bba2 N.T. bba3 W.A. bba4 N.S.W. bba5 N.T. bba6 N.S.W. bba7 N.T. bba8 W.A. bba9 N.T. bba10 N.T. bba11 W.A. bba12 Qld bba13 Singapore bba14 USA Kenya bba15 India bba16 America bba17 India bba18 bba19 South Africa Thailand bba20 btl S.A. bt2 W.A. bt3 N.T. bt4 N.T. bt5 W.A. bt6 N.T. bt7 W.A. bt8 W.A. bt9 Qld bt10 N.T. bde1 N.S.W. bde2 N.S.W. bde3 Qld bde4 Qld bde5 Qld Collector Clarke K.L 184, Bruhl J.1., Wilson K.L., Cowie J.D. Rice B.L. Clarke K.L 245, Bruhl J.J., Wilson K.L., Cowie J.D. Clarke K.L 241, Bruhl J.J., Wilson K.L., Cowie J.D. Bruhl J.J. 369A Clarke K.L 251, Bruhl J.1, Wilson K.L., Cowie J.D. Clarke K.L 239, Bruhl J.J, Wilson K.L., Cowie J.D. Wilson K.L. 5442 Beauglehole A.C. 26084 Mitchell A.S. 1150 Tindale M.D. 2058 Bruhl J.1., Hunter J.T., Egan J. 1269B Bell D.B. Latz P.K. 8263 Clarke K.L 160, Bruhl J.J, Wilson K.L. Knight F. 14185 Clarke K.L 221, Bruhl J.J, Wilson K.L., Cowie J.D. Clarke K.L 113, Bruhl J.J, Wilson K.L. Clarke K.L 100, Bruhl J.J. Burkill H.M., Shah M. HMB235 Hill S.R. 24361 Napper D.M., Kanuri 2079 Raizada M.B. Correll D.S. 52337 Rajn R.R.V. Polhill R. 847, Paulo S. Larsen K. 1299, Smitinand T., Warncke E. Cleland J.B. Mitchell A.A. 479 Latz P.K.7126 Latz P.K. 6339 Royce R.D. 1491 Latz P.K. 7087 George A.S. 820 Payne A.L. PRP 1854 Harris P.L. 342 Beaug1ehole A.C. 26568 Bruhl J.J., Quinn F.C. 1197 Hunter J.T., Hunter V. 2737 Forster P.I. PIF8482 Blake S.T. 21453 Bean A.R. 1570 Table 5.1 cont'd bde7 Qld Hubbard C.E. 3128 McKee H.S. 9317 bde8 Qld bde9 N.S.W. Williams J.B. Gray M. 3255 bdel0 N.S.W. bdell P.N.G. Croft 34706, Lelean bde12 Robbins R.G. 2660 P.N.G. bde13 Philippines Ramos M., Edaro G. bde15 Sri Lanka Davidse G. 7614 Eulbostylis sp. aff. baffdl Bean A.R. 3236 Qld baffd2 Field survey team 820 densa 1 China Eulbostylis sp. aff. bde16 South Africa Meeuse A.D.J. 10158 densa 2 bde17 South Africa Scheepers J.C. 1141 bde18 Swaziland Haines R.W. 7048 Eulbostylis bbul W.A. Hart R.P. 2092 burbidgeae bbu2 W.A. Carolin R.7640 bbu3 W.A. Mitchell A.A. 1929 bbu4 Burbidge N. 1102 W.A. bbu5 N.S.W. Payne A.L. PRP976 Eulbostylis sp. aff. baffbu N.T. Dunlop C.R. 4725 burbidgeae Latz P.K. 11364 Eulbostylis sp. aff. baffpu N.T. puberula Eulbostylis bpyl pyriformis N.S.W. Johnson L.A.S. bpy2 N.T. Latz P.K. 10622 bpy3 N.S.W. Hunter J.T., Bell D.B. bpy4 N.T. Latz P.K. 488? bpy5 N.S.W. Wilson K.L. 1479A bpy6 N.T. Latz P.K. 9852 bpy7 Sharpe P.R. 232 Qld bpy8 Qld Bean A.R. 4227 142 (Lye) R.W.Haines and B. hispidula subsp. senegalensis (Chenn.) Vanden Berghen) were restricted to use in cladistic analysis. Phenetic study As Bulbostylis was recovered as a distinct group in the main phenetic analysis perfonned in Chapter 3, only taxa from Bulbostylis were included in the phenetic analyses for this chapter. Pattern analyses Additional OTUs were added to the Bulbostylis OTUs in the main data set (in Chapter 3). A total of 70 specimens (OTUs) of Bulbostylis fonned the basis for the phenetic study (Table 5.1 see also Appendix 1 for full species list), where 20 quantitative and 89 qualitative morphological characters (Table 5.2) were analysed in PATN (Belbin 1993). Data were subjected to ordination, cluster and network analyses as detailed in Chapter 2, and the combined data set, analysed using the Gower Metric sin1ilaritycoefficient, is presented here. Groups that were clear-cut in the first analysis for the genus were relnoved and the data re-analysed as subsets to assess the remaining taxa. Two-dimensional scatter plots were used to present the ordination results. Boundaries of the 3-dimensional ordinations were outlined in the corresponding 2-dimensional scatter if the 2dinlensional groupings were indistinct. Table 5.2 Attribute codes and definitions used in the main phenetic analyses for the Australian Bulbostylis, including corresponding initial weight values. Weight values changed in subset analyses. Attribute Description charI Mean aerial spikelet width in mm (spikelets with mature fruit) at the widest point Mean aerial nut length in mm from base of stipe to nut apex (excluding persistent style base) Mean aerial nut width in mm at widest point Aerial nut length:width (ratio 1:W/L(x) (to decimall/x), ratio coefficient Mean aerial nut 'stipe' length in mm Stipe length/nut length (proportion) Mean aerial anther length in mm (including appendages) Mean aerial style length in mm (including style base to base of style arm junction) Mean aerial style width in mm (at mid third) Style length:width (1 :W/L(x) to decimal l/x), ratio coefficient Mean aerial stylebase length in mm (from base to constriction at style junction) Mean aerial stylebase width in mm (at widest point) Style base length:width (1 :W/L(x) to decimal I/x); ratio coefficient Mean aerial glume length in mm (from base of nerve to apical point) Mean aerial glume width in mm (at widest point) Glume length:width (1:W/L(x) to decimal l/x); ratio coefficient Mean leaf width in mm (at mid third) Mean culm width in mm (at mid third) Mean root width in mm (one em below plant base) Mean inflorescence-synflorescence length in mm (from base of main bract to furthermost point of spikelets) Basal spikelets O-absent: always only aerial; I-present: basal spikelets (morphologically distinct) as well as aerial spikelets Sub-radical spikelets (Wilson 1980), spikelets that are aggregated near the plant base that are morphologically similar to the aerial spikelets: the nuts are indistinct from aerial nuts Nut shape in transverse section is plano-convex; dorsal/ventral sides of a 3-angled fruit with the adaxial face distinct from the rest, being broader than the abaxial faces, often +/- rounded Nut shape in transverse section is strongly triqetrous with deeply concave faces Nut shape in transverse section is triquetrous, having 3-angles, with faces being concave Nut shape in transverse section is trigonous, 3-angles with faces somewhat flattened Nut shape in transverse section is rounded trigonously, with 3 equal sides but well rounded edges and faces (convex) Nut outline obovate (2:1 or 3:2) Nut outline widely obovate (6:5) Nut outline very widely obovate (1: 1) Nut outline pyriform (pear-shaped) Nut outline obtrullate Nut outline widely obtrullate (6:5) Nut outline very widely obtrullateI: 1) char2 char3 char4 char5 char6 char7 char8 char9 charlO charI 1 charI 2 char13 charI 4 charI 5 charI 6 charI 7 charI8 charI 9 char20 char21 char22 char23 char24 char25 char26 char27 char28 char29 char30 char31 char32 char33 char34 Weight 1 1 1 1 1 1 1 1 1 0.5 0.5 0.2 0.2 0.2 0.2 0.2 0.125 0.125 0.125 0.125 0.125 0.125 0.125 Table 5.2 (cont'd) char35 char36 Nut outline napiform Nut epidermis without protuberances (apparent at 50x magnification under a dissecting microscope Nut epidermis rugulose (minutely rugose) Nut epidermis rugose with rounded waves Nut epidermis rugose with acute waves (apex acute from a central raised silica body Nut epidermis sub-puncticulate, from single raised cells that are not prominent and are scattered over the surface Nut epidermis puncticulate, from prominent single cells raised evenly over surface Nut epidermis with rows of warts on face, usually 2 vertical rows on each face Nut epidermis reticulate, from distinct and raised cell walls Nut epidermis finely hexagonal, giving a honeycomb appearance, obvious at lOx magnification under a dissecting microscope Nut epidermal cells isodiametric; almost square to just rectangular Nut epidermal cells oblong longitudinally (2: 1) Nut epidermal cells narrowly oblong longitudinally (6; 1;3; 1) Stamen number: 1 Stamen number: 2 Stamen number: 3 Sheath glabrous (hairs absent) Sheath with short to medium hairs (60-100 /lm) Sheath with bristly hairs almost erect from surface (> 100-1000 /lm) Glume margins entire Glume margins ciliolate; small fine hair-like projections from the margins, sometimes only distally (5 div @ 50x - sometimes only distally) Glume margins fimbriolate; small, flattened projections from margins, sometimes only distally (100 /lm) Glume margins piliferous; fine, long, loose hairs arising from the margins (1 000 /lm) Glume apex rounded Glume apex acute (muticous) Glume apex sub-mucronulate Glume apex mucronulate Glume apex mucronate Glume apex acuminate Glume apex aristate Glume outline ovate Glume outline trullate (kite-shaped) Glume outline narrowly triangular Glume outline linearly triangular Glume apex reflexed at maturity Glume apex not reflexed at maturity Glume nerve muticous Glume nerve to a mucro point Glume nerve excurrent (greater than 0.5 mm) Glume abaxial surface glabrous char37 char38 char39 char40 char41 char42 char43 char44 char46 char47 char48 char49 char50 char51 char52 char53 char54 char55 char56 char57 char58 char59 char60 char61 char62 char63 char64 char65 char66 char67 char68 char69 char70 char71 char72 char73 char74 char75 0.125 0.125 0.125 0.125 0.125 0.125 0.125 0.125 0.125 0.2 0.2 0.2 0.2 0.33 0.33 0.33 0.33 0.33 0.33 0.25 0.25 0.25 0.25 0.143 0.143 0.143 0.143 0.143 0.143 0.143 0.25 0.25 0.25 0.25 0.5 0.5 0.33 0.33 0.33 0.167 Table 5.2 (cont'd) char76 char77 Glume abaxial surface with nerve only scabrid Glume abaxial surface scabrid over lower half of glume (even isolated, sparse, dense toothed hairs) Glume abaxial surface scabrid over most of the glume back (20-40 fim) Glume abaxial surface with short hairs (100 fim) Glume abaxial surface bristly, with erect hairs (> 1000 fim) Glume arrangement on the rachilla distichously spiral (glumes opposite each other and glume pairs ascending arranged spirally) Glume arrangement on the rachilla tristichous Non fertile glume number at the base of each spikelet: 0 Non fertile glume number at the base of each spikelet: 1 Leaf to culm ratio: 1to 1 Leaf to culm ratio: 2t03 Leaf to culm ratio: 1t02 Leaf to culm ratio: lt03 Leaf to culm ratio: 1 to 4 Culm surface glabrous Culm surface scabrid (includes distally) Culm surface with short hairs that are almost erect (c. 100 Jlm) Culm surface bristly with stiff erect hairs (includes distally) Leaf abaxial surface glabrous Leaf abaxial surface with scabrid margins Leaf abaxial surface scabrid over the abaxial surface Leaf abaxial surface with erect to ascending hairs (c. 100 fim) Leaf abaxial surface bristly/hispid (1000 Jlm - erect to outwardly ascending) Inflorescence-synflorescence mostly solitary (HF 1), or 1-2 coflorescences (Cofl) Inflorescence-synflorescence as anthelodia, main florescence (HF 1) plus multiple primary coflorescences (Cofl) on lengthened epipodia (rays); some coflorescences may be sessile, but not all Inflorescence-synflorescence as ramified (compound) anthelodia Inflorescence-synflorescence as 'heads' of 3-7 sessile spikelets Inflorescence-synflorescence hemispherical 'head' of> 7 sessile spikelets Inflorescence-synflorescence bracts present and glume-like Inflorescence-synflorescence bracts present and leaf-like Inflorescence-synflorescence bracts shorter than inflorescencesynflorescence length Inflorescence-synflorescence bracts equals the inflorescencesynflorescence length Inflorescence-synflorescence bracts longer than the inflorescencesynflorescence length Inflorescence-synflorescence prophyllar buds or spikelets present Inflorescence-synflorescence prophyllar buds or spikelets absent char78 char79 char80 char8l char82 char83 char84 char85 char86 char87 char88 char89 char90 char91 char92 char93 char94 char95 char96 char97 char98 char99 chariOO charlO 1 charl02 charl03 charI04 charl05 charI06 charl07 charl08 charl09 charI 10 0.167 0.167 0.167 0.l67 0.167 0.5 0.5 0.5 0.5 0.2 0.2 0.2 0.2 0.2 0.25 0.25 0.25 0.25 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.2 0.5 0.5 0.33 0.33 0.33 0.5 0.5 Cladistic analysis Ingroup Species of Crosslandia and Abildgaardia that were defined in Chapters 3 and 4 were combined with the terminal taxa of Bulbostylis as determined in the phenetic analyses of this chapter. Samples of Bulbostylis humilis, B. puberula, B. hispidula subsp. pyriformis and B. hispidula subsp. senegalensis were added to the ingroup data. Terminal taxa of Fimbristylis used in previous analyses were maintained in this cladistic analysis (Table 5.3). Outgroup The outgroup in the cladistic analysis for this chapter comprised Arthrostylis aphylla, provisional Actinoschoenus compositus, Trachystylis stradbrokensis (Donlin.) Klik., Schoenoplectus tabernaemontani (C.C.Gme!.) Palla (= S. validus Yahl), Schoenoplectiella lateriflora (J .F.Gmel.) Lye (= Schoenoplectus lateriflorus), and Schoenoplectiella laevis (S.T.Blake) Lye (= Schoenoplectus laevis) (Appendix 1). Characters and homology Guaglianone (1970) observed intraprophyllar buds within the inflorescence prophylls of species in Bulbostylis, and proposed the presence of intraprophyllar buds as a generic separator between Bulbostylis and Fimbristylis; intraprophyllar buds are absent in Fimbristylis. All specimens of Bulbostylis used in this study were examined for the presence of intraprophyllar buds or spikelets. Tablle 5.3 Taxa included in the cladistic analyses to assess the relationships of Australian species of Bulbostylis. Species fron1 Crosslandia and Abildgaardia included here were defined in Chapter 3 and 4 respectively. See Table 5.1 for Bulbostylis specimen list and Appendix 1 for specimen details. Taxa No. specimens sampled Ingroup Abildgaardia macrantha (provisional) Abildgaardia mexicana Abildgaardia odontocarpa (provisional) Abildgaardia ovata Abildgaardia oxystachya (provisional) Abildgaardia pachyptera (provisional) Abildgaardia schoenoides Abildgaardia triflora Bulbostylis barbata Bulbostylis burbidgeae Blllbostylis capillaris Bulbostylis densa Bulbostylis hispidula subsp. pyriformis Bulbostylis hispidula subsp. senegalensis Blllbostylis puberula Bulbostylis pyriformis Bulbostylis humilis Bulbostylis turbinata Bulbostylis sp. aff. barbata Bulbostylis sp. aff. burbidgeae Bulbostylis sp. aff. densa 1 Bulbostylis sp. aff. densa 2 Bulbostylis sp. aff. turbinata 1 Bulbostylis sp. aff. turbinata 2 Bulbostylis sp. aff. puberula Crosslandia anthelata (provisional) Crosslandia setifolia Crosslandia spiralis (provisional) Crosslandia vaginata (provisional) Fimbristylis bahiensis Fimbristylis blakei Fimbristylis cinnamometorum Fimbristylis depauperata Fimbristylis fimbristyloides Fimbristylis furva Fimbristylis hygrophila Fimbristylis microcarya Fimbristylis schultzii Fimbristylis sp L. Fimbristylis variegata 10 5 2 13 13 11 12 4 20 5 4 15 3 2 7 8 3 8 7 1 2 3 1 1 1 5 18 3 14 4 2 5 2 4 2 2 2 2 2 1 ｏｵｴｾｲｯｰ Actinoschoenus compositus (provisional) Arthrostylis aphylla Schoenoplectiella laevis Schoenoplectiella lateriflora Schoenoplectus tabernaemontani Trachystylis stradbrokensis 4 4 5 5 3 7 14S Embryo morphology Van der Veken (1965) and Goetghebeur (1986) sampled embryos of some species of Bulbostylis, reporting variation in general embryo shape and size, and primordial leaf development. Bulbostylis barbata was the only Australian species sampled in both studies. In this study, embryos from representatives of each of the species (and subspecies) were sampled and compared (Appendix 1). Second and third (ifpresent) prinliordial leaves were not scored due to difficulties in observing these structures in many of the embryos. Tissues in small embryos were much denser than in the larger embryos sampled, and I was unable to clear some small embryos sufficiently to define the inner layers (e.g. B. sp. aff. barbata, and B. sp. aff. densa l). Alternative methods in pre-treating the embryo before clearing, or using a different clearing medium, may be necessary in future work. Anatomy Leafblade and culm anatomy were sampled across the species of Bulhostylis studied to compare general shape and tissue arrangement. General anatomy (Metcalfe 1969, 1971) and photosynthetic pathway in Bulbostylis (Goetghebeur 1986; Bruhl 1995); has been reported to be the same as commonly found in Fimbristylis; sampling tested the uniformity in this study. PA up* analyses Parsimony analysis was performed on 47 terminal taxa and 155 characters using heuristic searches (hsearch swap=TBR addseq=rand nreps=l 000 hold=5 multrees=yes). Branch support was assessed using Bootstrap analysis (1000 bootstrap replications) because the computational time required to calculate the Bren1er support indices past 3 extra tree length steps was too protracted, even when limiting the addition-sequence replications to 10. Characters were traced in MacClade and the most relevant characters are presented in the cladogram. Results Phenetic study Representative OTUs for the genus Bulbostylis formed a distinct group in the initial main analysis (see Chapter 3), with some species groups of OTUs (B. sp. aff. barbata, B. barbata, B. burbidgeae and B. sp. aff. puberula) apparent in the Bulbostylis cluster at the broad level in 2-dimensional analysis (Figure 5.1). "Then additional samples of Bulbostylis were added to the first main analysis and re-analysed (see Materials and methods, this chapter), distinct species groups were fornled by the OTUs in the 2-dimensional ordination (stress = 0.18; Figure 5.2). Characters that were most strongly correlated with the groups formed within the ordination were consistent with the sYnf10rescence type, nut epidermal patterning, number of stamens, anther length, nut stipe length to nut length ratio, hairiness (or absence of) of culms and glumes, and shape of glumes (Figure 5.3). The hemispherical heads of sessile spikelets associated with B. barbata and B. sp. aff. barbata and the mostly primary rayed anthelodium (i.e. spikelets on lengthened epipodia) of B. pyriformis and B. turbinata were correlated with the separation of the groups. • Crosslandia III Abildgaardia l .. B. sp. aff. barbata X B. barbata :( B. turbinata • B. densa + B. burbidgeae <> B. sp. aft. puberula ' I' J I B. sp. aff. burbidgeae <> B. pyriformis o Fimbristylis • • •.#•• • ..,_ :;• •• •• • . ..... • # •• •• D D D D Den .. %{;} D 0 Figure 5.1. MDS ordination in 2-dimensions (stress = 0.17) from primary phenetic analysis (see Chapter 3) highlighting Bulbostylis. Species groups for B. sp. aff. barbata, B. barbata and B. burbidgeae are distinct in this broad level ordination. See Table 5.1 and Appendix 1 for OTU and specimen details. . n .. ·r· · • II , : • • Figure 5.2. MDS ordination for OTUs of Bulbostylis (stress = 0.18). Lines separating OTUs in B. densa (amphicarpic specimens) and B. sp. aff. densa indicate the clear-cut groups observed in 3dimensions (stress = 0.12). See Table 5.1 and Appendix 1 for OTU and specimens details. '.66.6 46,4307;:(3.100 +67 -gO -51 <>75 D1?j Stipe length/nut length Inflorescence: primary 'anthela' Glunne shape: ovate Nut width • • Culm glabrous Glume back glabrous Stamens: 3 o Anther length IT Inflorescence: hemispherical Glume shape: trullate Nut epidermis: reticulate Nut cell outline: isodiametric Figure 5.3 Characters that correlate (>80%) to group formation in the ordination shown in Figure 5.1. Inflorescence-synflorescence of many sessile spikelets forming a hemispherical 'head', glume shape: trullate, nut epidermis being reticulate with isodiametric cells, plus anther length separated Bulbostylis barbata and B. sp aff. barbata from the other OTU groups. Culm and glume backs glabrous contributed to separating the B. densa group and stamens numbering 3 correlated strongest with the B. burbidgeae OTUs. The group containing OTUs for B. turbinata and B. pyriformis were consistent with the correlated characters of inflorescence-synflorescence: primary 'anthela', highest stipe length-nut length ratio; B. pyriformis has the greatest nut widths. See Table 5.2 for attribute definitions. ＱｴｾＹｂ Bt2 Bt "'?';$l_ _ ＭＭＭｾ Bt7 Bt8 Bt1 I. I • B. aff. barbata B. barbata B. turbinata B. densa 1)( B. aft. densa • B. burbidgeae ;+ B. aff. puberula • B. aff. burbidgeae ! I B. pyriformis Figure 5.4. Minimum spanning tree (MST) for OTUs of Bulbostylis corresponding to ordination in Figure 5.2. Borders indicate greater separation seen in 3-dimensions (stress = 0.12). See Appendix 1 for specilnen details. 0.0359 baffba1 baffba4 baffba6 baffba7 baffba2 baffba5 baffba3 btl bt4 bt7 bt5 bt8 bt6 bt2 bt3 bt9 bt10 baffbu bbu1 bbu2 bbu3 bbu4 bbu5 baffpu bpy1 bpy2 bpy3 bpy7 bpy8 bpy4 bpy5 bpy6 bba1 bba2 bba3 bba8 bba4 bba6 bba5 bba10 bba9 bba11 bba20 bba12 bba7 bba13 bba14 bba15 bba16 bba17 bba18 bba19 bde1 bde2 bde8 bde10 bde9 bde13 bde15 bde12 bde3 bde4 bde7 bde6 bde5 baffd1 baffd2 bde16 bde17 bde18 0.1381 I 0.2403 1 1}_ 0.3426 1 1 0.4448 0.5470 I 1 4}_1_ 6}_ I 7}/_1_ 2} _ _ I 5}_1_1 3} _ _ 1 _ 28} _ 31} _ _ 34}_1_ I I 32} _ _ 1 _ _ 1_ I 35} 33} 1_1_ 29} 1 30) 1 36} 37} 62} 56}_ 57)1_ 58}_1 59}_11_ 60) _ _ 1 61 ) 63) _ 64}_ 65}1_ 69)_ I _ 1 1_1 _ _ 1 _ _ I 7o}_I_I_I_ 1 66} 67)_ II 68}_1 _ _ 11 - _ 8} _ _ 9}_1_ 10} _ _ I_ _ 15} 111) _ _ 13}_I _ 12}_ 17}_I _ _ 16} _ _ 1 _ _ 18}__ I 27} _ _ 1_ 1 19} _ _ 1_1 14} 20} 21}__ I _ 1_ 1 I 22}_1_1 1 __ 23} _ 1_ 24} 25) 1 1_1 26} 38)___ 39} _ _ 1____ 45}__ 47} _ _ 1_ 46) _ _ 1__ 49) I 50) 1_1_ 48} 1__ 40}______ 41}_ 44}_1__ 1_ 43} 1_____ 42} 51) I 152} 1 53}__ 1 54}_ I 1 55}_1_1 _ I 1 I I 1 1 1 I I 1 I 1 1 I 1 I 1 1 1 1 I 1 I I 0.0359 0.1381 0.2403 0.3426 0.4448 _ 0.5470 Figure 5.5. WPGMA Ｈｾ = -0.1) phenogram that corresponds with the ordination in Figure 5.2, for all OTUs of Bulbostylis. OTUs form six groups, the putative species B. sp. aff. barbata is clearly separated from B. barbata. See Table 5.1 and Appendix 1 for OTU and specimen details. 155 Species groups in the ordination were generally supported by network (Figure 5.4) and cluster analyses (Figure 5.5). Stronger grouping was obtained in the ordination for 3-dimensions (stress = 0.12) as indicated in the 2-dimensional scatter plot (Figure 5.2). Individual OTUs, baffbu (B. sp. aff burbidgeae), baffpu (B. sp. aff. puberula), bt9 and btl 0 (B. turbinata), were separated from other groups, and from each other, in the 3-dimensional ordination. Although the separation is evident in 2-dimensions for bafjbu and baffpu, this is not the case for OTUs bt9 and btl 0, which appear as if distinctly clustered with the other B. turbinata OTUs. Within the phenogranl (Figure 5.5), baffbu is clustered with OTUs of B. turbinata and baffpu is broadly included with the B. burbidgeae OTU group. However, the dissimilarity ofbaffpu to OTUs of B. burbidgeae is present in the phenogram. Discrete groups of OTUs were retrieved as the species groups B. sp. aff. barbara, B. barbata, B. pyriformis, B. burbidgeae and B. turbinata (excluding bt9, btl 0) within 3- and 2-dimensional ordinations (Figure 5.2), and generally in the phenogram (Figure 5.5). The remaining OTUs formed the B. densa group that included baffd 1,2 (B. sp. aff. densa) and bde16, 17, 18 (African amphicarpic samples of B. densa). Bulbostylis densa group In the subset analyses of the B. densa group, OTUs for the amphicarpic specimens colIected from Africa formed a group separate to the main B. densa OTUs, and to the OTUs of B. sp. aff. densa. The three groups are more robust in 3-dimensions (stress = 0.12), indicated by the drawn boundaries around the specific groups in the 2dinlensional scatter plot (Figure 5.6). There were 16 characters with greater than • • • • • • • •• • • • • • • B. densa • B. aft. densa B. densa amphicarpic - - - - Figure 5.6 MDS ordination in 2 dimensions (stress = 0.18) for the Bulbostylis densa group from the prirrlary Bulbostylis analysis (see Figure 5.2). OTUs for the amphicarpic material from Africa form a group separate to the main B. densa group and the separate OTUs of B. sp. aff. densa. The boundaries shown indicate the distinct OTU groups in the 3-dimensional ordination (stress =: 0.12). See Table 5.1 and Appendix 1 for OTU and specimen details. .7 .60 84 010 Anther L Basal spikelets: present Synflorescence: solitary or few rayed Glume apex: mucronate Glume margins: fimbriolate ::4( 56 .30 + 63 -21 ｾ ... 99 <> 57 44 659,46,36,43 28 Empty glumes: 1 Glume apex: rounded Nut epidermal sculpturing: reticulate Nut epidermal cells: isodiametric • o Nut outline: obovate Glume apex: acute • Nut outline: very widely obovate Glume apex: rounded Style L/W Nut epidermal cells: finely hexagonal Figure 5.7 Characters correlated (> 70 0/0) with the ordination in Figure 5.6 for OTUs of the ｳｩｬｹｾｯ｢ｵｂ densa group. Characters with >80 % correlation to the ordination were anther length, glume apex: acute, empty glume: 1, style length to width ratio, glume margins: ciliolate, and nut shape: obovate. L=length. See Table 5.2 for attribute definitions. Bd18 ＷＱ､ｂｾ Bd1 • Bd16 \Bd2 Bd15":7 • Bd10 Bd4 Bd13 ｾＲ､ｦ｡ｂ .B. densa • B. aff. densa Baffd1 B. densa amphicarpic Figure 5.8 Minimum spanning tree (MST) with linkages for the Bulbostylis densa group plotted onto the 2-dimensional ordination in Figure 5.6. See Appendix 1 for specimen details. 0.1600 bde1 bde2 bde3 bde4 bde7 bde5 bde8 bde9 bde10 bde13 bde15 bde12 bde16 bde17 bde18 baffdl baffd:2 1 1)_ 2) 1 3) 4)_ 6)_1 5) 7)_ 8)_1_ 9)_1 11) 12) 10) 15)_ 16}_ I 17} I_I 13} 14} 1 0.1600 0.2324 0.3048 I I 0.3772 1 0.4496 0.5220 I I 1 1 1 0.2324 I 0.3048 I 0.3772 1---I I 1 1 0.4496 0.5220 Figure 5.9 WPGMA (P = -0.1) phenogram for the Bulbostylis densa subset (see Figure 5.2 for all species of Bulbostylis) that best correlates with the ordination (Figure 5.6) and Minimum Spanning Tree (MST) (Figure 5.8). Operative Taxonomic Units (OTUs) for B. densa with amphicarpic nuts (bde16-18) are grouped together separate to the remaining OTUs for B. densa. Similarly, OTUs for B. sp. aff. densa (baffdl, baffd2) group separately. See Table 5.1 and Appendix 1 for OTU and specimen details. 160 ｾＧｊ＼ＰＷ correlation to the three groups seen in the ordination scatter plot (Figure 5.7). Characters with >800/0 correlation to the ordination were anther length, glume apex: acute, empty glume number: 1, style length to width ratio, glume margins: ciliolate, and nut shape: obovate. Linkages between OTUs in the minimum spanning tree (Figure 5.8) support the major groups (B. densa S.s., B. densa 'amphicarpic', B. sp. aff. densa) and the minor groups (within B. densa s.s.) observed in the phenogram (Figure 5.9). Bulbostylis turbinata-B. sp. aff. burbidgeae group Subset analysis of the B. turbinata-B. sp. aff. burbidgeae group of OTUs indicates that B. sp. aff. burbidgeae bt9 and btl a are discrete units from the main OTU group of B. turbinata in the 3-dimensional ordination (stress = 0.1), and to a lesser extent in 2-dimensions (Figure 5.10). There were 24 characters with >70% correlation to the ordination; characters with >80% correlation were inflorescence prophyllar branching: present, inflorescence prophyllar branching: absent, culm width, style length, glume width and style base width (Figure 5.11). The minimum spanning tree OTU linkages (Figure 5.12) correspond to the groups from the ordination and cluster analysis (Figure 5.13). Terminal taxa as recognised in the phenetic analyses, i.e. Bulbostylis sp. aff. barbata, B. barbata, B. turbinata, B. sp. aff. turbinata 1, (bt9), B. sp. aff. turbinata 2 (btl 0), B. pyriformis, B. burbidgeae, B. densa, B. sp. aff. densa 1, B. sp. aff. dellsa 2 (African amphicarpic), B. sp. aff. burbidgeae, and B. sp. aff. puberllla, were cornbined with samples from B. humilis, B. capillaris, B. hispidula subsp. senegalensis, and B. hispidula subsp. pyriformis for use in cladistic analysis. ' . 8 . turbinata .819 148t10 X 8. sp. aff. burbidgeae • • • • i • • • • Figure 5.10 MDS ordination in 2-dimensions (stress= 0.18) for OTUs of the B. turbinata group from Figure 5.2 The OTUs bt9, bt10 and B. sp. aff burbidgeae (baffbu) are separated from the main group of B. turbinata OTUs. The boundaries indicate the stronger group resolution seen in the 3-dimensional ordination (stress = 0.11). See Table 5.1 and Appendix 1 for OTU and specimen details. ,• 109 • 110 18 0 12 ;:( 8 • 15 Aerial style L Syn tlorescence prophyllar buds: present • • Syntlorescence prophyl1ar buds: absent CulmW o Aerial style base W • Aerial glume W Figure 5.11 Attributes correlated (>80%) with the ordination in Figure 5.10 for OTUs of the Bulbostylis turbinata group. Key attributes were style length, style base width, aerial glume width, culm width, synflorescences prophyllar bud, or growth, absent/present (polymorphic). L=length, W=width. See Table 5.2 for attribute definitions. I • B. turbinata I.Bt9 !.Bt10 I X B. sp. aft burbidgeae I Bt2 Bt1 Figure 5.12 Minimum spanning tree (MST) with linkages for the Bulbostylis turbinata group plotted onto the 2-dimensional ordination of Figure 5.10. See Appendix 1 for specimen details. 0.1480 btl bt2 bt3 bt6 bt4 bt7 bt5 bt8 bt10 bt9 baffbu 1) 1 0.2322 1 7) 0.4006 I 0.4848 1 --:- 1 11 _ _ I I T 5) 1 8) 1 _ _- . , . - 10) 1 __ 9) 11) 0.5690 I I --:-1 2) 3) 6) 4) 0.3164 1 _ ""--1----.,.-1-----:--1------,1,------------:1- I 1 0.1480 0.2322 0.3164 0.4006 0.4848 0.5690 Figure 5.13 WPGMA Ｈｾ］ -0.1) phenogram for the Bulbostylis turbinata subset (see Figure 5.2 for all OTUs of Bulbostylis) that fits the ordination (Figure 5.10) and MST (Figure 5.12). Bulbostylis OTUs group together and the OTUs bt9, btl0, and B. sp. aff. burbidgeae (baffbu) remain separate. See Table 5.1 and Appendix 1 for OTU and specimen details. 165 Cladistic analysis One hundred and twelve most parsimonious trees were retrieved (Tree length=1210, CI=0.4455, HI=O.5545, RI=O.5984, RC=O.2666) from a heuristic search. Tree 1 had similar topology to the tree obtained from strict consensus, and was selected to show branch support and character/branch associations (Figure 5.14). Two broad sister clades, the Bulbostylis-Fimbristylis clade (A) and the Abildgaardia-Crosslandia clade (B), form an internal clade sister to Fimbristylis bahiensis, which is, in tum, sister to Fimbristylis variegata (Figure 5.14). Fimbristylis continues to be retrieved as a non-monophyletic group (see also Chapters 3 and 4), seen by F. sp. Land F. blakei being placed sister to the Crosslandia-Abildgaardia clade. Crosslandia is not a monophyletic group in this analysis, as the provisional C. vaginata is placed sister to species of Abildgaardia. Within clade A, all taxa sampled from Bulbostylis formed a clade (Bootstrap=83%) sister to Fimbristylis fimbristyloides, and combined on branch C as a group sister to the clade Fimbristylis 1 (internal branch D, contains F. depauperata from the TYPE section of Fimbristylis section Fimbristylis). Both clades from C and D are nested within species assigned to Fimbristylis. The Bulbostylis clade (Bootstrap=83%) was formed by two main clades where B. humilis (B4) is sister to the remainder of the species (B3, B2 and B 1); the latter with moderate branch support (Bootstrap=74%). B. barbata (B3) was sister to the terminal groups B2 (B. pyriformis-B. turbinata-B. burbidgeae-B. hispidula clade) and B 1 (B. capillaris-B. densa-B. puberula clade). The strict consensus shows branch collapse for most of the terminal taxa within the Bulbostylis clades Bland B2; only the terminal B. puberula and B. capillaris (with weak branch support), and B. sp. aff. barbata-B. burbidgeae-B. sp. aff. burbidgeae branches persist. Moderate Figure 5.14 Cladogram for tree 1 of 112 shortest trees (tree length = 1490) in the assessment ofmonophyly for Australian species of Bulbostylis. Bulbostylis forms a monophyletic gTOUp sister to Fimbristylis fim bristyloides, fonning clade C, which is sister to Fimbristylis 1 in clade B. Fimbristylis depauperata that is from the TYPE section of the genus, is placed in clade B. Abildgaardia and Crosslandia form a broad group that is sister to Fimbristylis sp. L and F. blakei; all grouped in clade D. Crosslandia is not monophyletic in this analysis. Within the Bulbostylis clade four main groups frequently occur (B 1, B2, B3, B4), however, only 3 groups were retrieved from strict consensus of the 112 most parsimonious trees: B4, B3 and B 1-2. Bootstrap support values are given below the branches. A, B, C, and D indicate the intenlal branch for the main clades. Dashed lines indicate collapsed branches in the tree from strict consensus. See Appendix 1 for specimen details and Appendix 2 for characters. A ｲＭＧｈｦｉ！Ｋｆｩｭ｢ｳｴｹｬ variegata 153-3; 1-2; 14-2; 15-2; 28-2; 35-1 r--tlt-I- - - - - - - - - - - - - - - - - - - - - - - - - - - - - - 153-3 ［ｾＺｾＱＬＲ［ＵＭＱｾ［ＴＭＳＬ［ 24-2 Abildgaardia macrantha 35-1; 41-2 - - Abildgaardia pachyptera 15-1,2; 41-1 ＧｾＡＲＬＳ［ｾＱ 28-1 Crosslandia vaginata .Ill! 14-1,3; 1A-IP; 35-4; 73-1,3 1t;11-1 Fimbristylis hygrophila 28-2 35-1 Abildgaardia schoenoides ---l.I}I. f741l14-1,3,4; 15-1,2 Abildgaardia odontocarpa _ 41-2; Ｑｾ［ＳＹｾ 35-3 Abildgaardia oxystachya 1111111-1,1'+-'j,4; •• 150-2 86-4 115-1,2; 35-1; 41-1 ［ＳＭｾＱ Abildgaardia mexicana 681'"+++ I ＲｾＱＭ 15-3 Abildgaardia ovata 35-3; 41-1 "+-_ __.0011 135 -3 B _ Crosslandia spiralis 150-1 25-2 1ｾＱ Crosslandia anthelata 66 14-3 15-1 73-3 I Crosslandia setifolia 115- 1,2 III ""+ 14-34· 34-4 ﾷＱＭｉＱｾ ' 511 I " Fimbristylis furva 1111 14-3; 15-3; 24-1; 25-2 ［ＲＭＱｬｾ C I Fimbristylis fimbristyloides 153-2,3; 15-3; 24-1; 25-3 II r++IIII-·15--2-,-3;-7-3--3-------------- --- 35-1 F. cinnamometorum 15-3,4; 24-2; 25-2; 41-4; 57-5; 72-2 IIIII A Fimbristylis Sp. L. Fimbristylis blakei II 1-2; 14-1 15-2 r+-'------------I 153-2,3 83"4 ｾＺ . Bulbostylis barbata Bulbostylis sp. aft. turbinata 2 -.It---------- . • 74 Bulbostylis pyriformis .14-3 .! :,.! - + 1 - - - - : : 14-3,4 .ｾＵＳＭﾷＡ . •• '. Bulbostylis turbinata ｾＺ Bulbostylis sp. aft. barbata . 8_3_-1_2 _ Bulbostylis sp. aft. burbidgeae • 73·;.. _ _ ｾＺｩＵﾷ Bulbostylis hispidula subsp. senegalensis B. hispidula subsp. pyriformis Bulbostylis sp. aft. densa 2 .................. ... •• •• II . : . 153-23·15-3 ' Bulbostylis densa Bulbostylis sp. aft. densa 1 rtt-II--- : ｾＱＴＭＲＬ［ OJ N Bulbostylis burbidgeae 25-1 : Bulbostylis sp. aft. puberula -------- 86-1 +0 Bulbostylis puberula 15-1 Bulbostylis capillaris I Fimbristylis microcarya D Fimbristylis disticha 111111 II 15-4 24-1 25-3 41-2 57-5 Bulbostylis humilis Bulbostylis sp. aft. turbinata 1 150-3 11-3 24-2 25-1 - Fimbristylis schultzii +I 153-3"++111-- ---.1-5--2--11 2 I Fimbristylis depauperata ' rrtt:nfl'lll:1-'4-1--2- - - - - Actinoschoenus compositus 153-3 72 .,11------ Arthrostylis aphylla 15-2 1-2 11-1 28-2 Abildgaardia triflora 25-3,35-5,41-4 II ＱＭＲｾＭＭＭＭＭＭＭＭﾷＭＭｉＫＱＬＮ Fimbristylis bahiensis ｉｾ 153-1 141-1 III 15-4; 24-3; 35-1 Itl1-1; 72-1 1114-1; 41-4 +------- 11 15-397 1'1 50-291t11111-2; 41-1; 73-3 1-1 14-3 o Trachystylis stradbrokensis S- Schoenoplectus tabemaemontani oc Schoenoplectiella lateriflora Schoenoplectiella laevis 167 Bootstrap support (73%) for the terminal branch B. aff. burbidgeae and B. burbidgeae contradicts the branch collapse obtained from strict consensus (Figure 5.14). Observations Injlorescence-synjlorescence structure lV[ost species of the Bulbostylis included in the study possess prin1ary-rayed spikelets (coflorescences). Solitary spikelets rarely occur, and if spikelets are solitary then there are at least some rayed spikelets present within the plant (Figure 5.15 A). Secondary orders within the synflorescence were frequent in B. densa, and B. sp. aff. den.fJa 1, B. sp. aff. densa 2, B. puberula, B. pyriformis and B. hispidula subsp. senegalensis. Although intraprophyllar buds were often observed in the open rayed synflorescence, it was rare for the buds to mature and develop into spikelets. The 'head' of sessile spikelets in specimens of B. barbata and B. sp. aff. barbata (Figure 5.15 B, C) is formed from multiple 'branched' sessile primary coflorescences, plus sessile spikelets that develop from within the inflorescence prophylls of the coflorescences. Spikelets arising from the intraprophyllar growth are distinct and contribute to the density of the 'head' of spikelets that occur in the two species. This synflorescence type of terminal capitulum (56-10) was not a synapomorphy; B. sp. aff. barbata was grouped within the 'rayed' synflorescence types of reduced anthelodium and reduced ramified anthelodium seen in the B. pyriformis-B. turbinata-B. hispidula clade (Figure 5.14). Style bases in the Australian Bulbostylis pyriformis, and occasionally in B. turbinata, may persist on, or fall from the nut; the style always detaches from the Figure 5.15 Variation ofsynflorescence structure for some species of Bulbostylis. A. Most Bulbostylis in the study possess 2-4 primary rayed spikelets (primary coflorescences on lengthened epipodia) as seen in B. sp. aff densa 1 (baffd2 pictured); 1-2 secondary coflorescences on rays (lengthened epipodia) may be present in the B. densa group, including B. puberula. Scale=5 mm. B. Sessile spikelets plus spike1ets from prophyllar buds form a head in B. barbata and B. sp. aff. barbata (baffba1 pictured). Sca1e=5 lum. C. Representation of the prophyllar synflorescence structure seen in B. Sessile spike1ets arise from the axils within the prophyll and contribute to the ramification of the synflorescence 'head' as indicated by 2° and 3° orders. D. 'Subradical' spikelets (arrow) may be present in B. barbata (bba8 pictured), B. turbinata, and B. burbidgeae; these spike1ets have similar morphology to the aerial spikelets. Scale=10 mm. See Table 5.1 and Appendix 1 for OTU and specimen details. A 169 style base leaving the style base on the nut, even if only for a short time. This is in contrast to style bases in the B. Izispidula group, where the style may persist on the nut, or fall from the nut intact with the style. In some specimens of B. hispidula subsp. senegalensis, all the fallen styles observed had the style base intact. The large bulbous style base present in B. pyriformis and B. turbinata usually protrudes from the umbonate nut apex and tends to be easily removed. All other species scored in this study have smaller style bases that sit firmly at the apex of the nut. Amplzicarpy The spikelets observed at the base of the plant in B. Izumi/is show different morphology (73-3) to the basal spikelets of the African B. densa (B. sp. aff. densa 2). In B. Izumilis the basal spikelets are attenuate and florets may be bisexual. In the African B. densa (B. sp. aff. densa 2), the spikelets at the base of the plant are clustered in groups of 2 or 3 at the soil level (73-4) and have glumes that are much smaller than those in the aerial spikelets or in B. Izumi/is. The basal glumes in B. densa may fall early and leave the nut exposed. Both types of basal spikelets are amphicarpic, with nuts in the basal spikelets being larger than in the aerial spikelets; the glumes also differ. Subradical spikelets (73-2) in some B. barbata, B. turbinata, and B. burbidgeae differ from classic amphicarpic plants, as the nuts and glumes reselnble their aerial counterparts in size and shape; the spikelets occur on shortened culn1s (Figure 5.15 D). Nut sculpturing Nut epidermal shape and protuberances (or lack of) can be useful in identifying species (Figures 5.16-20), but were only broadly associated with the internal 170 Bulbostylis clades (Figure 5.14). Species with nuts that have papillate or granulate sculpturing (Figures 5.16-18) were split between the Bland B2 clades, so that nuts with vertically elongated epidennal cells (Figure 5.19) were interspersed. Bulbostylis burbidgeae nut epidennal cell walls are barely to mildly sinuose, as are the B. hispidula and B. pyriformis saJnples (Figure 5.19). In contrast, Bulbostylis barbata and B. sp. aff. barbata have epidennal cell walls that are very strongly sinuose (Figure 5.20). Samples from the Bulbostylis turbinata group fall between the two extremes (Figure 5.18). EmblYo morphology All species within this study, that fonned the clade Bulbostylis share the general Bulbostylis-type embryo (synapomorphy 150-3; Figure 5.14). Variation in the size and general shape of embryos was observed (Figures 5.21-22). The embryos from B. sp. aff. densa 1 and B. sp. aff. barbata were the smallest sampled, with the very dense cellular contents obscuring visibility of the primordial leaf or leaves (Figure 5.21 B, D). Embryos from B. hispidula subsp. pyriformis (Figure 5.22 A, B) and B. hispidula subsp. senegalensis (Figure 5.22 C, D) are conspicuously larger than embryos of the other species; the basal orientated shoot and root are prominent, and the second primordial leaf is well-developed and almost the same size as the first leaf. The embryo of Bulbostylis pyriformis is slightly smaller than that in B. hispidula and the second primordial leaf was visible, although not well-developed (Figure 5.22 E, F)" Bulbostylis humilis (Figure 5.22 G, H) has an embryo size and internal structure similar to that seen in the B. hispidula specimens. Figure 5.16 Scanning electron micrographs (SEM) showing the variation of nuts for some samples of the Bulbostylis densa group. A. B. densa (bde2) with B. epidermis at higher magnification; C. B. densa (bde3); and D. B. sp. aff. densa 1 (baffd1) with E. epidermis at higher magnification, showing epidermal cells with minute central silica body. Epidermal cell walls are sinuose. Scale bar for A, D=100 J.lm; B, E=50 J.lm; C=200 J.lm. See Table 5.1 and Appendix 1 for OTU and specimen details. Figure 5.17 Scanning electron micrographs (SEM) showing the variation of nuts for samples of the Bulbostylis burbidgeae group. A. B. burbidgeae (bbu5) with B. epidermis at higher magnification; C. B. sp. aff. burbidgeae (baffbu); with D. epidermis at higher magnification; E. B. sp. aff. puberula 1 (baffpu) with F. epidermis at higher magnification, showing epidermal cells forming angular ridges. Scale bars A, D=100 /-!m; B, E=50 /-!m; C=200 /-!m. See Table 5.1 and Appendix 1 for OTU and specimen details. Figure 5.18 Scanning electron micrographs (SEM) showing the variation of nuts for samples of the Bulbostylis turbinata group. A. B. turbinata (bt3) with B. epidermis at higher magnification; C. B. turbinata (bt?) with D. epidermis at higher magnification; E. B. turbinata (bt9) with F. epidermis at higher magnification; G. B. turbinata (btlO) with H. epidermis at higher magnification. OTUs bt9 and btlO formed a group separate to other B. turbinata samples (see Figure 5.2), consistent with the nut differences pictured. Scale bars for A, C, E, G=IOO Jim; B, D, F, G=50 Jim. See Table 5.1 and Appendix 1 for OTU and specimen details. Figure 5.19 Scanning electron micrographs (SEM) showing the variation of nuts for samples from Bulbostylis pyriformis and the B. hispidula complex. A. B. pyriformis (bpy5) with B. epidermis at higher magnification; C. B. hispidula subsp. pyriformis (M. Richards 23175B); with D. epidermis at higher magnification; E. B. hispidula subsp. senegalensis (J T Davey 10) with F. epidermis at higher magnification. All samples have vertically elongated epidermal cells that may be raised so that the nut sculpturing is rugose. Scale bars A=200 11m; C, E=500 11m; B, D, F=50 11m. See Table 5.1 and Appendix 1 for OTU and specimen details. Figure 5.20 Scanning electron micrographs (SEM) showing the differences between nuts of Bulbostylis barbata and B. sp. aff. barbata. A. B. sp. aff. barbata (baffba5) with B. epidermis at higher magnification; C. B. barbata (bba5); and D. epidermis at higher magnification, showing wax plates on the surface that break away to reveal strongly sinuose cell walls around the isodiametric cells, creating a reticulate pattern over the surface of the nut. Scale B, D=50 ｾｭＮ See Table 5.1 and Appendix 1 for OTU and specimen bar A, C=200 ｾｭ［ details. Figure 5.21 A. Embryo morphology in Bulbostylis. B. capillaris (G. Davidse) B. B. sp. aff. densa 1 (baffd1) C. B. sp. aff. barbata (baffba4) D. B. barbata (bba18) E. B. sp. aff. burbidgeae (baffbu) F. B. puberula (G. Davidse 9037). Collector and collection number, or specimen OTU label are given in brackets. Open arrow=shoot, closed arrow=root. Scale bar=100 J.!m (for all images). See Table 5.1 and Appendix 1 for specimen details. Figure 5.22 Variation in Bulbostylis embryo size, shape, and development of second primordial leaf. A. B. hispidula subsp.pyriformis (M Richards 23175B) and B. the welldeveloped second primordial leaf (long arrow). C. B. hispidula subsp. senegalensis (J T Davey 10) and D. the second primordial leaf is well-developed. E. B. pyriformis (bpy2) with a slightly smaller sized embryo and F. second primordial leaf visible but not well developed. G. B. humilis (C.P. Strong et al.) embryo with H. second primordial leaf welldeveloped. Open arrow=shoot, closed arrow=root. Collector and collector number, or OTU label are given in brackets. Scale bar=100 !-lm. See Table 5.1 and Appendix 1 for specimen details. A B c D E F H Figure 5.23 Culm and leaf blade transverse sections in Bulbostylis. A. Culm and B. leaf blade sections of B. capillaris (G. Davidse); C. culm and D. leaf blade sections of B. sp. aff. densa 1 (A.R. Bean 3236); E. culm and F. leaf blade B. sp. aff. densa 1 (Field survey team 820); G. culm and H. leaf blade sections of B. densa (bde2). Collector and collection number, or OTU label are given in brackets. Scale bar=100 !lm. See Table 5.1 and Appendix 1 for specimen details. A B c E F G H Figure 5.24 Culm and leaf blade transverse sections for Bulbostylis. A. Culm and B. leaf blade sections of B. burbidgeae (bbu5). Variation observed in C. culm and D. leafblade sections of B. barbata (K.L. Clarke 187 et al.), and E. culm and F. leaf blade for OTU bbal. G. Culm and H. leaf sections for B. sp. aff. barbata (baffba1). Collector and collection number, or OTU label are given in brackets. Scale bar=100 ｾｭＮ See Table 5.1 and Appendix 1 for specimen details. A B c D Figure 5.25 Culm and leaf blades transverse sections for Bulbostylis. A. Culm and B. leaf blade sections of B. puberula (G. Davidse 9037); C. culm and D. leaf blade sections of B. humilis (C.P. Strong et al.) Collector and collection number are given in brackets. Scale bar=100 J.lm. See Table 5.1 and Appendix 1 for full specimen details. 181 Vegetative anatomy The leaf blade anatomy of all species of Bulbostylis in this study conforms to the C4 fimbristyloid photosYnthetic pathway (Figures 5.23-25). Leaf blades in transverse section are either sub-triangular in outline, or channelled Ｈ｣｡ｮｬｩ｣ｵ｡ｴ･Ｉｾ usually with two shallow or acute ribs. Most species sampled have three vascular bundles within the leaf, occasionally five in B. barbata (Figure 5.24). A hypodermis is absent from the leaves of all sampled species and the adaxial epidermal cells are inflated roughly four times that of the abaxial epidermis. Culms are mostly regularly grooved and almost circular in outline, to irregularly circular and barely wavy (Figures 5.23--25). All species sampled have numbers of vascular bundles equal to the nun1ber of sclerenchyma strands. The vascular bundles form one concentric ring below the epidermal layer. Sclerenchyma strands are often bulbous, forming the ridges of the channels, but may be square to just rectangular (Figures 5.23-25). Discussion The group recovered in phenetic analyses as Bulbostylis sp. aff. barbata is a distinct new species, as indicated by its placement in the combined minor clades B 1 and B2, sister to B. barbata (see Figure 5.14). The variation observed in B. barbata is consistent across the global geographic range. I have not seen the TYPE specimen for B. barbata subsp. pulchella, but the representative samples from India included in this study (Appendix 1) do not differ greatly from the other sampled specimens (as defined in phenetic analyses). To assess the limits of the species and subspecies, it is recommended that B. barbata subsp. pulchella be compared with a wider sample. 182 High levels of homoplasy for many of the Bulbostylis sampled could explain the lack of overall terminal branch support within the cladistic analyses of this chapter. In general, species of Bulbostylis form a monophyletic group sister to the clade of main Fimbristylis species (Fl) and was in direct contrast to placement of Bulbostylis in Chapters 3 and 4, where the two species of Bulbostylis (B. barbata and B. dellsa) were nested within the same Fimbristylis species that form the Fl clade of this analysis. The consistency of the embryo type and vegetative anatomy appears to have been important in stabilising the results, despite extensive homoplasy across many of the morphological characters. The presence and development of the second and third primordial leaves in the embryo could provide a strong character for grouping species into sections if these structures could be viewed in all the sampled embryos. The sampled species of Bulbostylis with larger nuts (B. hispidula, B. humi/is) had larger embryos and a prominent second primordial leaf. Bulbostylis pi/osa falls into this category, having the largest embryo sampled in Van der Veken's (1965) study of BulbosZvlis, and was shown to have the second primordial leaf well-developed and almost as large as the first leaf. Bulbostylis breviculmis (a sYnonYm of B. striatella) was shown to have a poorly developed second leaf (Van der Veken 1965), differing from the embryo observed in B. humilis within this study. These differences question the SYnonon1Y of B. breviculmis, or the consistency, and therefore usefulness as a character, of the development of the second primordial leaf. The smaller embryo (although larger than the other Australian species), and nut size in general, plus the less developed second leaf in the Australian B. pyriformis contribute to maintaining species level status separate from B. hispidula. Clearly there are general similarities in nut shape, epidermal patterning (see Figure 5.19), and 183 synflorescence morphology between specimens of B. pyriformis and the B. hispidllla; however, a more comprehensive study is needed to assess the broad similarities between all entities of the B. hispidula complex and B. pyriformis. Those species with the smaller embryos, where the primordial shoot and root is less prominent, are mostly grouped in the B 1 clade (i.e. B. densa group, B. puberula, and B. capillaris). If the inner organs could be scored, however, the uncertainty with the placement of taxa that currently fall into the B2 clade may be resolved. These taxa, B. sp. aff. barbata, B. sp. aff. burbidgeae, B. sp. aff. puberula and B. burbidgeae, seem 'misplaced' due to general embryo morphology and nut characters. In samples with an abundance of fruits, sectioning embryos elnbedded in paraffin \vax, would allow the scoring of the internal organs to assess specific groups. Prophyllate spikelets, seen within the synflorescence for B. barbata and B. sp. aff. barbata, or prophyllate buds that were present in most of the Bulbostylis samples in the phenetic study, and as reported by Guaglianone (1970), have also been described for some species of Schoenoplectus (e.g. S. calfornicus) and Rhynchospora (e.g. R. corymbosa, R. brownii) (Kukkonen 1986; Vegetti 2003). Extending the sample across species in Bulbostylis is recommended to investigate the usefulness of the character more thoroughly. Spikelets found at the base of the plant have been recorded previously for B. humilis (syn. B. striatella), B. heterostachya Cherm. (Chermezon 1929), B. glaberrima Kilk. (Haines 1971), B. basilis Fosberg, B. schaJJneri (Boeck.) C.B.Clarke, B. sphaerocarpa (Boeck.) C.B.Clarke (Fosberg 1977), and B. funckii (Steud.) C.B.Clarke (Goetghebeur and Grager 1993). Amphicarpy occurs across a 184 number of genera and has been reported in Trianoptiles i.e. T capensis (Steud.) Harv., T solitaria (C.B.Clarke) Levyns (Levyns 1943; Haines and Lye 1977), Schoenoplectus i.e. S. erectus (Pair.) Palla ex J.Raynal subsp. raynalii (Schuyl,er) Lye, S. lateriflorus (J.F.Gmel.) Lye subsp. lateriflorus, S. microglumis Lye, S. articulatus (L.) Palla, S. senegalensis (Hochst. ex A.Rich.) Palla, S. leucanthus (Boeck.) J.Raynal, S. proximus (Steud.) J.Rayna1, Eleocharis i.e. E. minima Kunth (Browning 1992), E. caespitosissima Baker (Bruhl 1994), and Crosslandia (see Chapter 3). The African material of B. densa (baffd2: bde 16, 17, 18) has classic amphicarpic nuts, and when combined with the generally larger anther length, fimbriolate glume margins of the mostly mucronate glumes, and the inllorescence of mainly solitary or reduced anthelodia (anthela of 2-3 rays), forms a group separate to the other B. densa OTUs. It is necessary to compare the African amphicarpic material more broadly with other species of the B. densa complex as per Haines and (1983), and Gordon-Gray (1995) to fully assess the species boundaries. The variation in floret sex of the radical spikelets seen in B. humiliswas reported as common among species of Bulhastylis by Chermezon (1929). Bisexual or female florets nlay occur in the radical spikelets, and the stamen number may be variable compared to the more consistent numbers in aerial spikelets. Spikelets that may be present near the base of the plant as well as the aerial spikelets in B. barbata and B. turbinata do not exhibit amphicarpic features, and are tenned subradical by Wilson (1980). None of the examined overseas specimens of B. barbata have subradical spike1ets, the presence of which is a likely result of the extreme environmental conditions in which they grow. Most of the specimens that exhibit greatly reduced culms have been collected from the dry desert regions of central and north-western Australia (see Appendix 1). Subradical spikelets usually 185 produce fruit earlier than aerial spikelets, but this could be due to the later developlnent of aerial spikelets once conditions are more favourable. Experiments would be needed to test the effects of harsh environments on the production of subradical spikelets in Bulbostylis barbata. Further study could concentrate more specifically on the variation within B. barbata, by expanding the sample size and including molecular data within the study to fully explore the presence of subradical spikelets. RaYnal (1976) used the potential to develop amphicarpy in species of Schoenoplectus to redefine Schoenoplectus section Supini (Cherm.) J.RaYnal. Assessing all the species currently assigned to Bulbostylis could provide similar results, when used in conjunction with data from embryo morphology and anatomy, and general vegetative morphology. Using Clarke's (1908) classification of Bulbostylis, all the species known then to be amphicarpic, are found in sections I and II. The putative new species Bulbostylis sp. aff. barbata can now be named using the results of the phenetic and cladistic data as support. A distinctly smaller nut, di fferent from all other Australian species, and the hairy glume margins of the almost hyaline glumes, supports the recognition of this new species, known only from K.akadu National Park, Northern Territory. Given the limited sample size of the unknown identities, B. sp. aff. densa 1, bt9, btl 0, B. sp. aff. puberula and B. sp. aff. burbidgeae, and the broad variation within the Bulbostylis densa and B. hispidula groups, it seems necessary to explore the limits of taxa in these groups more broadly by increasing the sample size and the nunlber of species in subsequent analyses. Comparing the unknowns to other overseas species is also necessary to exclude the possibility of extended ranges, possibly introduced into Australia via human movement. A new combination is now provisionally put forward prior to valid publication. Nomenclature of Bulbostylis in Australia Genus: Bulbostylis Kunth (nom. cons.) Enumeratio Plantamm 2: 205 (1837) Bulbostylis capillaris (L.) Kunth ex C.B.Clarke in 1.D. Hooker, Fl. Brit. India 6:652 (1885) BasionYll1: Scirpus capillaris L. TYPE: 1. Bulbostylis barbata (RoUb.) C.B.Clarke BasionYll1: Scirpus barbatus Rottb. B. eustachyii Eardley 2. Blilbostylis bllrbidgeae K.L. Wilson 3. Bulbostylis dellsa (Wall.) Hand.-Mazz. BasionYll1: Scirpus densus Wall. Bulbostylis capillaris var. trifida (Nees) C.B.Clarke 4. Blilbostylis humilis (Kunth) C.B.Clarke BasionYll1: Isolepis humilis Kunth Fimbristylis arenaria Nees Isolepis breviculmis Kunth Scirpus arenarius (Nees) Boeck. Bulbostylis breviculmis (Kunth) C.B.Clarke Isolepis humillima Hochst. ex C.B.Clarke Bulbostylis striatella C.B.Clarke Abildgaardia humilis (Kunth) Lye Abildgaardia striatella (C.B.Clarke) Lye 5. Bulbostylis kakadu K.L.Clarke & 1.1.Bmhl sp. nov. ined. 6. Blilbostylis pyriforl11is S.T.Blake 7. Bulbostylis tllrbillata S.T.Blake Specimens with uncertain species limits, B. sp. aff. densa 1, B. sp. aff. densa 2 (African amphicarpic), B. sp. aff. turbinata 1, B.sp. aff. turbinata 2, B. sp. aff. burbidgeae, and B. sp. aff. puberula, need to be assessed against a broader sample of species from Bulbostylis prior to publication. 187 Species ofuncertain standing Bulbostylis pi/osa (Steud.) Beetle nom. illeg. Leaflets of Westem Botany 4: 45 (1944) Basionym: Isolepis pi/osa Steud. Type: Hrbr. Drummond IV nr. 360 (nisi schedula commutata) N. Holi. The TYPE specimen is not located in Australia, and until the TYPE sheet can be examined, placement of this taxon within Australian species cannot be determined. 188 Chapter 6 Testing monophyly of the tribe Abildgaardieae I.Jye Introduction The aim of this chapter is to test monophyly for the tribe Abildgaardieae Lye by subjecting representatives of all genera assigned to the tribe by Goetghebeur (1986, 1998) and Bruhl (1995) (Table 6.1) to cladistic analysis. The genera, Abildgaardia Vahl (the TYPE genus), Fimbristylis Vahl (including Tylocarya Nelmes (Kern 1974; Simpson 1993; Goetghebeur 1998), or Tylocarya treated as a distinct genus (Goetghebeur 1986; Bruhl 1995), Bulbostylis Kunth, Crosslandia W.Fitzg., Nemum Desv. ex Ham., and Nelmesia Van der Veken, are the focus of the study. The general history of the tribe Abildgaardieae was outlined in Chapter 1, as were problenl areas where disagreement on generic boundaries persists. The position of Abildgaardia, assigned as a section (or series) of Fimbristylis (Koyama 1961; Kern 1974; Simpson 1993): as Monostachyae Ohwi, or as a genus (Vahl1805; Kral 1971; Haines and Lye 1983; Goetghebeur 1986; Bruhl 1995; Gordon-Gray 1995; Goetghebeur 1998), is specifically relevant in this study because of the large number of species that occur in, and are endemic to, Australia (see Chapter 4). The status of Tylocarya is either as a monotypic genus (Bruhl 1995), or more generally accepted, as a species of Fimbristylis, F. nelmesii lKern (Kenl 1974; Simpson 1993; Goetghebeur 1998). 189 Although Bulbostylis currently has wide acceptance as a genus, separate fronl Fimbristylis, the distinction is tenuous. I(oyama (1961) classified Bulbostylis as a subgenus of Fimbristylis, and Lye (in Haines and Lye 1983) placed Bulbostylis as a subgenus within Abildgaardia, although both authors have since reverted to using Bulhostylis at the generic rank (Lye 1995; Simpson and Koyama 1998). The genus Nemum is thought to be close to Bulbostylis due to a similar embryo type and the presence of long coarse hairs that may be present at the mouth of the sheath-leafjunction (Raynal 1973); these two genera are otherwise quite different. Arthrostylis R.Br. and Actinoschoenus Benth. have been variously cOlnbined, as Arthrostylis (Kunth 1837; Bentham 1861; Thwaites 1864; Kiikenthal 1(44) within Fimbristylis (Boeckeler 1874; von Mueller 1875; Clarke 1893; Fitzgerald 1918; Ken1 1955; 1974; Latz 1990: recommending placement into Actinoschoenus). Rye (1992), in her treatment of the Kimberley Flora (Western Australia), placed the unnamed species into the reinstated genus Actinoschoenus. Trachystylis S.T.Blake has also been referred to Fimbristylis as F. stradbrokensis (Domin) J.Kern (Ken1 1959). Species and generic limits for Crosslandia (Chapter 3) and Abildgaardia (Chapter 4), and species limits for Bulbostylis in Australia (Chapter 5) were defined in the preceding chapters. A comprehensive assessment of species and generic limits for Bulhostylis and Fimbristylis was not possible because of the large number of species assigned to each genus, c. 200 and c. 300, respectively (World Checklist of Monocotyledons 2004). Representative samples from Bulbostylis and Fimbristylis were included in the cladistic study. To complete the cladistic data set for the tribe Abildgaardieae, data were collected from species of Nemum and from the monotypic genera Nelmesia and Tylocarya. 190 Materials and methods Ingroup To assess monophyly for the tribe, and therefore the relationships of genera within the tribe, all terminal taxa previously defined in the Crosslandia, Abildgaardia and Bulbostylis chapters (3, 4, and 5) were included in the tribal assessment. When combined with samples from Nemum spadiceum (Lam.) Desv.ex Ham., N. megastachyum (Cherm.) J. RaYn., N. equitans (Kuk.) J. RaYn., Nelmesia melanostachya Van der Veken, Tylocarya cylindrostachya Nelmes (= F. nelmesii), and selective species of Fimbristylis, the ingroup represented all genera currently accepted into the tribe Abildgaardieae. Previous analyses revealed that Fimbristylis is not a monophyletic group (see Chapters 3, 4, and 5), however, it was not possible to extend the sample species for this analysis due to time constraints. Representative taxa from Fimbristylis used in earlier work within this thesis were maintained for cladistic analysis in this chapter. A total of 52 species across 8 genera (or 7 if Tylocarya is excluded as a separate genus) fOlmed the basis for the final cladistic study (Table 6.1, see also Appendix I for specimen details). Outgroup Outgroup taxa used to polarise data were unchanged (Table 6.1; see also Appendix 1). The taxa comprised Arthrostylis aphylla, provisional Actinoschoenus compositus, Trachystylis stradbrokensis from the provisional Arthrostylideae (Goetghebeur 1986; Bruhl 1995) or Schoeneae (Goetghebeur 1998), plus Schoenoplectus tabernaemontani (C.C.Gme!.) Palla (= S. validus), Schoenoplectiella lateriflora Table 6.1 Taxa included in cladistic analysis to assess monophyly of the tribe Abildgaardieae. Species from Crosslandia, Abildgaardia, and Australian species of Bulbostylis included here were defined in Chapter 3, 4, and 5 respectively. See Appendix 1 for specimen details. Taxa Ingroup Abildgaardia macrantha (provisional) Abildgaardia mexicana Abildgaardia odontocarpa (provisional) Abildgaardia ovata Abildgaardia oxystachya (provisional) Abildgaardia pachyptera (provisional) Abildgaardia schoenoides Abildgaardia triflora Bulbostylis barbata Bulbostylis sp. aff. barbata Bulbostylis burbidgeae Bulbostylis sp. aff burbidgeae Bulbostylis capillaris Bulbostylis densa Bulbostylis sp. aff. densa 1 Bulbostylis sp. aff. densa 2 Bulbostylis hispidula subsp. pyriformis Bulbostylis hispidula subsp. senegalensis Bulbostylis humilis Bulbostylis puberula Bulbostylis sp. aff. puberula Bulbostylis pyriformis Bulbostylis turbinata Bulbostylis sp. aff. turbinata 1 Bulbostylis sp. aff. turbinata 2 Crosslandia anthelata (provisional) Crosslandia setifolia Crosslandia spiralis (provisional) Crosslandia vaginata (provisional) Fimbristylis bahiensis Fimbristylis blakei Fimbristylis cinnamometorum Fimbrisrylis depauperata Fimbristylisfimbristyloides Fimbristylis furva Fimbristylis hygrophila Fimbristylis microcmya Fimbristylis schultzii Fimbristylis sp L. Fimbristylis variegata Nelmesia melanostachya Nemum equitans Nemum megastachyum Nemum spadiceum Tylocarya cylindrostachya Outgroup Actinoschoenus compositus (provisional) ａｲｴｨｯｳｾｹｬｩ aphylla Schoenoplectiella laevis Schoenoplectiella lateriflora Schoenoplectus tabernaemontani Trachystylis stradbrokensis No. specimens sampled 10 5 2 13 13 11 12 4 20 7 5 1 4 15 2 3 3 2 3 7 1 8 8 1 1 5 18 3 14 4 2 5 2 4 2 2 2 2 2 1 1 2 2 4 1 4 4 5 5 3 7 192 (J.F.Gmel.) Lye (= Schoenoplectus lateriflorus) and Schoenoplectiella laevis (S.T.Blake) Lye (= Schoenoplectus laevis) in the tribe Scirpeae (Bruhl 1995) or Fuireneae, (Goetghebeur 1986, 1998) depending on the system of classification accepted. Characters and homology Additional characters were added to the cladistic data set when species from Nemum, Nelmesia and Tylocarya were sampled. These new characters were mainly associated with the spikelets, i.e. persistence of glumes to the rachilla in species of Nemum, presence of an intraspicular prophyll in Nelmesia, and variation in anatomical attributes (Appendix 2). Embryo morphology and anatomy Most specimens on loan for Nemum and Nelmesia (ISOTYPE!) could not be sanlpled for characters from leafblade and culm anatomy, or embryo morphology due to the limited amount of material available; missing data (mainly embryographical and anatomical data) were obtained from the literature (Van der Veken 1965; Metcalfe 1971; Rayna11973; Goetghebeur 1986). Prepared slides for leaf blade and culm anatomy were available for Tylocarya frOlTI a previous study by Bruhl (1990), however, embryo morphology for Tylocarya was obtained from Van der Veken (1965) and Goetghebeur (1986). Only material that was already loose on the sheets was used to obtain floret measurements, and for embryo morphology and SEM treatment (not TYPE sheets), ifneeded. Sampling across the taxa allowed leafblade and culm anatomy to be compared where possible. Scanning electron microscopy of the nut epidermis enabled the comparison of nut 193 characters, especially the micromorphological attributes e.g. epidermal cell shape and type 0 f protuberance, between taxa. Analyses Data for 55 terminal taxa and 152 characters were subjected to parsimony analsysis within PAUP* using heuristic techniques (hsearch swap=TBR addseq=rand nreps= 1000 hold=5 multrees=yes). Branch support was assessed using Bootstrap analysis (1000 bootstrap replications), as the computational time required to calculate the Bremer support indices past 3 extra tree length steps was protracted, even when limiting the addition-sequence replications to 10. Characters were traced in MacClade and the most relevant characters presented in the cladogram. Results Cladistic analysis A. heuristic search produced 91 most parsimonious trees (Tree length=1482, CI==0.3947, HI=0.6053, RI=0.5814, RC=0.2295). Tree 1 was one of the retrieved trees with similar topology to the strict consensus, and was selected to show branch support and character/branch associations. Taxa from the' Arthrostylideae', used with Schoenoplectus (including Schoenoplectiella) in the outgroup for Chapters 3, 4, and 5, violated the assunlption for monophyly of the ingroup in this analysis. All taxa from the ingroup, plus members of the provisional Arthrostylideae, formed a broad clade sister to the outgroup species of Schoenoplectus, with strong branch support indicated (Bootstrap=87%) (Figure 6.1). Arthrostylis aphylla and Actinoschoenus compositlls . 0 Schoenoplectu ｾ［ tabernaemontani 1' 109-2 III' 13-3; 28-2 44-1; 73-3 151-5; 152-12 11 11 -2; 74-3; 123-1,2 90 II 1 11 25-1; 73-4 II ＷｾＲ II F.mbristylis depauperata 1 11 11-2; 13-2 II ;j 0 ... '- C) ;j 0 Nemum megastachyum 88 • 151-4 C Q. SchoenoplectiE lIa lateriflora Schoenoplectiella laevis Nemum spadiceum "'11 111111 11I11 Tylocarya cylindrostachya 111122-1,2; 152-1 I F1 III Bulbostylis hurnilis 11119-1,2; 74-3; 123-1 II Bulbostylis barbata ｾ "19-1,2 74-1,2 Bulbostylis sp. aft. turbinata 2 11111111 '19-2 ｾＢｉＱｾ 81 .......· . : 1 ··. ·"0.. .... : Bulbostylis pyr,formis 19-2 • 0 • 0 Bulbostylis sp. aft. puberula ·.0 .. "r .. ' ｾＶ［ＹＱ I: .! Bulbostylis turlJinata 174 -12 o : .... Bulbostylis sp. aft. turbinata 1 119-2 25-1 86-2 100-2,5 l117-5 8 122-1 98-2 151-3 152-4 28-1 Bulbostylis kakadu Bulbostylis burbidgeae 74-12 A I/) 0 .0 :i B. hispidula sutsp. senegalensis 122-2 .1 74 -4 (Xl Bulbostylis sp. <1ft. densa 2 ···1 ............i :.1 19-1 Z B. hispidula subsp. pyriformis : ｾｉﾷ 19-2 .... . ·1 Ｎｾ .!!! Bulbostylis sp. aft. burbidgeae 19-2 0 Bulbostylis densa Bulbostylis sp. 0 : 641 3ft. densa 1 Bulbostylis capillaris B. puberula Fimbristylis fimlJristyloides II 11 Fimbristylis sctlultzii 151-1 152-2 III ｾ 12lt ＳＭＲＵＱｾ Fimbristylis furva Fimbristylis dis ticha 79 111 ＱｉＲＬｾＭ［ｭｩＬｾｲｯ｣｡ｹ Fimbristylis. bahiensis 152-1 ｾＧＱＳＭ［ 151-7 Firnbristylis "11-2; 13-2 II I F2 Flmbristylis cinnamometorum 13-2; 73-2; 152-2,6 5-L Fimbristylis variegata 152-6 M 6€ Actinoschoenus compositus X Arthrostylis aphylla X 28-2 73-1; 152-9 III 1 '11 19-2; 123-2; 152-1,7 I B 53 II '151-1 Fimbristylis blakei 1123-1 I III '25-2 Crosslandia vaginata 11174-1,3; 151-1; 152-6,7,8 II Abildgaardia macrantha Abildgaardia ovata p0.- I .. riJ" Abildgaardia oxystachya T152 -8 111111 .. 100 C2 Fimbristylis hygrophila 28-2 151-1 - --. Fimbristylis sp, L Nelmesia melanostachya 86-6 100-5; 151-3; 152-4 111 109-1' 130-5 61 _ ［ＲＭＶＢＵｾ * Abildgaardia schoenoides ' Abildgaardia sp. aft. schoenoides ｾ .... 86187130 ｾ 151 b .. 152 17 Ifo 64-1 Abildgaardia odontocarpa L.-4 1152-11 I rT137-2 55 •. .. '137-3 -'152-11 Abildgaardia mexicana ＱＬｾＡＮｉＺｬＭｴｩｪ 151-1 1526 14 I Nemum equitans 52-4 ｾｉｊＡｖ［ＱｫｩＺ '" Abildgaardia sp. aft. pachyptera III 25-3; 26-2 ........ "'74 3 Abildgaardia pachyptera Abildgaardia triflora 58 ., 1 1152-5 Abildgaardia sp. aft. odontocarpa Trachystylis stradbrokensis 2-1 Crosslandia s:Jiralis Crosslandia setifolia 75 Crosslandia anthelata X 195 were nested with the species F. variegata and the C 4 Fimbristylis bahiensis, and were sister to the F3-Abildgaardia-Crosslandia clade. Within this clade Trachystylts (also assigned to the outgroup) was paired with Nemum equitans as a group sister to the Abildgaardia clade. Two main clades, A and B, were sister to the clade Fimbristylis depauperata- Tylocarya cylindrostachya (clade C), and all were sister to the Nemum .spadiceumN. megastachyum clade that showed strong branch support (Bootstrap=87%). The species of Nemum sampled, therefore, did not form a monophyletic group. In clade A, all the species of Bulbostylis sampled formed a monophyletic group, with moderate branch support (Bootstrap=77%), sister to the clade Fimbristylis 2 (F. jimbristyloides, F. schultzii, F. cinnamometorum, F. furva, F. disticJza, F. microcarya). Clade B, however, contained the smaller F3 clade (F. s1'. L. and F. blakei - with weak support), which was sister to the Abildgaardia-Crosslandia clade, which included Fimbristylis hygrophila (= Abildgaardia hygrophila), Nelmesia, Trachystylis and Nemum equitans. The previously monophyletic Abildgaardia (Chapter 4) was rendered nonmonophyletic by the placement of Nelmesia melanostachya, which was nested within the Australian endemics. The Abildgaardia-Nelmesia branch showed weak Bootstrap support (67%) and the terminal arrangement of taxa did not collapse in the consensus tree, as indicated by solid lines in Figure 6.1. There are no obvious sYnapopmorphies that unite Nelmesia and Abildgaardia other than the large stipitate nut. 1% Species assigned to Crosslandia in Chapter 3 formed a monophyletic group that included C. setifolia, and the provisional C. anthelata and C. spiralis, and received moderate branch support (Bootstrap=73%). Crosslandia vaginata (= Ahildgaardia vaginata) was basal in the sister group to Crosslandia in this tribal analysis (Figure 6.1) and the Bu/bosty/is treatment (Chapter 5), although there was no internal support for the placement here. Fimbristylis hygrophila (= Abildgaardia hygrophila) persists as a sister to the species of Abildgaardia and Nelmesia melanostachya in the tribal analysis, however, internal branch support was absent for the placement, as was support for the placement of Nemum equitans and Trachystylis. Characters There are no unambiguous synapomorphies that clearly delimit the internal clades due to the poor tree resolution caused by the high level of homoplasy within the data set. Characters from embryo morphology, such as the Schoenoplectus-type embryo (151-5), the germination pore parallel to the first primordial leaf (154-1), plus anatomy of the culm (44-1, 51-1), separate the outgroup species of Schoenoplectus and Schoenoplectiella from the remaining taxa. The only strongly robust group, other than the outgroup (i.e. Schoenoplectus), is the clade of Bulbostylis. The synapomorphies for the Bulbostylis clade are: pilose hairs at the sheath-leafjunction (11-3); leaf vascular bundle number 5 or less (25-1); a minutely triangular style (117-5); style base persistent on the nut (always separates from the style) (122-1), although the specimens of B. hispidula may have deciduous style bases (it falls in tact with the style) (122-2); and Bulbostylis-type embryo (1514). 197 Observations Injlorescence-synjlorescence The inflorescence-synflorescence structure is very homoplastic, even when the structure of the head of sessile spikelets is broken down into different structural types. The simplest 'head' of3 sessile spikelets (57-4) occurs in Abildgaardia (A. mexicana), Fimbristylis bahiensis and Bulbostylis (B. humilis) (see Figure 4.12). A synflorescence head fonned as a compressed spike (multiple primary sessile coflorescences, of one spikelet per coflorescence, where the tenninal spikelet sits above the sessile coflorescences (57-5), was observed in Fimbristylis (F. schultzii) and Actinoschoenus. The multiple branched reduced anthelodia, where all spikelets are sessile (branching as rays) is highly reduced but discemable under the dissecting microscope (57-8), is seen in Crosslandia setifolia and Arthrostylis aphylla (see Figure 3.16). Crosslandia setifolia was the only taxa sampled that produced lateral heads (see Figure 3.17), where a primary coflorescence has developed into a secondary main florescence of a head of sessile spikelets. The 'prophyllar' head is a combination of the multiple branched reduced anthelodium and prophyllate branching from primary and sometimes secondary inflorescence branches (57-10), (see Figure 5.15 B, C); this type is restricted to Bulbostylis in this study (B. barbata and B. kakadu). The only other genus in the study with intraprophyllar growth within the inflorescence is Schoenoplectus-Schoenoplectiella, which differs in structure through the paniculodium base plan. In Schoenoplectiella laevis, prophylls were not restricted to the production of spikelets, as some prophylls were fertile, possessing a solitary nut in the axil without any other bract visible; unique for taxa within the study. The most common synflorescence type was the reduced anthelodium with a 198 sessile main primary florescence and primary coflorescences supported on rays (lengthened epipodia) (see Figures 3.14, 4.12, 4.15 A), which was found across most genera of the ingroup. Florescence ramification (57-7) within the synflorescence was common in members of Fimbristylis (F. blakei, F. sp. L., F. cinnamometorum; welldeveloped in F. microcarya, F. depauperata, F. furva and sometimes F. cinnamometorum; and in some species of Bulbostylis (e.g. B. densa, B. pyriformis, and may be well-developed in B. puberula and B. hispidula subsp. senegalensis). The solitary spikelet is the simplest of all the inflorescence-synflorescence structural types, and, within the study, is most common in Abildgaardia (see Figure 4.12 A). The spike in Nelmesia melanostachya superficially resembles a solitary spikelet, but the intraspicular prophyll within the solitary spike is exceptional for the tribe (Figure 6.2). The prohpyll is in place where a lateral branch arises as a solitary, sessile floret (i.e. single floret spikelet); this inflorescence type is not homologous with the solitary spikelet in Abildgaardia. Despite the difference in the structure of the inflorescence, Nelmesia was placed with species of Abildgaardia in many of the trees retrieved, although there was no support for the placement (Figure 6.1). Florets within all the studied taxa comply with the basic scirpoid floral arrangement: being tetracyclic, 2-3 carpels, 1-3 stamens, 0-6 perianth bristles enclosed by a glume (floral bract) (Vrijdaghs et a1. 2005). Perianth was absent frOlTI all the sampled ingroup taxa, with the exception of one collection of Abildgaardia schoenoides (see Chapter 4). Perianth may be present in SchoenoplectusSchoenoplectiella as bristles with retrorse barbs; Schoenoplectus tabernaemontani (= S. validus) has three perianth bristles present (see Figure 6.8 A). The unique perianth in the Abildgaardia schoenoides collection differed in having 2 perianth bristles with antrorse barbs (see Figure 4.13). Figure 6.2 Nelmesia melanostachya ISOTYPE showing general habit, including solitary spikes where the lateral spikelet is reduced to a single floret. The insert shows a single floreted spikelet bearing a mature nut; the large prophyll is obvious and sits between the nut and the rachis. Scale bar=2 mm. See Appendix 1 for specimen details. 200 Nut epidermal pattern The nut epidennis varies in the shape and orientation of epidennal cells, or sculpturing from single silica bodies (puncticulate to granulate), groups of multiple raised cells fonning various shapes (turbercules), cells raised in ridges that may be broken or continuous (rugose), raised cell walls (reticulate), or cells with a sunken lun1en (pitted), and is distinct at the species level. There is some consistency in the epidennal cell size and shape as seen in the group Abildgaardia - excluding Nelmesia, (see Chapter 4), where epidennal cells are distinctly rounded (A. ovata, A. oxystachya, A. schoenoides, A. sp. aff. schoenoides, A. odontocarpa, A. sp. aff. odontocarpa) or barely hexagonal in shape (A. pachyptera, A. macrantha, A. triflora and A. mexicana). Large tubercules are common across the species and vary in size and shape, and occasionally may be few or absent in A. pachyptera, A. macrantha and A. triflora. Cell walls are not sinuose. The Bulbostylis species sampled may have nut epidennal cells that are isodiametric or longitudinally rectangular, and barely rectangular to linearly rectangular in shape (see Chapter 5). Cellular protuberances may be absent (e.g. B. barbata, B. kakadu) or individual cells may have a central raised silica body producing a puncticulate or granulate surface, depending on the size of the silica body (e.g. B. densa, B. burbidgeae, B. turbinata). Alternatively, the longitudinal rectangular cells are raised to some extent, giving degrees of rugose patterning as transverse wrinkles that may be continuous or broken (e.g. B. puberula, B. pyriformis, B. hispidula). The cell walls are sinuose to some extent (finely-distinctly) in all but B. humilis. 201 Even in the limited Fimbristylis sample the variation in the nut epidetmis is evident. Epidermal cells vary from isodiametric hexagonal cells to barely circular, longitudinally rectangular, or transversely rectangular; cell walls may be straight, barely sinuous to sinuous. Protuberances, as single raised cells to multiple raised cells with various distribution patterns over the surface, occur frequently across the species sampled. Nut epidermal features do not seem to influence the group arrangement for the species of Fimbristylis F2 (Figures 6.3-4), although the poor resolution in the tree topology could mask the usefulness of the character at the sectional level. There are no similarities in the nut epidermal sculpturing for the paired Tylocarya and Fimbristylis depauperata (Figure 6.5). The nut in Tylocarya is smooth, with hexagonal shaped epidermal cells that have strongly sinuose walls, and contrasts with the striated epidermal cells in F. depauperata. The nut epidermis in Nemum spadiceum, N. megastachyum and N. equitans is distinct in the completely smooth nut surface (Figure 6.6) that is lustrous and coloured black, dark brown or grey-brown. Taxa from the Arthrostylideae (Arthrostylis, Actinoschoenus and Trachystylis) that fall with the ingroup in this analysis have variable nut characters (Figure 6.7). The most striking characters are the bulbous base of the nuts in the provisional Actinoschoenus compositus (= Fimbris(ylis composita) (Figure 6.7 C), and the minutely papillose epidermis (Figure 6.7 D). Some species of Fimbristylis have an external gynophore that is brown and attached at the base of the nut (e.g. F. depauperata, F. fimbristyloides, F. schultzii and F. bahiensis), however, none is as large as the brown spongy structure seen in Actinoschoenus compositus. The papillose nut epidermis also in A. compositus has not been seen among any of the other taxa studied. Figure 6.3 Scanning electron micrographs (SEM) showing the variation of nut outline and epidermal sculpturing in some species of Fimbristylis. A. Nut for F. furva (ff2) and B. epidermal sculpturing at higher magnification, with cells irregularly longitudinal and cell walls that are distinctly sinuose. C. Nut for F. microcarya (K.L. Clarke 319, L. Little) and D. at higher magnification, showing epidermal cells that are horizontally elongated and cell walls that are very fmely sinuose. The waxy covering is not plate-like but continuous over the surface. E. Nut for F. schultzii (K.L. Clarke 153 et al.) and F. at higher magnification, showing epidermal sculpturing and hexagonal cell shape. Scale bars A=100 Jlm; C, E=200 Jlm; B, D, F=50 Jlm. See Appendix 1 for OTU and specimen details. Figure 6.4 Scanning electron micrographs (SEM) showing the variation of nut outline and epidermal sculpturing in some species of Fimbristylis. A. Nut for F. disticha (fd2) and B. epidermal sculpturing at higher magnification, with cells mostly circular and straight cell walls. C. Nut for F. cinnamometorum (fc5) and D. at higher magnification, showing epidermal cells that are horizontally elongated and protuberances also elongated horizontally. The waxy covering is not plate-like but continuous over the surface. E. Nut for F. fimbristyloides (ffi3.) showing the truncate base and F. at higher magnification, epidermal sculpturing and hexagonal to circular cell outline. Scale bars A, C, E=100 11m; B, D, F=50 11m. See Appendix 1 for OTU and specimen details. Figure 6.5 Scanning electron micrographs (SEM) and light micrograph (LM) showing the variation of nut outline and epidermal sculpturing in species of Fimbristylis (including Tylocarya). A. SEM of nut for F. blakei (fbI) and B. epidermal sculpturing at higher magnification, with cells mostly hexagonal to circular in outline. C. LM of nut for F. depauperata (K.L. Clarke 305, L. Little) showing the square epidermal cells arranged in rows. D. SEM of nut for Tylocarya cylindrostachya (A.F.G. Kerr 21294) (= F. nelmesii) showing the nut outline and E. at higher magnification, the epidermal surface that is smooth with hexagonal cells with sinuose walls. OTU or collector and collection number are given in brackets. Scale bars A, D=200 /lm; C=20 /lm; B, E=50 /lm. See Appendix I for OTU and specimen details. Figure 6.6 Scanning electron micrographs (SEM) showing the nut outline and epidermal surface in two species of Nemum. A. Nut for N. spadiceum (E.A. Robinson 4677) and B. epidermal aurface at higher magnification. C. Nut for N megastachyum (Germain 4420) and D. at higher magnification. The nuts of both species are similar in outline and the epidermal surface is lineolate (marked with fme lines), as the cells are barely discemable at higher magnification. Collector and collection number are given in brackets. Scale bars A=lOO Jim; C=200Jim; B, D=50 Jim. See Appendix 1 for OTU and specimen details. Figure 6.7 Scanning electron micrographs (SEM) showing the variation of nut outline and epidermal sculpturing in species from the provisional tribe Arthrostylideae. A. Nut for Arthrostylis aphylla (G.N Batianoff10089) and B. epidermal sculpturing at higher magnification, with cells circular in outline. C. Nut for provisional Actinoschoenus composita (K.L. Clarke 178 et al.) and D. at higher magnification, showing the small papillae that cover the nut surface, which are unique among taxa within the study. E. Nut for Trachystylis stradbrokensis (E.J Thompson 78) showing the nut outline and E. at higher magnification, the epidermal cell pattern is reticulate-foveate. Collector and collection number are given in B, D, F=50 Jlm. See Appendix 1 for specimen details. brackets. Scale bars A, C, E=500 ｾｭ［ Figure 6.8 Scanning electron micrographs (SEM) showing the variation in nut outline and epidermal sculpturing for outgroup species of Schoenoplectus and Schoenoplectiella. A. Nut for Schoenoplectus tabernaemontani. (K.L. Wilson 4278) and B. epidermal surface at higher magnification, with cells narrowly elongated in outline. Perianth with retrorse barbs occur in this species. C. Schoenoplectiella lateriflora (P.K. Latz 3761) showing the rugose nut surface and D. the vertically linear epidermal cells at higher magnification. E. Nut for S. laevis (P.M Milthorpe 1777A, G. M Cunningham) showing the smooth epidermis and F. at higher magnification. Collector and collection number are given in brackets. Scale bars A=500 ｾ［ C, E=200 Jim; B, D, F=50 Jim. See Appendix 1 for OTU and specimen details. 208 In the outgroup species sampled for Schoenoplectus and Schoenoplectiella, the nut shows variable nut sculpturing (Figure 6.8). Schoenoplectus tabernaemontani and Schoenoplectiella laevis have smooth nuts, while the nuts in S. lateriflora are tightly rugose. All nuts for the three species have vertically linear cells, which are raised in S. lateriflora. Embryo In species of Fimbristylis, the Fimbristylis-type embryo, although consistent in the orientation of the primordial shoot and root, is variable in size and shape across the species sampled (Figure 6.9). Tylocarya cylindrostachya (or ｆｩｭ｢ｲｳｾｶｬｩ nelmesia) was shown to have a variant of the Fimbristylis-type embryo (Goetghebeur 1986) that is a sY11apomorphy for the Tylocarya-F. depauperata clade (Figure 6.1). Species of Crosslandia (including C. vaginata) also share the Fimbristylis-type embryo and are placed separate to species currently assigned to Fimbristylis (see also Chapter 3). The embryo type in species of Nemum is not typical and varies between the Abildgaardia-type and Bulbostylis-type (Figure 6.10) (see also Chapters 4 and 5). The primordial shoot and root are of roughly equal size, or the shoot may be slightly larger (as in the Abildgaardia-type); the embryo size itself is closer to the Bulbostylis-type. In Nemum equitans the embryo is similar in outline to the Bulbostylis-type and is trigonous from the top view of the embryo (Figure 6.10), although the root is not prominent and the second leaf not detectable, possibly obscured by the cellular contents surrounding the organs. The elliptic rather than rounded or trigonous outline in the top view of the embryo in sampled Ii. spadiceum and N. megastachyum specimens coincides with the di-stigmatic style of both species; Nemum equitans has tri-stigmatic styles. The variation of the eJnbryo 209 features seen in Nemum was reflected in the analysis, as Nemum equitans and Trachystylis stradbrokensis fonned a minor clade, well removed from the other species of Nemum, despite having different embryo types - the Nemum-type and Carex-type (Figure 6.10) respectively. Nelmesia melanostachya is shown to have a variation of the Abildgaardia-type embryo (Van der Veken 1965; Goetghebeur 1986) (Figure 6.10, see also Chapter 4), where the primordial shoot is basal and larger than the parallel root. The Inain difference between the Nelmesia-type and Abildgaardia-type embryos is the size, although the embryo of Nelmesia in this study has been extrapolated from Van der Veken (1965) and Goetghebeur (1986) and, therefore, may not be a true representation of the embryo size. The Nelmesia-type embryo is an autapomorphy for Nelmesia and disrupts the Abildgaardia-type embryo sYnapomorphy for the species otherwise grouped as Abildgaardia (see also Chapter 4). The Bulbostylis-type embryo was one of the sYnapomorphies for the Bulbostylis clade (also see Chapter 5), in contrast to the Fimbristylis-type embryo, which was homplastic across clades C, A, and B (Figure 6.1) and variable across the sampled taxa. Taxa in the Arthrostylis-Actinoschoenus s.l. clade share the Schoenus-type embryo (Figure 6.10) as a sYnapomorphy on the internal branch that unites the four taxa (Figure 6.1). A F G H Figure 6.9 Light micrographs of whole cleared embryos showing the variation in shape and size for some species assigned to Fimbristylis, plus schematic embryos for Tylocarya and Nelmesia. Fimbristylis depauperata (KI. Clarke 305, I. Little) A. side view, B. frontal view, and C. top view of shoot with the second primordial leaf in view (thin arrow) directly behind the first leaf. D. F shultzii (KI. Clarke 108 et al.), E. Ffurva (KI. Clarke 210 et al.), F. F disticha (fdl) and G. F cinnamometorum (fc2) share the Fimbristylis-type embryo. H. The embryo for Tyolocarya is a variant of the Fimbristylis-type. I. In Nelmesia the embryo is a variation of the Abildgaardia- and Bulbostylis-types. Scale bars=lOO Ｎｭｾ Solid arrow=root, open arrow=shoot, thin arrow=second primordial leaf. OTU label or collector and collection number are given in brackets. See Appendix 1 for specimen details. Embryo schematics H and I are adapted from Vander Veken (1965) and Goetghebeur (1986). Figure 6.10 Light micrographs of whole cleared embryos for some species from the outgroup used in cladistic analyses: Actinschoenus, Trachystylis and Schoenoplectiella. A. Actinoschoenus composita (K.L. Clarke 178 et al.) has the Schoenus-type embryo (indicated by the arrows) with the embryo outline wide and saucer shaped in side view, B. Trachystylis stradbrokensis (S T Blake 13201) has a Carex-type embryo with a widened cotyledon. C. The distinctive Schoenoplectus-type embryo is shown for Schoenoplectiella laevis (K.L. Wilson 8041 et al.), where the cotyledon extends past the primordial shoot, and D. at higher magnification showing the genn pore parallel to the first primordial leaf. Scale Solid arrow=root, open arrow=shoot. Collector and collection number are bars=100 ｾｭＮ given in brackets. See Appendix 1 for specimen details. 212 Anatomy Leafblade and culm anatomy show 47 of the 55 taxa sampled, all from the ingroup, share the C 4 fimbristyloid photosynthetic pathway. The C 3 photosynthetic pathway arises several times amid the ingroup in the tribal analysis, and taxa with C 3 anatomy (Arthrostylis aphylla, Actinoschoenus compositus, Fimbristylis variegata and Trachystylis stradbrokensis (Figure 6.11) are placed with C 4 species (Fimbristylis bahiensis and Nemum equitans, respectively). Despite variation in the general shape of transverse sections of leafblade and culrn, in the number of vascular bundles, the shape and number of sclerenchyma, and the shape and arrangement of parenchyma among the C 4 species, there was a general consistency with all having only sclerenchyma strands in leaf blades and culms. The exception is seen in the sections of Tylocarya (Figure 6.12). Leafblade anatomy in Tylocarya shows similar structure to some of those seen in Fimbristylis and Abildgaardia. In addition to the usual abaxial row of sclerenchyma strands that occur below the epidermis, Tylocarya varies in that adaxial strands of sclerenchyma are present and associated with the largest vascular bundles (four in this sample, excluding the usual comer support) (Figure 6.12). Adaxial strands were observed in only one other species in this study, and that was F. fimbristyloides (Figure 6.12). Culm anatomy in Tylocarya, however, is distinct from any of the other specimens sampled; there are many layers of vascular bundles arranged in rough concentric rings (3 developed and the 4th newly formed), decreasing in size as the newer bundles develop below the outermost tissue layers. The large dome-shaped bundles of support sclerenchyma are in direct contact with many of the newest vascular bundles in the outermost ring, and are clearly girders and not strands. The vascular 213 bundles have extra sclerenchyma support, with an inner cap of approximately six strands, with son1e extra rows of sc1erenchyma supporting the largest bundles that have been pushed inwards (Figure 6.12 B). Nemum spadiceum was the only species of Nemum sampled for anatomy. The culIn is highly sc1erified, showing an almost continuous undulating band of fibres. The central pith is absent and prominent air spaces occur between most of the vascular bundles. The number of vascular bundles arranged in the single ring corresponds to the crescentiform section of the undulations, with stomata protruding through and above the sclerenchyma fibres in the narrow fibre regions (Figure 6.12 E). The leaf blade outline in transverse section is almost elliptic, possessing only four vascular bundles (not presented). The hypodermis is restricted to a couple of cell layers and is three to four cells wide in the adaxial central region. The leaf margins in both Nemum and Nelmesia are folded in and joined at the leaf sheath junction (see Metcalfe 1971). The transverse culm outline in Arthrostylis aphylla is four-sided. Tannins are present within the epidermal cells, forming a broken line between the strands of sclerenchyma. The vascular bundles are found around the perimeter of the culm, and do not correspond in number to the many small mounds of sclerenchyma strands (Figure 6.11 A, B). Actinoschoenus compositus is the only sampled species with sclerenchyma girders and strands present within the culms. The prominent ribs have equally prominent thickly V-shaped or crescentiform strands adjacent to the smaller vascular bundles (Figure 6.11 C, D). The deep channels have twin stomata at the sides near the base of the channel, and hairs are prominent near the outer margin. The epidermal layer is A c Figure 6.11 Culm and leaf blade transverse sections for some species from the provisional tribe 'Arthrostylideae' selected as outgroup taxa for use in cladistic analyis, showing the typical outlines, arrangement of sclerenchyma strands per vascular bundle, and C3 anatomy. A. Arthrostylis aphylla (A. Gunness AGL1965) 4-sided culm and B. at higher magnification, showing the greater number of small mounded sclerenchyma strands than vascular bundles. C. Actinoschoenus composita (K.L. Clarke 178 et at.) culm that is regularly, deeply ribbed, with vascular bundles alternately large and small; at higher magnification D. thin rectangular girders are associated with the larger bundles and large crescentiform sclerenchyma strands are opposite the smaller vascular bundles. Twin stomata oppose each other at the base of each channel formed by the rib (indicated by the arrow heads). E. In Trachystylis stradbokensis (s. T Blake1320l) the culm is distinctly triangular in outline and F. the leaf blade is subtriangular, with vascular bundles completely immersed within the chlorenchyma. Scale bars A, C=500 Jim; B, E, G=20 Jim; D, F=50 Jim. Collector and collection number are given in brackets. See Appendix 1 for specimen details. B G Figure 6.12 Culm and leaf blade transverse sections for Fimbristylis, Tylocarya, and Nemum. A. Culm and B. leaf blade sections of Fimbristylis depauperata (K.L. Clarke 305, L. Little). Tylocarya cylindrostachya (A.G.F. Kerr 21294) (= F. nelmesii) culm sections at C. low magnification and D. at high magnification, plus leaf blade sections at E. low magnification and F. at higher magnification, showing the variation and detail of vascularisation. G. Culm section for Nemum spadiceum (E.A. Robinson 4676) showing the continuous ring of undulating sclerenchyma. Collector and collection number are given in brackets. See Appendix 1 for specimen details. Scale bars A, D=20 Jlm; B, E= 500 Jlm; C, F, G=50 Jlm. 216 densely stained with tannins, seen as a continuous line, and has a thick cuticle layer (Figure 6.11 E). In Trachystylis, both the leaf and culm are triangular in outline, and both are dense with tannin-filled cells. Some tannin is deposited in the epidennal tissues in both culms and leaves, although not so densely. The three vascular bundles of the leaf blade are completely surrounded by the chlorenchYma (Figure 6.11). Tannin deposits in the epidennallayer do not occur in any of the other taxa sampled, including the outgroup, and were only found within the members of the provisional Arthrostylideae. In all of the other taxa studied, the tannin deposits were mostly observed within the chlorenchYma tissue. Discussion It is not possible to draw substantial conclusions from the main cladistic analysis, other than that the tribe Abildgaardieae, as it is currently accepted by Bruhl (1995) or Goetghebeur (1998), does not fonn a monophyletic group. The weak branch support in earlier analyses (see Chapters 3, 4, 5) for Arthrostylis aphylla, ActinoschoeJlllS compositus and Trachystylis stradbrokensis with the outgroup clade gave an indication of the instability of the outgroup. Including Eleocharis within the current outgroup could resolve this instability, however, the lack of leafblade characters and the solitary spikelets limit the usefulness of the genus as an outgroup here. Ghamkar et a1. (2006, in press), in a molecular study of the tribe, found that samples from Actinoschoenus (Fimbristylis composita Latz), K.L. Clarke et al. 214, K.L. Clarke et al. 213 (NE and NSW) and Arthrostylis, K.L. Clarke et al. 212, K.L. Clarke et al. 183 (NE and NSW), all collected from Northern Territory, were nested within Fimbristylis; surprising considering the major ditTerences seen in this study for 217 vegetative anatomy and embryo morphology bet\veen the sampled Actinoschoenus and Arthrostylis, and the species of Fimbristylis (see Figures 6.9-10 and 6.11-] 2). Clearly, Arthrostylis, Trachystylis, plus overseas and Australian Actinoschoenlls need to be assessed more thoroughly to fully determine their position, especially considering the taxonomic history of these taxa as species of Fimbristylis. Species of Bulbostylis and Fimbristylis in clade A of this study are all C 4 taxa, however, any conclusions drawn regarding uniformity of the photosynthetic pathway across these genera are limited by the small samples sizes used in this study. The C 3 taxa that occur within the predominantly C 4 clade 13 of the Abildgaardia-Crosslandia group do not show any relative grouping patterns to explain their positioning within the cladogram. Stock et al. (2004) demonstrated a connection between phylogeny and geographical distribution in the tribes Cypereae, Scirpeae and Schoeneae; the sample size for the Abildgaardiaeae was too small to be informative. Schoenoplectus has both C 3 and C 4 species in the genus (Stock et al. 2004), however, Bruhl and Wilson (2005, in press) suggest that the C4 S. pulchella sampled by Stock et al. may be rnisidentified, as all other species of Schoenoplectus are reportedly C 3 . The species of Shoenoplectus and Schoenoplectiella sampled in this study i.e. Schoenoplectus tabernaemontani (= S. validus), Schoenoplectiella lateriflora (= Schoenplectus lateriflorus) and Schoneoplectiella laevis (= Schoenoplectus laevis) are all C 3 taxa. Alternatively, Schoenoplectus pulchella may be misplaced. The study by Bruhl and Wilson (2005, in press) reports the presence of C 3 and C 4 species in Abildgaardia. However, A. hygropyhila is the only C 3 species currently accepted in the genus, and is misplaced in Abildgaardia (see Chapter 4). Future studies in Fimbristylis could determine if C 3 species such as Fimbristylis variegata (with a Schoenus-type embryo and a prior history with Abildgaardia) should be removed 218 frOlTI Fimbristylis that is currently accepted as containing both C 3 and C 4 species (Bruhl and Wilson 2005, in press). The placement of Tylocarya with the representative species for the ｆｩｭ｢ｲｳｾｹｬｩ TYPE section, Fimbristylis section Fimbristylis (F. depauperata) supports the current acceptance of Tylocarya as a species of Fimbristylis (F. nelmesii) (Kern 1974; Sinlpson 1993; Goetghebeur 1998). The differences in the culm anatomy and the variation in the Fimbristylis-type embryo need to be resolved against a broader Fimbristylis sample, especially when these two species have separated from the other species of Fimbristylis to be sister to most of the ingroup taxa. The remaining species of Fimbristylis did not conform to Kern's (1974) sections. The only terminal branch in Fimbristylis with moderate support (Bootstrap=70%) was the paired F. disticha (section Fuscae) and F. microcarya (section Trichelostylus), which are clearly classified in separate sections. A similar result was obtained in the combined trnL-F and ITS regions data sets of Ghamkhar et a1. (2005, in press), where a larger sample of species of Fimbristylis were included across the analysis, but sectional groups for the genus were not retrieved. A broader sampling of the embryos across the species assigned to Fimbristylis could provide more natural sectional groups, as there were distinct differences between the Fimbristylis embryos observed in this study and those by Van der Veken (1965) and Goetghebeur (1986). Although intraprophyllar buds were observed in the Bulbostylis species studied (excluding B. striatella), their presence was not captured in the overall inflorescence-synflorescence structure; the buds can remain dormant and appear to be absent in some specimens. The difficulty also lies in the amount of material 2\9 available for examination, as some specimens with only buds may be damaged or destroyed during examination. Nevertheless, the presence or absence 0 f intraprophyllar buds, or growth, in species assigned to Bulbostylis could be a useful distinguishing character at the sectional level within Bulbostylis, if not at the higher rank of genus. There is potential for future work on this. It is not surprising that Lye (in Haines and Lye 1983) used the appearance of the embryo to place Bulbostylis as a subgenus of Abildgaardia. The arrangement of the shoot and root primordia positioned basally, and the well-developed second primordial leaf are strikingly similar in all species of Abildgaardia and some species of Bulbostylis (B. hispidula, B. striatella and B. pilosa). However, the overall size of the embryo in Abildgaardia is consistently larger and the shoot is always more prominent than the root, while the reverse is true in all of the Bulbostylis species sampled in this study and those studied by Van der Veken (1965) and Goetghebeur (1986). The species of Bulbostylis examined by Van der Veken and Goetghebeur were: B. caespitosa Peter (= B. oritrephes (Rid!.) C.B.Clarke), Fimbristylis cioniana Savi (= B. cioniana (Savi) Lye, B. coleotricha (Hochst. ex A.Rich) C.B.Clarke, B. conifera (Nees) Kunth, B. fendleri C.B.Clarke, B. lanifera (Boeck.) Kilk., B. pringlei (Britt.) Beetle (= B. schaffneri (Boeck.) C.B.Clarke), B. vanderystii Cherm., B. melanocephala (Rid!.) C.B.Clarke, and B. oligostachya (Hochst. ex A.Rich.) C.B.Clarke. The Bulbostylis-type embryo united the species of Bulbostylis, however, there are many species placed in this genus that require sampling; the embryo size, and the number and development of primordial leaves have potential for assessing the sectional limits of the genus. The perianth bristles observed in Abildgaardia schoenoides (see Chapter 4 and Figure 6.3) are similar to those in Eleocharis, as both may have antrorse barbs 220 (Dahlgren et al. 1985; Wilson 1993), in contrast to the retrorse barbs in the species of Schoenoplectus with a perianth (see Figure 6.3). Many genera are composed of species with or without perianth (e.g. Schoenoplectus, Schoenus and Rhynchospora) and considering the rarity of the perianth within the tribe, the novel observation seems merely to be a remnant feature. The fact that the bristles were not welldeveloped in every floret (although many florets had aborted) and that the specin1en is distinctly grouped with the other samples for A. schoenoides, adds support to the remnant hypothesis; it is less likely to me that the presence of perianth in this material is a reversal. The surprise placement of Nelmesia within the Abildgaardia clade is not such a surprise when these results are compared to the systematic study of Goetghebeur (1986), where Nelmesia was placed in the same clade as Abildgaardia. The sample size of one collection, missing data, and the many autapomorphies associated with Nelsmesia, could have contributed to the placement. Nelmesia is known only from the Belgian Congo and placement within the Australian species of Abildgaardia is not a likely scenario. The intraspicular prophyll (cf. Haines 1967) present in Carex L., Kobresia Willd., Schoenoxiphium Nees (Snell 1936; Kern 1958; Tilllonen 1998; Starr et al. 2004), and Lipocarpha R.Br. (Goetghebeur 1998), although, the prophyll is modified into a utricle in genera of Carieaceae and the mixed floret sex, differs from the bisexual florets of Nelmesia with the non-modified prophyll. Lateral branches are consistently one-flowered spikelets in Nelmesia while the taxa shown in Kern (1958), Timonen (1998) and Starr et al. (2004) may have lateral branches with varying numbers of florets within the spikelet. Expanding the sample size to include taxa with features similar to Nelmesia and more collections of the species, could resolve some of the problems in assessing monophyly. 221 \Vith Nelmesia excluded from the analysis, as occurred in Chapter 4, the species that form Abildgaardia are monophyletic and in a clade separate to most species of Fimbristylis, including F. depauperata, the representative for the TYPE section, Fimbristylis section Fimbristylis. Despite the problems within Fimbris(ylis, in the full analysis, Abildgaardia does not form a clade with the species of Fimbristylis (excluding F. blakei and F. sp. L (Kimberley Flora). If Abildgaardia is considered as a section in Fimbristylis, then my study shows that Bulbostylis and Crosslandia would need to be demoted from the generic rank to the rank of section as well. Actinoschoenus and Trachystylis would also need to be reassigned as sections of Fimbristylis, if the results from my analysis are interpreted as sections. The Carex-type embryo is considered to be closest to the ancestral form (Goetghebeur 1998), and is unique to Trachystylis in this study. Actinoschoenlls thouarsii Benth., A. filiformis and A. repens were shown to have the Carex-type embryo (Van der Veken 1965; Goetghebeur 1986), however Actinoschoenus compositus and Arthrostylis aphylla (see also Goetghebeur 1986) share the Schoenus-type embryo. It is worth noting that Trachystylis with the Carex-type embryo and C 3 anatomy, both considered as ancestral features, was not placed in any of the basal positions within the clades, but was among taxa with the derived embryo types and anatomy. I cannot see why Trachystylis stradbrokensis and Nemum equitans were placed together within the Abildgaardia-Crosslandia clade, other than the lack of informative characters from the many autapomorphies present in both taxa, inhibiting assessment of the relationships. Expanding the sample to include more species of Nemum and Actinoschoenus, plus other taxa from the tribe Schoeneae Dumort., the alternative valid tribe in which Trachystylis, Arthrostylis and AClinoschoenus are placed (Goetghebeur 1998), may assist in resolving the lack of 222 monophyly achieved in this analysis. In addition, selecting a broader outgroup sample to include other closely related genera and so aid optimisation 0 f the tree (Grandcolas et al. 2004), starting with Eleocharis R.Br., is recommended. There is still much work to be done to assess the limits of the genera with the largest number of species, i.e. Fimbristylis and Bulbostylis, and the tTIonotypic genera that have only minimal collections, i.e. Nelmesia and Tylocarya. The lack of resolution for relationships in the current study is due in part to problems at both ends of the sampling spectrum, confounded by high levels of hOlTIoplasy and the difficulty in defining adequately characters for the cladistic analyses. Problematic data is not new, and many articles have been written about the use of characters in cladistic studies in an attempt to work through some of the lack of cladistic or phylogenetic resolve in analysis (Rieger 1979; Scotland and Williams 1993; Thiele 1993; Donoghue and Ackerly 1997; Poe and Wiens 2000; Wiens 2000; Desutter-Grandcolas et al. 2005). The next step that could be taken is the merging of the plant morphology, embryo morphology and anatomical data from this study with the nlolecular study for the tribe, and then seeing if the combined data set is more stable. 223 General conclusions The tribe Abildgaardieae, as currently delimited, does not form a monophyletic group. The data analysed were obtained from morphology, vegetative anatomy and en1bryo morphology. Monophyletic groups were retrieved for some genera in Chapters 3, 4, and 5, where Crosslandia, Abildgaardia, Fimbristylis, and Bulbostylis fanned the 'ingroup'. Species of Crosslandia formed a monophyletic group: C. setifolia, C. anthelata ined., C. spiralis ined. (Fimbristylis spiralis) and C. vaginata ined. (Abildgaardia vaginata). Crosslandia vaginata, although consistently retrieved, had little support and in the Bulbostylis analysis (Chapter 5) and the 'whole tribe' analysis (Chapter 6) was placed as sister to the Crosslandia clade; it is important to note that Crosslandia vaginata did not fall within the Fimbristylis or Abildgaardia s.s. clades (although it was placed in the broad Abildgaardia-Crosslandia clade). The variation in embryo morphology (Fimbristylis-and Schoenus-types) and inflorescence-synflorescence structure indicates that the sample size needs to be increased to fully define the limits before validly publishing new combinations. Extending the molecular sample is also recommended to explore the genetic variability across the geographical range. The remaining three species of Crosslandia have support for their placement as separate species within Crosslandia. There is no evidence to support maintaining Abildgaardia as a section of Fimbristylis, as the species of Fimbristylis did not form a monophyletic group in any of the analyses, and Bulbostylis was placed more closely to Fimbristylis than were the species of Abildgaardia. A well-supported, monophyletic group was formed by 224 species of Abildgaardia in the analyses in Chapters 3, 4, and 5, but not in the final analysis in Chapter 6, where Nelmesia melanostachya rendered the group nonmonophyletic. Nevertheless, the Abildgaardia clade did not fall within the species of Fimbristylis, although two species of Fimbristylis (F. blakei and F. sp. L) were grouped in the same broad clade as Abildgaardia and Crosslandia. Abildgaardia hygrophila was not supported as a species of Abildgaardia, even though it is placed near the provisional Crosslandia vaginata and the other species of Abildgaardia. Similarities between A. hygrophila (= Fimbristylis hygrophila) and species of Abildgaardia, as defined here, are purely superficial. The fact that this species was not grouped with any of the species of Fimbristylis begs for further investigation into the correct placement of this C 3 species bearing a Fimbristylis-type embryo. There was no support for the inclusion of Abildgaardia baeothryon within Abildgaardia. A study of the embryo and anatomy of Abildgaardia papillosa is recommended because of the findings for Fimbristylis bahiensis (= A. baeothryon) in this study and the affinity between the two species. Broader sampling to capture the variation between Abildgaardia oxystachya and A. pachyptera to define their limits is needed prior to publishing. In contrast, species status for the samples A. sp. aff. odontocarpa and A. sp. aff. pachyptera is merited and descriptions are being prepared. Meanwhile, detelmining where the name A. schoenoides R.Br. should be applied and if Fimbristylis squarrulosa (TYPE) is a synonym of A. schoenoides must be sought by exanlining the A. schoenoides TYPE specimen held at BM. Comparing the TYPES assigned to both names with the groups retrieved from phenetic analyses is necessary to determine the correct application of the names. Species of Bulbostylis formed a monophyletic group that was well supported in Chapters 5 and 6. Bulbostylis kakadu ined. is a distinct species separate to B. barbata 225 and a description can now be prepared for valid publication.. Any relationship between the Australian B. pyriformis and the B. hispidula group was inconclus lve. A more thorough study is recommended to explore fully the relationship between all the entities of the B. hispidula complex and the Australian B. pyriformis. No sections from Kern's (1974) classification of Fimbristylis were retrieved in this study, however, the placement of Tylocarya with Fimbristylis depauperata (from the TYPE section, Fimbristylis section Fimbristylis) supports Kern's (1958) decision to place Tylocarya in Fimbristylis as F. nelmesii. ｾｬ･ｭ｢ｲｳ of the provisional Arthrostylideae loosely formed the outgroup beside species of Schoenoplectus and Schoenoplectiella. However, the shift of Actinoschoenus, Arthrostylis and Trachystylis into the ingroup when Nemum, Nelmesia and Tylocarya were added, reflected the tenuous support for the outgroup placement in previous chapters (Chapters 3, 4, and 5). Actinoschoenus and Arthrostylis require further investigation to explore the species and generic limlts - a study is currently underway. Expanding the study to include other melllbers of the Schoeneae, considered close to Actinoschoenus and Arthrostylis, is necessary. Better tree topology may result by adding these taxa and a broadened sample of species fronl Fimbristylis and Bulbostylis, where work is also required to assess species and generic limits. The tribe Abildgaardieae may need to be reclassified in the near future. Expanding Abildgaardieae to include Actinoschoenus, Arthrostylis and Trachystylis is one option, or defining smaller tribal groups where the name Abildgaardieae is applied to taxa in the broad Abildgaardia-Crosslandia clade, and Fimbristylideae Cherm ex Raynal reinstated to accommodate taxa in the Fimbristylis-Bulbostylis clade is a 226 second option. However, further cladistic studies to resolve monophyletic groups are needed. Combining the morphological, anatomical, and embryographic data from this study with the molecular data from a sister study by Ghamkar (2004), to assess congruence and monophyly of the combined data, may move towards resolving monophyletic groups. In the tribe Abildgaardieae and the Cyperaceae in general, the search for monophyletic groups to develop natural systems of classification may well rely on more collaborative projects, especially where genera with large numbers of species cover vast areas globally - and all of us with limited resources. 227 References Adams CD (1994) Cyperaceae. In 'Flora Mesoamericana'. (Eds GD Davidse, l\![ Sousa Sand AO Chater). (Ciudad Universitaria : Universidad Nacional Aut6noma de Mexico, Instituto de Biologia, Missouri Botanical Garden: Mexico) Belbin L (1993) PATN pattern analysis package. (Division of Wildlife and Ecology, CSIRO: Canberra) Bentham G (1861) 'Flora hongkongensis: A description of the Flowering Plants and FenlS of the island of HongKong.' (L. Reeve: London) Bentham G (1878) Cyperaceae. In 'Flora Australiensis: A Description of the Plants of the Australian Territory'. (L. Reeve & Co.: London) Bentham G, Hooker 1D (1880) Cyperaceae. In 'Genera Plantarum' pp. 1037-1073. (L. Reeve & Co.: London) Blake ST (1940) Monographic studies in the genus Fimbristylis (Cyperaceae). Proceedings ofthe Royal Society ofQueensland 13, 1-14. Blake ST (1941) Notes on Australian Cyperaceae, V. Proceedings ofthe Royal Society of Queensland 52, 55-61. Blake ST (1947) Notes on Australian Cyperaceae, VIII. Proceedings ofthe Ro.val Society of Queensland 58, 35-50. Blake ST (1969) Studies in Cyperaceae. Contirbutions from the Queensland Herbarium 8, 1-48. Boeckeler 0 (1874) Fimbristylis planiculmis. Linnaea 38, 391. BreIner K (1994) Branch support and tree stability. Cladistics 10, 295-304. Brown R (1810) 'Prodromus Florae Novae Hollandiae.' (Hafner Publishing Co.: New York) Browning 1 (1992) HyogYnous bristles and scales in basal florets in amphicarpous Schoenoplectus species (Cyperaceae). Nordic Journal ofBotany 12, 171-175. Bruhl 11 (1990) Taxonomic relationships and photosYnthetic pathways in the Cyperaceae. PhD thesis, Australian National University. Bruhl 11 (1994) Amphicarpy in the Cyperaceae, with novel variation in the wetland sedge Eleocharis caespitosissima Baker. Australian Journal ofBotany 42, 441-448. Bnlhl 11 (1995) Sedge genera of the world: relationships and a new classification of the Cyperaceae. Australian Systematic Botany 8, 125-305. 228 Bruhl JJ, Perry S (1995) Photosynthetic pathway-related ultrastructure of C 3 , C 3 -like C 3 -C 4 intennediate sedges (Cyperaceae), with special reference to Eleocharis. Australian Journal ofPlant Physiology 22, 1-10. ('4 and Bruhl JJ, Stone NE, Hattersley PW (1987) C4 acid decarboxylation enzymes and anatomy in sedges (Cyperaceae). Australian Journal ofPlant Physiology 14, 719728. Bruhl JJ, Wilson KL (2005, in press) Towards a comprehensive survey ofC 3 and C4 photosynthetic pathways in Cyperaceae. In 'Monocots: comparative biology and evolution'. (Eds JT Columbus, EA Friar, CW Hamilton, JM Porter, Lrvl Prince and MG Simpson) pp. xx-xxx. (Rancho Santa Ana Botanic Garden: Claren10nt) Chennezon H (1929) Sur quelques Scirpus a epillets basicaules. Archives de Botanique, Bulletin Mensuel, Caen 3, 193-197. Clarke CB (1893) Cyperaceae 2. In 'The Flora of British India'. (Ed. JD Hooker) pp. 585-672. (L. Reeve and Co.: London) Clarke CB (1900) Cyperaceae. In 'Flora Capensis: A systematic description of the Plants of the Cape Colony, Caffraria, and Port Natal (and neighbouring territories)'. (Eel. SWT Thiselton-Dyer) pp. 149-310. (Lovell Reeve & Co. Ltd: London) Clarke CB (1908) New genera and species of Cyperaceae. Bulletin ofAfiscellaneous Information 8, 1-196. Crisp MD, Weston PH (1993) Geographic and ontogenetic variation in morphology ofAustralian \varatahs (Telopea: Proteacae). Systematic Botany 42, 49-76. Dahlgren RMT, Clifford HT, Yeo PF (1985) 'The Families of the Monocotyledons.' (Springer-Verlag: Berlin) Dallwitz MJ, Paine TA, Zurcher EJ (1999) DELTA Editor. (CSIRO Entomology: Canberra) Davis GL (1966) 'Systematic Embryology of the Angiospenns.' (John Wiley and Sons, Inc.: New York) Desutter-Grandcolas L, Legendre F, Grandcolas P, Robillard T, Murienne J (2005) Convergence and parallelism: is a new life ahead of old concepts? Cladistics 21, 5161. Donoghue MJ, Ackerly DD (1997) Phylogenetic uncertainties and sensitivity analyses in comparative biology. In 'Plant Life Histories: Ecology, Phylogeny, and Evolution'. (Ed. J Silvertown, Franco, M., and Harper, J.L.) pp. 17-35. (Cambridge University Press: Cambridge) Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39, 783-791. Fitzgerald WV (1918) The Botany of the Kimberleys, North-West Australia. Journal and Proceedings ofthe Royal Society of Western Australia 3, 102-224. Fosberg FR (1977) Miscellaneous notes on the flora of Aldabra and neighbouring islands: IV. A new Bulbostylis and observations on Cyperus (Cyperaceae). Kew Bulletin 31, 829-835. Ghamkhar K, Marchant AD, Wilson KL, Bruhl JJ (2006, in press) Phylogeny of Abildgaardieae (Cyperaceae) inferred from ITS and trnL-F data. In'Monocots: cOIYlparative biology and evolution'. (Eds JT Columbus, EA Friar, CW Han1ilton, JM Porter, LM Prince and MG Simpson) pp. xx-xxx. (Rancho Santa Ana Botanic Garden: Claremont) Gilliland MG, Gordon-Gray KD (1978) Kranz and non-Kranz cells in Cyperaceae. Proceedings of the Electron Microscopy Society of Southern Africa 8, 85-86. Goetghebeur P (1986) Genera Cyperacearum. Een bijdrage tot de kennis van de morfologie, systematiek en fylogenese van de Cyperaceae-genera. Doctoral thesis, Rijksuniversiteit Gent. [Flemish] Goetghebeur P (1998) Cyperaceae. In 'The Families and Genera of Vascular Plants'. (Ed.. K Kubitzki) pp. 141-190. (Springer-Verlag: Berlin) Goetghebeur P, Coudijzer J (1984) Studies in Cyperaceae 3. Fimbristylis and Abi/dgaardia in Central Africa. Bulletin Jardin Botanique National Belgium 54, 6589. Goetghebeur P, Coudijzer J (1985) Studies in the Cyperaceae. 5: The genus Bulbostylis in Central Africa. Bulletin Jardin Botanique National Belgium 55, 207259. Goetghebeur P, GrageI' A (1993) Studies in Cyperaceae II. The unispiculate species of Bulbostylis in the Americas. Biologisch Jaarboek 60,64-75. Gonzalez-Elizondo MS, Peterson PM, Granzow-de La Cerda I (1997) A cladistic and phenetic analysis of the Pauciflorae group of Eleocharis (Cyperaceae). BioLlania, Edicion Especial; Monografias Cientificas del Museo de Ciencias Naturales de la UNELLEZ Guanare 6, 341-368. Gordon-Gray KD (1971) Fimbristylis and Bulbostylis: generic limits as seen by a student of southern African species. Mitteilungen (aus) der Botanischen Staatssammlung Miinchen 10, 549-574. Gordon-Gray KD (1988) Studies in Cyperaceae in southern Africa: 13. The Bulbostylis schoenoides complex (Cyperaceae) in Africa, south of the Sahara. South African Journal ofBotany 54, 565-570. Gordon-Gray KD (1995) Cyperaceae in Natal. Strelitzia 2. 230 Grandcolas P, Guilbert E, Robillard T, D'Haese CA, Murienne J, Legendre F (2004) Mapping characters on a tree with or without the outgroups. Cladistics 20, 579-582. Greuter W, Brummitt RK, Farr PM, Kilian N, Kirk PM, Silva PC (Eds) (1993) 'Names in Current Use for Extant Plant Genera.' Regnum Vegetabile (Koeltz Scientific Books: Konigstein, Germany) Guaglianone ER (1970) Un nuevo caracter, util en la distinci6n generica entre Fimbristylis Vahl y Bulbostylis Kunth. (Cyperaceae). Darwiniana 16, 40-48. Haines RW (1967) Prophylls and branching in Cyperaceae. Journal ofthe East Africa Natural History Society and National Museum 26, 51-70. Haines RW (1971) Amphicarpy in east African Cyperaceae. Mitteilungen (aus) der Botanischen Staatssammlung Miinchen 10, 534-538. Haines RW, Lye KA (1977) Studies in African Cyperaceae. 15. Amphicarpy and spikelet structure in Trianoptiles solitaria. Botaniska Notiser 130, 235-240. Haines RW, Lye KA (1983) 'The Sedges and Rushes of East Africa.' (East African Natural History Society: Nairobi) Hennig W (1979) 'Phylogenetic Systematics.' (University of Illinois Press: Urbana) Holmgren PK, Holmgren NH, Barnett LC (1990) 'Index Herbariorum. Part I: The Herbaria of the World.' (New York Botanical Garden: Bronx) Hooper SS (1973) Cyperaceae. In 'The Families of Flowering Plants'. (Ed. J Hutchinson) pp. 863-871. (Clarendon Press: Oxford) Hutchinson J (1934) 'The Families of Flowering Plants II. Monocotyledons.' (Macmillan and Co. Ltd: London) Hutchinson J (1959) 'The Families of Flowering Plants.' (Clarendon Press: Oxford) Johansen DA (1940) 'Plant Microtechnique.' (McGraw-Hill Book Co.: New York) Johri BM, Ambegaokar, K.B., Srivastava, P.S. (1992) 'Comparative En1bryology of Angiosperms. ' (Springer-Verlag: Berlin) Keams CA, Inouye DW (1993) 'Techniques for Pollination Biologists.' (University Press of Colorado: Niwot, Colorado) Kern JH (1955) Florae Malesianae precursores X. Notes on Malaysian and SOllle S.E. Asian Cyperaceae 3. Blumea 8,110-169. Kern JH (1958) Florae Malesianae Precursors XXI. Notes on Malaysian and some S.E. Asian Cyperaceae VII. Acta Botanical Neerlandica 7, 786-800. 231 Kern JH (1959) Florae Malesianae precursores XXII. Cladium and Machaerina (Cyper.). Acta Botanica Neerlandica 8, 263-268. Kern JH (1963) Acriulus once more reduced Ｈｃｹｰｾｲ｡｣･ＩＮ Blumea 12,41-44. Kern JH (1974) Cyperaceae. In 'Flora Malesiana'. (Ed. CGGJ van Steenis) pp. 435753. (Noordhoff International Publishing: Leyden) Koyama T (1961) Classifification of the family Cyperaceae: 1. Journal ofthe Faculty ofScience 8, 37-148. Kral R (1971) A Treatment of Abildgaardia, Bulbostylis and Fimbristylis (Cyperaceae) for North America. Sida 4,57-227. Kral R (2002) Abildgaardia. In 'Flora of North Anlerica: North of Mexico'. (Ed. FoNAE Committee). (Oxford University Press: New York) Kral R, Strong MT (1999) Eight novelties in Abildgaardia and Bulbostylis (Cyperaceae) from South America. Sida 18, 837-859. Kraus JE, de Sousa HC, Rezende MH, Castro NM, Vecchi C, Luque R (1998) Astra Blue and Basic Fuchsin double staining of plant m.aterials. Biotechnic and Histochemistry 73, 235-243. Kiikenthal G (1944) Vorarbeiten zu einer Monographie der Rhynchosporideae. 15. Feddes repertorium specierum novarum regni vegetabilis 53, 186-197. Kukkonen I (1986) Special features of the inflorescence structure in the family Cyperaceae. Annales Botanici Fennici 23, 107-120. Kukkonen I (1991) Problems in Carex section Physodeae and Cyperus congolmeratus within the Flora Iranica area. Flora et Vegetatio Mundi 9, 63-73. Kunth CS (1837) 'Enumeratio Plantarum. 2.' (Stutgardiae & Tubingae) Latz PK (1990) Taxonomic studies of Fimbristylis (Cyperaceae) in Northern Australia. Nuytsia 7,161-182. Le Roux LG, Kellogg EA (1999) Floral developm.ent and the formation of unisexual spikelets in the Andropogoneae (Poaceae). American Journal ofBotany 86,354-366. Levyns MR (1943) A revision of Trianoptiles Fenzl. Journal ofSouth African Botany 9, 21-26. Lye KA (1971) The generic concept of Bulbostylis Kunth ex C.B. Clarke. Mitteilungen (aus) del' Botanischen Staatssammlung Munchen 10,539-547. Lye KA (1973) Studies in African Cyperaceae VIII. The taxonomic position of Abildgaardia Vahl and Nemum Hamilton. Botaniska Notiser 126, 325-329. Lye KA (1995) Cyperaceae. In 'Flora of Somalia'. (Ed. sLN Herbariunl)) 232 Lye KA (1996) A new species of Bulbostylis hispidula (Cyperaceae) frOlTI Somalia. Willdenowia 25, 595-600. Lye KA (2000) Achene structure and function of structure in Cyperaceae. In 'Monocots II: Systematic and Evolution'. Sydney. (Ed. KL Wilson, and Morrison, D.A.) pp. 615-628. (CSIRO) Maddison WP, Maddison DR (1992) MacClade: Analysis of phylogeny and character evolution. In. (Sinauer Associates, Inc.: Sundlerland, Massachusetts) Maheshwari P (1964) 'Embryology in relation to taxonomy.' (Pergamon Press Limited: Oxford) Metcalfe CR (1969) Anatomy as an aid to classifying the Cyperaceae. American Journal ofBotany 56, 782-790. Metcalfe CR (1971) Cyperaceae. In 'Anatomy of the Monocotyledons'. (Ed. CR Metcalfe). (Clarendon Press: Oxford) Microsoft (1999) Microsoft Excel 2000. Mitchell CH, Diggle PK (2005) The evolution of unisexual flowers: morphological and functional convergence results from diverse developmental transitions. American Journal ofBotany 92, 1068-1076. Muasya AM, Bruhl JJ, Simpson DA, Culham A, Chase MW (2000) Suprageneric phylogeny of Cyperaceae: a combined analysis. In 'Monocots: Systematic and Evolution'. (Eds KL Wilson and DA Morrison) pp. 593-601. (CSIRO Publishing: Melbourne) Nelrnes E (1949) Notes on Cyperaceae: XIX. A new genus of Scirpeae. Kew Bulletin 4,139-140. Page RDM (1996) TREEVIEW: An application to display phylogenetic trees on personal computers. Computer Applications in the Biosciences 12, 357-358. Penet L, Nadot S, Ressayre A, Forchioni A, Dreyer L, Gouyon PH (2005) Multiple developmental pathways leading to a single morph: monosulcate pollen (examples frorn the Asparagales). Annals ofBotany 95, 331-343. Poe S, Wiens JJ (2000) Character selection and the methodology ofmorphologlcal phylogenetics. In 'Phylogenetic Analysis of Morphological Data'. (Ed. JJ Wiens) pp. 20-36. (Smithsonian Institution Press: Washington) Ramlrez MJ (2005) Resampling measures of group support: a reply to Grant and Kluge. Cladistics 21, 83-89. Raynal J (1973) Notes Cyperologioques: 19. Contribution a la classification de la Sous- famille des Cyperoideae. Adansonia 13, 145-171. 2.3.3 Raynal J (1976) Notes Cyperologioques: 26 Le Genre Schoenoplectus 11. L'arnphicarpie et la sect. Supini. Adansonia 16, 119-155. Rieger RaT, S. (1979) The homology theorem in ultrastructural research. American Zoologist 19, 655-664. Robyns W, Tournay R (1955) 'Flore des Spermatophytes du Parc National Albert.' Rye BL (1992) Cyperaceae. In 'Flora of the Kimberley Region'. (Ed. JR Wheeler) pp. 1035-1107. (Western Australian Herbarium, Department of Conservation & Land Management: Como, W .A.) Scotland RW, Williams DM (1993) Multistate characters and cladograms: when are two stamens more similar to three stamens than four? A reply to Lipscomb. Cladistics 9, 343-350. Sharpe PR (1986) Keys to the Cyperaceae, Restionaceae and Juncaceae of Queensland. Queensland Botany Bulletin 5, 1-34. Simpson DA (1992) 'A revision of the genus Mapania.' (Royal Botanic Gardens, Kew: Surrey, UK) Simpson DA (1993) Genera of Cyperaceae recognised at Kew. Cyperaceae Newsletter 12, 8-12. Simpson DA, Koyama T (1998) Cyperaceae. Flora of Thailand 6,247-485. Simpson DA, Muasya AM, Alves M, Bruhl JJ, Dhooge S, Chase MW, Furness CA, Ghamkhar K, Goetghebeur P, Hodkinson TR, Marchant AD, Reznicek AA, Nieuwborg R, Roalson EH, Smets E, Starr JR, Thomas WW, Wilson KL, Zhang X (2005, in press) Phylogeny ofCyperaceae based on DNA sequence data - a new rbcL analysis. In 'Monocots: comparative biology and evolution'. (Eds JT Columbus, EA Friar, CW Hamilton, JM Porter, LM Prince and MG Simpson) pp. xx-xxx. (Rancho Santa Ana Botanic Garden: Claremont) Snell RS (1936) Anatomy of the spikelets and flowersof Carex, Kobresia and Uncinia. Bulletin ofthe Torrey Botanical Club 63, 277-295. Soros CL, Bruhl JJ (2000) Multiple evolutionary origins of C 4 photosynthesis in the Cyperaceae. In 'Monocots: Systematics and Evolution'. (Eds KL Wilson and DA Morrison) pp. 629-636. (CSIRO: Melbourne) Soras CL, Dengler NG (2001) Ontogenetic derivation and cell differentiation in photosynthetic tissues of C 3 and C4 Cyperaceae. American Journal ofBotany 88, 992-1005. Starr JR, Harris SA, Simpson DA (2004) Phylogeny of the unispicate taxa in Cyperaceae tribe Cariceae I: Generic relationships and evolutionary scenarios. Systematic Botany 29, 528-544. 234 Statsoft I (1996) Statistica for Windows. (Statsoft Inc.: Tulsa) Steudel EG (1855) Cyperaceae Juss. In 'Synopsis Plantarum Cyperacearum'. (lB. Metzler: Stuttgartine) Stock WD, Chuba DK, Verboom GA (2004) Distribution of South African C3 and C 4 species of Cyperaceae in relation to climate and phylogeny. Austral Ecology 29, 313319. Stuessy TF (1990) 'Plant Taxonomy.' (Columbia United Press: New York) Swofford DL (2001) PAUP*: Phylogenetic analysis using parsimony (*and other methods). In. (Sinauer Associates: Sunderland, Massachusetts) Thiele K (1993) The holy grail of the perfect character: the cladistic treatment of morphometric data. Cladistics 9, 275-304. Th\\Taites GHK (1864) 'Enumeratio Plantarum Zeylaniae.' (Dulau: London) Timonen T (1998) Inflorescence structure in the sedge tribe Cariceae (Cyperaceae). Publications in Botany from the University ofHelsinki 26, 1-35. Vahl M (1805) 'Enumeratio Plantarum. 2.' (Hauniae : Impenis auctoris, & prostat apud J.H. Schubothe: Copenhagen) Van der Veken P (1955) Trois taxa nouveaux de Cyperacees du Congo BeIge. Bulletin du Jardin botanique de Etat a Bruzelles 25, 143-147. Van der Veken P (1965) Contribution a l'embryographie systematiques des Cyperaceae: Cyperoideae. Bulletin du Jardin botanique de Etat a Bruzelles 35, 285354. Vegetti AC (2003) Synflorescence Typology in Cyperaceae. Annals Botanici Fcnnici 40,35-46. von Mueller F (1875) 'Fragmentia Phytographiae Australiae.' (Melbourne) Vrijdaghs A, Caris P, Goetghebeur P, Smets E (2005) Floral ontogeny in Scirpus, Eriophorum and Dulichium (Cyperaceae), with special reference to the perianth. Annals ofBotany 95,1199-1209. Weberling F (1989) 'Morphology of flowers and inflorescences.' (Cambridge University Press: Cambridge) Wiens JJ (2000) Coding morphological variation within species and higher taxa for phylogenetic analysis. In 'Phylogenetic Analysis of Morphological Data'. (Ed. ] ] Wiens) pp. 115-145. (Smithsonian Institution Press: Washington) Wilson KL (1980) Notes on some Australian species ofCyperaceae. Telopea 1,457467. 23S Wilson KL (1991) Systematic studies in Cyperus section Pinnati (Cyperaceae). Telopea 4, 361-559. Wilson KL (1993) Cyperaceae. In 'Flora of New South Wales. Vol. 4.' (Ed. GL Harden) pp. 293-396. (New South Wales University Press: Kensington) World Checklist of Monocotyledons (2004) The Board of Trustees of the Royal Botanic Gardens, Kew. In 'Published on the Internet; http://w\vw.kew.org/nl0nocotChecklist/'. 236 Appendix 1. List of all specimens used in phenetic and/or cladistic analyses. The OTU label corresponds to the code for each specimen used in phenetic analyses. Herbarium codes (Herb. Code.) are provided for all sampled specimens followed by the sheet number where available. Specimens collected in Australia show the state in which they were collected, if collected overseas then the country or continent is given. Specimens used for Scanning electron microscopy (SEM=*), embryo morphology (embryo=#) or leaf blade and/or culm anatomy (anatorny=+) are indicated in the phenetic code column next to the OTU label, if given. N.T.=Northem Territory, W.A.=Westem Australia, Qld=Queensland, Vic=Victoria. (JJB)=prepared sections provided by J.J. Bruhl. Names given here are prior to analyses and are based on Abildgaardia and Tylocarya as genera. Generic groups studied and species names OTU label Herb. code State or Collector Country Ingroup Cross/audia W.Fitzg. Crosslandia anthelata nom. C18 provo Goetgh. ined. C19# C20# C21 C22 C23 Crosslandia setifolia W.Fitzg. C1 C2 C3 C4 C5 C6 C7 C8 C9 C10 C11 C12 C13 C14 MEL 1590282 BRI 313723 NE 61439 NSW 452330 NSW 452331 CANE as CBG 8309188 BRI 313715 MEL 2048535 NSW 452305 MEL 2048435 BRI 313716 NSW 460200 CANB 264103 NE(exNSW 416310) NSW 303698 NSW 460198 MEL 2048448 BRI 313711 MEL 2048434 NE N.T. Dunlop C.R. 6854, Wightman G. N.T. N.T. N.T. Dunlop C.R. 3446 Bruhl J.J., Hunter Y., Egan J. 1268 Dunlop C.R. 3408 N.T. Wilson K.L. 5150, Dunlop C.R. N.T. Thompson H.S. 403 N.T. Chippendale G. 1268 W.A. Poulton G. 5 W.A. Wilson K.L. 4885 N.T. Craven L.A. 7928, Whitbread G. N.T. N.T. Blake S.T. 17420 Cowie J.D. ill 4639 W.A. Pullen R. N.T. Wilson K.L. 5260 W.A. Wilson K.L. 4859 W.A. Wilson K.L. 4803 N.T. Lazarides M. 8, Adams L.G. W.A. N.T. Burbidge N. 5703 Dunlop C.R. 6789 W.A. Clarke K.L. 166, Bruhl J.J., Wilson K.L. 237 C15 C16 C17 Fimbristylis spiralis R.Br. Fl# F2 F3 Abildgaardia vaginata R.Br. Avl Av2# Av3# Av4 Av5 Av6 Av7+ Av8 Av9# Avl0 Avl1 Av12 Av13 Av14 N.T. Blake S.T. 16585 W.A. Van Rijn P.l. 19 N.T. N.T. Clarke K.L. 155, Bruhl 1.1., Wilson K.L., Cowie I.D. Specht R.L. 235 N.T. N.T. Dunlop C.R. 2957 Leach G. 3601, Cowie I.D. Qld N.S.W. Blake S.T. 15540, Webb LJ. Floyd A.G.F. AGF2205 Qld N.S.W. Qld Brass L.l. 18362 O'Hara 1. 3472 and Coveny R. Blake S.T. 8598 N.T. Cowie I.D. 6801 N.T. N.S.W. Qld Brennan K. 2588 Bell D.M. Forster P.I. PIF9732, Machim P. Qld Qld Forster P.I. PIF16257 Blake S.T. 8222 Qld Blake S.T. 22499 Qld Qld Brass L.l. 1924 Sharpe P.R. 5299 and Bird L. K 2525 Brazil Mattes Silva L.A. 394, Ribeiro AJ., da S. Brito H. K2528 Brazil Almeida de 1esus 1. 1466 CANB 159880 CANB 78686 NE MEL 2048472 DNA 36442 NSW 422184 BRI300861 NSW 285362 BRI300844 BRI 300865 MEL 2048450 DNA 128735 DNA 72733 NE 54172A MEL 716909 BRI 601359 MEL 2048451; BRI300850 CBG 8309188 CANB 506846; BRI300856 BRI 300848 BRI 549964; CANB 505073 Abildgaardia Vahl Abildgaardia baeothryon A.St.-Hil. *#+ (all as Fimbristylis bahiensis #+ Steud.) KEW 2529 Brazil Brazil K 2527 Abildgaardia hygrophila (Gordon-Gray) Lye Abildgaardia macrantha (Boeck.) comb. provo Goetgh. * NE 71642 #+ NU AmI CANB 280535 South Africa South Africa W.A. Bento Pickel D. 3140 Mori S. 10389, dos Santos S., White I. Ward C.l. 2794 Tinley K.L. 307 Hartley T.G. 14405 238 NSW 454230 Am3* NSW 452333 BRI301881 Am4 MBA Am5 NSW Am6 338994 Am7*# BRI437357 DNA Am8 122604 CANB Am9 242626 Aml0 NE N.T. Cowie J.D. 6202 & Booth R. N.T. Wilson K.L. 4971 N.T. Qld Qld Qld N.T. Dunlop C.R. 4102 Clarkson J. 8324 Wilson K.L. 8073, Clarkson J., Jacobs S.W.L. Clarkson J. 6624 Cowie J.D. 5260, Taylor S. N.T. Dunlop C.R. 3453 N.T. *#+ Mexico Clarke K.L. 249, Bruhl J.1., Wilson K.L., Cowie J.D. Pringle C.G. 3127 Mexico Breedlove D.E. 54895, Davidse G. Mexico Kral R. 25115 Mexico Pringle C.G. 9294 Mexico GomilezS.1116 Mexico Arsene G. Qld Blake S.T. 13582 (HOLOTYPE) Am2 Abildgaardia mexicana (Palla) Kral Fimbristylis odontocarpa S.T.Blake F. sp. aff. odontocarpa Abildgaardia ovata (Burm.f.) Kral MEL 268631 MO 3524080 MO 1917406 MO 3058759 MO 3632024 MEL 2050681 Aodl BRI341121 MEL 2048459 MEL 2048468 Aod2* BRI574804 #+ Aaffod BRI 302127 *#+ Aovl BRI 476426 (ISOTYPE) (ISOTYPE) Qld Turpin G.P., Thompson EJ. W.A. Carey J. Qld Specht R.L. 408, Reeves R.D. Aov2* BRI457532 Qld Aov3 MEL Qld 2048512 N.S.W. Aov4 NSW 468324 Aov5 NSW 87198 N.S.W. Aov6 NSW N.S.W. 468325 N.S.W. Aov7 MEL 2048525 Aov8* NSW N.S.W. 468326 Aov9 BRI384713 Qld Aov10 BRI 591266 Qld Aovll NE Qld * Batianoff G.N. 11056 O'Shanessy P.A. 1656 Wilson K.L. 5818 Johnson L.A.S. Rodd A.N. 2277 Mueller F. Rodd A.N. 2434 StanleyT.8019 Neldner V.J. 3905 Clarke K.L. 99, Bruhl J.J., Wilson K.L. 239 A. ovata (cont'd) Abildgaardia oxystachya (F.Mue:ll.) comb. provo Goetgh. CANB MEL 2050881 MEL 2050886 MEL 2050863 MEL 2050864 Aox1* NSW 452336 Aox2 Aox3* # Aox4+ Aox5* + Aox6+ Aox7 Aox8 Aox9 Aox10 Aox11 Aox12 + Abildgaardia pachyptera (S.T.Blake) comb. provo Goetgh. Aox13 ApI Ap2 Ap3 Ap4 Ap5 Ap6 Ap7 Ap9+ A. sp. aff. pachyptera Abildgaardia schoenoides R.Br. Timor India Wiriadinata H. 449 Mueller F. U.S.A. Medley Wood J. 4918 India Thomson G. India Unknown W.A. Latz P.K. 4038 BRI302157 Qld NE 61223 N.T. Blake S.T. 15725, Webb L.J. Bruhl J.J. 1252 NE NSW 452337 NE N.T. N.T. Latz P.K. 8667 Wilson K.L. 5369 W.A. BRI336290 BRI 386514 CANB 280532 CANB 353598 DNA 123491 CANB 505055; BRI203628 DNA 21755 NE 61427A MEL 1615159 MEL 252996 NSW 452342 NSW 452335 NSW 452334 MEL 2048462 NE Qld W.A. W.A. Clarke K.L. 124, Bruhl LT., Wilson K.L. Blake S.T. 19620 Cane S. 53 Hartley T.G. 14357 W.A. Carr G.W. 4377, Beauglehole A.C. N.T. Booth R. 618K.L. Qld Blake S.T. 13611 N.T. N.T. N.T. Wightman G. 424 and Dunlop C.R. Hunter J.T. 1547, Bruhl J.1. Dunlop C.R. 9041 N.T. Jones M., Booth R. 24 W.A. Dunlop C.R. 5339 N.T. Wilson K.L. 5109, Taylor S. N.T. Wilson K.L. 5207 N.T. Chippendale G N.T. Clarke K.L. 253, Bruhl LT., Wilson K.L., Cowie I.D. Clarke K.L. 181, Bruhl LT., Wilson K.L., Cowie I.D. Adams L.G. 1715 Ap10+ NE N.T. Ap11 N.T. CANB 166722 Aaffpa NE ch As1 NSW 468344 N.T. W.A. Clarke K.L. 201, Bruhl J.1., Wilson K.L., Cowie I.D. Wilson K.L. 4888 240 A. schoenoides (cant' d) Abildgaardia sp. aff. schoenoides Abildgaardia triflora (L.) Abeywickr. As2 BRI 480019; NSW 468346 As3 NSW 468347 As4 NSW 468343; MEL 1601269 As7+ BRI 533192 As8 NSW 229593 As9*+ NE W.A. Dunlop C.R. 7838 W.A. Mitchell A.A. 2129 N.T. Dunlop C.R. 865], White N.G. Qld Qld Bruhl 1.1. 487 lacobs S.W.L. 5903 Qld N.T. Clarke K.L. 70, Bruhl 1.1., Wilson K.L. Clarke K.L. 157, Bruhl 1.1., Wilson K.L. Clarke K.L. 216, Bruhl 1.1., Wilson K.L., Cowie J.D. Clarke K.L. 120,1 Bruhl 1.1., Wilson K.L. Clarke K.L. 230, Bruhl 1.1., Wilson K.L., Cowie J.D. Perry R. 222. N.T. Bruhl 1.1. 1261, Hunter 1.1'., Egan 1. N.T. Zaire Dunlop C.R. 5863, Craven L.A. Malaisse F. 400, Goetghebeur P. AsI0* NE W.A. Asll+ NE N.T. As12+ NE W.A. As13* NE + As14 CANB 16035 As5*# NE 61432 + As6*# BRI329001 * MO 4579353 #+ N.T. Tanzania Greenway P.l. 1859 South Ward C.l. 1708 Africa Sri Lanka Koyama T. 13910, Koyama M. Fimbristylis VahI Fimbrislylis blakei Latz fbI *#+ DNA N.T. D0120436 fb2 BRI435191 N.T. (as Fimbristylis sp.) #+ Fimbrislylis cinnamometorum (Yahl) Kunth fcl+ DNA A0086876 DNA 134230 NE fc2#+ NE N.T. fc3# fc4 NE NE Qld N.T. fc5*#+ NE Dunlop C.R. 10015, Latz P.K. Latz P.K. 10375 N.T. Latz P.K. 11214 N.T. OrrT.M.442 Qld Clarke K.L. 61, Bruhl 1.1., Wilson K.L Clarke K.L. 132, Bruhl 1.1., Wilson K.L., Cowie J.D. Clarke K.L. 276 Clarke K.L. 228, Bruhl 1.1., Wilson K.L., Cowie I.D. Clarke K.L. 139, Bruhl 1.1., Wilson K.L. W.A. 241 Fimbristylis depauperata #+ NE Qld Clarke K.L. 305, Little L. Qld Clarke K.L. 263, Bruhl J.1., Wilson K.L. Dallachy R.Br. NE Fimbristylis fimbristyloides ffil (F.Muell.) Druce Fimbristylis furva R.Br. ffi2+ ffi3# ffl MEL 269463 BRI336296 QRS 43342 NE f£1*#+ NE #+ NE Fimbristylis fusca (Nees) Qld Qld N.T. Qld N.T. Blake S.T.18678 Flecker H. Clarke K.L. 267, Bruhl J.1., Wilson K.L., Cowie J.D. Clarke K.L. 52, Bruhl ｊＮＱｾ Wilson K.L Clarke K.L. 210, Bruhl J.1., Wilson K.L., Cowie J.D. Dunlop C.R. 5922 Craven L.A. ffu1 BRI329000 N.T. ffu2 fsL1 BRI 301745 PERTH 2272911 PERTH 2272962 PERTH 2272911 N.T. W.A. Blake S.T. 16571 Kenneally K.F. 11171 W.A. Kenneally K.F. 11168 W.A. Kenneally K.F. 11167 NE W.A. NE N.T. NE NE Qld W.A. NE W.A. NE 65205 South Africa Clarke K.L. 131, K.L. Clarke K.L. 268, K.L. Clarke K.L. 319, Clarke K.L. 108, K.L. Clarke K.L. 153, K.L. Browning J. 834 Benth. Fimbristylis sp. L (Kimberley flora) fsL2+ # Fimbristylis microcarya #+ F.Muel1. Fimbristylis schultzii Boeck. * Fimbristylis variegata *# Gordon-Gray Bruhl J.J., Wilson Bruhl J.J., Wilson L. Little Bruhl J.J, Wilson Bruhl J.J., Wilson Bulbostylis Kunth Bulbostylis barbata (Rottb.) bba1+ DNA 24124 Qld Wilson K.L. 5442 C.B.Clarke bba2 bba3 bba4 bba5 bba6# DNA 49868 DNA 62627 CANB 410874 NE 61441 NE 60472 N.T. W.A. N.S.W. Beauglehole A.C. 2608 Mitchell A.S. 1150 Tindale M.D. 2058 N.T. N.S.W. Bruhl J.J., Hunter J.T, Egan J. 1269B Bell D.B. bba7 bba8 BRI 316049 N.T. W.A. NE bba9 N.T. bba10 CANB 325418 NE bba11 NE W.A. N.T. Latz P .K. 8263 Clarke K.L. 160, Bruhl J.J., Wilson K.L. Knight F. 14185 Clarke K.L. 221, Bruhl K.L., Cowie J.D. Clarke K.L. 113, Bruhl K.L. ｊＮＱｾ Wilson ｊＮｾ Wilson 242 B. barbata (cont'd) bba12 NE bba13 bba14 K MO 4280627 bba15 EA bba 16 NSW 468581 bba17 NSW bba18# BRI 512812 Qld Clarke K.L. 100, Bruhl J.J., Wilson K.L. Singapore Burkhill H.M., Shah M. HMB235 U.S.A. Hill S.R. 24361 Kenya India Napper D.M., Kanuri 2079 Raizada M.B U.S.A. India Correll D.S. 52337 Rajn R.R.V Po1hill R. 847, Paulo S. + bba19 EA bba20 L 2623 South Africa Thailand NE N.T. + Bulbos(ylis sp. aff. barbata Bulbostylis burbidgeae K.L.Wilson Bulbostylis sp. aff. burbidgeae Bulbostylis capillaris (L.) Kunth ex C.B.Clarke baffba1 NE N.T. + baffba2 DNA 22621 N.T. baffba3 NE N.T. baffba4 NE # baffba5 CANB 421033 baffba6 NE N.T. baffba7 NE N.T. bbu1 W.A. Clarke K.L. 251, Bruhl J.1., Wilson K.L., Cowie I.D. Clarke K.L. 239, Bruhl J.1., Wilson K.L., Cowie I.D. Hart R.P. 2092 W.A. Carolin R. 7640 W.A. Mitchell A.A. 1929 W.A. W.A. Burbidge N. 1102 Payne A.L. PRP976 N.T. Dunlop 4725 U.S.A. Davidse G. U.S.A. Curtiss A.H. PERTH 4275098 bbu2 PERTH 1083007 bbu3 PERTH 5223741 bbu4 BRI 311667 bbu5#+ NSW 452309 baffbu DNA 14302 NSW 452328 MO + N.T. U.S.A. Styains A.H. # bde1 MEL 268550 MEL 268552 L 65104 NE 60798 U.S.A. N.S.W. Horr W.H. E170 Bruhl J.J. 1197, Quinn F.e. bde2 NE 63629 N.S.W. Hunter J.T., Hunter V. 2737 + Bulbostylis densa (Wall.) Hand.-l'vlazz. N.T. Larsen K. 1299, Smitinand T., Wamc1 E. Clarke K.L. 187, Bruhl J.1., Wilson K.L., Cowie I.D. Clarke K.L. 184, Bruhl J.1., Wilson K.L., Cowie I.D. Rice B.L. Clarke K.L. 245, Bruhl J.1., Wilson K.L., Cowie I.D. Clarke K.L. 241, Bruhl J.1., Wilson K.L., Cowie I.D. Bruhl J.J. 369A 243 B. densa (cont'd) Bulbostylis sp. aff. densa 1 BulbostyUs sp. aff. densa 2 bde3 NSW 468227 bde4 BRI 304640 bde5 BRI 472840 CANB bde7 50320 BRI311651 bde8 CANE 117835 BRI407433 bde9 bdel0 CANB 282965 bdell CANB 410867 bde12 CANB 87831 bde13 MEL 268539 bde15 BRI 342062 baffd 1* BRI 506795 #+ baffd2 MO + 4501576 bde16 PRE bde17 PRE bde18 Bulbostylis hispidula subsp. *#+ pyriformis (Lye) R.W.Haines *#+ PRE K Bulbostylis hispidula subsp. *#+ senegalensis (Chenn.) Van den Berghen Bulbostylis puberula C.B.Clarke #+ Qld Forster P.I. PIF8482 Qld Qld Qld Blake S.T. 21453 Bean A.R. 1570 Hubbard C.E. 3128 Qld McKee H.S. 9317 N.S.W. N.S.W. Williams J.B. Gray M. 3255 P.N.G. Croft, Lelean 34706 P.N.G. Robbins R.G. 2660 Philippine Ramos M., Edaro G. s Sri Lanka Davidse G. 7614 Bean A.R. 3236 Qld China Field survey team 820 South Africa South Africa Swaziland Kenya Meeuse A.D.l. 10158 Scheepers J.C. 1141 Haines R.W. 7048 McCallum-Webster K NSW 452317 K Tanzania Richards M. 23175B Eritrea Pappi A. Sudan Davey J.T. 10 MO 5018124 K Senegal Vanden Berghen C. 7484 K CANB 67185 L 65128 CANB 216795 K K DNA 70798 Bulbostylis sp. aff. puberula baffpu NSW 452329 Bulbostylis pyriformis bpy1 NSW 400826 S.T.Blake Sri Lanka Davidse G. 9037, Sumithraarachchi D.B. Jacobs M. 5691 Borneo Singapore Sinclair J. Sri Lanka Clayton D. 5112 Thailand Brunei Indonesia N.T. van Beusekom C.F. 2247, Smitinand 1 Coode M.J.E. 774, Kirkup D.W. Elsol J.A. Latz P.K. 11364 N.S.W. Johnson L.A.S. 244 bpy2#+ NSW 452308 NE 60921A bpy3 BRr 311664 bpy4 NSW bpy5 452307 NSW bpy6 400827 BRr 9336 bpy7 BRr 542784 bpy8 PRE Bulbostylis Izumi/is (Kunth) C.B.Clarke 2827DD PRE 2927BC NE 58064 #+ bt2 DNA 62665 Blllbostylis turbinata S.T.Blake DNA 51665 bt3 DNA 72393 bt4 PERTH bt5 2091526 bt7 PERTH 2073552 PERTH bt8 5221005 BRr 476931 bt9 btl0 MEL 1620712 Ne/mesia Van der Veken Nelmesia melanostachya NY Van der Veken Nemum Desv. ex Hamilt. Nemum equitans (Kilk.) # NU J.Raynal EA EA NemUln spadiceum (Lam.) Desv. #+ NU * NU * K Nemum megastachyum (Cheml.) J.Raynal EA Ty/ocarya Nelmes Tylocarya cylindrostachya *+(JJB L 65198 Nelmes ) Outgroup Actinoschoenus compositus *#+ NE (Latz) nom. provo ined. NE B. pyriformis (cont' d) N.T. Latz P.K. 10622 N.S.W. N.T. N.S.W. Hunter J.T., Bell D.B. Latz P.K. 488? Wilson K.L. 1479A N.T. Latz P.K. 9852 Qld Qld South Africa South Africa N.S.W. W.A. Sharpe P.R. 232 Bean A.R. 4227 Jarman N. 134 N.T. N.T. W.A. Latz P.K. 7126 Latz P.K. 6339 Royce R.D. 1491 W.A. George A. S. 820 W.A. Payne A.L. PRP 1854 Qld N.T. Harris P.L. 342 Beauglehole A.C. 26568 Congo Gerard 57 (ISOTYPE) Zambia Robinson E.A. 2681 Zambia Zambia Zambia Robinson E.A. 3912 Greenway P.J. Robinson E.A. 5165 Zambia Zambia Congo Robinson E.A. 4676 Robinson E.A. 4677 Germain 4420 Thailand Kerr A.F.G. 21294 N.T. Clarke K.L. 178 Bruhl J.J., Wilson K.L., Cowie I.D. Clarke K.L. 231 Bruhl J.1., Wilson K.L., Cowie I.D. Clarke K.L. 235 Bruhl J.1., Wilson K.L., Cowie I.D. Clarke K.L. 211 Bruhl J.1., Wilson K.L., Cowie I.D. Cowie I.D. 5643, Brennan K. N.T. NE N.T. *#+ NE N.T. * NE N.T. Ruch M. 9 Strong C.P., Fletcher J.R., Sharp G.C. Mitchell A.A. 479 245 A. compositus (cont'd) NE Arthrostylis aphylla R.Br. NSW N.T. 196791 MEL N.T. 1619328 BRI 311099 Q1d Clarke K.L. 227 Bruhl J.1., Wilson K.L., Cowie J.D. Wilson K.L. 7520 Cowie J.D. 4643 Brass L.J. 18669 BRI429959 BRI 399429 NSWex Herbario Kewensis 24110 NSW 259584 NSWex AD 75517 NSW NE 30785 NE 58440A NSW BatianoffG.N.10089 Qld Gunness A. AG 1937 Q1d McKee H.S. RSNH 24110 New Hebrides * NSW NSW N.T. Qld * NSW * *+ Schoenoplectus tabernaell10ntani (C.C.Gmel.) Palla (=S. validus Vahl) Schoenoplectiella lateriflora (J.F.Gmel.) Lye (=s. lateriflorus G.F.Gme1) NSW NSW Schoenoplectiella laevis (S.T.Blake) Lye (=s. laevis S.T.B1ake) * *#+ Trachystylis stradbrokensis (Domin.) Kuk. N.T. * #+ * * NSW NSW 251326 NSW NSW 338962 BRI 156925 BRI 156923 BRI 399437 BRI 10875 MEL 716466 MEL 2048446 BRI227559 BRI541420 BRI 156931 CANB 193493 Vic Wilson K.L. 6804 S.A. Hunt D. 1788 Q1d N.S.W. N.S.W. W.A. Wilson K.L. 4278 Wallace B.J. Bruhl J.J. 150 Jacobs S.W.L. 4226 Latz P.K. 3761 Wilson K.L. 3389, Sharpe P.R., Johnson L.A.S., B1axell D. Philippine Ramos M. s Unknown India Dunlop C.R. 5405 W.A. W.A. N.S.W. N.T. Qld Q1d Qld Q1d Q1d Q1d Brennan K. Milthorpe P.L. 1777A, Cunningham G.M. Wilson K.L .5399 and Scarlet C. Wilson K.L. 8041, Clarkson J. and Jacobs S.W.L. White C.J. Q1d Blake S.T. 13201 Gunness A. Ag 1965 Durrington L 3 12 Sharpe P.R. 5199, Wilson K.L., S.W.L. Blake S.T. 23328 Q1d Q1d Q1d Qld Sharpe P.R. 2280, Dowling R. Thompson E.1. 78 Blake S.T. 15945 Brass L.J. 18660 ｊ｡｣ｯ｢ｾ 246 Appendix 2 Full character list used in the assessment of monophyly for the tribe Abildg,aardieae. All characters used in cladistic analyses in Chapters 3, 4, and 5 are subsets from this list. Characters presented in the cladograms from subset analyses are given in square brackets after the tribal character name, with the related chapter indicated as: Chapter 3==!, Chapter 4=/\ and Chapter Ｕ］ｾ following the subset character number. 1. Longevity whether 1. annual 2. perennial (with remains of old sheaths and or culms) Some young perennial plants may appear to be annual therefore care must be taken when scoring this character - if possible check plants in the area when collecting and note 2. Perennial rhizome whether 1. caespitose - indistinct due to compaction giving clumped base, detectable by persistent sheaths from last years growth 2. base clumped but rhizome visible sometimes growing vertically 3. base not distinctly clumped but has obvious 'running rhizome' giving smaller clumps spread 4. distinct thick horizontal rhizome with many scales, not spreading widely 5. very distinct and elongated horizontally - base widely spreading 3. Sheath surface cover 1. glabrous 2. scabrid backs 3. scabrid margins 4. medium to dense cover of short to medium hairs 5. mixture of short and long hairs 6. short plus or minus horizontal hairs 60-100 ,. lIn 7. sparsely bristly (hispid - hairs horizontal or nearly so) 8. densely bristly 9. sparse to medium distribution oflong flexuose hairs (c. 300 Ilm, as in Arthrostylis aplzyl/a) 10. dense matt oflong hairs 4. Sheath fitting whether at maturity 1.. tight fitting around culm, especially evident at sheath apex - sheath closed (as in Sclzoenoplectus mucronatus) 2. fits against culm but not really tight and not loose 3. sheath reduced to near base of plant and is open - recognised from sheath margins (as in F. fimbristyloides) 4. seems absent and reduced to extreme culm base (leaf blade seems to go all the way to the base of the plant) 5. open at apex but not loose (as in Abildgaardia oxystachya) 6. sheath loose evenly around the culm and the sheath length, not more so around the apex - sheath closed (as in Schoenoplectus tabernaemontani) '7. loose and open around culm, especially at sheath apex (as in Crosslandia setifolia and Abildgaardia vaginata) 8. sheath apparently open along it's length, at least at maturity - if fused then only in young culms 5. Sheath margins texture 1. sheath margins barely discemable 2. hyaline (thin and translucent (transmits light - very easily damaged)) 3. membranous (thin and semi-translucent (like frosted glass), membrane-like) 4. thinly chartaceous (thinner than chartaceous) S. chartaceous (papery, opaque (light not transmitted) and thin) 6. subcoriaceous (thickish and strong) 7. coriaceous (thick and leathery but flexible) 6. Sheath bases whether 1. sheath base intact as interveinal tissue persists 2. breaking down to fibres from remaining nerves - distinct 7. Sheath backs texture 1. hyaline (thin and translucent) 2. membranous (thin and semi-translucent - membrane-like) 3. thinly chartaceous (papery, opaque (not transparent, dull not shining) and thin) damaged with forceps not flexible 4. distinctly chartaceous (colour and feel of thin papyrus) 247 5. scarious (thin and dry appearing shrivelled) 6. subcoriaceous (thick and leathery) not easily damaged 7. cartilaginous (hard and tough but flexible) 8. fibrous (having loose woody fibres) 9. pannose (with a felty texture) 8. Sheath apex shape excluding hair extensions 1. pointed (apex longest on margin receding back to culm giving pointed triangular affect) 2. truncated (margins abrubtly end and are the same width the length of the sheath) 3. rounded (margins rounded and are or at least close to the full width of margins along rest of sheath) 4. tapered (margins narrowed at leaf junction gradually widening to full margin width of sheath along rest of length) 5. auriculate (sheath tapering then with rounded ends forming auricle) 6. extended (extends beyond sheath-blade junction abaxially) 9. Sheath orifice adaxial 1. open when mature 2. deep V 3. fused at apex 10. Sheath colour in dried material 1. cream 2. straw coloured (golden) 3. yellow-brown 4. light brown 5. dark golden brown 6. pale orange brown 7. orange brown 8. pink-brown 9. red brown J[ O. mid brown 1l1. dark red brown 12. dark brown (at least nerves are very dark) 11. Sheath leaf junction whether [121\] 1. glabrous 2. short hairs unrelated to ligule (hairs restricted to sheath apex margin and not across the width of the blade junction) 3. pilose (at sheath apex but not continuing across the full width of blade junction) 4. long coarse hairs 12. Leaf to culm ratio (mature culms and leaves, with or without leaf blade) 1.1:4 2. 1:3 3. 1:2 4.2:3 :5. 1: 1 13. Ligule (whether present) 1. absent 2. as a fringe of stout hairs (near the sheath apex adaxial across the blade or subulate point as in Fimbristylis depauperata) 3. as a membranous flap (formed as a continuation of the sheath margins adaxial across the blade or subulate point as in Schoenoplectus) 14. Leafblade whether present or absent 1. absent (binseniform) 2. reduced to subulate points only 3. mixture (of absent or reduced and well developed blades within an individual) 4. present (always) 15. Leaf number per culm (includes bladeless sheaths associated with an individual culm) 1. one per culm 2. two to three per culm 3. greater than three and up to five per culm 4. greater than 5 less than 10 5. greater than 15 248 doesn't include open sheaths that are restricted to base of 'groups' of culms 16. Leafblades shape (see Radford 1974 p. 129) 1, narrowly elliptic (with widest axis at midpoint of structure and with margins synunetrically curved; more then L W 6:1-3:1) 2. linear (with widest axis at midpoint of structure and with margins essentially parallel; more than L W 12:1) 3. linear-Ianceolate 4. ovate (with widest axis below middle and with margins symmetrically curved; L W 2: 1-3:2) 5, ovate-Ianceolate 6, lanceolate (with widest axis below middle and with margins symmetrically curved; more than L W 6:1-3:1) 7, obovate (inversely ovate) 8. falcate (broad blade that is sickle-shape, arcing back from centre of plant) 17. Leafblades habit whether 1. erect (follows line of and is usually intermingled with culms) 2. erect then recurved near apex 3. curly ascending (slightly horizontal then upright) 4. ascending (slightly horizontal and then upright from mid ofleafusually at side of culms) 5. ascending spreading 6. loosely ascending as leaves all 'mishappened' 7. strongly falcate (leaves bent backwards from near base of plant) 18. Leafblade shape in transverse section (along mid-third) 1. sub-triangular (adaxially concave and abaxial midrib distinct as a point with convex sides) 2. concave triangular (abaxial sloping concave faces) 3. shallow channel (crown-like abaxial central rib as point and concave sides giving shallow channelled appearance from the tlrree points adaxial side usually concave as in B. sp aff. barbata.) 4. strongly channelled (with deep channels as in B. turbinata) 5. thickly crescentiforrn 6. v-shaped 7. thickly v-shaped 8. U-shaped 9. crescentiform 10. half-circular or obliquely so 11. depressed elliptic 12. thinly crescentiforrn (as in F. schultzii) 13. broadly linear (almost flat - horizontal and thin 14. fused at the margins and is almost subcylindrical 15. fused at the margins and is almost triangular 19. Leafblade width (at midpoint of blade) 1. to 0.3 mm 2. greater than 0.3 to 0.5 mm 3. greater than 0.5 to 0.73 mm 4.0.75 to 0.8 mm 5.0.85 to 1.4 mm 6.1.5 to 2.0 mm 7.2.1 to 3.0 mm 8. greater than 3.0 to 4.8 mm 20. Leafblade vestiture 1. glabrous 2. minutely scabrid margins 3. scabrid margins 4. scabrid over abaxial surface 5. hairy not scabrid (hairs almost horizontal to slightly antorse) 6. densely hirsute backs 7. sparsely bristly 8. bristly hispid 21. Leaf anatomy sclerenchyma presence 1. apparently absent 2. in abaxial area only 249 3, in abaxial and to a lesser extent adaxially (not including sclerenchyma at leaf margins) 22. Leaf anatomy sclerenchyma 1. present as strands (not in contact with vascular bundle but adjacent to) 2. present as girders (in contact with vascular bundle) 3. present as cap above the phloem 4. present as a cap on the inner side of the vascular bundle 23. Leafblade anatomy sclerenchyma shape for abaxial surface 1. square to square with concave sides 2. rhombic (with upper epidermal edge shorter than inner one) 3. reverse rhombic (with upper epidermal edge longer than the inner edge) 4. low mound 5. dome (wide base towards VB) 6. high dome 7. oval-elliptic 8. circular-rounded 9. crescentiform 10. pulviniform (rounded rectangular) 11. rectangular 12. triangular (point towards VB) 24. Number of sclerenchyma strands or girders (compared to vascular bundle number) 1. less than the number of vascular bundles 2. equals the number of vascular bundles 3. greater than number of vascular bundles 25. Leaf anatomy vascular bundle number 1. less than five 2. five to seventeen 3. greater than twenty 26. Leaf anatomy vascular bundles, whether 1. vascular bundles form one layer below the abaxial epidermis 2. vascular bundles form a partial or complete second row as new VB's form near the abaxial epidermal region 27. Leaf anatomy size of vascular bundles., whether 1. same size 2. 2 sizes with midrib VB being the larger 3. varying sizes with largest bundle at midib 28. Leaf and culm anatomy type of vascularisation [27'''; 26!] 1. C4 fimbristyloid (primary and secondary bundles have PCR tissue interrupted laterally by the metaxylem vessel elements, the mestome sheath complete and surrounded by PBS) 2. C3 type (having two sheath layers, formed by the mestome sheath which is surrounded by large achlorenchymatous parenchyma sheath) The inner border parenchyma cells are large and chlorenchymatous, constituting the PCR tissue, and interrupted laterally by the metaxylem vessel elements; the mestome sheath of small, achlorenchymatous, thick-walled cells; and a complete (unless interrupted by sclerenchyma) PBS, which is usually smaller and less chloroplast laden than the surrounding PCA tissue (a PBS also surrounds the secondary bundles). Definitions from Bruhl (1990). The C4 anatomical types are described in terms of primary vascular bundles, the latter being recognised by the possession of meta- and proto-xylem, often associated with a protoxylem lacuna. C4 has either fimbristyloid, chlorocyperoid, eleocharoid, or rhynchosporoid type Fimbristyloid C 4 comprises three bundle sheaths in primary and secondary bundles: the inner border parenchyma cells are large and chlorenchymatous (=PCR tissue) and is interrupted laterally by the metaxylem vessel elements; the mestome sheath of thick-walled, achlorenchymatous cells; and a complete PBS of smaller cells that are less chloroplast laden than the surrounding PCA tissue. Chlorocyperoid C4 anatomy is essentially similar, but here the PBS is restricted to one or a few cells lateral to the metaxylem vessel elements, or sometimes completely absent (being always absent from secondary bundles). The border parenchyma cells also constitute the PCR tissue in eleocharoid C4 anatomy, but are usually not interrupted by the metaxylem vessel elements, and the PBS is absent. The mestome sheath constitutes the PCR site in rhynchosporoid C4 species, and the PBS is present but irregularly incomplete. 250 29. Leaf anatomy parenchymatous bundle sheath (peA) [27!] 1. colourless (no chlorophyll present) as seen in Schoenoplectlls and Arthrostylis 2. contains chlorophyll 30. Leaf vascular bundles whether 1. embedded within chlorenchymatous tissue 2. not embedded within chlorenchymatous tissue but intrudes into the clear parenchyma tissue 3. chlorenchymatous tissue apparently absent, or at least highly reduced and undiscemable (tannin cells prevalent) 31. Leaf chlorenchyma shape 1. globular parenchyma surrounding VB 2. elongated parenchyma near epidermal area above VB and globular around lower part of VB 3. elongated cells surround VB 4. obvious rectangular palisade surrounding the VB 32. Leaf anatomy mesophyll between vascular bundles, whether 1. completely radiate (chlorenchyma arranged in a distinct ring around all major and minor vascular bundles) 2. incompletely radiate (individual vb's partially encircled by chlorenchymatous cells that are radiately arranged - no radial vb's occur at the xylem end of the vb, radiate around the phloem end) 33. Leaf anatomy bulliform cells, whether 1. absent 2. present 34. Leaf anatomy epidermal cells, whether 1. epidermal cells the same size or only slightly larger than abaxial counterparts 2. adaxial epidermal cells about twice as large as abaxial epidermis 3. adaxial epidermal cells mostly three to four times as large as abaxial 4. adaxial epidermal cells about 6 times larger than abaxial cells 5. central adaxial epidermal cells three to four times the size of abaxial epidermis 6. central adaxial epidermis cells about six times the size of abaxial epidermis 1'. central adaxial epidermal cells about 10 times larger than abaxial epidermal cells Adaxial epidermal cells situated at the midrib that are much larger than the neighbouring epidermal cells - scored as present when obviously enlarged compared to abaxial epidermal cells 35. Leaf anatomy adaxial hypodermis, whether 1. absent, as epidermal cells only present 2. present, as 1 row often incomplete 3. present, as 2 rows (sometimes 2nd row incomplete) 4. present, as 3 rows (sometimes incomplete) :;. many rows present ie greater than four rows 36. Leaf anatomy air cavities, whether ]. absent 2. present 37. Leaf anatomy stomata 1. raised (protrude above cuticle layer - above leaf surface) 2. flush (with leaf surface - discrete between epidermal cells) 3. sunken (into leaf surface at bottom of dissecting ribs or pits etc) 38. Culm whether noded vertically 1. not stalked (no obvious ascending nodes present) 2. stalked (node is obvious and culms appear stalked) 39. Culm outline in transvers section, whether 1. narrowly elliptic 2. elliptic no dissections or wavy margins 3. elliptical smooth dissected (deep dissections around the girth of the culm) 4. elliptic wavy no dissections 5. elliptical wavy or undulating with dissections evenly around the margin 6. elliptical deeply undulating 7. elliptic with distinct ribs (grooved) 8. irregularly elliptic with wavy margins 9. irregularly fusiform shaped (waves caused by protruding sclerenchyma bundles as in Fimbristylis microcarya) 251 40. 41. 42. 43. 44. 10. irregular quadrangle 4 sides, with the parallel sides having one shorter than the other (see Arthrosytlis aphylla) 11. transversly oblong 12. irregular no definite shape 13. irregularly 7-ribbed 14. irregularly 9-ribbed 15. sub-symmetrical 6-ribbed 16. sub-symmetrically 7 ribbed, distinct 17. sub symmetrically 8-ribbed 18. circular 19. circular undulating 20. circular with distinct ribs (see Actinoschoenus composita) 21. pentagonal (having 5 sides) 22. acutely hexagonal 2:3. irregular hexagon with ribs (6 main points with 6 subpoints forming ribs) 24. depressed triangular with convoluted margins 2: 5. triangular 26. triquetrous (3-angled with concave sides (acutely triangular Metcalfe 1971) Culm width (at mid third section) 1. to 0.3 mm 2. 0.31 to 0.8 mm 3.0.85 to 1.1 mm 4.1.15 to 1.3 mm 5. 1.5 to 1.6 mm 6. 1.7 to 2.4 mm 7.2.5 to 3.0 mm 8.3.2 to 4 mm 9. greater than 4 mm Culm cover t. glabrous (often scabrid at inflorescence junction) 2. scabrid (minute prickle hairs isolated to sparse cover) 3. scabrid (minute prickle hairs with dense cover) 4. hairy not scabrid 5. toothed ascending hairs (40-60 flm) 6. finely pubescent (short interlocking hairs) '7. hairs mixed (short and long ?toothed) 8. bristly (distally) 9. bristly along the culm length 10. long hairs (c. 600 flm) dense - A<B (when A is distance between trichomes and B is trichome height); sparse - A=B to 5xB isolated - A>5xB Culm anatomy, total number of vascular bundles is [41-] 1. less than fifteen 2. greater than fifteen less than thirty 3. greater than thirty less than thirty-five 4. greater than forty Culm anatomy, size of vascular bundles 1. all same size 2.2 sizes alternating 3. 2 sizes not alternating 4. varying sizes alternating evenly 5. varying sizes not alternating evenly Culm anatomy, number of rings of vascular bundles (VB's) 1. not arranged in rings apparently 'unorganised' within the culm 2. ring of vascular bundles around the outer culm edge and dispersed throughout the culm 3. single ring of bundles all of similar size or alternating size 4. one complete ring and a second semi ring (with the inner VB's being primary and pushed inward when secondary VB's form in the outer region) 252 45. 46. 47. 48. 49. 5. two complete rings of vascular bundles (the inner ring formed from a lesser number of primary VB's when secondary VB's develop in the outer ring) 6. multiple rings (older VBs pushed inward while newest VBs are small and numerous near the culm margin) Culm anatomy sc1erenchyma [421\] 1. present as strands (not in direct contact with vascular bundle) 2. present as girders (in contact with vascular bundle) 3. present as a cap to four cells thick above the phloem 4. present as a cap to 9 cells thick at the base of the vascular bundle 5. present as multiple layers around the vascular bundle See Metcalfe 1971 Culm anatomy sc1erenchyma strand number per VB I . less than number of VB in first ring 2. equals number of vascular bundles in first ring (or equals the number in both rings) 3. greater than the number of vascular bundles in first ring Culm anatomy sc1erenchyma shape 1. square strand 2. rhombic (bottom wider than epidermal section) 3. reverse rhombic (upper edge wider than inner one) 4. low mound to dome shaped strand (wide base towards VB) 5. high dome strand 6. reverse high dome strand (widest part on epidermis) 7. pulviniform strand (rounded rectangluar) 8. elliptical strand 9. circular strand la. bulbiform strand 11. crescentiform strand 12. rectangular strand 13. stilted rectangular strand (edges with legs protruding into the parenchyma as in Arthrostylis planiculmis) 14. thickly v-shaped strand (see Actinoschoenus composita) 15. triangular strand (point towards VB) 16. continuous around the culm 17. reverse high dome girder (widest at the VB) 18. roughly circular girder 19. pulviniform girder 20. rectangular girder (see Actinoschoenus composita) 21. triangular girder (point to VB apex) 22. crescentiform girder on inside of VB (as seen in Schoenoplectus tabernaemolltani and Eleocharis) 23. cap above phloem to four cells thick 24. cap at base of vascular bundle to 9 cells thick follows Metcalf 1971 Culm anatomy sc1erenchyma cap on vascular bundle 1. absent 2. present up to 4 cells Culm anatomy photosynthetic parenchyma shape 1. elongate rounded rectangular cells usually arranged roughly in two to four rows beneath the epidermis 2. roughly three to four rows of rounded cells sometimes stretched and packed tightly 3. irregularly shaped shorter parenchyma stacked like brickwork in alternating rows 4. shorter rounded irregularly rectangular single upper row and rounded cells beside and below 5. single row of distinct palisade upper and rounded cells beside and below 6. rounded cells packed tightly in a single row above vascular bundle, but more may be present between the bundles (sometimes slightly stretched but not palisade) 50. Culm vascular bundles 1. not fully immersed within the chlorenchyma tissue as all protrude into the pith (especially evident in C 3 culms) 253 51. 52. 53. 54. 55. 56. 57. 2. younger vascular bundles immersed within the chlorenchYma tissue while mature bundles protmde into the pith (especially evident in C 4 culms) 3. younger vascular bundles partly immersed while mature bundles fully immersed (see Metcalfe p: 394) 4. chlorenchYma apparently absent or highly reduced Culm anatomy central clear parenchYma present as medulla ｛ＵＰｾ｝ 1. absent (no pith cells visible) 2. absent, breaks down to strands between vascular bundles only 3. present and is distinct 4. present but breaking down in the centre Culm anatomy stomata when sunken 1. single stomata at bottom of groove or dissection 2. twin stomata near on side walls of groove or dissection Culm anatomy stomata 1. none apparent or at least very few 2. raised 3. flush (located between sclerenchYma) 4. sunken (at base of dissection) Root width ｛ＵＳｾ｝ 1. to 0.13 mm 2.0.15 to 0.45 mm 3. 0.46 to 0.55 mm 4.0.56 to 0.8 mm 5. 0.81 to 0.85 mm 6. 0.89 to 1.0 mm 7. 1. 1 to 2.0 mm 8.2.1 to 3.0 mm Root colour I. pale cream yellow brown 2. distinctly yellow 3. light brown (straw coloured) 4. orange brown .5. mid brown 6. red brown 7. grey brown 8. dark brown to black Root cover 1. glabrous 2. few hairs 3. villous (many long hairs not matted) 4. tomentose (thickly matted) Inflorescence-synflorescence stmcture 1. spike (intraspicular prophylls indicate lateral branches with a solitary floret, as in Nelmesia) 2. solitary (main florescence only) 3. highly reduced anthelodium (main florescence plus one coflorescence either rayed or sessile) 4. reduced anthelodium (main florescence plus multiple 'rayed' coflorescences - usually 2 to 3) 5. sessile reduced anthelodium (main florescence plus sessile coflorescences - usually 2 giving 3 spikelets) 6. compressed reduced paniculodium forming a 'head' (compressed spike ie main florescence plus multiple lateral primary coflorescences ie reduced paniculodium as in Fimbristylis schultzii) 7. highly reduced secondary anthelodium (with one second order main florescence (HF2), either sessile or on lengthened epipodia (ray) 8. ramified reduced anthelodium (spikelets of second order or higher branching that are mostly 'rayed' or sometimes sessile) 9. many spikeleted 'head' of sessile spikelets formed by primary secondary growth and their secondary lateral growth i.e. HF 1, Cofl + Cof2 (anthela type florescences with reduced epipodia as seen in Crosslandia WA) 10. lateral hemispherical 'head' on long mesopodia (one or two primary 'rays' supporting lateral 'heads' formed from secondary main florescence (HF2) plus sessile secondary coflorescences (Cofl2) and their sessile lateral branches (Cofl3) 254 11. terminal 'head' from many sessile spikelets formed from lateral rays (Cofl) and ramification from intraprophyllar growth (as in Bulbostylis barbata) 12. pseudolateral digitate sessile or on lengthened epipodia (reduced paniculodium) 13. second order pseudolateral digitate lateral and intraprophyllar growth (paniculodium) Synflorescence structures in this study are based on the premise of consistent reduction ie paracladia are absent from the main florescence therefore are also absent from the coflorescences; branches arising are considered as secondary lateral branches (ie ramification) 58. Inflorescence-synflorescence number of usual primary coflorescence branches 'rayed' or sessile ｛ＵＷｾ［ 54"'] 1. absent 2. one 3. two to four 4. five to eight 5. eight to fourteen 6. greater than fifteen 59. Inflorescence (maximum number of orders consistently present) 1. primary (HF 1 and Cofll includes solitary and simple anthelas) 2. secondary (HF2 lateral florescence growth arising from primary florescence parts with its own lateral growth) 3. secondary intraprophyllar (growth from within primary prophylls) 4. tertiary (HF3 lateral florescence growth arising from secondary florescence parts with its own lateral growth) 5. tertiary intraprophyllar (growth arising from with secondary prophylls) 6. fourth (HF4 lateral florescence growth arising from tertiary florescence parts) Orders were determined using compound rayed specimens and then extrapolated to taxa with congested 'heads'. Orders were determined using the terminal spikelets on each culm or ray. For example if a rayed spikelet (i.e. Cofl) developed lateral spikelets (either sessile or on rays) then that spikelet became the terminal spikelet (HF2) for that ray and the lateral spikelets are CoO. Care is required when assessing congested 'heads' of spikelets to ensure that congestion is from the same florescence order. See Crosslandia Figures 3.12-3.17 and Bulbostylis Figure 5.15 for synflorescence type detail. 60. Inflorescence-synflorescence position whether [56"'] 1. terminal (and ascending on the culm) 2. pseudolateral (pushed laterally by the main bracts that usually continue to ascend in line with the culm, although when large spikelet numbers present (as in Schoenoplectus) the main bract is reflexed. In Trachystylis the coflorescence appears in the terminal growing position and the main florescence is pushed sideways) 61. Inflorescence synflorescence, whether open or contracted due to ray length 1. consistently open (solitary spikelet or spikelet 'rays' long and spreading - looks gangly) 2. open (rays lengthened, but not extremely long - as in Bulbostylis turbinata) 3. consistently contracted (spikelets on short rays that can be easily seen giving shortened appearance but not capitate; as in Fimbristylis complanata) 4. mixed (some single spikelets on 'rays' while others may be sessile or on shortened rays) 5. congested (multiple sessile spikelets on severly restricted epipodia, forming heads or finger clusters; see inflorescence-synflorescence structure for different types) 62. Inflorescence-synflorescence length 1. to 35 mm 2.38 to 60 mm :.. greater 60 to 75 mm 4.80 to 100 mm 5. 105 to 120 mm 6. 125 to 170 mm 64. Inflorescence-synflorescence bracts whether [60"'] ]. absent 2. present 65. Inflorescence-synflorescence bracts (when present) 1. g1ume-like and clearly associated with inflorescence when not solitary (may have an apiculate extension as seen in some lower glumes on spikelets) 2. leaf-like and ascending (growing upright at roughly 45 degrees or sometimes loosely erect) 3. leaf-like and spreading (between 45 and 90 degrees separation from culm) 255 66. 67. 68. 69. 70. 71. 72. 4. leaf-like and distinctly erect 5. leaf-like and reflexed downwards due to many aggregated spikelets (as in ｆｩｭ｢ｲｳｾｶｬ scllllltzii) 6. culm-like and continuing in line with the culm (or sometimes obviously bent backwards due to many aggregate spikelets) Inflorescence-synflorescence bracts length 1. shorter than inflorescence 2, equals inflorescence 3. longer than inflorescence 4. much longer than inflorescence Inflorescence-synflorescence primary bract number [63"'] 1. one 2. two (closely alternately opposite) 3. three (ascending alternately of roughly equal length) 4. many (of roughly equal length) Spikelet axis whether 1. monopodial (rachilla of spikelet has one axis ie growth continuous from one growing point) 2. sYmpodial (rachilla formed from multiple reduced axes) Crosslandia with sYmpodial growth has distichous glumes which when glumc pulled away breaks the section of rachilla away as well. The nut is partly surrounded by the glume margins of the opposite (lower) glume. The nutlet is buried deep within the rachilla section. Basal and aerial spikelets show the same pattern even though the spikelets are morphologically different. Aerial spikelet prophylls whether 1. present (distinct bract-like extension between nut and rachilla) 2. absent Aerial spikelet rachilla shape of wings [641\] 1. wingless or minute 2. reduced and rounded (due to compaction of the rachilla) 3. reduced and truncate (due to compaction of the rachilla) 4. reduced and tapered to a point 5. distinct (elongated) rounded - broad with rounded apex (rachilla expanded where fertile) 6. distinct oblong - narrow with curved apex (rachilla expanded where fertile) 7. distinct truncate (apex terminates abrubtly) 8. distinct narrowly triangular - narrow with pointed apex Spikelet sex in aerial spikelets (excluding lowest empty glumes) 1. hermaphrodagamous (bisexual florets only spikelet) 2. hermaphrodandrous (male florets proximal, bisexual florets distal in spikelet) 3. gynehermaphroditic (bisexual florets proximal, female florets distal in the spikelet) 4. gynagamous (female florets only in spikelet) 5. gynecandrous (male florets proximal, female florets distal in spikelet) 6. androgynous (male florets only in spikelet) 7. gynehermaphrodandrous (male, bisexual mid, female distal) Aerial spikelet outline 1. glumes angular - widely dome shaped (truncated proximally and broadly rounded at the apex) 2. elliptic (2: 1 to 3:2 widest point at centre) 3. loosely narrowly elliptic (6: 1 to 3: 1 narrow at base and apex widest point at centre - glumes not tightly imbricate or angular) 4. angularly narrrowly elliptic (6: 1 - glumes somewhat reflexed) 5. obliquely ovate (curving on one side of spikelet due to rachilla twisting) 6. smoothly ovate (2: 1 to 3:2 wider at the base narrowing towards apex) 7. loosely ovate 8. angularly ovate (glumes apex reflexed) 9. cylindrical (as in Schoenoplectiella laevis) 10. smoothly narrowly cylindrical (glumes tightly imbricate - 6: 1 as in Nelmesia melanostachya) 11. loosely narrowly cylindrical 12. loosely lanceolate (glumes not tightly imbricate 3: 1 to 6: 1 wider at the base narrowing towards the apex) 13. loosely obliquely lanceolate (curving on one side due to twisting rachilla) 14. angularly lanceolate 15. narrowly oblong (3: 1 to 6: 1 base and apex same width) 16. triangular (2: 1 to 3:2 wide at base to pointed apex) 256 17. narrowly triangular (6: 1 to 3: 1 wide at base to pointed apex) 18. smoothly linear (glumes tightly imbricate - 12: 1 narrower than narrowly oblong sides even) 19. angularly linear (glume nerves long and recurved) 73. Number of fertile florets (male or female) per aerial spikelet ｛ＷＲｾ｝ 1. 1 to 2 (as in Actinoschoenus, Arthrostylis, Trachystylis) 2. greater than 2 to 4 3. many (greater than 4 up to 25 sometimes many glumes but few nuts as in Abildgaardia) 4. numerous greater than 25 (as in Nemum) 74. Spikelets whether morphologically different to aerial spikelets ｛ＷＳｾ［ 66!] 1. aerial only (always on lengthened culms) 2. aerial plus subradical (subradical culms distinctly shortened and spikelets near base, but spikelets otherwise identical to aerial counterparts) 3. Aerial plus basal (basal spikelets absent or very highly reduced and with different morphology and floret sex to aerial counterparts) 4. aerial plus subterranean (spikelets reduced to one or two nuts close to or below ground level amphicarpy) Basal spikelets are those that differ in morphology and frequently sexuality, from aerial spikelets, usually maturing before their aerial counterparts i.e. amphicarpic. As seen in Crosslandia, Fimbristylis spiralis and occasionally in Abildgaardia vaginata, plus some species of Bulbostylis and Schoenoplectiella. 75. Basal spikelet sex (excluding lowest empty glumes whether) 1. hermaphrodagamous (bisexual [perfect] florets only in spikelet) 2. gynehermaphroditic (bisexual florets [perfect] proximal, female florets distal in the spikelet) 3. gynagamous (female florets only in spikelet) 76. Basal spikelet (shape) 1. irregularly widely ovate (due to extreme reduction of number of florets - usually one to two) 2. narrowly elliptic (6: 1 to 3: 1 narrow at base and apex widest point at centre) 3. lanceolate (3: 1 to 6: 1 wider at the base narrowing towards the apex) 4. narrowly oblong (3: 1 to 6: 1 base and apex same width) 5. oblanceolate (6: 1 to 3: 1 narrow at base and wider at apex due to spreading glumes) 6. narrowly triangular (6: 1 to 3: 1 wide at base to pointed apex) 7. linear triangular (12: 1 wider at base than the tapered apex) 8. linear (12: 1 narrower than narrowly oblong with sides even) 77. Basal spikelet (floret numbers) 1. one to two 2. greater than two but less than four 3. greater than four to many 78. Basal spikelets whether 1. Basal spikelets always sessile (no culm present) 2. Basal spikelets mostly sessile although some with very short culms present (less than 3 mmlong) 3. Basal spikelets mostly on reduced culms (at first appearing sessile and clumped at plant base although rarely some longer culms with gynagamous spikelets present) 4. Basal spikelets mostly on shortened culms greater than 5mm long (often c. 15 mm) and restricted to plant base 79. Spikelet glumes whether 1. always falls with mature nuts 2. persists on the spikelet after nuts mature 80. Aerial glume length 1. to 2.12 mm 2. 2.2mm to 3.5 mm 3.3.6 to 3.8 mm 4.3.9 to 4.1 mm 5.4.2 to 4.55 mm 6.4.6 to 4.85 mm 7. 4.9 to 5. 1 mm 8.5.2 to 5.75 mm 9.5.8 to 5.95 mm 10. 6.0 to 6.25 mm 11..6.3 to 6.55 mm 12. 6.6 to 6.8 mm 257 13.6.9 to 7.1 mm 14.7.2 to 8.4 mm 15.8.5 to 10.1 nun 16. 10.2 to 11.6 mm 17. 12 to 13.7 mm 18. 14 to 15 mm 81. Aerial glume width [74/\] 1. to 0.7 mm 2.0.75 to 0.9 mm 3. greater than 0.9 to 1.46 mm 4. 1.5 mm to 1.7 mm 5. greater than 1.7 mm to 1.85 mm 6. greater than 1.85 mm to 2.05 mm 7.2.05 mm to 2.4 mm 8. 2.45 mm to 2.55 mm 9. 2.6 mm to 2.9 mm 10. greater than 2.9 mm 82. Aerial glume back colour (excluding tannins) 1. no real colour as is translucent (some tannin may be present mainly near the glume base) 2. cream 3. straw 4. yellow brown 5. pale orange brown 6. orange brown 7. light brown 8. pink-brown 9. mid brown 10. red brown 11. deep burgundy (as in Nemum) 12. burgundy black (very dark burgundy appearing almost black) 83. Aerial glume margin colour whether 1. darker than gume back 2. same colour as glume back and sides 3. colour lighter than glume back 4. consistantly colourless and distinct 84. Aerial glume margins ｛ＸＳｾ｝ 1. entire (without indentations, incisions, or trichomes along margins) 2. lacerate (margins irregularly cut, appearing torn) 3. with antrorse prickle hairs pointing towards apex 4. minutely ciliolate «20 11m) 5. ciliolate (with tiny or small trichomes protruding from margins c. 20 11m) 6. short hairs 7. hispid or almost so (as a continuation of glume back indumentum) 8. fimbriolate (minutely fimbriate flattened projections) 9. fimbriate (fringed margins with flattened processes) 10. ciliate at apex only 11. loosely ciliate (long lax hairs that look mishappen) 12. piliferous (with long conspicuous trichomes c. 200 11m, that are lax or flexuose, and protruding from margins) 85. Aerial glume whether margin in transverse section 1. is continuing in line with the glume sides, not inrolled or splayed 2. is inrolled between 1/2 and upper 1/3 of glume 3. margins only slightly curved backwards or flattened with glume still generally boat shaped 4. is splayed so that the margin is between 45 and 90 degrees - flattening out giving a narrow keel with splayed sides 5. is almost flattened (only the nerve is raised; the glume sides are flattened against the glume below, although the basal area around the nutlet may be boat shaped) 6. is revolute (margin strongly recurved and bending backwards) 86. Glume texture whether [77!] 1. hyaline (thinner than membranous and very delicate, usually colour is absent) 258 87. 88. 89. 90. 2. membranous (almost transparent and usually colourless nut can be seen through the glume as in Bulbostylis sp. aff barbata) 3. finely chartaceous (thicker than membranous light may be seen but is dulled and papery, has some flexibility but easily damaged with forceps, as in Fimbristylis blakei) 4. tougher than chartaceaous as is more flexible but damaged with forceps (as in Actinoschoen/ls) 5. chartaceous (thicker than finely chartaceous as light does not pass through but may still be damaged, as in Crosslandia anthelata) 6. fine leathery (not overly thickened but is quite tough and flexible not easily damaged unless pulled, as in some Fimbristylis) 7. subcoriaceous (quite tough and hard to bend, thick and not at all transparent, as in species of Abildgaardia ) Aerial glume margins ｛ＸＶｾ［ 80"']] 1. margins hyaline (light passes directly through) 2. membranous (light is opaque) 3. finely chartaceous and indistinct, or almost so, from rest of glume 4. indistinct from rest of glume (with same texture as glume backs which are subcoriaceous as in Abildgaardia) Aerial glume apex outline 1. rounded (margins and apex forming a smooth arc) 2. retuse (lobe rounded; sinus depth to 1/16 distance to midpoint of blade; margins convex) 3. emarginate (lobe rounded; sinus depth 1/16 to 1/8 distance to midpoint ofblade;margins straight or convex) 4. obtuse (margins straight to convex, forming a terminal angle more than 90 degrees) 5. acute (base cuneate - margins straight to convex forming a terminal angle 45-90 degrees; muticous) 6. acuminate (base narrowly cuneate - margins straight to convex forming a terminal angle of less than 45 degrees; muticous) 7. sub-mucronate (nerve less than 0.1 mm, but not muticous) 8. mucronulate (1: 1 I w nerve 0.1 mm) 9. mucronate (with nerve less than 3: 1 length/width, straight and stiff between 0.1 and 0.3 mm) 10. apiculate (more than 3: 1 length/width, usually slightly curled and flexuous; used for nerve extension greater than 0.3mm but equals or less than 0.7 mm) 11. aristate (more than 3: 1 length/width, usually prolonged, straight and stiff; used here for excurrent nerve forming awn-like projection greater than 0.7 mm long) Aerial glume general shape [80!] 1. oblong (2: 1 to 3:2 with widest axis at midpoint of structure and with margins essentially parallel) 2. narrowly oblong (6:1 to 3:1 with widest axis at midpoint of structure and with margins essentially parallel) 3. spathulate 4. linear (more than 12: 1 with widest axis at midpoint of structure and with margins essentially parallel) 5. ovate 6. widely ovate 7. very widely obovate (1: 1 with apex curving in a wide arc) 8. narrowly emarginate glume apex narrowing but often rounded at end with nerve extending and is usually reflexed (see Fimbristylis schultzii) 9. emarginate (glume apex almost as wide as base and rounded, dipping into but not attached to nerve) 10. obtuse (nerve terminates below glume apex and apex is broad and round) 1 Jl. trullate (2: 1 to 3:2 kite shaped with widest point near base) 12. lanceolate (more than 6: 1 to 3: 1 with widest axis below middle and with margins symmetrically curved) 13. oblanceolate (more than 6:1 to 3:1 reverse of laneeolate) 14. narrowly trullate (6:1 to 3:1 with widest axis below middle and with straight margins, trowel shaped) 15. triangular (2:1 to 3:2 with 3 sides and 3 angles) 16. narrowly triangular (6: 1 to 3: 1 with 3 sides and 3 angles) 17. linear triangular (more than 12: 1 with 3 sides and 3 angles) Aerial glume apex whether 1. not recurved at maturity (usually looks quite sleek and neat) 2St) 2. distinctly straight - not bent backwards or curving forwards 3. slightly reflexed backwards, but not strongly recurved 4. strongly and consistently recurved at maturity (as in Abildgaardia schoenoides) 91. Aerial glume nerve whether 1. muticous (nerve does not extend pass glume apex, frequently finishes abrubtly beneath glume apex) 2. submruco point (c.0.05 to 0.1 mm) 3. nerve to mucro point (to 0.1 to 0.5 mm) 4. nerve excurrent (greater than 0.5 mm) 92. Aerial glume abaxial surface l. glabrous 2. nerve only scabrid 3. sparsely scabrid 4. scabrid over most of the surface :5. short antrorse appressed hairs 6. antrorse hairs (c. 100um long at 45 degrees to glume back) 7. bristly 8. tomentose 93. Glume epidermal cells shape in glume sides (at 50 x magnification) 1. indistinct 2. rectangular 1:2 to 1:3 longitudinally lengthwise with strongly sinuose walls 3. rectangular 1:2 to 1:3 longitudinally lengthwise with straight walls 4. linearly rectangular 1:6 longitudinally 5. linear longitudinally 6. irregularly elongate 94. Glume epidermal cells whether 1. tanin idioblasts absent 2. tanin idioblasts present (cells filled with tanins but not raised) 95. Glume epidermal tanins shape 1. present as cellular 'dots' 2. present as squares 3. present as rectangular c. 3: 1 4. minute striations which are very faint 5. present as long striations (vertical lines along length of glume) 6. present as joined striations forming almost continuous lines 7. forming continuous 'colour' 96. Aerial glumes tanin cover 1. restricted mainly to apex 2. sparse and restricted mainly to glume base 3. sparse and restricted to glume backs and sides, not on margins 4. sparse mostly occurring near outer margins only 5. sparse mostly occurring on glume sides and margins (rarely in nerve area) 6. evenly over glume but not all cells gives speckled appearance 7. dense over glume sides and margins 8. dense and continuous over glume back and thinning on sides 97. Glume epidermal cells whether 1. raised' gland' cells (as seen in Fi11lbristylis cinna11lo11letoru11l) 2. devoid of any obvious raised gland-like cells 98. Aerial glume shape in cross-section [91/\] 1. single highly thickened nerve giving distinct keel (see Fi11lbristylis schultzit') 2. narrowly keeled (as in Bulbostylis barbata, narrow V of 3-nerves forming a keel) 3. narrow U of 3 to 5 nerves 4. broad V (from multiple nerves but forming a distinct V as in Abildgaardia vaginata) 5. nerve area broad 5 to 7 nerves forming U but margins continuous texture with sides (sometimes glume flattened but nerves distinct) 6. nerve area broad (forming U with distinct margins from glume sides, usually consists of 5 to 7 nerves forming rounded V-bottom) 7. nerves indistinct (glume forms wide, shallow V with margins not flattened as in 2; nerves usually indistinct or only 1 fine nerve visible) 8. nerve distinct and broad with glume broad and shallow curved or almost flat 260 9. nerve area often indistinct or as apparent single slim nerve glume relatively flat at least above 99. Aerial glumes glandular cover 1. mostly restricted to apex 2. mostly restricted to margins 3. over most of surface 100. Aerial glume arrangement 1. distichous (attached directly oppositely ascending, sometimes rachilla twists to give spiraldistichous impression but glumes in definite rows) 2. sub-distichous (glumes at least distichous at first then twisting, but not fully spiral spirodistichous) glumes appear spirally arranged and not in rows but spikelet is slightly depressed in cross section 3. almost alternately opposite (decussuate but ascending minutely acropetally as seen in Bulhostylis barbata) 4. opposite decussate (as in Trachystylis where empty glumes are paired and 90 degrees from distal fertile pair) 5. tristichous where glumes attached ascending in a tristichous spiral 6. spiral (glumes attached in a close ascending spiral acropetally) 101. Aerial glumes (number neutral (empty) per spikelet) 1. none 2. one 3. two 4. three 5. four 6. five 7. six or seven 102. Basal glume (length) 1. less than 2 mm 2. greater than 4 mm 103. Basal glumes (margins) 1. entire 2. ciliate 3. ciliolate 4. fimbriate 5. fimbriolate 6. involute 7. lacerate 104. Basal glumes (general outline) 1. linear (12: 1 - widest axis at midpint of structure and margins essentially parallel) 2. narrowly oblong (6:1 to 3:1) 3. lanceolate (more than 6: 1 to 3: 1 - widest axis below middle and margins symmetrically curved) 4. ovate (2:1 to 3:2 - see lanceolate) 5. widely obliquely ovate 6. narrowly trullate (more than 6: 1 to 3: 1 - with widest axis below middle and with straight margins) 7. triangular (2:1 to 3:2 - with 3 angles and 3 sides) 8. narrowly triangular (6: 1 to 3: 1 - with 3 sides and 3 angles) 9. linear-triangular (more than 12:1 - with 3 sides and 3 angles) 105. Basal glumes abaxial surface 1. glabrous 2. nerve scabrid 3. back scabrid 106. Basal glume epidermal cells whether 1. tanin idioblasts absent 2. tanin idioblasts present 107. Basal glume epidermal cells tanin cover 1. restricted to glume base 2. sides of glumes 3. over entire glume (giving striated appearance) 108. Perianth whether present 1. absent 261 2. present as bristles Hypogynous bristles or scales are absent in members of the Abildgaardieae, however, there is one specimen of Abildgaardia schoenoides collected from Kakadu NP that has perianth present. 109. Bristle hairs, whether 1. antrorse 2. retrorse 110. Perianth number when present 111. Stamen number in aerial male or bisexual florets 1. one 2. two 3. three 4. four 5. five 6. six 112. Anther length, including apiculum 1. to 0.15 mm 2.0.2 to 0.75 mm 3.0.8 to 0.9 4. 0.95 to 1.50 mm 5. 1.55 mm to 1.65 6. 1.70 to 1.85 7. 1.9 to 3.0 8. 3.0 to 3.50 9. 3.60 to 4.2 10.4.3 to 6.7 mm 113. Anther apiculum, whether connective tissue extends past the antheridium 1. indistinct (either absent or to 0.05 mm in length) 2. distinct (greater than 0.05 mm to 0.2mm) 3. prominent (greater than 0.2 mm) 114. Aerial style length (measured from base of stylebase to base of stigmas) 1. to 1.25 mm 2. 1.30 to 2.75 mm 3. 2.80 to 3.35 mm 4. 3.4 to 3.8 mm 5. 3.9 to 4.30 6.4.40 to 4.9 7. 5.0 to 5.50 mm 8. 5.6 to 6.2 mm 9.6.5 to 7.9 mm 10.8.0 to 12.60 mm 115. Aerial style width [106!] I. to 0.1 mm maximum measurement 2. greater than 0.1 to 0.15 mm (0.1 is minimum measurement) 3. greater than 0.15 mm to 0.2 mm 4. greater than 0.2 mm to 0.25 mm 5. greater than 0.25 mm to 0.30 mm 6. greater than 0.30 mm to 0.34 mm 7.0.35 mm to 0.45 mm 116. Aerial style surface cover, (excludes style-base) 1. glabrous 2. isolated fimbriola 40-80um 3. sparse fimbriola 40-80um 4. dense fimbriola 40-80um 5. isolated fimbria 100-140um, scattered along style often nearer the base and missing from apex 6. sparse fimbria 100-140um, with the distance between each process greater than the length of each process 7. dense fimbria 100-140um, with the distance between each process less than the length of the process 8. coarsly fimbriate 220 ｾｭ long and 100 ｾｭ apart dense becoming very dense towards base 9. ciliate 262 10. densely matted with long hairs excluding style-base 117. Aerial style outline in transverse section 1. flattened and broad 2. distinctly flattened (strap-like, as seen in many Fimbristylis species) 3. terete (apparantly, as seen in Trachystylis) 4. finely triangular (as seen in Crosslandia) 5. minutely triangular (as seen in Bulbostylis barbata) 6. distinctly triangular (sometimes slightly flattened but with 3rd angle distinct) 7. broadly triangular with distinct flattened face giving flat appearance 118. Aerial style base length (in millimetres) 1. from 0.05 to 0.2 mm 2. to 0.5 mm 3. greater than 0.5 and less than 1.5 mm 4. greater than 1.5 to 3.5 mm 119. Aerial style base width (in millimetres) 1. to 0.15 mm 2. 0.2 to 0.5 mm 3. greater than 0.5 to 1.0 mm 4. greater than 1.0 mm 120. Style-base shape 1. not widened or distinct 2. globular (as seen in Bulbostylis barbata) 3. bulbous 4. narrowly triangular (as in Crosslandia and Abildgaardia vaginata) 5. regularly triangular (no face distinctly concave or convex) 6. triangular (with adaxial faces concave and abaxial face flat as in Abildgaardia macrantha) 7. conical 8. broadly triangular (somewhat rounded not with distinct triangular edges) 9. broad and convex (on abaxial side away from rachilla giving a depressed appearance) 10. squarish to barely triangular and depressed (as seen in Fimbristylis depauperata continues in line with style but is distinct) 121. Aerial style-base surface cover [110!] 1. glabrous 2. sparse rounded fimbriola (20 11m in length) 3. isolated to sparse fimbriola (40-60 11m) 4. dense fimbriola (40-60 11m) 5. fimbriate (100-140 11m) 6. coarsely fimbriate (c. 220 11m and 100 11m apart thick coarse looking very dense on style base) 122. Aerial or basal style-base, whether persistent or deciduous [116/\] 1. persistant (style-base always separates from style and often persists on nut apex, but not always) 2. deciduous (style-base remains connected to style when abscissed from nut apex) 123. Aerial style stigma number 1. two 2. three 124. Aerial style stigmas relative length, when compared with style length I. stigmas (less than length of style) 2. approximately equals (stigmas=style L) 3. stigmas greater than style length 125. Aerial style stigmatic processes observed at lOx magnification 1. minute papillae 2. small fine papillae 3. fimbriolate 4. fimbriate 5. woolly (distinctly) 126. Aerial style stigma colour 1. white 2. golden brown 3. red brown 4. deep red-brown 127. style (surface cover, excludes style-base) 263 1. glabrous 2. occasional fimbriola 3. sparse 4. hairs denser at base and sparse or absent towards style apex 128. Basal style (shape in transverse section) 1. ligulate 2. terete 3. narrowly triangular 4. minutely triangular 129. Basal stylebase (indumentum cover) whether 1. absent as stylebase is glabrous (at least appearing so) 2. occasional fimbriola 3. sparse fimbriola occuring near style base to nut abscission zone 4. dense fimbriola 40-120 ｾｭ 130. Aerial nutlet length [1241\] 1. less than 0.65mm 2. 0.65 to 1.3mm 3. 1.35 to 2.0mm 4. greater than 2 and less than 3mm 5. greater than 3 less than 4.5mm 131. Aerial nutlet width [1251\] 1. to 0.60 mm 2. 0.61 to 1.15 mm 3. greater than 1.15 mm to 1.34 mm 4. 1.35 mm to 1.50 mm 5. greater than 1.50 mm to 1.75 mm 6. greater than 1.75 mm less than 1.90 mm 7. 1.90 mm to 2.50 mm 132. Aerial nutlet stipe length 1. zero as no constriction observed 2. to 0.2 mm 3. greater than 0.2 to 0.28 4. 0.3 to 0.4 5. greater than 0.4 to 0.8 6. greater 0.8 to 1.0 mm 7. greater than 1.0 to 1.10 133. Aerial nut hypogynophore [127 /\] 1. apparently absent or highly reduced enclosed within the fruit wall (may appear as a 'button') 2. distinct as a stalk not enclosed by the fruit wall of the nut (usually brownish) with filaments attached at the base 3. completely enclosed within the 'stipe' of the nut fruit wall and not apparent from the outside (with filaments attached at the base of the stipe) Hypogynophore is a stalk directly below the ovary and may be apparent as separate from the nut proper or enclosed within the stipe of the nut. On some nuts the hypogynophore may be highly reduced or appear absent. Stamens and perianth parts are attached at the base of the hypogynophore. In Abildgaardia odontocarpa the hypogynophore is present in the stipe of the nut and not evident from the outside - separating from the seed proper when the wet nut is opened. The hypogynophore adheres to the inner area of the nut that encased it. 134. Aerial nutlet shape in transverse section (at mid-third of organ) [123!] 1. biconvex (lenticular depressed due to two faces) 2. cylindrical (without distinct sides appearing rounded - ribs not evident) 3. rounded trigonous 4. distinctly plano-convex (distinct dorsal ventral sides in 3-sided fruit ie has definite face where fruit sits against the rachilla evenly-flatly - usually slightly larger than the other 2 faces that form an almost single convex face eg A macrantha) 5. sub-trigonous 6. trigonous (3-sided, of roughly equal size, faces not concave) 7. triquetrous 8. strongly triquetrous 264 135. Aerial nutlet nut apex (excluding any persistent stylebase) 1. nut apex with a distinct extension or point formed from style base (as in Schoenoplectus) 2. nut apex umbonata (sometimes nut apex may have a small point but it is not formed from the stylebase and is usually no more than 0.1 nun in length) 3. nut apex generally rounded 136. Aerial nutlet outline excluding external gynophore 1. ovate 2. elliptic (2: 1 to 3:2 as in Crosslandia) 3. widely elliptic (6:5) 4. obovate (2: 1 to 3:2) 5. widely obovate (6:5 as in Bulbostylis barbata) 6. very widely obovate (1: 1 as seen in Schoenoplectus) 7. pyriformis (pear shape - nut with truncate base with style-base protruding, apex umbonate) 8. obampulliformis (compressed in 3-dimensions - apex somewhat elliptic with slight contriction near base but not like stipe in Abildgaardia) 9. obcordate (as in Abildgaardia pachyptera nut wings give the distinct shape with wings pronounced or not, sometimes entire or with notches) 10. obtrullate (3:2) 11. widely obtrullate (6:5) 12. very widely obtrullate (1: 1) 13. napiform (more tapered than obtrullate with smoother lines) 14. widely napiform (starting as widley obovate but with tapered base) ] 5. clavate (club shaped as in A macrantha as stipe forms club handle) 16. capitate (head like as in A ovata as stipe forms neck) 17. strongly capitate (where nut severly restricted half way forming tight head at apex on a long narrow stalk or stipe) 1. 8. mace shaped (from pronounced horns from deep notches on strongly constricted head with extended stipe) 137. Aerial nutlet wings, whether present or absent on the nut (see Lye 2000 p:625) 1. absent as there are no pronounced projections from face edges 2. present (in the horizontal plane ie not on the convex face sometimes greatly reduced and notched) 3. as three distinctly broad notched protusions from the face edges and top (looks mace·-like) 138. Aerial nutlet colour 1. white 2. pale pink, with white 'bloom' on outer surface 3. cream 4. straw :5. yellow 6. dark golden brown 7. port wine (may have cream-pink face and port wine ribs as seen in Bulbostylis barbata) g. light grey 9. dark grey 10. light brown 11. grey brown 12. dark brown 13. black 139. Aerial nutlet surface whether (follows Radford et al. 1974) 1. glaucous (covered with a bloom or smooth waxy coating) 2. glaucescent (sparingly or slightly glaucous; does not include waxy coating removed or damaged) 3. dull (light not reflected back; surface not coating with wax but not highly lustrous) 4. lacquered (nut appearing as iflacquered; some light reflected but not lustrous) 5. glistening (especially evident in white nuts) 6. shining (nitid or laevigate; appears lustrous or polished) 140. Aerial nutlet epidermal cell outline (at 50x magnification) 1. indistinct 2. minutely ovate (need measurements) 3. appearing circular to hexagonal (c. 20 )lm in size and at 500x are actually hexagonal) 4. widely elliptic to roughly circular (40-80 )lm sometimes squarish, at 500x are actually hexagonal) 265 5. 6. 7. 8. 9. distinctly hexagonal (giving nut surface a honecombe appearance) cells almost square to just rectangular transverely 1 by 1(or 1 by 2) transversly narrowly oblong (20 x 60 /lm to 20 x 120 /lm) transversely rectangular 40 x 60 /lm (in distinct longitudinal rows) transversly rod-shaped having tapered ends (walls straight) 10. longitudinally oblong (40 x 20/lm) II. longitudinally narrowly oblong 6: 1 (120 x 20 /lm), 3: 1 (60 x 20 /lm) 12. aciculated (marked with very fine longitudinal irregular streaks, as if produced by the point of a needle) 141. Aerial nutlet surface patterning (including protuberances follows Lye 2000) 1. smooth (no apparent pattern on nutlet surface, epidermal cells indistinct at lOx magnification) 2. subpuncticulate (some single papillae raised but not over all of surface or not prominent) 3. puncticulate (single papilla 5-15 urn diameter raised to form minute bumps prominent in cells over the surface of the nutlet) 4. subpusticulate (1-3 cells raised in groups forming low mounds and are not prominent) 5. small tubercules (from 1-3 cells prominent) 6. pusticulate (large rounded tubercules) 7. tuberculate (from multiple cells - prominent greater than 4, protruding conical outgrowths or papillae formed over more than one epidermal cell, usually 20-100 urn diameter) 8. verrucate (usually flat topped and very distinct and upright) 9. continuously rugulose 10. discontinuously mildly rugose 11. continuously mildly rugose over nut excluding stipe 12. rugose (longitudinally elongate epidermal cells that are raised to form prominent undulating transverse wrinkles) 13. continuously acutely rugose 14. longitudinally grooved striated (grooves in prominent longitudinal rows) 15. transversly oblong cells in indistinct longitudinal rows 16. transversly interlocking rod-shaped in roughly longitudinal rows 17. reticulate ('netted' epidermal cells with defined walls but not in distinct rows) 18. scalariform (ladderlike and almost in rows) 19 mostly individual cells sunken 20, individual epidermal cells raised (not forming tubercules and not puncticulate) 21. reticulate-foveate (cell walls raised and thickened) 22. alveolate (cell depression but cell walls not raised and distinct) 142. Aerial nutlet protuberance or pattern distribution 1. absent 2. occasional 3. uneven distribution (sparse) 4. restricted to vertical rows (usually two) down the fruit wall face and nut trigonous ribs 5. bordering margins of nutlet (i.e. along face ribs) 6. sparse distribution even over upper three quarters of area of nutlet (no constriction) 7. sparse distribution spread evenly over all of nutlet (no constriction) 8. sparse distribution over nutlet surface excluding stipe (constriction) 9. dense distribution over upper three quarters of nutlet (not constricted towards base) 10. dense distribution spread evenly over nutlet surface excluding stipe (constriction) 11. dense distribution evenly of nutlet surface (constriction not present or at least minimal) 143. Basal nut length 1. to 1.2 mm 2. from 1.3 to 2.1 mm 144. Basal nut width 1. from 0.75 to 1.25 mm 145. Basal nutlet, whether distinctly beaked at nut apex 1. nut with a distinct beak formed from style base (as in Schoenoplectus) 2. distinct beak absent (sometimes nut apex may have a small point but it is not formed from the stylebase and is usually no more than 0.1 mm in length) 146. Basal nutlet shape in transverse section 1. cylindrical (dorsal ventral sides not easily determined) 2. plano convex (dorsal ventral sides obvious in fruit) 3. sub trigonous to trigonous 26() 147. Basal nutlet outline shape 1. elliptic (2:1 to 3:2) 2. narrowly elliptic (3: 1) 3. widely elliptic (6:5) 4. obovate (2:1 to 3:1) 5. widely obovate 148. Basal nutlet colour 1. cream 2. golden 3. dark golden brown 4. dark grey 5. dark brown 6. black 149. Basal nutlet surface protuberances (follows Lye 2000) [138!] 1. absent 2. rugose 3. puncticulate (each cell with a raised silica body) 4. subpusticulate (individual epidermal cells raised -not forming tubercules and not puncticulate as seen in Crosslandia nutlets) 5. groups of 1-3 cells raised but not prominent (giving 'chequered appearance of raised areas over entire nut surface) 6. cells walls raised and thickened over nut surface 7. small tubercules (from 1-3 cells) raised evenly over nutlet surface 150. Basal nutlet surface, whether [139!] 1. glaucous (covered with a bloom or smooth waxy coating) 2. glaucescent (sparingly or slightly glaucous) 3. dull 4. lacquered 5. shining (nitid, laevigate, lustrous or polished) 151. Embryo, general type ｛ＱＵＰｾ［ 145"\] 1. Fimbristylis-type (root orientation lateral and smaller than basal orientated shoot) 2. Abildgaardia-type (root orientation basal and smaller than basal orientated shoot, but larger than Fimbristylis-type) 3. Bulbostylis-type (root orientation basal and the same size or larger than the basal orientated shoot) 4. Nemum-type (root orientation basal and smaller in size to the basal orientated shoot) 5. Schoenoplectus-type (root orientation lateral distal (apical) beneath mushroom shaped cotyledon and and smaller than basally pointing shoot but midway along cotyledon apical extension) 6. Carex-type (root terminal and larger than the inconspicuous lateral shoot) 7. Schoenus-type (shoot and root distinctly sub basal) 152. Embryo cotyledon outline 1. narrowly top-shaped (wide at the apex and gradually narrowing to a 'point' as in Fimbristylis Atype embryo) 2. reverse dome shaped (with base rounded and almost parallel sides as in Fi1llbristylis cinna1llo1lletorum Fimb B type) 3. roughly reverse dome shaped with cotyledon having a saddle (as in Fimbristylis disticha Fimb C type) 4. broadly top-shaped with base widely rounded due to shoot and root size (as in Bulbostylis-type embryo) 5. very widely top shaped with base wide due to shoot root (as in Ne1llum megastaclzYllln) 6. inversely bell shaped (as in Crosslandia type embryo Fimb D type embryo) 7. broadly inverse bell-shaped (as in Abildgaardia-type embryo) 8. very broadly inversely bell shaped (cotyledon very wide brimmed and not deep almost hat-like as in Abildgaardia oxystachya base rounded) 9. saucer shaped (as in Actinoschoenus broad and compressed at poles) 10. ellipsoid (as in Carex-type embryo - seen in Actinoschoenus, see Kern 1974) 11. subpyramidal (distinctly 3-sided not rounded and sharply pointed at the base as in Abildgaardia mexicana) 12. mushroom shaped with cotyledon distal extension (as in Schoenoplectus-type embryo) 267 The outline of the embryo can depend on the view i.e. from the side the embryo may look inversely bell shaped but from the front position may look ellipsoid. 153. Cotyledon shape from distal or proximal view 1. narrowly elliptic (cotyledon appearing somewhat flattened on the sides) 2. elliptic 3. circular 4. cotyledon almost triangular-trigonous 5. triangular 154. Embryo morphology, orientation of the germ pore compared to the first leaf primordia ｛ＱＵＳｾ｝ 1. parallel with the first leaf primordia 2. perpendicular 3. distinctly open and circular in the centre (as seen in Fimbristylis schllltzii) 268 Appendix 3 Glycerin jelly for semi-permanent slides (Kearns and Inouye 1993). Ingredients Quantities Distilled water 35 mL Glycerin 30mL Gelatin 10 g Phenol -- crystalline (preservative) 1g N.B. phenol is a known carcinogen. Method: In a beaker, dissolve the gelatin in distilled water by heating gently. Add the glycerin and phenol, stirring while on low heat. Avoid creating bubbles in the mixture. When dissolved pour slowly into two new Petri dishes and allow to cool. If preferred the phenol can be omitted, but the jelly will need to be kept refrigerated to prevent mould growing. Preparing semi-permanent slides: Take a cleaned microscope slide and place onto it a small cube of glycerin jelly c. 5 x 5 mm. Place the prepared sections onto the top of the jelly in a small amount of distilled water. Gently place a cover slip so that it balances on top of the jelly. Heat the slide simply by placing onto a dissecting microscope slide that has under lighting and heat until melted and the coverslip is sitting flat on the slide. Allow to cool before storing in slide boxes.

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