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Turkish Journal of Agricultural and Natural Sciences Special Issue: 2, 2014 TÜRK TARIM ve DOĞA BİLİMLERİ DERGİSİ TURKISH JOURNAL of AGRICULTURAL and NATURAL SCIENCES www.turkjans.com A comparative morphological characteristics of Chenopodium album L., C. missouriense Aellen and C. probstii Aellen Neli GROZEVA Department of Biology and Aquaculture, faculty of Agriculture, Trakia University, Stara Zagora, Bulgaria Corresponding author: grozeva@uni-sz.bg Abstract A comparative morphological characteristics of two North American invasive species – C. probstii and C. missouriense and the representative of genus Chenopodium in Bulgarian flora closest to them – C. album L. have been made. А total of 18 quantitative and 11 qualitative features are included in the morphological analysis. For more detailed study of generative organs the Scanning electron microscope (SEM) method has been used. Information about the chorology and ecological preferences of C. probstii and C. missouriense, possible ways of their penetration into Bulgarian flora, as well as the reasons for their later discovery have been given. Results showed that the differentiation of C. album, C. missouriense and C. probstii is possible in all phases of their development. Key words: Chenopodium album, Chenopodium missouriense, Chenopodium probstii, morphology, Bulgaria, distribution Materials and methods The object of the study are 2 populations of C. missouriense and 3 of C. probstii (Table 1). Еighteen quantitative characters have been included in the morphological analysis: 1. Height of stem; 2. Length of basal leaf; 3. Width of basal leaf; 4. Length/width ratio; 5. Length of basal leaf petiole; 6. Length of upper leaf; 7. Width of upper leaf; 8. Length/width ratio; 9. Length of upper leaf petiole; 10. Length of inflorescence; 11. Diameter of bisexual flower; 12. Length of flower petiole; 13. Diameter of female flower; 14. Length of flower petiole; 15. Length of seed; 16. Width of seed; 17. Length/width ratio; 18. Thickness of seed. A total of 11 qualitative traits was also recorded: 1.Colour of stem; 2. Shape of leaf lamina; 3. Colour of leaf lamina; 5. Apex of leaf lamina; 6. Margin of leaf lamina; 7. Colour of perianth; 8. Presence of keel on perianth lobes (petals); 9. Contour of perianth lobes (petals); 10. Colour of seed; 11. Edge of seed. The summarized results for these 2 species have been compared to the data for C. album, published by Grozeva and Cvetanova (2008). For more detailed study of flowers and seeds the Scanning electron microscope (SEM) method has been used. The electron microscope tests have been conducted at the laboratory for X-ray analysis of the Faculty of Chemistry and Pharmacy of Sofia University. Introduction Globalization and growth of trade relationships worldwide in recent decades have resulted in the advent of many new plant species in European countries. The representatives of Chenopodium incl. studied species are some of the most aggressive and quickly seize new territories. The most widespread is C. album. It is almost cosmopolitan, avoiding cold and tropical regions and an extremely common weed in all kinds of cultivated land (Uotila, 2001). C. probstii Aellen invaded Scotland through the import of wool in 1913 (Aellen, 1931) and for several decades it has spread to become part of the flora of 20 European countries reaching Romania to the south-east (Dostálek and Jehlík, 2004). C. missouriense Aellen was introduced with the transportation of grain from North America to Sweden in 1926 (Aellen, 1928) and by 2003 it has invaded 13 European countries reaching Russia and Ukraine to the east (Dostálek and Jehlík, 2004). As a result of a complex population study of Chenopodium in Bulgaria, C. probstii and C. missouriense have been established in our country as well (Grozeva, 2010). The aim of this research is to compare morphologically C. missouriense and C. probstii to the most similar to them species from Bulgarian flora – C. album, in order to establish their environmental requirements and to seek possible ways of their penetration in the country. 1949 Turkish Journal of Agricultural and Natural Sciences Special Issue: 2, 2014 Table 1. Studied populations of C. probstii and C. missouriense. Locality Ecological conditions C. missouriense Aellen Black Sea coast (North), Albena Flat region, regularly flooded by sea sands, resort area, between the sea 199 m a.s.l., ruderal areas with C. album coast and the resort area. domineering and Corispermum nitidum and C. glaucum as co-dominants. Tundzha hilly plain, Nova Zagora, Flat region, alluvial soil, 131 m a.s.l., ruderal the west end of the town. community with C. album domineering. C. probstii Aellen Sofia region, city of Sofia, ruderal in the area behind the Pliska Hotel. Thracian plain, town of Merichleri, around the railway station. Thracian plain, town of Stara Zagora, around bus stop in “Mityo Stanev-west” area. Flat region, leached humus soil, 550 m a.s.l., ruderal community with C. probstii domineering. Flat region, leached humus soil, 123 m a.s.l., ruderal community with Urtica dioica domineering. Flat region, maroon forest soil, 196 m a.s.l., ruderal community with C. polyspermum domineering. Results Table 2. Studied quantitative features of C. probstii, C. missouriense and C. album Species C. probstii C. missouriense Character (min, x, max) (min, x, max) No. (150) 174,73 (200) cm (5,7) 6,82 (9,5) cm (3,8) 4,41 (6,5) cm (1,2) 1,56 (2,5) (5,1) 5,14 (5,21) (2,2) 3,17 (4,5) cm (0,5) 1,53 (1,2) cm (1,8) 2,11 (3,5) (3,05) 3,13 (3,21) cm (23) 52,78 (75) cm (0,49) 0,5 (0,52) (0,8) 0,85 (1) mm (0,9) 1,05 (1,3) mm (0,4) 0,52 (0,7) mm (1,2) 1,23 (1,3) mm (114) 1,21 (1,3) mm (1,07) 1,08 (1,2) (0,53) 0,54 (0,55) mm Diffuse spatial structure, number - 17 specimens, area 25 m2. Diffuse spatial structure, number - 22 specimens, area 70 m2 Diffuse spatial structure, number - 32 specimens, area 30 m2. Diffuse spatial structure, number - 15 specimens, area 75 m2. Diffuse spatial structure, number - 13 specimens, area 77 m2. missouriense and C. probstii are presented in Plate 1, and of the seeds – in Plate 2. The ecological requirements and locality of studied populations of C. missouriense and C. probstii are described in Table 1. Values for each of the 18 quantitative characters are presented in Table 2. Data about the quantitative features are summarized in Table 3 and Figure 1. The results from the Scanning electron microscope study of the flowers of C. album, C. 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. Population (75) 118,93 (170) cm (4,2) 6,37 (4,8) cm (2,5) 3,25 (4,5) cm (2,04) 2,06 (3,5) (2,8) 2,98 (3,13) (2,4) 3,53 (4,5) cm, (0,5) 1,60 (2,5) cm (1,8) 2,18 (2,4)) (0,5) 0,88 (2) cm (34,8) 37,84 (113,5) cm (0,2) 0,24 (0,27) (0,7) 0,87 (1) cm (0,6) 0,82 (1) mm (0,5) 0,61 (0,62) mm (0,7) 0,81 (1,1) mm (0,8) 0,81 (1,0) mm (0,99) 1,01 (1,2) (0,3) 0,37 (0,5) mm 1950 C. album (min, x, max) (35) 64,26 (100) cm (1,5) 3,07 (8) cm (1) 1,58 (3,5) cm (1,2) 1,99 (2,8) (0,52) 1,43 (2,47) (0,7) 1,35 (3,5) cm (0,2) 0,54 (1,2) cm (1,5) 2,78 (5,4) (0,35) 0,65 (0,8) (12) 37,78 (74) cm (0,00) 0,05 (0,3) (0,8) 1,16 (1,5) cm (0,9) 1,11 (1,3) mm (0,3) 0,57 (0,7) mm (1,1) 1,26 (1,4) mm (1,15) 1,23 (1,3) mm (1,02) 1,09 (1,25) (0,3) 0,34 (0,5) mm. Turkish Journal of Agricultural and Natural Sciences Special Issue: 2, 2014 Table 3. Studied quanlitative features of C. probstii, C. missouriense and C. album feature C. probstii C. missouriense 1. Color of brown-green to dark-green, striped striped with green to stem with red spots on the base of side yellow-green, rarely with branches reddish spots bluish-green when young dark-green, often tinged brown, red 2. Leafto dark pure green after near the margin, red-purple lamina flowering; margin pigmentation usually after flowering color, coarsely serrate with margin, diffused to the entire leaf; margin acute teeth, apex acute; apex and serrate to irregularly dentate, apex leaf blade rhombic-elliptic shape acute; leaf blade 3-lobed broadly ovate, to oval-rhombic and elongated- to rhombic in basal and rhombic in basal and middle leaves to middle leaves and lanceolate in upper. lanceolate, almost complete in upper part. oval, light-green to green, 3.Perianth triangular to broadly lanceo-late, light-green to reddish-green, keeled, keeled, with narrow lobes with a wide light margin and slight membranous margin and (petals) – clear cloud- and net-like colour, keel cloud- and net-like nervation, nervation. and contour triangular to broadly lanceolate,. 4. Color, seed coat brown to black, opaque seed coat black, glossy, coat and with edge of seed enlarging at the edge slightly keeled edge of area of the seedling root. evenly wide along its seed entire length Figure 1. Variation of shape of: А) C. probstii; B) C. missouriense; C) C. album 1-2 basal leaves; 3-5 middle leaves; 6-7 upper leaves. 1951 C. album striped with greyish to bluish green and red greyish green to green, sometimes after flowering reddish green; margin shallowly and irregularly serrate to dentate; apex acute; leaf blade rhombic to elliptic in basal and middle leaves and lanceolate to elongated-lanceolate almost complete in upper part. elongated-elliptic, green, keeled, with a wide light margin and slight cloud- and net-like nervation. seed coat black, glossy, edge fairly acute evenly wide along its entire length. Turkish Journal of Agricultural and Natural Sciences Special Issue: 2, 2014 Discussion The results of our studies reveal that the morphological characteristics of C. missouriense, C. probstii and C. album coincide with the data by Aellen (1960); Dvořák (1987, 1994); Dostálek et al. (1990); Аkеroyd (1993), Uotila (2001), Dostálek and Jehlík (2004). The authors who had studied the above species think that their distinction before flowering is almost impossible. The main distinctive features in their opinion are the seed characteristics complemented by those of the leaves. The present study revealed that the distinction of C. missouriense, C. probstii and C. album by using the morphological characteristics of stems and leaves is also possible before the generative phases. C. album plants are up to 1 m high with alternating reddish, white and green stripes on stems (Tables 2, 3). Plants of C. probstii and C. missouriense are significantly taller but in the first species stems are dark green with more or less reddish tinge and in the second they are light green (Tables 2, 3). From the leaf characteristics the most significant for the distinction of the three species are color, shape and margin. C. probstii is the easiest to be distinguished. Its leaf-blade is dark green to reddish-violet, with unevenly dentate margin, for basal and middle leaves – 3-lobed, oval-rhombic to rhombic, for the upper ones – elongated rhombic to lanceolate (Table 3, Fig. 1A). For C. missouriense the leaf-blade is bluish-green to green, rhombic-elliptic to rhombic in basal and middle leaves and lanceolate, almost entire in upper and margin is coarsely serrate with acute teeth (Table 3, Fig. 1B). C. album has green to dark green leaf-blade; rhombic to elliptic in basal and middle leaves and lanceolate to elongated-lanceolate almost entire in upper leaves with shallowly and irregularly serrate to dentate margin (Table 3, Fig. 1C). It is well distinguished from the other two species by its smaller leaves, too (Table 2). During flowering and fruiting stems of all three species become ligneous, usually the basal and some of the middle leaves fall off. That is why during the generative phases mainly the characteristics of flowers and seeds are used for distinction purposes. C. probstii can be distinguished from the other two species by a stronger and marked keel of perianth lobes (Plate I, Fig. 1). In C. missouriense the cloud- and net-like nervation of perianth lobes is very pronounced (Plate I, figs 2,3), while in the other two species it is slightly noticeable (Plate I, Fig. 4). Plate I. Scanning electron micrographs of flowers: Fig. 1. C. probstii, Figs 2-3. C. missouriense; Fig. 4. C. album. 1952 Turkish Journal of Agricultural and Natural Sciences Special Issue: 2, 2014 The greatest is the area of occurrence of C. album (Aellen, 1960; Dostálek et al., 1990; Uotila, 2001). The geographic area of C. probstii comprises of North America, Central and South-East Europe, North Australia, East Asia (North Korea), North Africa (Dostálek et al., 1990; Uotila, 2001; Dostálek and Jehlík, 2004). The most restricted is the area of C. missouriense comprising North America, North and Central Europe (Dostálek et al., 1990; Uotila, 2001; Dostálek and Jehlík, 2004). By the size of seeds C. probstii and C. album are quite similar, while C. missouriense has smaller seeds (Table 2, Plate II, Fig. 4). The perfect distinction of the first two species is by the colour and the edge of seeds – in C. probstii these are brownish-black, opaque with typical broadening of the edge in the area of the seedling root (Plate II, Figs 1-3), while in C. album they are black, glossy with a narrower edge of an even width along its entire length (Plate II, Figs 5, 6). Plate II. Scanning electron micrographs of seeds: Figs 1-3. C. probstii, Fig. 4. C. missouriense; Figs 5-6. C. album. Both in our country (Fig. 2) and in other European countries C. probstii is distributed and adapted more rapidly in comparison with C. missouriense. One of the most probable causes for Most probably the advent of both North American species in our country is related to the import of plant materials and the carrying of their tiny seeds by air currents. 1953 Turkish Journal of Agricultural and Natural Sciences Special Issue: 2, 2014 Republik and neighbouring countries. Feddes Repert. 115 (5-6): 483-503. Dvořák, F., 1987. Study of Chenopodiumacerifolium Andrz. and C. missouriense Aellen.Feddes Repert. 98 (11-12): 561-582. Dvořák, F., 1994. Study of some species subsumed under Chenopodium probstii Аellen and on C. purpurascens B. de Juss. ex Jacq. Feddes Repert. 105 (3-4): 113-139. Grozeva, N.2007.C. pumilio (Chenopodiaceae): a new species to the Bulgarian flora. Phytol. Balkan. 13(3): 331-334. Grozeva, N., 2010. Reports 1709-1714. – In: Kamari, G., Blanchė, C., Siljak-Yakovlev (eds). Mediterranean chromosome number reports 20. Flora medit. 20: 266-272. Grozeva, N., Cvetanova, Y., 2008. Variability among the populations of two Chenopodium species – In coll.: Proceedings of the VІІth scientific and technical conference with international participation “Ecology and health – 2008”, 10 Apr 2008: 135-140. (in Bulgarian) Homsher, P. J., 1963. Cytogenetic studies in Chenopodium. – Amer. J. Bot. 50(6): 621-622. Kjellmark, S., 1934. Einige neue Chromosomenzahlen in der Familie Chenopodiaceae. Bot. Notiser 87: 136-140. Keener, C., 1970. Documented plant chromosome numbers 70 (1) – Sida, 3: 533-536. Schwarzova, T. 1986. Chromosome numbers of some species of the genus Chenopodium L. from localities in Czechoslovakia. Acta. Fac. Rer. Nat. Univ. Comen. Bot.: 32-33 Uotila, P., 2001. Chenopodium L. – In: Jonsell, B. (ed.), Flora Nordica. Vol. 2, pp. 4-31.N. A.S., Stockholm. Zosimovich, B., 1965. Life forms, polyploidy and evolution of families from Magnoliopsida. 5-38. Cytology and cenetics, Kiev. (in Russian) that is the more abundant fruiting and mass ripening of the seeds of the species found out in the present study. C. missouriense is quite rare which probably results from the weaker fruiting and the failure of a considerable part of its seeds to ripen. It may well be that the reference data about its more restricted distribution results from its wrong classification to C. album due to the difficult distinction between both species. The results of this study confirm the statement of Dostálek and Jehlík (2004), that C. probstii and C. missouriense have similar ecological requirements – they occur mainly in ruderal communities. Their ecological preferences (Table 1) are quite similar to the ones found for the Bulgarian populations of C. album (Grozeva and Cvetanova 2008). Their probable finding among weeds in cultural communities is an issue of the near future. Conclusion The distinction of C. album, C. missouriense and C. probstii is possible both before flowering, and during flowering and fruiting. The main distinctive features before flowering are colour and height of stem; colour, margin and shape of the leaf blade. During flowering the three species can be diagnosed by keel, contour and nervation of perianth lobes (petals). During fruiting of paramount importance for their differentiation and distinction are colour, size and edge of seed. C. album, C. missouriense and C. probstii occur most often in ruderal communities. Basic mechanisms for their distribution are anthropochoria (through imported plant materials) and anemochoria. References Aellen, P., 1928. Neue adventive Chenopodien aus Schweden. Bot. Not. 1928: 203-210. Aellen, P., 1931. Die wolladventiven Chenopodien Europas. Verh. Naturforsch. Ges. Basel 50: 151-152. Aellen, P., 1960. Chenopodiaceae – In: G. Hegi, Illustrierte flora von Mitteleuropa, ed. 2, 3/2: 533-692. Berlin-Hamburg. Akeroyd, J. 1993. Chenopodium L. – In: Tutin et al. (eds), Flora Europaea Ed. 2, vol. 1, 11-14. Cambridge Univ. Press., Cambridge. Dostálek, J., Hejný, S., Husák, Š., Schwarzová, T., Dvořák, F., 1990. Chenopodium L. – In: Hejný, S., Slavík, B. (eds), Flora of Czech Republic. Vol. 2, 223265. Acad, Praha. Dostálek, J., Jehlík, V., 2004. Chenopodium probstii and C. missouriense: two North American plant species in the Czech Republic, Slovak 1954