Egypt. J. Bot. Vol. 59, No. 2, pp. - 1254 (2019) Auto-taxonomy of Brassica nigra (L.) Koch (Brassicaceae) in Egypt Wafaa Amer(1), Mahmoud Shoulkamy(2), Ahmed Faried(3) and Hadeer Abd ElBaset(2)# (1) Botany and Microbiology Department, Faculty of Science, Cairo University, Giza, Egypt; (2)Botany and Microbiology Department, Faculty of Science, Minya University, Minya, Egypt; (3)Botany and Microbiology Department, Faculty of Science, Assiut University, Assiut, Egypt. B RASSICA L. is one of the most economically important genera within family Brassicaceae, it includes approximately 80 species worldwide. In Egypt, the genus represented by five species. Among them, Brassica nigra (L.) Koch, which grown as a weed in field crops as well as roadsides of Mediterranean region. The field and herbarium observations reflected the notable morphological diversity within the species populations. Accordingly, morphological and palynological studies for the different geographical populations of Brassica nigra were carried out to trace the species diversity and helps for identification the infra-species taxa. The taxonomic revision of the species in Egypt, was carried out on the herbarium specimens as well as fresh materials represented by 26 populations, distributed along the Nile Valley and the Nile Delta. The results revealed the presence of two varieties namely var. bracteolata and var. nigra. The fruit peak and trichomes are the differential characters delimiting the two varieties. Moreover, the results showed also presence of var. nigra in two different biotypes. SEM of the seed coat and pollen grains showed the presence of notable infra-specific diversity. For pollen grains, the size, apertures and exine ornamentation confirming this diversity. Photographs and taxonomic key for varieties and forms will be addressed. Keywords: Biotypes, Brassica, Egyptian Flora, Forms, var. bracteolata, var. nigra, SEM, Pollen grains. Introduction Brassicaceae (Cruciferae) is a monophyletic group, which currently includes approximately 3709 species and 338 genera (Al-Shehbaz et al., 2006). It is one of the most agronomical important plant families, it includes vegetables, ornamental and crop species (Kasem et al., 2011). Nearly all members of this family are Mediterranean, Irano-Turanian and Saharo-Sindian zones except Antarctica region (Hedge, 1976 and Lysak & Koch, 2011). Genus Brassicais one of the most agriculturally important genera of Brassicaceae family, it includes about 80 accepted species worldwide,with high morphological diversity and wide-ranging utility (Song et al., 1988 and The Plant List, 2013). In Egypt, Brassicaceae is one of the four largest families, represented by about 103 species and 53 genera (Boulos, 1999). It showed the presence of high degree of phenotypic variations # with and within populations. Taxonomic structure of the whole family is characterized by a large number of monotypic and small genera, mostly with clearly defined taxonomic limits. Due to its great economic importance, crops of Brassica received the attention of taxonomists from the earliest times (Warwick, 2011). Genus Brassica represented in Egypt by five species, namely: B. rapa L., B. tournefortii Goauan, B. deserti Danin & Hedge, B.nigra (L.) Koch and B. juncea (L.) Czernj. & Coss. Brassica nigra is an annual herbaceous plant that originated in the Middle East, its seed has long been used in central and southern Europe, North Africa and Asia for cooking oil and medicine (Tsunoda, 1980 and Vaughan, 1977). Now it is widely cultivated as a primary source of the mustard seeds used in making the condiment sauce, table mustard, and others. Brassica nigra is known to exhibit considerable morphological variations: It has been divided taxonomically into Corresponding author email: hadeer_dahy@yahoo.com DOI: 10.21608/ejbo.2019.6375.1254 Edited by: Prof. Dr. Fawzy M. Salama, Faculty of Science, Assuit University, Assuit, Egypt. ©2019 National Information and Documentation Center (NIDOC) 2 WAFAA AMER et al. varieties. The morphological diversity of B. nigra in Egypt still unresolved issue where, Täckholm (1974) reported the occurrence of three varieties belonging to B. nigra, namely v. brcteolata (Fisch. & Mey.) Spach,v. torulosa Alef. and v. turgida Alef. Later, Boulos (1999, 2009) considered these varieties as synonyms to B. nigra (L.) Koch. Although pollen morphologyis useful in the separation of closely related genera within angiosperm families (Rollins, 1979 and AlShehbaz, 1989), most members of Brassicaceae are considered to be stenopalynous with a uniform pollen shape (Al-Shehbaz et al., 2006 and Erdtman, 1972). However, numerous studies (e.g. Erdtman, 1972; Jonsell, 1979; Rollins & Banerjee, 1979; Lahham & Al-Eisawi, 1987 and Anchev & Deneva, 1997) have demonstrated that pollen characters were useful for assessing phenetic relationships and resolving taxonomic problems within different taxa of Brassicaceae. Seed coat ultrastructural pattern using SEM are very significant for assessing phenotypic relationships and resolving taxonomic problems (Kasem et al., 2011 and Heywood, 1971). The importance of seed coat sculpturing in discrimination and identification of Brassica was performed by numerous taxonomists (among them: Musil, 1948; Berggren, 1962; Mulligan & Bailey, 1976; Stork et al., 1980; Buth & Roshan, 1983; Fayed & El Naggar, 1988; Setia et al., 1989; Ren & Bewley, 1998; Koul et al., 2000 and Kasem et al., 2011). However, little is known about the extent and distribution of genetic variation in B. nigra in Egypt. Brassica nigra is reported to have little morphological variation compared with the major Brassica crops (Prakash & Hinata, 1980). Therefore, in this study we aimed to study the morphological and palynological and seed sculpture traits of different populations of Brassica nigra representing its geographic range in Egypt to provide detailed information about the species diversity and for identification of the infra-species taxa (species auto-taxonomy). Materials and Methods Morphological data of Brassica nigra were achieved by examination of fresh specimens belonging to 26 populations collected from Egypt. J. Bot. 59, No.2 (2019) different geographical regions during flowering and fruiting seasons. Localities of the sampled populations are given in Table 1 and mapped in Fig. 1. However, the floras mentioned that the species traced in Mediterranean region (Boulos 2009), the last specimens deposited in Cairo University Herbarium dated back to 1964 and no specimens were traced in the field trips. Fresh samples were collected and directly fixed in FAA fixative (50ml ethyl alcohol, 10ml formaldehyde, 5ml glacial acetic acid, and 35ml distilled water) and voucher samples were dried and deposited in herbaria of both of Assiut (ASTU) and Minya Universities. All the distribution localities were studied for morphological diversity using different morphological criteria of stem, leaves, inflorescence and fruit characters (Table 2). Stem, leaves and fruits were examined and photographed by Olympus SZ61 stereomicroscope provided with a digital Olympus camera SC100. Sample preparation for SEM For pollen SEM study, representative population from each variety and biotype was chosen. Fresh anthers were collected from floral buds and photographed by Scanning Electron Microscope (SEM) using Joel 1200 EXІІSEMat 20KV. Measurements of size, polar axis, equatorial axis and colpus length and width were recorded in Table 3. For seed sculpture, representative clean, full ripened seeds were selected in triplicate and fixed on aluminium stumb using double stick adhesive and then coated and photographed using Joel 1200 EX ІІ SEM at 20KV. The terminology of Punt et al. (1994) and Koul et al. (2000) are used for describing the pollen morphological characters as well as seed coat sculpturing, respectively. Results Species morphology Brassica nigra (L.) Koch, is annual herb 0.4–1.5 (2)m, glabrous to sparingly hispid with stiff hairs. Stem erect branched from upper and lower parts. The radicle leaves lyrate- pinnatisect (in different forms), up to 75cm long, petiolate serrate–double serrate margin. Cauline leaves ovate with 18– 20×2.0–2.5cm and serrate margin. Inflorescence richly branched. Flowers, with yellow, claw-shaped 3 AUTO-TAXONOMY OF BRASSICA NIGRA (L.) KOCH (BRASSICACEAE) IN EGYPT. petals and 6 stamens (tetradynamous). Fruit siliqua up to 40mm length, its peak ranges from 1/4 to 1/2 of the fruit length. Seeds numerous, globose, dark brown, 0.2mm in diameter. Infra-specific diversity Based on macromorphological traits The present study carried out on 26 Brassica nigra populations in addition to the herbarium specimens deposited in Cairo, Assiut and Minya herbaria. 50 morphological traits are used to revealthe infra specific diversity in the studied populations grouped these populations under two varieties. The radical leaves showed clear variations in size (up to 75cm in var. nigra and up to 50cm in var. bracteolata) as shown in Table 2. The leaf at the species level is lyrate, with pinnatisect lobes (Fig. 2). Terminal lobe in var. bracteolata is broad ovate with clear basal collar lobes not-reached to the midrib (Fig. 2 C), where this basal collar lobes reach to the midrib in var. nigra (Fig. 2 A and B). Moreover, var. nigra showed triangular terminal lobe in form 1 and elliptical in form 2, the both forms provided with irregular serratedentate margin, while var. bracteolata showed clear dentate margin. The siliqua fruit ranges from up to 35mm in var. nigra to up to 40mm in var. bracteolata (Table 2). The later also distinguished by its longer fruit peak which extended to half of the fruit length. The taxonomic study of the studied Brassica nigra populations using 50 morphological characters outlined in Table 2 showed the presence of high degree of phenotypic variations among the studied 26 populations and revealed the presences of distinct three infraspecific taxa namely var. bracteolata, var. nigra form 1and var. nigra form 2. TABLE 1. The phyto-geographical locations of the studied Brassica nigra populations. St. No. Localities GPS coordinates N E 27° 34’ 3.348” 30° 48’ 7.085” 27° 2721.438” 30° 49’ 24.584” 27° 24’ 22.361” 30° 52’ 10.707” 27° 21’ 32.429” 30° 54’ 59.543” 27° 925.341” 31° 12’ 40.289” 27° 4’ 20.880” 31° 15’ 10.959” 27° 2’35.92” 31°17’45.25” 27° 0’ 19.712” 31° 20’ 26.911” 26° 53’ 41.863” 31° 26’ 33.989” 26° 49’ 11.029” 31° 30’ 9.114” 26° 46’ 32.078” 31° 31’ 3.658” 26° 44’ 8.908” 31° 33’ 7.876” 26° 41’ 4.925” 31° 36’ 44.159” 28° 17’ 35.856” 30° 42’ 55.315” 28° 19’ 40.130” 30° 42’ 9.552” 28° 23’ 58.125” 30° 46’ 14.230” 28° 30’ 57.945” 30° 47’ 31.618” 28° 36’ 14.588” 30° 48’ 46.146” 28°37’38.189” 30°49’8.758” 28° 1’ 48.513” 30° 48’ 1.002” 27° 52’ 3.226” 30° 48’ 0.528” 27° 46’ 55.051” 30° 50’ 28.675” Associated field 11 12 13 14 15 16 17 18 19 20 21 22 23 1 2 3 4 5 6 7 8 9 Dirout – Assuit South to Qousia – Assuit Bani Zeid , Qousia – Assuit Bani Rafea , Manflout – Assuit Abo teeg – Assuit 5 Km to Assuit El Mouteaa – Sohag Sedfa – Sohag Tma – Sohag Shatora – Sohag Tehta – Sohag Banawit – Sohag El Maragha – Sohag 5 Km to Samalot - Minya Samalot – Minya Matai – Minya El Gharabawi – Minya Dahrout – Minya Maghagha – Minya El Howslia – Minya Abo Qorkas – Minya Malawy – Minya Alfalfa Wheat Alfalfa Alfalfa Alfalfa Alfalfa Alfalfa Cabbage Wheat Alfalfa Wheat Wheat Wheat Alfalfa Alfalfa Lentil Alfalfa Alfalfa Alfalfa Wheat Wheat Wheat 10 Dermawas – Minya 27° 37’ 12.988” 30° 50’ 40.104” Wheat 24 25 26 Beni Suef El Giza Ashmoon – al-Minufiyah 29° 04’ 22.45” 30°18’ 33.14 30°12’ 39” 31° 05’ 27.33 31°10’ 91.44 30° 57’ 23” Wheat Wheat Wheat Egypt. J. Bot. 59, No.2 (2019) 4 Egypt. J. Bot. 59, No.2 (2019) WAFAA AMER et al. Fig. 1. Distribution map of Brassica nigra in Egypt. 5 AUTO-TAXONOMY OF BRASSICA NIGRA (L.) KOCH (BRASSICACEAE) IN EGYPT. TABLE 2. Morphological features of the studied Brassica nigra taxa. Characters var. nigra form 1 var. nigra form 2 var. bracteolata Hairy all over Pinnatisect Irregularly serrate-dentate Petiolate Up to 75cm Oblong ovate Kidney shape stomata Acute 3–7 lobes 1.5–5cm Midrib only Pinnatisect Irregularly serrate-dentate Petiolate Up to 30cm Oblong ovate Kidney shape stomata Acute 2–5 lobes 2–5cm Glabrous Pinnatisect Irregularly dentate Petiolate Up to 50cm Oblong ovate Kidney shape stomata Obtuse 2–6 lobes 1.5–4cm Serrate denticulate Petiolate 1–4 lobes Denticulate Petiolate 1–2 lobes Denticulate Petiolate 1–3 lobes Up to 2.5×20 Up to 1.5×15 Up to 2×18 Ovate Acute Ovate Acute Ovate Acute 4-7×0.5-1.5 Linear Glabrous 3 Obtuse Entire 5-6×1-1.5 Linear Glabrous 3 Obtuse Entire 4-6×0.5-1.5 Linear Glabrous 3 Acute Entire 9-14×3-5.5 L of claw 3-5 L of blade 6-9 9-12×4-6 L of claw 3-5 L of blade 6-9 9-11×4-6 L of claw 3-5 L of blade 5-7 Basal (radical) leaf: 1-Hairs 2-Shape 3-Margin 4-Petiole 5-Length 6-Shape 7-Stomata 8-Apex 9-Segmentation 10-Width of the upper lobe Upper (cauline) Leaf: 11-Margin 12-Petiole 13-Segmentation 14-Upper lobe: L ×Wcm 15-Shape 16-Apex Flower: Sepals: 17-(L×W)mm 18-Shape 19-Surface 20-No. of main veins 21-Apex 22-Margin Petals: (divided into claw and blade) 23-(L×W)mm 24-Shape of claw Filiform Filiform Filiform 25-Surface Glabrous Glabrous Glabrous 5-7 Obtuse Entire 5-9 Obtuse Entire 5-9 Obtuse Entire 5-7×0.3-0.5 Linear Glabrous 4-7×0.3-0.5 Linear Glabrous 5-7×0.2-0.5 Linear Glabrous 1.5-3×0.5-1 Oblong Normal Obtuse Capitate bi-lobed 1.5-3×0.5-1 Oblong Normal Obtuse Capitate bi-lobed 1.5-2×0.5 Oblong Normal Obtuse Capitate bi-lobed 26-No. of main veins 27-Apex 28-Margin Stamens: 6 (2+4) Filaments: 29-L×Wmm 30-Shape 31-Surface Anthers: 32-L×Wmm 33-Shape 34-Base of attachment 35-Apex 36- Stigma Egypt. J. Bot. 59, No.2 (2019) 6 WAFAA AMER et al. TABLE 2. Cont. Characters Pedicle: 37-Length Flower bract: 38-Hairs 39-Vennation 40-Margin 41-Apex 42-Shape 43-Length Fruit: 44-Length of the whole fruit 45-Length of the fruiting part 46-Length of peak 47-Ratio (Fp/Flp) 48-Number of seeds/ fruit 49-Surface 50-Shape of receptacle var. nigra form 1 var. nigra form 2 var. bracteolata Up to 10mm Up to 7mm Up to 15mm Hairy Pinnate reticulate Dentate Acute Ovate Up to 3.5cm Glabrous Pinnate reticulate Dentate Acute Ovate Up to 3cm Glabrous Pinnate reticulate Almost entire Acute Obovate Up to 3cm Up to 35mm Up to 20mm Up to 15mm 10–25 6-12 Glabrous Flat Up to 35mm Up to 22mm Up to 15mm 27-50 8-12 Glabrous Flat Up to 40mm Up to 35mm Up to 12mm (7:1) – (4:1) 8-20 Glabrous Flat TABLE 3. Pollen features of the studied B.nigra taxa, ( ): mean value. Characters Pollen Shape Pollen aperature Pollen sculpture Endexine sculpture Muri-wall Muri-pattern Polar Axis (P)µm Equatorial Axis (E)µm P/E (µm) Colpus length (L) var. nigra Form 1 Form 2 Oblate Suboblate Tricolpate Tricolpate Reticulate Reticulate Sparsely-warty Densely warty Warty Sparsely warty Regular-heterobronchate Regular-heterobronchate 20.50–21.0 25.50–26.20 (20.75) (25.85) 40.50–41.50 30.90–31.70 (41.0) (31.30) 0.506 0.825 35.70 21.40 var. bracteolata Oblate Tricolpate Reticulate Smooth warty irregular-heterobronchate 19.20–20.0 (19.60) 29.20–30.70 (29.95) 0.654 26.35 Fig. 2. Radical leaves diversity in Brassica nigra, A: var. nigra form 1 , B: var. nigra form 2 and C: var. bracteolata . Egypt. J. Bot. 59, No.2 (2019) 7 AUTO-TAXONOMY OF BRASSICA NIGRA (L.) KOCH (BRASSICACEAE) IN EGYPT. The taxonomic differentiation was based on the 50 morphological characters including features of basal leaves, cauline leaves, inflorescence as well as fruit characters, seeds and flower structure (Table 2), claimed the following differential key: Key 1 -Leaves glabrous, fruit peak up to 1/4 fruit length …..….............… var. bracteolata -Leaves hairy, fruit peak up to 1/2 fruit length ………..................…………...........(2) 2 -Basal leaves up to 30 cm, hairs on all over the surface .…............................ var. nigra form 1 -Basalleaves up to 75cm, hairs found only on mid rips of leaf lower surface................. var. nigra form 2 Based on micromorphological traits using SEM Stem Scanning of the stem epidermal system in the studied B. nigra represented in Fig. 3. This figure reflecting a clear striate epidermal sculpture in var. nigra (Fig. 3 A and B) and incontinuous ridges in var. bracteolata (Fig. 3 C), the later also characterized by high density of verrucate cuticular deposits of varying –size, where it appears uniform and sparse in var. nigra as outlined in Fig. 3 A. Leaf The scanning of the leaf upper surface outlined in Fig. 4, confirming the distinctive differential micromorphological characters. Among them the presence of abundant stomata on the same level in epidermal cells in var. nigra form 1 (Fig. 4 A) and it appeared on raised ridges in var. nigra form 2 (Fig. 4 B). On the other hand, stomata in case of var. bracteolata appeared sparse and in lower level than epidermal cells. Moreover, var. bracteolata, this variety characterized by abundant verrucate– shaped cuticular deposits, which appeared sparse in var. nigra (Fig. 4 C). Seed coat The micromorphological features of the seed coat of the studied B. nigra populations as seen by SEM showed common reticulate seed coat pattern (Fig. 5). Notable infra-specific variations noticed in the periclinal wall; as it appears elevated wavy and folded in var. nigra form 1, elevated and straight in form 2, while it is grooved in var. bracteolata (Figs. 5 A, B & C). The anticlinal surface is straight with faint papillae in form 1, straight, striate in form 2 and papillate in var. bracteolata (Fig. 5 A, B & C). Pollen grains The SEM of the studied Brassica taxa showed that the pollen grains are radially symmetric, isopolar, with oblate-suboblate shapes. Apertures are tricolpate with syncolpi in both form 1 and 2 while in var. bracteolata the pollens have narrow apocolpi. Evine surface is widely reticulate in forms 1 and 2, while it is narrowly reticulate in var. bracteolate (Fig. 6 A, B & C). The measurements and descriptions based on 30 pollen samples are summarized in Table 3. Fig. 3. Stem sculpture in Brassica nigra; A, B: var. nigra (A: form 1 and B: form 2) and C: var. bracteolata. Egypt. J. Bot. 59, No.2 (2019) 8 WAFAA AMER et al. I II Fig. 4. Leaf surface sculpture of B. nigra using SEM; I- Stomata pattern, II- Magnified part. A and B: var. nigra with notable stomata and sparse cuticular deposits (A: form 1 and B: form 2) and C: var. bracteolata, with sparse stomata and dense cuticular deposits. Fig. 5. Seed coat sculpture of B. nigra A & B: var. nigra (A: form 1, with elevated wavy-folded periclinal walls & B: form 2, with elevated-straight periclinal walls) and C: var. bracteolata , with grooved periclinal walls. Egypt. J. Bot. 59, No.2 (2019) 9 AUTO-TAXONOMY OF BRASSICA NIGRA (L.) KOCH (BRASSICACEAE) IN EGYPT. Size: The pollen grain diameter of the studied B. nigra taxa ranges from 29.95–41µm (Table 3), var. bracteolata is the smallest (P×E: 19.6 × 29.95), as showed in Table 3 and Fig. 6 C. Shape: The pollen according to P/E ratio (Table 3), distinguished two types oblate shape (P/E: 0.50.75) in var. bracteolata and var. nigra form 1. While form 2 is suboblate (P/E: 0.76–0.88). Pollen apertures: Tricolpate pollen in B. nigra with shortest colpus (21.40µm) in var. nigra form 2 and the largest (35.70µm) in var. nigra form 1, while var. bracteolata was intermediate in colpus length (Fig. 6). Exine sculpture: It is reticular, with regular heterobronchate muri-pattern in var. nigra (Fig. 6 A & B), to irregular heterobronchate in var. bracteolata (Fig. 6 C). While, the endexine sculpture appeared warty in var. nigra and being smooth in var. bracteolata (Fig. 6). Discussion The taxonomic identity of the B. nigra (L.) Koch at the infraspecific level was subjected to debates long time ago since 1966 (Zohary & FeinbrunDothan, 1966), who classified B. nigra (L.) Koch into two varieties var. bracteolata (Fisch. & Mey.) Spach ex cross. and var. nigra, based mainly on the presence of epidermal trichomes and the length of fruit peak. In flora of Egypt, Täckholm (1974) mentioned the presence of B. nigra in three varieties namely var. bracteolata (Fisch. & Mey.) Spach ex Cross., var. torulosa (Pers.) Alef. and var. turgida(Pers.) Alef. Later, all these infra-specific taxa were grouped as synonyms for B. nigra species by Bolous (1999 and 2009). Worldwide, the “Plant list” and the “IUCN Red List” (Korpelainen et al., 2011), also grouped all the infra-specific taxa as synonyms to the species. In Egypt, the earlier works were at the flora and not in taxonomic levels, and no detailed taxonomic study was carried out at the infra-specific level, in addition to the notable morphological diversity observed during field trips and herbarium specimens, which enhance authors to carry the species auto-taxonomy. The study based on 50 macro-morphological characters (Table 2), revealed the presence of two distinctive varieties, var. bracteolata with glabrous plant surface and fruit peak not exceeding the quarter of the fruit length and var. nigra with trichomes and fruit peak extended to half of the whole fruit, in addition to the rest of the 50 characters outlined in Table 2 and Fig. 2, which outlined the notable variation in the terminal lobe of the basal leaf between the two varieties. This achieved classification is supported by the earlier grouping by Zohary & FeinbrunDothan (1966). Fig. 6. Pollen grains with reticulate-heterobronchate sculpture of the studied B. nigra (A & B: var. nigra with wide, regular muri-pattern and C: var. bracteolata, with narrow irregular muri-pattern). Egypt. J. Bot. 59, No.2 (2019) 10 WAFAA AMER et al. The identified varieties (var. nigra and var. bracteolata) showing no distinctive geographical distribution (Fig. 1 and Table 1) for each variety, both are co-distributed sometimes in mixed populations along the cultivated area in Nile Delta and Nile valley, this is an evidence for the absence of the environmental influence in the varieties distribution within the Egyptian borders. The retrieved micromorphological traits using SEM on stem (Fig. 3) and leaf (Fig. 4) showed distinctive differences between the studies 26 B. nigra populations, confirming grouping of them into two varieties. However, this grouping is not supported earlier by Boulos (1999 and 2009) and Korpelainen et al. (2011), works in which these varities treated as synonyms to B. nigra. The data retrieved from seed coat (Fig. 4), confirming our grouping of B. nigra into two varities, and the observed variations supported the potential use of seed coat pattern as a parameter for grouping to varity level. The underspecific variations in seed coat was supported by Song et al. (1988), Delseny et al. (1990), Prakash & Hinata (1980) and Koul et al. (2000), who reported a wide range of morphotypes in diploid Brassicaceae species, in which, Brassica nigra among them. The pollen grains of B. nigra possess reticulate, tricolpate type (Fig. 6), this reported by Abdel Khalek et al. (2002). The observed pattern regular heterobronchate in var. nigra and irregular heterobronchate in var. bracteolata, also the warty endexine in var. nigra compared to the smooth one in var. bracteolata (Table 3 and Fig. 6). These variations can give a phylogenetic point of view that B. nigra is more primitive than B. bracteolate. This phylogenetic opinion can be supported by the syncolpate aperture in the former versus the noticeable apocolpi in the later. The seed coat characters and the macromorphological characters of both stem and leaves confirming the presence of two distinct varieties for B. nigra in Egypt. The high taxonomic significance of pollen morphology at the infra-specific level was confirmed earlier by Amer & Abdo (2014). Conclusion Finally, we conclude that the autotaxonomy of B. nigra in Egypt based on the macro- and micromorphological characters confirming the presence of two distinctive varieties var. nigra and var. Egypt. J. Bot. 59, No.2 (2019) bracteolata with a trend in phylogeny within them and these two varieties should adopted taxonomically. References Abdel Khalek, K., Van Der Berg, R.G., Van Der Maesen, L.J.G. and El Hadidi, M.N. (2002) Pollen morphology of some tribes of Brassicaceae from Egypt and its systematic implications. Feddes Repert, 133(3-4), 211-223. Al-Shehbaz, I.A. 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Ren, C. and Bewley, J.D. (1998) Seed development, testa structure and precocious germination of Chinese cabbage (Brassica rapa subsp. pekinensis). Seed Science Research, 8(3), 385-398. Rollins, R.C. (1979) "Dithyrea and a Related Genus (Cruciferae)". Publ. Bussey Inst. Harvard Univ. 1979, pp. 3-20. Rollins, R.C. and Banerjee, U.C. (1979) "Pollen of the Cruciferae". Publ. Bussey Inst. Harvard Univ. 1979, pp. 33-64. Setia, R.C., Richa, S.N. and Malik, C.P. (1989) Phenolic acid induced shattering resistance in siliquae of Brassica juncea (cv RLM 514)-anatomical evaluation. Phytomophology, 39, 195-8. Song, K.M., Osborn, T.C. and Williams, P.H. (1988) Brassica taxonomy based on nuclear restriction fragment length polymorphisms (RFLPs). Theoretical and Applied Genetics, 75 (5), 784-794. Stork, A.L., Snogerup, S. and Wuest, J. (1980) Seed characters in Brassica section Brassica and some related groups. Candollea, 35, 421-50. Täckholm, V. (1974) "Students’ Flora of Egypt". Egypt: Cairo University, Egypt. The Plant List (2013). Version 1.1. URL: http://www. theplantlist.org Tsunoda, S. (1980) Eco-physiology of wild and cultivated forms in Brassica and allied genera. In: "Brassica Crops and Wild Allies: Biology", S. Tsunoda K.H.C., Gómez-Campo (Eds.). Vaughan, J.G. (1977) A multidisciplinary study of the taxonomy and origin of Brassica crops. Bio Science, 27(1), 35-40. Musil, A.F. (1948) Distinguishing the species of Brassica by their seed (No. 643). US Dept. of Agriculture. Warwick, S. (2011) Brassicaceae in agriculture. In "Genetica and genomics of the Brassicaceae". Shmidit, R. and Boncroft, I. (Eds.), pp. 34-65. Springer Publication. Prakash, S. and Hinata, K. (1980) Taxonomy, cytogenetics and origin of crop Brassicas, A review. Opera Bot. 55, 1-57. Zohary, M. and Feinbrun-Dothan, N. (1966) "Flora Palaestina", Jerusalem: Israel Academy of Sciences and Humanities. Punt, W., Blackmore, S., Nilsson, S. and Le Thomas, A. 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Bot. 59, No.2 (2019) ‫‪12‬‬ ‫‪WAFAA AMER et al.‬‬ ‫التصنيف الذاتي للمسترده السوداء (الفصيلة الصليبية) في مصر‬ ‫)‪(2‬‬ ‫وفاء عامر)‪ ،(1‬محمود شلقامي )‪ ،(2‬أحمد فريد )‪ ،(3‬هدير عبد الباسط‬ ‫)‪ (1‬قسم النبات والميكروبيولوجي كلية العلوم – جامعة القاهرة – الجيزة – مصر‪(2) ،‬قسم النبات والميكروبيولوجي‬ ‫كلية العلوم – جامعة المنيا – المنيا – مصر‪ (3)،‬قسم النبات والميكروبيولوجي كلية العلوم – جامعة أسيوط –‬ ‫أسيوط – مصر‪.‬‬ ‫جنس المسترده هو أحد أهم األجناس األقتصادية في الفصيلة الصليبية‪ ،‬ويحتوي علي ما يقرب من ‪ 80‬نوع‬ ‫علي مستوى العالم‪ .‬والمسترده السوداء هو أحد هذه األنواع وينمو بريا في حقول المحاصيل والطرق في منطقة‬ ‫المتوسط‪ .‬وقد أثبتت المالحظات الحقلية والمعشبية لتجمعات المسترده السوداء تنوع ملحوظ في الشكل الظاهري‪.‬‬ ‫ومن هنا كان لزاما ً إجراء دراسة تشمل الشكل الظاهري وحبوب اللقاح للتجمعات المختلفة بالمناطق الجغرافية‬ ‫المختلفة لهذا النوع للتعرف علي الوضع التصنيفي للوحدات تحت هذا النوع‪.‬‬ ‫وقد شملت الدراسة العينات المعشبية و ‪ 26‬تجمع للنبات موزعة على طول وادي النيل والدلتا‪ .‬وأثبتت‬ ‫الدراسة وجود صنفان األول بركتيوالتا والثاني نجرا‪ ,‬وقد ميزهما صفات الثمرة والشعيرات‪ .‬كما أوضحت‬ ‫المراجعة وجود صنف النجرا في نوعين بيولوجيين )بيوتيب(‪.‬‬ ‫كما أثبت المسح السطحي لقصرة البذرة بالميكروسكوب الماسح اإلليكتروني )‪ (SEM‬وكذلك حبوب اللقاح‬ ‫أن نوع المسترده السوداء يحتوي على وحدات تصنيفية واضحه تحت النوع وظهر هذا جليا ً من خالل حجم وشكل‬ ‫وفتحات األنبات لحبوب اللقاح‪ .‬وخلصت هذه الدراسة إلى وضع مفتاح تصنيفي للتمييز بين األصناف واألشكال‬ ‫البيولوجية لهذا النوع )المسترده السوداء(‪.‬‬ ‫)‪Egypt. J. Bot. 59, No.2 (2019‬‬