POLLEB Bangladesh J. Plant Taxon. 30(1): 1-19, 2023 (June) DOI: https://doi.org/10.3329/bjpt.v30i1.67023 © 2023 Bangladesh Association of Plant Taxonomists POLLEN MORPHOLOGY OF TWENTY THREE SPECIES OF ALLIUM L. (AMARYLLIDACEAE) FROM TURKEY ÜMMÜGÜLSÜN YILDIZ* Republic of Turkey, Ministry of Agriculture and Forestry, General Directorate of Nature, Conservation and Nationals Parks, Karşıyaka-İzmir-Turkey Keywords: Allium; Amaryllidaceae; Pollen morphology; LM; SEM; TEM; Flora of Turkey. Abstract The pollen morphology of 23 Allium L. species, grown in Turkey, belonging to the sections; Rhizirideum, Schoenoprasum, Cepa, Molium, Brevispatha, Scorodon, Acanthoprason and Melanocrommyum, were investigated by LM (light microscopy), TEM (transmission electron microscope), and SEM (scanning electron microscopy). This paper presents detailed pollen morphological features are given for these taxa. Common characteristics of pollen of Allium species investigated are ellipsoidal and heteropolar, bilateral symmetry; pollen grains are shed as monads. They are monosulcate (extended sulcate types) and monosulcate-operculate. The sculpturing of the exine, pollen membrane ornamentation, lumina shape, and sulcus membrane with a fragmented operculum are found to be characteristic features for separating species. Based on these properties three main pollen types were determined with three different exine sculpturing. The characteristic structures of pollen ornamentation, observed in SEM micrographs, are striate­perforate, striate­regulate-perforate and regulate-perforate. Sulcus membrane ornamentations are psilate, psilate-perforate, rugulate and regulate-perforate. The exine is semitectate and the tectum perforate. Columellae were found to be simplicolumellate. Introduction The genus Allium L. is one of the most diverse and taxonomically difficult groups of the monocots. The most recent classifications are based on morphological characters and molecular data, recognized more than 920 species in 15 subgenera and 80 sections (Friesen et al., 2006). Allium is a taxonomically quite complex and has unresolved nomenclatural problems. In the last decades, many Allium taxa were newly described for Turkey and the certain number of Allium species and subspecies raised the number to about 220, classified into 15 sections, ca. one-third of which are endemic to this territory, demonstrating that Turkey is a prominent part of the southeastern Asian center of Allium diversity (Kollmann, 1984; Davis, 1984, 1988; Güner et al., 2000; Friesen et al., 2006; Koyuncu and Eker 2011; Celep et al., 2012; Güner et al. 2012; Özhatay and Genç, 2013 and Govaerts et al., 2013; Ekşi et al., 2015, 2016; Duman et al., 2017; Fırat et al., 2018; Govaerts et al., 2019; Özdöl et al., 2022). Cytological and data on pollen morphology of representatives of Allium genus has been subjected to earlier investigations (Nair and Sharma, 1965; Kuprianova, 1967; Radulescu, 1973; Kuprianova and Aliev, 1979; Schulze, 1980, 1980a; Pastor, 1981; Diez et al., 1987; El-Sadek et al., 1994; Kosenko and Kudryashova, 1995; Tolgor, 1995; Hosseinzadeh et al., 2009; Namin et al., 2009; Neshati et al., 2009, Maassoumi et al., 2014; Wrońska-Pilarek et al., 2016; Perveen and Qaiser, 2015; Hosseini, 2018). Palynological studies on Allium in Turkey; detailed studies of the pollen morphology of 14 species of Allium were made by Güler and Pehlivan (2006), 23 species of Allium were examined by Özhatay and Koçyiğit *Corresponding author: ugyildizz@gmail.com; ummugulsun.yildiz@tarimorman.gov.tr https://doi.org/10.3329/bjpt.v30i1.67023 mailto:ugyildizz@gmail.com mailto:ummugulsun.yildiz@tarimorman.gov.tr 2 YILDIZ (2009) and 16 species of Allium were examined by Özler and Pehlivan (2010), 12 taxa belonging to Sect. Codonoprasum were examined by Koçyiğit (2014) and later, 10 species Allium L. taxa, 6 of which are endemic to Turkey, were examined by Başer et al. (2019), respectively. In literature on the Allium genus, the following pollen grain features are considered as having the highest diagnostic value. Turkey Allium an extremely polymorphous and taxonomically difficult genus species show a great morphological diversity and has unresolved nomenclatural problems therefore many taxonomical problems remain unsolved (Kollmann, 1984; Gurushidze et al., 2007). To solve the systematic problems of these polymorphic groups further cytological, anatomical and palynological studies are urgently needed. In order to obtain more morphological data to solve taxonomical problems, detailed study of pollen morphology in Alliaceae family always has been suggested and pollen structure of some species of the genus Allium has been subjected to earlier investigations. In literature on the Allium genus, the following pollen grain features are considered as having the highest diagnostic value. Pollen information of some species from Turkey Güler and Pehlivan (2006) and Özler and Pehlivan (2010) suggested the usefulness of pollen characteristics for the systematics of the genus, and observed the possible use of these characters for solving certain taxonomical problems as well. Güler and Pehlivan (2006) recognized the sulcus type, sulcus structure, exine ornamentation, ekzin structure, presence or absence of operculum and operculum type, presence or absence of perforations on the pollen surface, density of perforations, size of perforations, and size of the pollen grains characteristics, which are also important for separating taxa at different taxonomic ranks. The present study adds pollen morphological data for a further 23 species (including 14 sections and to compare the details of sculpturing), again using light, scanning electron microscopy and transmission electron microscope. An overview of pollen characteristics for all the Turkish representatives of the genus Allium, thus concluding a series of studies aimed at describing the characteristics of all the Turkish Allium genera, and enabling an overall review of the pollen data both from this study and from our previously published studies (Güler and Pehlivan 2006). Material and Methods The pollen grains were obtained from the Faculty of Pharmacy of Ankara University (AEF) herbarium. The complete list of the investigated taxa with sample provenance is reported in Table 1. Pollen morphological terminology of Walker (1974a, 1974b), Faegri and Iversen (1989), Punt et al. (1994, 2007); El-Sadek et al. (1994) and Hesse et al. (2009) was followed. LM investigation Samples were taken from herbarium specimens. For LM observations, the pollen was first treated with 70% ethyl alcohol to remove oily substances, and then embedded in glycerine-jelly, stained with safranin. For morphological analysis, pollen grains of specimens for LM investigations were prepared according to the methods of Wodehouse (1935) and Erdtman (1960). Pollen dimensions of all species were measured in such amounts that the resulting data followed Gaussian curves these measurements which were made in acetolysed, nonacetolysed pollen. In each sample, 50 pollen grains were measured in order to obtain the maximum and average value of the size. LM studies were made using a Nikon Alphaphot-2 YS2 microscope, under (E40, 0.65) and oil immersion (E100, 1.25), using 15 eye piece (ocular) and the following paremeters, as which pollen size i.e. long axis (LA) and short axis (SA), length of the sulcus (slg), width of the sulcus (slt) sulcus width for all types and operculum type, exine and intine thickness for all types. POLLEN MORPHOLOGY OF 23 SPECİES OF ALLIUM 3 The used eyepiece and lens scales require a conversion of the measurement results to micrometers (μm). These measurements which were made in acetolysed, nonacetolysed pollen and on LM micrographs are given in (Figs. 67-98). The LM photomicrographs were taken with an Orthomat- w camera connected to a Carl Zeiss-photo binocular light microscope. SEM investigation For SEM investigations, the pollen was first treated with 70% ethyl alcohol then air-dried before being mounting on SEM specimen stubs subsequently coated with gold plate, and examined under a Jeol JSM-840A (Turkish Petroleum Corporation, TPAO, Turkey) scanning electron microscope. The clearest SEM photographs representing each pollen type and the main pollen features were selected for this paper (Figs 1-24, Figs 25-48, Figs 49-56). SEM micrographs were used mainly for studying the overall shape, type of sculpturing, and more detailed information on the sculptures. Figs 1­12: SEM photographs of the pollen grains of Allium species, showing pollen grain in distal view. 1. A. scabriscapum: Pollen grain in distal view, SEM x 2500; 2. A. szovitsii: Pollen grain in distal view, SEM x 1500; 3. A. cepa: Pollen grain in distal view with psilate-perforate of the sulcus membrane, SEM x 4000; 4. A. schoenoprasum: Pollen grain in distal view with operculum, SEM x 2500; 5. A. cassium: Pollen grain in distal view, SEM x 2000; 6. A. zebdananse: Pollen grain in distal view, SEM x 2000; 7. A. subhirsutum: Pollen grain in distal view, perforate-striate ornamentation, SEM x 2200; 8. A. subhirsutum: Pollen grain in distal view, SEM x 1800; 9. A. cupani subsp. hirtovaginatum: Pollen grain in distal view with rugulate-perforate of the sulcus membrane, SEM x 2500; 10. A. longisepalum: Pollen grain in distal view, SEM x 2000; 11. A. callidictyon: Pollen grain in distal view, SEM x 2500; 12. A. callimischon subsp. haemostictum Pollen grain in distal view, SEM x 1500; Scale bar = 10 µm (1,2,4,5,6,7,8,9,10,11,12); 1 µm (3). 4 YILDIZ Figs 13-24: 13. A. frigidum: Pollen grain in distal view with psilate-perforate of the sulcus membrane, SEM x 2500; 14. A. kossoricum: Pollen grain in distal view with psilate ornamentation of the sulcus membrane, SEM x 2700; 15. A. akaka: Pollen grain in proximal view with sulcus extending to the proximal face and with broad and rounded ends view with rugulate-perforate ornamentation, SEM x 2000; (16) A. akaka: Pollen grain in distal view SEM x 1900; 17. A. akaka: Pollen grain in distal view with striate-rugulate-perforate ornamentation, SEM x 2300; 18. A. chrysanterum: Pollen grain in distal view, SEM x 2500; 19. A. cardiostemon: Pollen grain in distal view with operculum, SEM x 2500; 20. A. colchicifolium: Pollen grain in distal view, SEM x 2500; 21. A. decipiens: Pollen grain in distal view with psilate ornamentation of the sulcus membrane SEM x 3500; 22. A. kharputense: Pollen grain in distal view, with operculum, SEM x 2500; 23. A. noëanum: Pollen grain in distal view with rugulate ornamentation of the sulcus membrane SEM x 2500; 24. A. lycaonicum: Pollen grain in lateral view, SEM x 2500 with rugulate ornamentation. Scale bar = 10 µm. TEM investigation Acetolyzed pollen grains were stained with 2% OsO4 and uranyl acetate, dehydrated and embedded in epon araldite according to the method described by Skvarla and Turner (1966). Ulthrathin sections of the pollen grains were obtained with a glass knife in a Reichert Supernova microtome (Gazi University, Faculty of Medicine, Ankara Turkey). Post-staining was done with POLLEN MORPHOLOGY OF 23 SPECİES OF ALLIUM 5 lead citrate for 5 minutes (Reynolds, 1963), and the sections were examined under a Zeiss EM9 (Figs 57-66). TEM micrographs were used mainly for studying the overall shape, pollen walls type of sculpturing, and more detailed information on the sculptures. Figs 25-39: Ornamentation of Allium pollen grains. 25. A. scabriscapum: Striate-perforate ornamentation, SEM x 10000; 26. A. szovitsii: Striate-perforate ornamentation, SEM x 10000; 27. A. shoenoprasum: Striate-perforate ornamentation, SEM x 10000; 28. A. subhirsutum: Striate-perforate ornamentation, SEM x 10000; 29. A. cassium: Striate-rugulate-perforate ornamentation SEM x 10000; 30. A. cupani subsp. hirtovaginatum: Striate-rugulate- perforate ornamentation SEM x 10000; 31. A. zebdananse: Striate-rugulate-perforate ornamentation SEM x 10000; 32. A. longisepalum: Distal face showing perforate-rugulate ornamentation, SEM x 10000; 33. A. longisepalum: Proximal face showing perforate-rugulate ornamentation, SEM x 10000; 34. A. callidictyon: Perforate-striate-rugulate ornamentation, SEM x 10000; 35. A. callimischon subsp. haemostictum: Perforate-striate-rugulate ornamentation, SEM x 10000; 36. A. frigidum: Rugulate-perforate ornamentation SEM x 10000; Perforate-striate-rugulate ornamentation, SEM x 10000; 37. A. kossoricum: Striate-perforate ornamentation, SEM x 10000; 38. A. akaka: Striate-rugulate-perforate ornamentation SEM x 10000; 39. A. cardiostemon: Perforate-striate ornamentation, SEM x 10000; Scale bar = 10 µm. 6 YILDIZ Figs 40-48: Ornamentation of Allium pollen grains. 40. A. colchicifolium: Striate-perforate ornamentation, SEM x 10000; 41. A. decipiens: Rugulate-perforate ornamentation SEM x 10000; 42. A. orientale: Distal face showing Striate- rugulate-perforate ornamentation SEM x 3500; 43. A. orientale: Proximal face showing Striate-rugulate-perforate ornamentation SEM x 3500; 44. A. kharputense: Striate-perforate ornamentation, SEM x 10000; 45. A. noëanum: Striate-perforate ornamentation, SEM x 10000; 46. A. lycaonicum: Proximal face showing perforate-rugulate ornamentation, SEM x 10000; 47. A. lycaonicum: Distal face showing sitriate-perforate ornamentation, SEM x 10000; 48. A. hirtifolium: Distal face showing sitriate-perforate ornamentation, SEM x 10000; Scale bar = 10 µm. Results and Discussion All studied pollen grains are medium-sized (25-50 μm) with more or less bilateral symmetry. The sulcus is either as long as the half of the circumference of the pollen grain (longest axis), or much longer and extends to the proximal face. The main palynogical features of Turkish Allium taxa (and specimens) examined are summarized in Table 2. According to LM and SEM investigations, the pollen grains are monad, monosulcate, monosulcate-operculate, ellipsoidal, heteropolar with more or less bilateral symmetry; 25.98 to 50.32 µm long axis (LA) and 16.26 to 32.10 µm short axis (SA), the form was prolate (mean of LA/SA ratio 1.12 to 1.69). The pollen shapes (based on long axis (LA)/ short axis (SA) ratio) prolate or subprolate, outline more or less circular in polar view and boat-shaped. Pollen morphological parameters are given in Table 2. Medium to large in size with LA 25-50 µm and SA 17-36 µm. The sulcus extends from distal to proximal in all species. Sulcus ends are sharp, blunt, broad and rounded. Sulcus membrane ornamentations were rugulate or psilate. In most of the species with operculum, there are fragmented operculum within the sulcus of A. szovitsii, A. shoenoprasum (Fig. 69), A. cupani subsp. hirtovaginatum (Fig. 80), A. callidictyon (Fig. 82), A. POLLEN MORPHOLOGY OF 23 SPECİES OF ALLIUM 7 callimischon (Fig. 86), A. akaka (Fig. 89), A. chrysantherum (Fig. 91), A. cardiostemon (Fig. 19), A. decipiens (Fig. 94), A. orientale (Figs 95-96), A. kharputense (Fig. 22), A. lycaonicum (Fig. 97), and A. hirtifolium (Fig. 98). Sulcus membrane ornamentations are psilate, psilate-perforate, rugulate and and rugulate-perforate (Figs 49-56). Figs 49-56: Ornamentation of the sulcus membrane of Allium pollen grains. SEM photographs of the pollen grains of Allium species, showing pollen grain in distal view. 49. A. szovitsii: rugulate-perforateof the sulcus membrane SEM x 7000; 50. A. subhirsutum: Psilate-perforate of the sulcus membrane, SEM x 5000; 51. A. zebdananse: Rugulate- perforate ornamentation of the sulcus membrane, SEM x 5000; 52. A. longisepalum: Psilate-perforate of the sulcus membrane, SEM x 5000; 53. A. cardiostemon: Rugulate-perforate ornamentation of the sulcus membrane, SEM x 8000; 54. A. colchicifolium: Rugulate-perforate of the sulcus membrane, SEM x 8000; 55. A. kharputense: Psilate of the sulcus membrane, SEM x 6500; 56. A. lycaonicum: Rugulate-perforate ornamentation of the sulcus membrane, SEM x 6000 Scale bar = 1 µm. 8 YILDIZ Figs 57-66: TEM photomicrographs of exine structure (Pollen wall stratification and morphology of Allium pollen grains). 57. A. scabriscapum: TEM x 10000; 58. A. schoenoprasum: TEM x 30000; 59. A. cepa: TEM x 10000; 60. A. subhirsutum: TEM x 20000; 61. A. cupani subsp. hirtovaginatum: TEM x 20000; 62. A. callidictyon: TEM x 20000; 63. A.: TEM x 20000; 64. A. frigidum: TEM x 12000; 65. A. cardiostemon: TEM x 30000; 66. A. decipiens: TEM x 20000. POLLEN MORPHOLOGY OF 23 SPECİES OF ALLIUM 9 Figs 67-82: LM photographs of selected Allium species. 67. A. szovitsii: Proximal view with ornamentation LM x 1000 (A); 68. A. szovitsii: oblique polar proximal view, showing sulcus border, LM x 1000 (A); 69. A. schoenoprasum: oblique polar proximal view, showing operculum, LM x 1000 (A); 70. A. schoenoprasum: Pollen grain in proximal view, Striate-perforate ornamentation, LM x 1000 (A); 71. A. cepa: Pollen grain Equatorial distal view LM x 1000 (A); 72. A. cepa: Proximal view LM x 1000 (A); 73. A. cepa: oblique polar proximal view, LM x 1000 (N); 74. A. subhirsutum: Proximal view with ornamentation LM x 1000 (A); 75. A. cassium: Pollen grain Equatorial distal view LM x 1000 (N); 76. A. cassium: Pollen grain in distal view, striate-rugulate-perforate ornamentation LM x 1000 (A); 77. A. cassium: oblique polar proximal view, LM x 1000 (A); 78. A. cassium: Equatorial distal view LM x 1000 (A); 79. A. zebdananse: oblique polar proximal view, LM x 1000 (N); 80. A. cupani subsp. hirtovaginatum: Pollen grain in proximal view with sulcus extending to the proximal face, LM x 1000 (A); 81. A. cupani subsp. hirtovaginatum: Pollen grain in distal view with fragmented operculum, LM x 1000 (N); 82. A. callidictyon: Pollen grain in distal view with fragmented operculum, LM x 1000 (N); Scale bar = 1µm (40-52). (A) = acetolysed, (N) = not acetolysed. 10 YILDIZ Figs 83-98: LM photographs of selected Allium species. 83. A. callidictyon: pollen grain in proximal view with sulcus extending to the proximal face, LM x 1000 (N); 84. A. callidictyon: oblique polar proximal view, LM x 1000 (N); 85. A. callimischon subsp. haemostictum: Pollen grain in distal view LM x 1000 (N); 86. A. callimischon subsp. haemostictum: Pollen grain in distal view with fragmented operculum, LM x 1000 (N), 87. A. callimischon subsp. haemostictum: Equatorial distal view with operculum, LM x 1000 (N); 88. A. frigidum: Proximal view with ornamentation LM x 1000 (A); 89. A. akaka: Pollen grain in distal view with fragmented operculum, LM x 1000 (N); 90. A. akaka: Pollen grain in distal view with fragmented operculum, LM x 1000 (N); 91. A. chrysantherum: oblique polar proximal view, view showing operculum, LMx1000 (N); 92. A. cardiostemon: oblique polar proximal view, LMx1000 (N); 93. A. decipiens: Pollen grain in equatorial distal view, LMx1000 (N); 94. A. decipiens: Pollen grain in distal view with fragmented operculum, LM x 1000 (N); 95. A. orientale: Pollen grain in distal view with operculum, LM x 1000 (N); 96. A. orientale: Pollen grain in equatorial distal view with fragmented operculum, LM x 1000 (N); 97. A. lycaonicum: Pollen grain in distal view with operculum, LM x 1000 (N); 98. A. hirtifolium: Pollen grain in distal view showing sulcus, LM x 1000 (N). Scale bar = 1 µm (83-98). (A) = acetolysed, (N) = not acetolysed. POLLEN MORPHOLOGY OF 23 SPECİES OF ALLIUM 11 The exine, is mainly composed of columellae covered by the tectum which contains perforations (eutectate and microperforate pollen grains). These microperforations were only visible on SEM micrographs. There are also differences in the number of perforation, diameter of perforation and thickness of lirae. Fine perforation is getting bigger through sulcus side (Figs 13, 14, 21), (The perforations small ca. 0.08-0.35 µm the width of the intervening tectum). The most typical (separate species) have structured muri, simplicolumellate muri and were formed by lirae. The lumina are almost circular in Allium species (Figs 25-48). Schulze (1980) and Pınar et al. (2009) have shown that muri and lumen shapes of the pollen are taxonomically important characters. Small convex sculpture elements (supratectal muri) on the surface are more or less irregularely arranged and short (rugulate to rugulate-perforate pattern, sometimes muri very shallow) with perforations of variable diameter in between, rarely long and parallel (transitions to striate condition) These were formed by striae of different length and orientation and detail of the exine showing irregular striate-perforate ectexine (perforation only on the outside of the tectum) are presented. The number of perforation in 1µm² is 4-14, the diameter of perforation on average is 0.08-0.35 µm and the thickness of lira on average is 0.16-0.33µm (Figs. 25-48). The diameter of perforation was observed to be the highest in A. frigidum (Fig. 36). Intine is 0.52-0.96µm thick. The A. longisepalum and A. subhirsutum had thickest intine while A. schoenoprasum, A. kossoricum, had thinnest one (Table 2). According to SEM survey, exine ornamentation can be described best as the exine sculpture was striate-perforate, striate-rugulate-perforate and rugulate-perforate (Figs 25-56). However, in A. longisepalum, one of the lateral surfaces of the pollens is striate-rugulate-perforate and the other surface of the pollen grains was striate-perforate (Figs 10, 32-33); A. akaka (Figs 15-17, 38), A. orientale (Figs 42-43), A. lycaonicum (Figs 24, 46-47), one of the lateral surfaces of the pollens are striate-perforate and the other surface of the pollen grains was rugulate-perforate Therefore, these pollens grains are para-isopolar. Perforate-striate, perforate-rugulate-perforate and perforate- striate-rugulate exine structure have been reported in previous investigations (Pastor 1981; Diez et al., 1987; El-Sadek et al., 1994; Güler and Pehlivan, 2006; Namin et al., 2009; Neshati et al., 2009; Özhatay and Koçyiğit, 2009; Özler and Pehlivan, 2010; Koçyiğit, 2014; Maassoumi et al., 2014; Wrońska-Pilarek et al., 2016; Hosseini, 2018 and Başer et al., 2019). A rugulate-perforate ornamentation was observed in A. frigidum (Fig. 36) and A. decipiens (Fig. 41), while striate- rugulate-perforate ornamentation was seen in A. cepa (Fig. 3), A. cassium, (Fig. 29) and A. zebdananse (Fig. 31), A. longisepalum (Fig. 32) A. cupani subsp. hirtovaginatum (Fig. 30), A. callidictyon (Fig. 34), A. callimischon subsp. haemostictum (Fig. 35), A. akaka, A. orientale (Fig. 42), A. lycaonicum (Fig. 47) Whereas in the other species striate-perforate ornamentation was observed. The present study revealed that Allium taxa possess three types of ornamentational characteristics as follows: (i) Striate-perforate: A. scabriscapum, A. szovitsii, A. schoenoprasum, A. subhirsutum, A. kossoricum, A. chrysanterum, A. cardiostemon, A. colchicifolium, A. kharputense, A. noëanum, A. hirtifolium. (ii) Striate-regulateperforate: A. cepa, A. cassium A. zebdananse, A. longisepalum, A. cupani subsp. hirtovaginatum, A. callidictyon, A. callimischon subsp. haemostictum, A. akaka, A. orientale, A. lycaonicum. (iii) Rugulate-perforate: A. frigidum, A. decipiens. 12 YILDIZ POLLEN MORPHOLOGY OF 23 SPECİES OF ALLIUM 13 14 YILDIZ POLLEN MORPHOLOGY OF 23 SPECİES OF ALLIUM 15 In SEM photomicrographs, sulcus membranes are psilate in A. scabriscapum, A. schoenoprasum, A. kossoricum (Fig. 14), A. decipiens (Fig. 21), A. orientale and A. kharputense (Fig. 55); and psilate - perforate sulcus membrane ornamentation was seen in A. cepa (Fig. 3), A. subhirsutum (Fig. 50), A. cassium, A. longisepalum (Fig. 52), A. callidictyon, A. frigidum (Fig. 13) and A. colchicifolium (Figs. 20, 54); Rugulate sulcus membrane ornamentation was found in A. chrysanterum and A. noëanum (Fig. 23), and rugulate-perforate sulcus membrane ornamentations are seen in A. szovitsii (Fig. 49), A. zebdananse (Fig. 51), A. cupani subsp. hirtovaginatum (Fig. 9), A. callimischon subsp. haemostictum, A. decipiens (Fig. 21), A. akaka (Figs 15, 16), A. cardiostemon (Fig. 53), A. lycaonicum (Fig. 56) and A. hirtifolium similarly, Güler and Pehlivan (2006); Özler and Pehlivan (2010); Maassoumi et al. (2014) and Başer et al. (2019) reported that sulcus membrane ornamentations were psilate, striate-reticulate, rugulate and regulate-perforate in Allium taxa. Several researchers have emphasized that the sulcus membrane sculpturing may be a taxonomic value in some families (Kosenko, 1999; Güler and Pehlivan, 2006; Özler and Pehlivan, 2010). The genus studied includes the sections; Rhizirideum, Schoenoprasum, Cepa, Molium, Brevispatha, Scorodon, Acanthoprason and Melanocrommyum. We aimed to elucidate the infra- familial positions of the Allium. On the basis of the pollen structure of Allium within Alliaceae it found to be homogenous genus from the pollen point of view and a heterogenous family from the morphological point of view. Pollen morphology of 23 taxa of Allium was investigated under LM (Figs 67-98), SEM (Figs 1-56) and TEM (Figs. 57-66). The common characteristics of pollen grains were monad, monosulcate (extended sulcate types), and monosulcate-operculate, ellipsoidal, heteropolar with bilateral symmetry. However, pollen of A. longisepalum, A. akaka, A. orientale and A. lycaonicum are paraisopolar. This study shows that pollen characters have been significant value in classification of Allium. The main palynological differences have been found at the section level, especially in the sulcus membrane and the presence of an operculum. A distal fragmented operculum and extended sulcate type are typical for Allium of Alliaceae. Sulcus long, getting the ends of the pollen grain or spreading to the proximal side. In the present study, the biggest pollen size was found in A. longisepalum, whereas the smallest was found in A. cardiostemon. There is a decrease in pollen size in the sequence of the subsections Rhizirideum, Schoenoprasum, Cepa, Molium, Brevispatha, Scorodon, Acanthoprason, Melanocrommyum .The sulcus extends from distal to proximal and the sulcus ends are braod and rounded only in A. akaka (Figs 15, 16) while they were sharp in other species such as A. subhirsutum (Figs. 8, 74), A. zebdanense (Fig. 6), A. longisepalum (Fig. 10), A. cupani subsp. hirtovaginatum (Fig. 80), A. callidictyon (Figs. 11, 83), A. callimischon (Fig. 87), A. chrysantherum (Fig. 18) and A. orientale and it’s blund in other ones. The longest length dimension of sulcus was seen in in section Molium are A. subhirsutum, A. cassium, A. zebdanense and A. longisepalum. Figs 5, 6, 8, 10). The widest sulcus dimension was seen in A. subhirsutum (Figs 8, 74). It was recognized that the sulcus extends from distal to proximal end in all the taxa investigated (Table 2). The operculum was found to be fragmented on the sulcus membrane (Figs 69, 81, 82, 86, 87, 89, 90, 91, 94, 95, 96, 97) or sometimes completely covering it. Three are in our previous unreported from this genus, the operculum was determined only in A. pallens subsp. pallens, A. bassitense and A. hirtovaginum under the Section Codonoprasum (Güler and Pehlivan 2006). In the other studies of this genus, the operculum was determined only in A. albidum subsp. caucasicum (Section Rhizirideum), A. rupicola (Section Codonoprasum), A. asperiflorum under the Section Allium (Özler and Pehlivan, 2010), 12 taxa belonging to Section Codonoprasum 16 YILDIZ (Koçyiğit, 2014) and A. arlgirdense under the Section Scorodon (Başer et al., 2019). According to Kosenko (1992), a non-operculate exine is a plesiomorphic peculiarity. Several researchers have emphasized that the sulcus features and the presence of operculum may be a taxonomic value in some families (Chanda et al., 1979; Halbritter and Hesse, 1993; Güler and Pehlivan 2006; Özler and Pehlivan, 2010). The advantage of a monosulcate aperture (extended sulcate) in monocotyledons with the inclusion of Allium, is underlined by Harley and Zavada (2000); Güler and Pehlivan (2006); Namin et al., 2009; Neshati et al. (2009); Özhatay and Kocyigit (2009); Özler and Pehlivan (2007, 2010); Maassoumi et al. (2014) and Başer et al. (2019). Palynological data related to exine ornamentation indicate the heterogeneous characters of this genus. In this study we have determined that there are intraspecific variations among studied species as well which are based on exine sculpturing, and sulcus ornamentation. Allium species divided into 4 pollen types according to sulcus membrane sculpturing; among Allium species a psilate, psilate-perforate, rugulate and rugulate-perforate sulcus membrane is distinctive. In the present investigated taxa such as A. scabriscapum (Fig. 57), A. schoenoprasum, (Fig. 58) and A. cepa (Fig. 59). The exine is semitectate and the tectum perforate while the tectum is with intervals and is formed by simple columella. Exine thickness 1-2 µm. Ectexine is thicker than endexine and that endexine exhibits a very thin continuous structure. Tectum is thicker than foot layer with intervals (Figs 57-66). The results show that there were several pollen characters of taxonomic significance in the genus Allium. There are also differences in the size of the pollen, exine sculpturing, ornamentation of sulcus membrane and lumen shape, size and murus size, exine thickness and number of perforation, diameter of perforation and thickness of lira. We recognized 3 main types, distinct by pollen sculpturing, lumina shape and sulcus membrane ornamentation. The main palynological differences have been registered at the section level. These results are similar to the earlier studies (Güler and Pehlivan, 2006; Özler and Pehlivan, 2010; Neshati et al., 2009; Özhatay and Kocyigit, 2009; Başer et al., 2019; Table 2; Figs 25-48). Acknowledgements Special thanks are due the Council of Highre Education (YÖK), the Scientific and Technological Research Council of Turkey (TUBITAK). The authors are also grateful to Prof. Dr. Mehmet Koyuncu, Ankara University-Department Biology. We also would like to thank to ANK herbarium for their kind support. I would like to express my gratitude to the Turkish Petroleum Corporation (TPAO) access to their scanning electron microscope and technical support. Special thanks are due to TEM studies Gazi University, Faculty of Medicine, Ankara. References Başer, B., Fırat, M. and Binzet, R. 2019. 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