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Pollen morphology of certain species of the family Lamiaceae in Saudi Arabia
Accepted Manuscript Pollen Morphology of certain species of the Family Lamiaceae in Saudi Arabia Abdullah R. Doaigey, Mohamed El-Zaidy, Ahmad Alfarhan, Abd El-Salam Milagy, Thomas Jacob PII: DOI: Reference:
S1319-562X(17)30084-0 http://dx.doi.org/10.1016/j.sjbs.2017.03.001 SJBS 918
To appear in:
Saudi Journal of Biological Sciences
Received Date: Revised Date: Accepted Date:
11 May 2016 23 February 2017 1 March 2017
Please cite this article as: A.R. Doaigey, M. El-Zaidy, A. Alfarhan, A.E-S. Milagy, T. Jacob, Pollen Morphology of certain species of the Family Lamiaceae in Saudi Arabia, Saudi Journal of Biological Sciences (2017), doi: http:// dx.doi.org/10.1016/j.sjbs.2017.03.001
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Pollen Morphology of certain species of the Family Lamiaceae in Saudi Arabia
Abdullah R. Doaigey Botany & Microbiology Dept., College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia. Tell. :0096614675879 e-mail: [email protected] Mohamed El-Zaidy Botany & Microbiology Dept., College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia. Tell. : 0096614675878 e-mail: [email protected] Ahmad Alfarhan Botany & Microbiology Dept., College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia. Tell. :0096614675808 e-mail: [email protected] Abd El-Salam Milagy Botany & Microbiology Dept., College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia. Tell. :0096614675878 e-mail: [email protected] Jacob Thomas Botany & Microbiology Dept., College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia. Tell.: 0096614675807 e-mail: [email protected]
Pollen Morphology of certain species of the Family Lamiaceae in Saudi Arabia Abdullah R. Doaigey ⃰, Mohamed El-Zaidy, Ahmad Alfarhan, Abd El-Salam Milagy and Thomas Jacob Botany & Microbiology Dept., College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia.
⃰ E-mail: [email protected].
ABSTRACT The intention of the present wok is to provide an account on the pollen morphological features of Lamiaceae in Saudi Arabia as a basis for future studies of Lamiaceae pollens in the region. Pollen morphology of 20 species belong to 16 genera of the Lamiaceae has been investigated using Light Microscope (LM) and Scanning Electron Microscope (SEM). The study revealed that the pollen grains were characterized by 3-zonocolpate or 6-zonocolpate. Size of the pollen is variable between the genera but not among the species of the same genus. The surface pattern of the exine varies from fine reticulate, rough reticulate, mega-reticulate, reticulateperforate, bireticulate- perforate or granulate, leading to 6 types of pollen grains. These variations revealed by this study implies that pollen Morphology may be of significant value sharing in solving problems in the classification of Lamiaceae members. A Key to the species, based on the morphological features of pollen grains, is also provided.
KEY WORDS: Colpi, Lamiaceae, Pollen grains, Pollen Morphology, Saudi Arabia.
INTRODUCTION Lamiaceae is represented in Saudi Arabia by 26 genera and about 70 species (Migahid, 1989 and Collenette, 1999). 14 species are endemic to the Kingdom of Saudi Arabia, while 27 species are endangered, vulnerable or rare (Collenette, 1998). Pollen morphology including aperture number, shape and tectum ornamentation were studied in different members of the family Lamiaceae and found these features were helpful in
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its taxonomy (Erdtman, 1945; Abu-Asab and Cantino 1994; Raj, 1974; Rudall, P. 1980; Bazarragchaa, et al., 2012). However, Perveen and Qaiser (2003) and Al-Watban, et al. (2015) were studied pollen morphology of several species in different genera of the Family Lamiaceae and suggested that it is useful in grouping and identifying the species. The genus Salvia was studied by several investigators (Jafari and Nikian, 2008; Yildiz et al, 2009; Kahraman et al. 2009 a, b, 2010 a, b); Kahraman and Doghan, 2010) who reported the pollen characters of species were useful for their identification. Celenk, et al. (2008 a, b), indicated that the pollen characters of the genus Mentha were valuable for taxonomic applications. Erkara and Koyuncu (2007), Salmaki, et al. (2008) and Boi et al. (2013) indicated in their study on Stachys species that the pollen morphological characters were useful in the identification of certain sections and species. Akgul, et al. (2008) reported in their study of the genus Marrubium in Turky that morphological characters of pollens are of taxonomic significance in the genus. Harley, et al. (1992) found in their studies that most of Ocimum species pollens are with bireticulate- perforate tectum sculpture and suggested a re-evaluation in their classification. As well as Arogundade and Adedeji (2009) in their study of Ocimum L. species concentrated on pollen colpi reporting different types of pollens acolpate to octacolpate pollens which reveal that there are significant differences among the species. Moreover, Dinç et al, (2009) reported in their study on Lallemantia species growing in Turkey, the exine sculpture of pollen had taxonomic value. Kose, et al. (2011) reported that the granulate or reticulate type of exine ornamentation in Ajuga species may be is useful in segregating the species within the genus. Hasani-Nejad et al. (2011) and Jamzad and Hasani-Nejad (2013) showed that the exine ornamentation could be used as a taxonomic feature below generic level classification of Scutellaria. Osman (2012) in his study of Ballota species in Egypt mentioned that pollen characters were very useful in the separation of the species. Pollen grains of Leucas indica (L) R.Br.Ex Vatke., were investigated by Prabhakar and Ramakrishna (2014) and suggested that pollen shape, aperture and tectum sculpture may help in identification of the species. Navarro, et al. (2004) mentioned in Teucrium, pollen tectum surface ornamentation types and pollen size seem to have a taxonomic value. The forgoing investigations indicate that there is no definitive agreement for which level of classification can be useful of using pollen morphological characters in Lamiaceae. The
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intention of the present work is to provide an account of pollen morphological features of Lamiaceae members in Saudi Arabia as a basis for future studies.
MATERIALS AND METHODS Pollens were excised from either fresh and preserved
samples or from
herbarium specimens deposited either at the herbarium of King Saud University (KSU) or at the National Herbarium (RIY) of Saudi Arabia. (Table,1). Five samples of defferent collections were prepared for each species to confirm the stability of pollen characters within a species, by using the usual acetolysis method (Erdman, 1971). Measurements were made within two to four days period to avoid the possiblility of error introduced by differential swelling. Twenty pollen grains were measured using an ocular micrometer (Zeiss Photometric microscope III) and the mean was taken. Measurements include polar length, equatorial length and the length of the colpi. The shape category is, basically, determined by dividing the equatorial width into the polar length of the pollen grain (P/E) Erdtman (1945). Preparation for scanning electron microscopic studies were carried out by dusting the treated pollen onto specific Aluminium stubs coated with silver. The stubs were then coated with gold and observed and photographed in JEOL T-20 Scanning Electron Microscope. Descriptive terms were according to Moore and Webb (1978) and Punt et al. (2007). Size measurements for the pollen grains were taken according to Erdtman (1971) [very small < 10 μm in dimensions; small 10-25 μm; medium (25-50) μm; large (50-100) μm; very large (100-200) μm; huge > 200 μm].
RESULTS AND DISCUSSION
This study concentrated on shape, number of colpi, size and surface characters of pollen grains of 20 species. The main features of the pollen are illustrated in Figures 1-32 and summarized in Table 2. The study shows that the shape of pollen grains is prolate in one species, spheriodal in ten species and sub-spheroidal in one species under 6 zonocolpate group (Table 2). However, the shape of the pollen grains is spheriodal in seven species and sub-spheroidal in one species under 3 zonocolpate group (Table 2),
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which revealed no significant differnces in pollen shapes of studied species. The study shows that these pollen can be broadly divided into two groups, based on the number of colpi, viz.: 1) Hexa-zonocolpate group with 14 species (Table 2 ; Figs. 1-8 and Figs. 18-26). And 2) tri-zonocolpate group comprises 8 species (Table 2 and Figs.9-17). The heterocolpate character in the pollen of the investigated species coincides with the results obtained by earlier workers (Erdtman, 1971; Raj, 1974). Pollen grains show great variations in size among species. The largest size of pollen (56.63 x 55.81µm) was recorded in Ocimum basilicum, and the smallest one (18.32 x 11.66µm) in Leucas inflata.), this result agrees with that reported by Raj (1974) of pollen grains in Ocimum spp. belonging to the Indian region. These variations in size is perhaps due to the ecophysiological variations in these two countries. However, the size of pollen grains is slightly different in Ocimum species, Lavandula species and Leucas species of the present study (Table 2) this revealed that size variation may be used in generic level.The tectum surface ornamentation of the pollens which is one of the readily distinguishing characters, is found to be not constant among genera with more than one species studied (Table2). Fine reticulate is the common type of tectum surface ornamentation among the species studied, since, it occurs in 6 species of 5 genera, Ajuga arabica (Table2, Fig.12), Lallemantia royleana (Table2, Figs., 22 and 29), Plectranthus asirensis (Table2, Figs. 5 and 18) and Scutellaria arabica (Table2, Fig.17 ) Lavandula dentata and Lavandula pubescnes (Table2, Figs.3 and 8 , examined by LM) The results of the first three species agree with that mentioned by Kose, et al. (2011), Dinç et al, (2009) and Jamzad and Hasani-Nejad (2013). The surface ornamentation of the last two species is mentioned as reticulate under SEM (Evert. and Eichhor, 2013) which also confirmed with our findings in this tudy. However, these 6 species belong to two different groups, Ajuga arabica and Scutellaria arabica pollen with tri-zonocolpate, while Lallemantia royleana, Plectranthus asirensis, Lavandula dentata and Lavandula pubescnes with hexa-zonocolpate pollens. Pollen with megareticulate tectum surface occurs in 3 species of Ocimum (Ocimum basilicum, O. filamentosum and O. forsskaolii) (Table2, Figs.2,6,7,24 and 32) . Pollen surface with bireticulate perforate exine present in two species of two genera Ballota undulata and Endostemon gracile, each of them in different group, since, Ballota undulata pollen with tri-zonocolpate (Table 2, Figs.14 and 28 ) and Endostemon gracile hexa-
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zonocolpate (Table 2, Figs.1 and 21 ). Pollen with rough reticulate surface observed in Leucas alba (Table 2, Figs. 15 and 31) this result in general agree with the findings of Prabhakar and Ramakrishna (2014) in the pollen of Leucas indeca. Because of smallest size of Leucas inflata pollen, its exine surface not obvious under LM (Fig.11). Granulate pollen surface occurs in Teucrium oliverianum (Table 2, Figs. 16 and 30 ) which agree with that reported by Navarro et al, (2004). Pollen with reticulate-perfotate surface present in 6 species of 6 genera, Marrubium vulgare (Table 2, Fig. 26) which agree with that reported by Akgul, et al. (2008) and Stachys aegyptiaca (Table 2, Figs, 13 and 30) which match with that mentioned by Boi et al. (2013), both species with trizonocolpate pollens while Origanum syriacum pollen ( Table 2, Fig.19) which correspond with that exhibited by Yildiz et al, (2009). Mentha longifolia ( Table 2, Fig.23) which agree whith that stated by Celenk et al. (2008a), Micromeria imbricata ( Table 2, Fig.25) which in general agreement with those observed by Kremer, et al (2014) and Thymus decussatus ( Table 2, Fig. 20) which coincide with that reported by Satil, et al (2005), all these species have pollens with heixa-zonocolpate (Table 2). There are a few variations in the colpi lengths at least on generic level which depends on the pollen size of each species (Table 2). They are generally narrow with tapering ends. Broad colpi with acute ends are also observed in many species. (e.g. Origanum syriacum and Ocimum spp.). However, based on colpi numbers, the species studied can be divided into two groups viz.: Group1, pollen with 6-zonocolpate and Group 2, pollen with 3 zonocolpate (Table 2) and this agrees with the division of the Lamiaceae into two groups (Erdtman, 1945). As well as, according to type of tectum surface ornamentation: Group1 divides into 4 subgroups including 12 species in 10 genera and Group 2 divides into 5 subgroups which includes 8 species in 7genera (Table 3). The study of the available pollen material of the 20 species of the family Lamiaceae based on tectum ornamentation revealed the presence of 6 pollen types; including genera divided into 6 groups (Table 3). Moreover, a key based on overall morphological features of pollen grains for the studied species where can be classified into different categories is proposed.
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Key. Based on the morphological features of pollen grains of the species investigated: Group I. Pollen grains with 6-zonocolpate Subgroup a. Tectum surface fine reticulate 1 - Pollen shape spheriodal …………………………… Lavandula dentata …………………………….……………………….Lavandula pubescence ……………………………… ….…………………...Plectranthus asirensis 2- Pollen shape subspheriodal ……………………Lallemantia royleana Subgroup b.Tectum surface mega-retic.…………………. Ocimum basilicum …………………….………………………….. Ocimum filamentosum …………………………….…………………….. Ocimum forsskaolii
Subgroup c.Tectum surface reticulate - perforate 1 - Pollen shape prolate……………………………….Origanum syriacum 2 - Pollen shape spheriodal………………………. Marrubium vulgare ………………………………………………… Micromeria imbricata ……………………..…………………………… Mentha longifolia ……………………………………………………Thymus decussatus Subgroup d.Tectum surface bireticulate- perforate. …..Endostemon gracile Ggroup II. Pollen grains with 3-zonocolpate Subgroup a. Tectum surface fine reticulate Pollen shape spheriodal ……. ….……………………………..……………............ Ajuga arabica …..…………………………………………………….. Scutellaria arabica Subgroup b. Tectum surface rough reticulate pollen shape spheriodal ………. ……….. ….. ….. ……. ….. ….. ….. …… ….. ……….. Leucas alba ……………………………………………………………...Leucas inflata Subgroup c. Tectum surface reticulate- perforate, pollen shape spheriodal ………………………………………………... Stachys aegyptiaca Subgroup d. Tectum surface bireticulate- perforate pollen shape subspheriodal …………………………………………………… Ballota undulata Subgroup e. Tectum surface granulate Pollen shape spheriodal ......................................................................... Teucrium oiverianum
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ACKNOWLEDGEMENTS This project was supported by king saud university, deanship of scientific research, college of science researh center. The authors would like to thank the curators of the National Herbarium (RIY) and the Herbarium (KSU) in the Botany & Microbiology dept. of King Saud University for providing herbarium specimens. Thanks are also extended to the staff at the SEM, Structural Studies Research Service at the Reading University, U.K. for providing the facilities.
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Kahraman, A. Celep, F. and Dogan, M. (2009b). Comparative Morphology, Anatomy and Palynology of two Salvia L. species (Lamiaceae) and their taxonomic Implications. Bangladesh j. Plant Taxon, 16, 73 - 82. Kahraman, A. Celep, F. and Dogan, M. 2010a. Anatomy, trichome morphology and palynology of Salvia chrysophyllaStapf. (Lamiaceae). South African Journal of Botany, 76 187 -195. Kahraman, A.,Celep, F. and Dogan, M. 2010b. Morphology, Anatomy, Palynology and Nutlet Micromorphology of Salviamacrochlamys (Labiatae) in Turkey. Biologia. 65/2: 219-227. Kahraman, A., Doghan, M. 2010. Comparative study of Salvia limbata C.A. and S. palaestina Bentham (sect. Aethiopis Bentham, (Labiatae) from East Anatolia, Turkey. Acta Bot. Croat. 69(1):47-64. Kose, Y. B., Erkara, I. P. And Alan,S. 2011. Pollen Morphology of Some Turkish Ajuga L. (Lamiaceae) and Its Taxonomic Value. Bangladesh J. Bot. 40(1): 2933. Kremer. D, Dunkić, V., Stešević, D., Kosalec, I., Ballian, D., Bogunić, F., Bezić. Nada and Stabentheiner, E. 2014. Micromorphological traits and essential oil of Micromeria longipedunculata Bräuchler (Lamiaceae). Central European Journal of Biology, 9(5), 559-568. Migahid, A.M., 1989. Flora of Saudi Arabia. Vol. II. King Saud University Libraries, Riyadh. Moore, P. D, Webb, J. A 1978. An illustrated Guide Pollen Analysis. Hodder and Stoughton, London. Navarro, T., Oualidi, J. E., Trigo, M. D. M. 2004. Pollen morphology of Teucrium (Labiatae) and its taxonomic value. Belgian Journal of Botany Volume 137, Number 1, July pp. 70-84(15). Osman A.K. 2012. Comparative anatomical and palynological studies on genus Ballota (Lamiaceae) from Egypt. Journal of Medicinal Plants Research Vol. 6(47), pp. 5797-5812. Perveen, A., Qaiser, M. 2003. Pollen Morphology of the Family Lamiaceae from Pakistan. Pak. J. Bot. 35(5):671-693.
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Prabhakar, R. and Ramakrishna, H. 2014. Pollen diversity of Ethnomedicinal Plants of Nirmal Forest Division in Adilabad District, Telangana State, India. World Journal of Pharmaceutical ReseaRch. Volume 3, Issue 8, 220-237. Punt W., Hoen, P. P, Blackmore, S., Nilsson, S., Thomas, A. L. 2007 .Glossary of pollen and spore terminology. Rev. Palaeobot. Palynol. 143:1-81. Raj, B. 1974. Pollen morphology of Indian Labiatae, 1- Tribe Ocimoideae and Satureneae, Jour. of Palynology,Vol. 10. (2): 89-105. Rudall, P. 1980. Pollen morphology in the subtribe Hyptidinae (Labiatae), Kew Bull. 35(3): 453-458. Salmaki, y., Jamzad z., zarre s., Braüchler c.2008. pollen morphology of Stachys (Lamiaceae) in iran and its systematic implication. flora 203, 627-639. Satil, F., Kaya, A., Biçakci, A., Özatli, S. and Tümen, G. 2005. Comparative morphological, anatomical and palynological studies on Thymus migricus Klokov & Des.-Shost and T. Fedtschenkoi Ronniger Var. Handelii (Ronniger) Jalas grown in East Anatolia. Pak. J. Bot., 37(3): 531-549. Yildiz, K., Gücel, S. and Dadandi, M. Y. 2009. A palynological investigation of endemic taxa from Northern Cyprus. Pak. J. Bot., 41(3): 991-1007.
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Figure Captions Figures,1-8. LM micrographs showing hexacolpate pollen grain shapes in polar view. 1. Endostemon gracile pollen grain, spheroidal (x 800); 2. Ocimum filamentosum, spheroidal (x 800); 3. Lavandula dentata pollen grain, spheroidal (x800) 4. Thymus decussatus pollen grain, spheroidal (x 800); 5. Plectranthus asirensis pollen grain, spheroidal (x 800); 6. Ocimum forsskaolii pollen grain, spheroidal (x 800); 7. Ocimum basilicum pollen grain, spheroidal (x 800); 8. Lavandula pubescens , par = 50 µm.
Figures, 9-11. LM micrographs showing tricolpate pollen shapes in polar view. 9. Ajuga arabica , spheroidal (x 800); 10. Stachys aegyptiaca, spheroidal , (x 800); 11. Leucas inflata spheroidal (x 800).
Figures, 12-17. SEM micrographs showing tricolpate pollen grains, tectum surface; 12. Ajuga arabica, fine reticulate (x 2000); 13. Stachys aegyptiaca, reticulate perforate (x 3500);; 14. Ballota undulata, bireticulate-perforate (x 3500); 15. Leucas alba, rough reticulate (x 3500); 16. Teucrium oliverianum, granulate (x 2000). 17. Scutellaria arabica, fine reticulate (x 3500).
Figures, 18-26. SEM micrographs showing hexacolpate pollen grains, tectum surfaces. 18. Plectranthus asirensis pollen grain in equatorial view, fine reticulate (x 2000); 19. Origanum syriacum pollen grain in equatorial view, reticulateperforate (x 3500); 20. Thymus decussatus pollen grain in polar view, perforate (x 2000); 21. Endostemon gracile pollen grain in polar view, bireticulate perforate (x 2000); 22. Lallemantia royleana pollen grain in equatorial view, fine reticulate (x 2000); 23. Mentha longifolia pollen grain in polar to equatorial
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view, perforate (x 3500); 24. Ocimum forsskaolii pollen grain in polar view, mega- reticulate (x 2000). . 25. Micromeria imbricata pollen grain in polar view perforate (x 2000); 26. Marrubium vulgare pollen grain in polar view, reticulate - perforate (x 2000).
Figures 27-32. SEM micrographs enlarged from originals showing
examples of
terminology used to describe tectum surfaces ornamentations of pollen grains of studied species , 27. Reticulate-perforate,
Micromeria imbricata; 28.
Bireticulate – perforate, Ballota undulata; 29. Fine reticulate, Lallemantia royleana; 30. Granulate, Teucrium oliverianum; 31. Rough reticulate, Leucas alba; 32. Mega -reticulate, Ocimum forsskaolii.
12
13
14
15
16
17
Table (1): List of voucher specimens used in this studyand their locations and collectors. Species
Locality
Date
1
Ajuga arabica P. Davis
Hair, Riyadh
1981
2
Sameera, Hail Road Wadi Raida
8
Ballota undulata (Fres.) Benth. Endostemon tereticaulis (Poir.) Ash. Lallemantia royleana Benth. Lavandula dentata L. Lavandula pubescens Decne. Leucas alba (Vahl) R.Br. Leucas inflata Benth.
9
Marrubium vulgare L.
3 4 5 6 7
10 Mentha longifolia (L.) Huds. 11 Micromeria imbricata (Forssk.) C. Chr. 12 Ocimum basilicum L. 13 Ocimum filamentosum Forssk. 14 Ocimum forsskaoleii Benth. 15 Origanum syriacum L. 16 Plectranthus asirensis Wood 17 Scutellaria arabica Jaub. & Spach. 18 Stachys aegyptiaca Pers. 19 Teucrium oliverianum Ging. ex Benth. 20 Thymus decussates Benth.
Jabal Eja, Hail Abha Abha
Collector
S. Chaudhary 1985 S. Collenette 1985 S. Collenette 1986 S. Collenette 1402H. A. Doaigey 1402H. A. Doaigey
RIY- 9947 RIY- 9975 RIY - 10314 906 (KSU) 905 (KSU)
Dharb-Abha Road Bisha, Baha Road Abha
1982
1403H. A. Doaigey
1004(KSU)
Taif
1970
RIY-13170
Abha
1403H. A. Doaigey
1015(KSU)
Riyadh Jabal Fayfa
1205(KSU) RIY-8665
Alkharj Road Riyadh
1404H. A. Doaigey 1983 S. Chaudhary 1981 S. Chaudhary 1980 Abulfatih 1989 S. Chaudhary 1982 S. Chaudhary 1404H. A. Doaigey 1401H. A. Doaigey
East of Sakaka
1981
RIY- E877
Jabal Fayfa Abha Jabal Fayfa Jabal Soodah
18
1978
S. Chaudhary Megahid
Herbarium / jar No. RIY-1041
De Marco
S. Chaudhary
RIY-3923 KSU-14639
RIY-3817 RIY-7923 RIY-5203 RIY-3937 1207(KSU) 41 (KSU)
Table (2): Chief characteristics of the Lamiaceae pollen examined. Name of the Species 1
Ajuga arabica
2
Ballota undulata
3
7
Endostemon gracile Lallemantia royleana Lavandula dentata** Lavandula pubescnes** Leucas alba
8
Leucas inflata**
9
Marrubium vulgare Mentha longifolia
4 5 6
10 11 12 13 14 15 16 17 18 19 20
Micromeria imbricata Ocimum basilicum** Ocimum filamentosum** Ocimum forsskaolii Origanum syriacum Plectranthus asirensis Scutellaria arabica Stachys aegyptiaca Teucrium oliverianum Thymus decussatus
Size of pollen Mean
Colpi Length
Shape*
Tectum Surface Fine reticulate
P
E
P/E
40.39
37.05
1.09
26.23
Spheroidal
25.40
24.57
1.33
19.99
Subspheroidal
34.14
32.99
1,03
12.49
Spheroidal
31.64
27.07
1.16
14.99
Subspheroidal
37.09
34.15
1.08
24.15
Spheroidal
39.56
38.32
1.03
23.74
Spheroidal
19.15
18.74
1.02
13.33
Spheroidal
18.32
11.66
0,97
14.58
Spheroidal
28.32
27.09
1.04
18.74
Spheroidal
28.74
28.32
1.01
20.41
Spheroidal
34.57
31.33
1,10
23.32
Spheroidal
56.63
55.81
1.01
38.73
Spheroidal
44.97
43.31
1.03
33.73
Spheroidal
54.15
49.98
1.08
28.32
23.74
17.19
1.38
47.06
43.73
22.91
Type of Colpi
BireticulatePerforate BireticulatePerforate
3zonocolpate 3zonocolpate 6zonocolpate
Fine reticulate
6-zonocolpat
Fine reticulate (LM) Fine reticulate(LM)
Not examine (SEM) Reticulateperforate Reticulate perforate Reticulateperforate Mega-reticulate (LM) Mega- reticulate (LM)
6zonocolpate 6zonocolpate 3zonocolpate 3zonocolpate 3zonocolpate 6zonocolpate 6zonocolpate 6zonocolpate 6zonocolpate
Spheroidal
Mega- reticulate
6-zonocolpat
14.99
Prolate
Reticulate perforate
1.07
34.57
Spheroidal
Fine reticulate
21.24
1.07
15.82
Spheroidal
Fine reticulate
22.91
20.41
1.12
18.74
Spheroidal
Reticulate perforate
41.64
39.98
1.04
26.65
Spheroidal
granulate
27.09
27.07
1.00
19.16
Spheroidal
Reticulate perforate
6zonocolpate 6zonocolpate 3zonocolpate 3zonocolpate 3zonocolpate 6zonocolpate
Rough reticulate
*P/E ratio: Prolate =1.33- 2.00; Prolate- Spheroidal =1.00–1. 14; Spheroidal = 0.88-1.14; sub-spheroidal = 0.751.33 (Punt et al., 2007). E, equatorial axis; P, polar axis. All measurements in µm. **Pollen morphology of these species were only examined by light microscope (LM).
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Table (3): Pollen types of the 20 species and their genera of the family Lamiaceae based on their tectum ornamentation. Pollen types
Type I
Tectum ornamentation
Genera
fine reticulate
Ajuga Scutellaria Lallemantia Lavandula Plectranthus Stachys Origanum Marrubium Mentha Micromeria Thymus Leucas
Type II
fine reticulate-perforate
Type III
rough reticulate
Type IV
mega -reticulate
Ocimum
Type V
bireticulate – perforate
Type VI
granulate
Ballota Endostemon Teucrium
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Species Ajuga arabica Lallemantia royleana Scutellaria arabica Lavandula dentata Lavandula pubescnes Plectranthus asirensis Stachys aegyptiaca Origanum syriacum Marrubium vulgare Mentha longifolia Micromeria imbricata Thymus decussatus Leucas alba Leucas inflata Ocimum basilicum Ocimum filamentosum Ocimum forsskaolii Ballota undulata Endostemon gracile Teucrium oliverianum
S1319-562X(17)30084-0 http://dx.doi.org/10.1016/j.sjbs.2017.03.001 SJBS 918
To appear in:
Saudi Journal of Biological Sciences
Received Date: Revised Date: Accepted Date:
11 May 2016 23 February 2017 1 March 2017
Please cite this article as: A.R. Doaigey, M. El-Zaidy, A. Alfarhan, A.E-S. Milagy, T. Jacob, Pollen Morphology of certain species of the Family Lamiaceae in Saudi Arabia, Saudi Journal of Biological Sciences (2017), doi: http:// dx.doi.org/10.1016/j.sjbs.2017.03.001
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Pollen Morphology of certain species of the Family Lamiaceae in Saudi Arabia
Abdullah R. Doaigey Botany & Microbiology Dept., College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia. Tell. :0096614675879 e-mail: [email protected] Mohamed El-Zaidy Botany & Microbiology Dept., College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia. Tell. : 0096614675878 e-mail: [email protected] Ahmad Alfarhan Botany & Microbiology Dept., College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia. Tell. :0096614675808 e-mail: [email protected] Abd El-Salam Milagy Botany & Microbiology Dept., College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia. Tell. :0096614675878 e-mail: [email protected] Jacob Thomas Botany & Microbiology Dept., College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia. Tell.: 0096614675807 e-mail: [email protected]
Pollen Morphology of certain species of the Family Lamiaceae in Saudi Arabia Abdullah R. Doaigey ⃰, Mohamed El-Zaidy, Ahmad Alfarhan, Abd El-Salam Milagy and Thomas Jacob Botany & Microbiology Dept., College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia.
⃰ E-mail: [email protected].
ABSTRACT The intention of the present wok is to provide an account on the pollen morphological features of Lamiaceae in Saudi Arabia as a basis for future studies of Lamiaceae pollens in the region. Pollen morphology of 20 species belong to 16 genera of the Lamiaceae has been investigated using Light Microscope (LM) and Scanning Electron Microscope (SEM). The study revealed that the pollen grains were characterized by 3-zonocolpate or 6-zonocolpate. Size of the pollen is variable between the genera but not among the species of the same genus. The surface pattern of the exine varies from fine reticulate, rough reticulate, mega-reticulate, reticulateperforate, bireticulate- perforate or granulate, leading to 6 types of pollen grains. These variations revealed by this study implies that pollen Morphology may be of significant value sharing in solving problems in the classification of Lamiaceae members. A Key to the species, based on the morphological features of pollen grains, is also provided.
KEY WORDS: Colpi, Lamiaceae, Pollen grains, Pollen Morphology, Saudi Arabia.
INTRODUCTION Lamiaceae is represented in Saudi Arabia by 26 genera and about 70 species (Migahid, 1989 and Collenette, 1999). 14 species are endemic to the Kingdom of Saudi Arabia, while 27 species are endangered, vulnerable or rare (Collenette, 1998). Pollen morphology including aperture number, shape and tectum ornamentation were studied in different members of the family Lamiaceae and found these features were helpful in
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its taxonomy (Erdtman, 1945; Abu-Asab and Cantino 1994; Raj, 1974; Rudall, P. 1980; Bazarragchaa, et al., 2012). However, Perveen and Qaiser (2003) and Al-Watban, et al. (2015) were studied pollen morphology of several species in different genera of the Family Lamiaceae and suggested that it is useful in grouping and identifying the species. The genus Salvia was studied by several investigators (Jafari and Nikian, 2008; Yildiz et al, 2009; Kahraman et al. 2009 a, b, 2010 a, b); Kahraman and Doghan, 2010) who reported the pollen characters of species were useful for their identification. Celenk, et al. (2008 a, b), indicated that the pollen characters of the genus Mentha were valuable for taxonomic applications. Erkara and Koyuncu (2007), Salmaki, et al. (2008) and Boi et al. (2013) indicated in their study on Stachys species that the pollen morphological characters were useful in the identification of certain sections and species. Akgul, et al. (2008) reported in their study of the genus Marrubium in Turky that morphological characters of pollens are of taxonomic significance in the genus. Harley, et al. (1992) found in their studies that most of Ocimum species pollens are with bireticulate- perforate tectum sculpture and suggested a re-evaluation in their classification. As well as Arogundade and Adedeji (2009) in their study of Ocimum L. species concentrated on pollen colpi reporting different types of pollens acolpate to octacolpate pollens which reveal that there are significant differences among the species. Moreover, Dinç et al, (2009) reported in their study on Lallemantia species growing in Turkey, the exine sculpture of pollen had taxonomic value. Kose, et al. (2011) reported that the granulate or reticulate type of exine ornamentation in Ajuga species may be is useful in segregating the species within the genus. Hasani-Nejad et al. (2011) and Jamzad and Hasani-Nejad (2013) showed that the exine ornamentation could be used as a taxonomic feature below generic level classification of Scutellaria. Osman (2012) in his study of Ballota species in Egypt mentioned that pollen characters were very useful in the separation of the species. Pollen grains of Leucas indica (L) R.Br.Ex Vatke., were investigated by Prabhakar and Ramakrishna (2014) and suggested that pollen shape, aperture and tectum sculpture may help in identification of the species. Navarro, et al. (2004) mentioned in Teucrium, pollen tectum surface ornamentation types and pollen size seem to have a taxonomic value. The forgoing investigations indicate that there is no definitive agreement for which level of classification can be useful of using pollen morphological characters in Lamiaceae. The
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intention of the present work is to provide an account of pollen morphological features of Lamiaceae members in Saudi Arabia as a basis for future studies.
MATERIALS AND METHODS Pollens were excised from either fresh and preserved
samples or from
herbarium specimens deposited either at the herbarium of King Saud University (KSU) or at the National Herbarium (RIY) of Saudi Arabia. (Table,1). Five samples of defferent collections were prepared for each species to confirm the stability of pollen characters within a species, by using the usual acetolysis method (Erdman, 1971). Measurements were made within two to four days period to avoid the possiblility of error introduced by differential swelling. Twenty pollen grains were measured using an ocular micrometer (Zeiss Photometric microscope III) and the mean was taken. Measurements include polar length, equatorial length and the length of the colpi. The shape category is, basically, determined by dividing the equatorial width into the polar length of the pollen grain (P/E) Erdtman (1945). Preparation for scanning electron microscopic studies were carried out by dusting the treated pollen onto specific Aluminium stubs coated with silver. The stubs were then coated with gold and observed and photographed in JEOL T-20 Scanning Electron Microscope. Descriptive terms were according to Moore and Webb (1978) and Punt et al. (2007). Size measurements for the pollen grains were taken according to Erdtman (1971) [very small < 10 μm in dimensions; small 10-25 μm; medium (25-50) μm; large (50-100) μm; very large (100-200) μm; huge > 200 μm].
RESULTS AND DISCUSSION
This study concentrated on shape, number of colpi, size and surface characters of pollen grains of 20 species. The main features of the pollen are illustrated in Figures 1-32 and summarized in Table 2. The study shows that the shape of pollen grains is prolate in one species, spheriodal in ten species and sub-spheroidal in one species under 6 zonocolpate group (Table 2). However, the shape of the pollen grains is spheriodal in seven species and sub-spheroidal in one species under 3 zonocolpate group (Table 2),
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which revealed no significant differnces in pollen shapes of studied species. The study shows that these pollen can be broadly divided into two groups, based on the number of colpi, viz.: 1) Hexa-zonocolpate group with 14 species (Table 2 ; Figs. 1-8 and Figs. 18-26). And 2) tri-zonocolpate group comprises 8 species (Table 2 and Figs.9-17). The heterocolpate character in the pollen of the investigated species coincides with the results obtained by earlier workers (Erdtman, 1971; Raj, 1974). Pollen grains show great variations in size among species. The largest size of pollen (56.63 x 55.81µm) was recorded in Ocimum basilicum, and the smallest one (18.32 x 11.66µm) in Leucas inflata.), this result agrees with that reported by Raj (1974) of pollen grains in Ocimum spp. belonging to the Indian region. These variations in size is perhaps due to the ecophysiological variations in these two countries. However, the size of pollen grains is slightly different in Ocimum species, Lavandula species and Leucas species of the present study (Table 2) this revealed that size variation may be used in generic level.The tectum surface ornamentation of the pollens which is one of the readily distinguishing characters, is found to be not constant among genera with more than one species studied (Table2). Fine reticulate is the common type of tectum surface ornamentation among the species studied, since, it occurs in 6 species of 5 genera, Ajuga arabica (Table2, Fig.12), Lallemantia royleana (Table2, Figs., 22 and 29), Plectranthus asirensis (Table2, Figs. 5 and 18) and Scutellaria arabica (Table2, Fig.17 ) Lavandula dentata and Lavandula pubescnes (Table2, Figs.3 and 8 , examined by LM) The results of the first three species agree with that mentioned by Kose, et al. (2011), Dinç et al, (2009) and Jamzad and Hasani-Nejad (2013). The surface ornamentation of the last two species is mentioned as reticulate under SEM (Evert. and Eichhor, 2013) which also confirmed with our findings in this tudy. However, these 6 species belong to two different groups, Ajuga arabica and Scutellaria arabica pollen with tri-zonocolpate, while Lallemantia royleana, Plectranthus asirensis, Lavandula dentata and Lavandula pubescnes with hexa-zonocolpate pollens. Pollen with megareticulate tectum surface occurs in 3 species of Ocimum (Ocimum basilicum, O. filamentosum and O. forsskaolii) (Table2, Figs.2,6,7,24 and 32) . Pollen surface with bireticulate perforate exine present in two species of two genera Ballota undulata and Endostemon gracile, each of them in different group, since, Ballota undulata pollen with tri-zonocolpate (Table 2, Figs.14 and 28 ) and Endostemon gracile hexa-
4
zonocolpate (Table 2, Figs.1 and 21 ). Pollen with rough reticulate surface observed in Leucas alba (Table 2, Figs. 15 and 31) this result in general agree with the findings of Prabhakar and Ramakrishna (2014) in the pollen of Leucas indeca. Because of smallest size of Leucas inflata pollen, its exine surface not obvious under LM (Fig.11). Granulate pollen surface occurs in Teucrium oliverianum (Table 2, Figs. 16 and 30 ) which agree with that reported by Navarro et al, (2004). Pollen with reticulate-perfotate surface present in 6 species of 6 genera, Marrubium vulgare (Table 2, Fig. 26) which agree with that reported by Akgul, et al. (2008) and Stachys aegyptiaca (Table 2, Figs, 13 and 30) which match with that mentioned by Boi et al. (2013), both species with trizonocolpate pollens while Origanum syriacum pollen ( Table 2, Fig.19) which correspond with that exhibited by Yildiz et al, (2009). Mentha longifolia ( Table 2, Fig.23) which agree whith that stated by Celenk et al. (2008a), Micromeria imbricata ( Table 2, Fig.25) which in general agreement with those observed by Kremer, et al (2014) and Thymus decussatus ( Table 2, Fig. 20) which coincide with that reported by Satil, et al (2005), all these species have pollens with heixa-zonocolpate (Table 2). There are a few variations in the colpi lengths at least on generic level which depends on the pollen size of each species (Table 2). They are generally narrow with tapering ends. Broad colpi with acute ends are also observed in many species. (e.g. Origanum syriacum and Ocimum spp.). However, based on colpi numbers, the species studied can be divided into two groups viz.: Group1, pollen with 6-zonocolpate and Group 2, pollen with 3 zonocolpate (Table 2) and this agrees with the division of the Lamiaceae into two groups (Erdtman, 1945). As well as, according to type of tectum surface ornamentation: Group1 divides into 4 subgroups including 12 species in 10 genera and Group 2 divides into 5 subgroups which includes 8 species in 7genera (Table 3). The study of the available pollen material of the 20 species of the family Lamiaceae based on tectum ornamentation revealed the presence of 6 pollen types; including genera divided into 6 groups (Table 3). Moreover, a key based on overall morphological features of pollen grains for the studied species where can be classified into different categories is proposed.
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Key. Based on the morphological features of pollen grains of the species investigated: Group I. Pollen grains with 6-zonocolpate Subgroup a. Tectum surface fine reticulate 1 - Pollen shape spheriodal …………………………… Lavandula dentata …………………………….……………………….Lavandula pubescence ……………………………… ….…………………...Plectranthus asirensis 2- Pollen shape subspheriodal ……………………Lallemantia royleana Subgroup b.Tectum surface mega-retic.…………………. Ocimum basilicum …………………….………………………….. Ocimum filamentosum …………………………….…………………….. Ocimum forsskaolii
Subgroup c.Tectum surface reticulate - perforate 1 - Pollen shape prolate……………………………….Origanum syriacum 2 - Pollen shape spheriodal………………………. Marrubium vulgare ………………………………………………… Micromeria imbricata ……………………..…………………………… Mentha longifolia ……………………………………………………Thymus decussatus Subgroup d.Tectum surface bireticulate- perforate. …..Endostemon gracile Ggroup II. Pollen grains with 3-zonocolpate Subgroup a. Tectum surface fine reticulate Pollen shape spheriodal ……. ….……………………………..……………............ Ajuga arabica …..…………………………………………………….. Scutellaria arabica Subgroup b. Tectum surface rough reticulate pollen shape spheriodal ………. ……….. ….. ….. ……. ….. ….. ….. …… ….. ……….. Leucas alba ……………………………………………………………...Leucas inflata Subgroup c. Tectum surface reticulate- perforate, pollen shape spheriodal ………………………………………………... Stachys aegyptiaca Subgroup d. Tectum surface bireticulate- perforate pollen shape subspheriodal …………………………………………………… Ballota undulata Subgroup e. Tectum surface granulate Pollen shape spheriodal ......................................................................... Teucrium oiverianum
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ACKNOWLEDGEMENTS This project was supported by king saud university, deanship of scientific research, college of science researh center. The authors would like to thank the curators of the National Herbarium (RIY) and the Herbarium (KSU) in the Botany & Microbiology dept. of King Saud University for providing herbarium specimens. Thanks are also extended to the staff at the SEM, Structural Studies Research Service at the Reading University, U.K. for providing the facilities.
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Collenette, S., 1998. A checklist of botanical species in Saudi Arabia, International Asclepiad Soc., West Sussex. Collenette, S., 1999. Wild flowers of Saudi Arabia, National Commission for wildlife conservation and development, Riyadh. Dinç, M., Pinar, N.M., Dogu, S., and Yildirimli, S. 2009. Micromorphological Studies of Lallemantia L. (Lamiaceae) Species Growing in Turkey. Acta Biologica Cracoviensia Series Botanica 51/1: 45–54. Erdtman, G., 1945. Pollen morphology and plant taxonomy, 4, Labiatae. Verbanaceae and Aviceniaceae, Svensk, Bot. Tidskr. 39: 279-285. Erdtman, G. 1971. Pollen morphology and plant Taxonomy of Angiosperms. Angiosperms. Hafner Publishing Co., New York. Erkara, İ. P. and Koyuncu . O. 2007, A Study of the Anatomy and Pollen Morphology of Two Economically Important Species of Stachys L. (Lamiaceae) in Turkey. Journal of Applied Biological Sciences 1 (3): 49-56. Evert. R. F. and Eichhor S. E. 2013. Raven Biology of Plants. Eighth Edition p.467. W. H. Freeman and Company Publishers. Harley, M. M., Paton, A., Harley, R. M. and Cade, P. G. 1992. Pollen marphological studies in tribe Ocimeae (Nepetoideae: Labiatae): I. Ocimum L. Grana, 31: 161-176. Hasani- Nejad, M., Jamzad Z. and Uosofi, M. 2011. A palynological study of Scutellaria L. (Lamiaceae) in Iran.TBJ, 3(7): 33-44. Jafari, A. and Nikian, M. 2008. Micromorphological, anatomical and pollen ornamentation study of four desert species of Salvia in Central Iran. Asian Journal of Plant Sciences. Vol. 7(8), 736–741. Jamzad, Z.and Hasani-Nejad, M. 2013.Taxonomic implications of pollen exine morphology in infrageneric classification of Scutellaria (Lamiaceae). Nordic Journal of Botany.Volume 32, (2), 233–244, Kahraman, A. Celep, F. and Dogan, M. 2009a. Anatomy, trichome morphology and palynology of Salvia chrysophylla Stapf. (Lamiaceae). South African Journal of Botany, 76 187 -195.
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Kahraman, A. Celep, F. and Dogan, M. (2009b). Comparative Morphology, Anatomy and Palynology of two Salvia L. species (Lamiaceae) and their taxonomic Implications. Bangladesh j. Plant Taxon, 16, 73 - 82. Kahraman, A. Celep, F. and Dogan, M. 2010a. Anatomy, trichome morphology and palynology of Salvia chrysophyllaStapf. (Lamiaceae). South African Journal of Botany, 76 187 -195. Kahraman, A.,Celep, F. and Dogan, M. 2010b. Morphology, Anatomy, Palynology and Nutlet Micromorphology of Salviamacrochlamys (Labiatae) in Turkey. Biologia. 65/2: 219-227. Kahraman, A., Doghan, M. 2010. Comparative study of Salvia limbata C.A. and S. palaestina Bentham (sect. Aethiopis Bentham, (Labiatae) from East Anatolia, Turkey. Acta Bot. Croat. 69(1):47-64. Kose, Y. B., Erkara, I. P. And Alan,S. 2011. Pollen Morphology of Some Turkish Ajuga L. (Lamiaceae) and Its Taxonomic Value. Bangladesh J. Bot. 40(1): 2933. Kremer. D, Dunkić, V., Stešević, D., Kosalec, I., Ballian, D., Bogunić, F., Bezić. Nada and Stabentheiner, E. 2014. Micromorphological traits and essential oil of Micromeria longipedunculata Bräuchler (Lamiaceae). Central European Journal of Biology, 9(5), 559-568. Migahid, A.M., 1989. Flora of Saudi Arabia. Vol. II. King Saud University Libraries, Riyadh. Moore, P. D, Webb, J. A 1978. An illustrated Guide Pollen Analysis. Hodder and Stoughton, London. Navarro, T., Oualidi, J. E., Trigo, M. D. M. 2004. Pollen morphology of Teucrium (Labiatae) and its taxonomic value. Belgian Journal of Botany Volume 137, Number 1, July pp. 70-84(15). Osman A.K. 2012. Comparative anatomical and palynological studies on genus Ballota (Lamiaceae) from Egypt. Journal of Medicinal Plants Research Vol. 6(47), pp. 5797-5812. Perveen, A., Qaiser, M. 2003. Pollen Morphology of the Family Lamiaceae from Pakistan. Pak. J. Bot. 35(5):671-693.
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Prabhakar, R. and Ramakrishna, H. 2014. Pollen diversity of Ethnomedicinal Plants of Nirmal Forest Division in Adilabad District, Telangana State, India. World Journal of Pharmaceutical ReseaRch. Volume 3, Issue 8, 220-237. Punt W., Hoen, P. P, Blackmore, S., Nilsson, S., Thomas, A. L. 2007 .Glossary of pollen and spore terminology. Rev. Palaeobot. Palynol. 143:1-81. Raj, B. 1974. Pollen morphology of Indian Labiatae, 1- Tribe Ocimoideae and Satureneae, Jour. of Palynology,Vol. 10. (2): 89-105. Rudall, P. 1980. Pollen morphology in the subtribe Hyptidinae (Labiatae), Kew Bull. 35(3): 453-458. Salmaki, y., Jamzad z., zarre s., Braüchler c.2008. pollen morphology of Stachys (Lamiaceae) in iran and its systematic implication. flora 203, 627-639. Satil, F., Kaya, A., Biçakci, A., Özatli, S. and Tümen, G. 2005. Comparative morphological, anatomical and palynological studies on Thymus migricus Klokov & Des.-Shost and T. Fedtschenkoi Ronniger Var. Handelii (Ronniger) Jalas grown in East Anatolia. Pak. J. Bot., 37(3): 531-549. Yildiz, K., Gücel, S. and Dadandi, M. Y. 2009. A palynological investigation of endemic taxa from Northern Cyprus. Pak. J. Bot., 41(3): 991-1007.
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Figure Captions Figures,1-8. LM micrographs showing hexacolpate pollen grain shapes in polar view. 1. Endostemon gracile pollen grain, spheroidal (x 800); 2. Ocimum filamentosum, spheroidal (x 800); 3. Lavandula dentata pollen grain, spheroidal (x800) 4. Thymus decussatus pollen grain, spheroidal (x 800); 5. Plectranthus asirensis pollen grain, spheroidal (x 800); 6. Ocimum forsskaolii pollen grain, spheroidal (x 800); 7. Ocimum basilicum pollen grain, spheroidal (x 800); 8. Lavandula pubescens , par = 50 µm.
Figures, 9-11. LM micrographs showing tricolpate pollen shapes in polar view. 9. Ajuga arabica , spheroidal (x 800); 10. Stachys aegyptiaca, spheroidal , (x 800); 11. Leucas inflata spheroidal (x 800).
Figures, 12-17. SEM micrographs showing tricolpate pollen grains, tectum surface; 12. Ajuga arabica, fine reticulate (x 2000); 13. Stachys aegyptiaca, reticulate perforate (x 3500);; 14. Ballota undulata, bireticulate-perforate (x 3500); 15. Leucas alba, rough reticulate (x 3500); 16. Teucrium oliverianum, granulate (x 2000). 17. Scutellaria arabica, fine reticulate (x 3500).
Figures, 18-26. SEM micrographs showing hexacolpate pollen grains, tectum surfaces. 18. Plectranthus asirensis pollen grain in equatorial view, fine reticulate (x 2000); 19. Origanum syriacum pollen grain in equatorial view, reticulateperforate (x 3500); 20. Thymus decussatus pollen grain in polar view, perforate (x 2000); 21. Endostemon gracile pollen grain in polar view, bireticulate perforate (x 2000); 22. Lallemantia royleana pollen grain in equatorial view, fine reticulate (x 2000); 23. Mentha longifolia pollen grain in polar to equatorial
11
view, perforate (x 3500); 24. Ocimum forsskaolii pollen grain in polar view, mega- reticulate (x 2000). . 25. Micromeria imbricata pollen grain in polar view perforate (x 2000); 26. Marrubium vulgare pollen grain in polar view, reticulate - perforate (x 2000).
Figures 27-32. SEM micrographs enlarged from originals showing
examples of
terminology used to describe tectum surfaces ornamentations of pollen grains of studied species , 27. Reticulate-perforate,
Micromeria imbricata; 28.
Bireticulate – perforate, Ballota undulata; 29. Fine reticulate, Lallemantia royleana; 30. Granulate, Teucrium oliverianum; 31. Rough reticulate, Leucas alba; 32. Mega -reticulate, Ocimum forsskaolii.
12
13
14
15
16
17
Table (1): List of voucher specimens used in this studyand their locations and collectors. Species
Locality
Date
1
Ajuga arabica P. Davis
Hair, Riyadh
1981
2
Sameera, Hail Road Wadi Raida
8
Ballota undulata (Fres.) Benth. Endostemon tereticaulis (Poir.) Ash. Lallemantia royleana Benth. Lavandula dentata L. Lavandula pubescens Decne. Leucas alba (Vahl) R.Br. Leucas inflata Benth.
9
Marrubium vulgare L.
3 4 5 6 7
10 Mentha longifolia (L.) Huds. 11 Micromeria imbricata (Forssk.) C. Chr. 12 Ocimum basilicum L. 13 Ocimum filamentosum Forssk. 14 Ocimum forsskaoleii Benth. 15 Origanum syriacum L. 16 Plectranthus asirensis Wood 17 Scutellaria arabica Jaub. & Spach. 18 Stachys aegyptiaca Pers. 19 Teucrium oliverianum Ging. ex Benth. 20 Thymus decussates Benth.
Jabal Eja, Hail Abha Abha
Collector
S. Chaudhary 1985 S. Collenette 1985 S. Collenette 1986 S. Collenette 1402H. A. Doaigey 1402H. A. Doaigey
RIY- 9947 RIY- 9975 RIY - 10314 906 (KSU) 905 (KSU)
Dharb-Abha Road Bisha, Baha Road Abha
1982
1403H. A. Doaigey
1004(KSU)
Taif
1970
RIY-13170
Abha
1403H. A. Doaigey
1015(KSU)
Riyadh Jabal Fayfa
1205(KSU) RIY-8665
Alkharj Road Riyadh
1404H. A. Doaigey 1983 S. Chaudhary 1981 S. Chaudhary 1980 Abulfatih 1989 S. Chaudhary 1982 S. Chaudhary 1404H. A. Doaigey 1401H. A. Doaigey
East of Sakaka
1981
RIY- E877
Jabal Fayfa Abha Jabal Fayfa Jabal Soodah
18
1978
S. Chaudhary Megahid
Herbarium / jar No. RIY-1041
De Marco
S. Chaudhary
RIY-3923 KSU-14639
RIY-3817 RIY-7923 RIY-5203 RIY-3937 1207(KSU) 41 (KSU)
Table (2): Chief characteristics of the Lamiaceae pollen examined. Name of the Species 1
Ajuga arabica
2
Ballota undulata
3
7
Endostemon gracile Lallemantia royleana Lavandula dentata** Lavandula pubescnes** Leucas alba
8
Leucas inflata**
9
Marrubium vulgare Mentha longifolia
4 5 6
10 11 12 13 14 15 16 17 18 19 20
Micromeria imbricata Ocimum basilicum** Ocimum filamentosum** Ocimum forsskaolii Origanum syriacum Plectranthus asirensis Scutellaria arabica Stachys aegyptiaca Teucrium oliverianum Thymus decussatus
Size of pollen Mean
Colpi Length
Shape*
Tectum Surface Fine reticulate
P
E
P/E
40.39
37.05
1.09
26.23
Spheroidal
25.40
24.57
1.33
19.99
Subspheroidal
34.14
32.99
1,03
12.49
Spheroidal
31.64
27.07
1.16
14.99
Subspheroidal
37.09
34.15
1.08
24.15
Spheroidal
39.56
38.32
1.03
23.74
Spheroidal
19.15
18.74
1.02
13.33
Spheroidal
18.32
11.66
0,97
14.58
Spheroidal
28.32
27.09
1.04
18.74
Spheroidal
28.74
28.32
1.01
20.41
Spheroidal
34.57
31.33
1,10
23.32
Spheroidal
56.63
55.81
1.01
38.73
Spheroidal
44.97
43.31
1.03
33.73
Spheroidal
54.15
49.98
1.08
28.32
23.74
17.19
1.38
47.06
43.73
22.91
Type of Colpi
BireticulatePerforate BireticulatePerforate
3zonocolpate 3zonocolpate 6zonocolpate
Fine reticulate
6-zonocolpat
Fine reticulate (LM) Fine reticulate(LM)
Not examine (SEM) Reticulateperforate Reticulate perforate Reticulateperforate Mega-reticulate (LM) Mega- reticulate (LM)
6zonocolpate 6zonocolpate 3zonocolpate 3zonocolpate 3zonocolpate 6zonocolpate 6zonocolpate 6zonocolpate 6zonocolpate
Spheroidal
Mega- reticulate
6-zonocolpat
14.99
Prolate
Reticulate perforate
1.07
34.57
Spheroidal
Fine reticulate
21.24
1.07
15.82
Spheroidal
Fine reticulate
22.91
20.41
1.12
18.74
Spheroidal
Reticulate perforate
41.64
39.98
1.04
26.65
Spheroidal
granulate
27.09
27.07
1.00
19.16
Spheroidal
Reticulate perforate
6zonocolpate 6zonocolpate 3zonocolpate 3zonocolpate 3zonocolpate 6zonocolpate
Rough reticulate
*P/E ratio: Prolate =1.33- 2.00; Prolate- Spheroidal =1.00–1. 14; Spheroidal = 0.88-1.14; sub-spheroidal = 0.751.33 (Punt et al., 2007). E, equatorial axis; P, polar axis. All measurements in µm. **Pollen morphology of these species were only examined by light microscope (LM).
19
Table (3): Pollen types of the 20 species and their genera of the family Lamiaceae based on their tectum ornamentation. Pollen types
Type I
Tectum ornamentation
Genera
fine reticulate
Ajuga Scutellaria Lallemantia Lavandula Plectranthus Stachys Origanum Marrubium Mentha Micromeria Thymus Leucas
Type II
fine reticulate-perforate
Type III
rough reticulate
Type IV
mega -reticulate
Ocimum
Type V
bireticulate – perforate
Type VI
granulate
Ballota Endostemon Teucrium
20
Species Ajuga arabica Lallemantia royleana Scutellaria arabica Lavandula dentata Lavandula pubescnes Plectranthus asirensis Stachys aegyptiaca Origanum syriacum Marrubium vulgare Mentha longifolia Micromeria imbricata Thymus decussatus Leucas alba Leucas inflata Ocimum basilicum Ocimum filamentosum Ocimum forsskaolii Ballota undulata Endostemon gracile Teucrium oliverianum