Pollen morphological variation of berberis l. from pakistan and its systematic importance
1 like128 views
This research article examines pollen morphological variation in 10 Berberis species from Pakistan using light and scanning electron microscopy. A variety of qualitative and quantitative pollen characteristics were measured, including shape, size, presence/absence of colpi, colpi length and width, exine thickness, ornamentation, aperture type, and polar-equatorial ratio. Results showed variation in all characteristics studied. Five species had pantocolpate pollen while five had spiroaperturate pollen. Pollen size ranged from 29-35 μm to 60-65 μm. Ornamentation included psilate-regulate and foveolate-fossulate. This pollen morphological data provides useful information for taxonomic identification of
![FIGURE 3 Comparative number of
colpi in different pollen species [Color
figure can be viewed at
wileyonlinelibrary.com]
FIGURE 4 Comparative
length and width of colpi in
different pollen species [Color
figure can be viewed at
wileyonlinelibrary.com]
FIGURE 5 Comparative
polar–equatorial length in
different pollen species [Color
figure can be viewed at
wileyonlinelibrary.com]
UR RAHMAN ET AL. 1597](https://image.slidesharecdn.com/pollenmorphologicalvariationofberberisl-200821075505/85/Pollen-morphological-variation-of-berberis-l-from-pakistan-and-its-systematic-importance-5-320.jpg)
![4 | DISCUSSION
In this study, it was intended to assess Berberis pollen flora from
Pakistan using LM and SEM for its taxonomic importance in Berberis
species identification. Using different observed palyno-morphological
features, a taxonomic key was proposed for fast and correct identifi-
cation of collected Pakistani Berberis species. Because of species
identification problems in the field, the pollen morphology of family
Berberidaceae is still poorly understood. Our study sought to provide
important and useful palynological information and is the first report
of the characters studied in some species. In this part, we provide a
discussion on various variable characters observed during our study
which has implications in palynology. Price and Ayers (2008) stated
that pollen shape can be influenced by various factors such as drying
FIGURE 6 P/E ratio of different
pollen species. P/E ratio, polar/
equatorial ratio [Color figure can be
viewed at wileyonlinelibrary.com]
FIGURE 7 Variation in exine
thickness in different pollen species [Color
figure can be viewed at
wileyonlinelibrary.com]
1598 UR RAHMAN ET AL.](https://image.slidesharecdn.com/pollenmorphologicalvariationofberberisl-200821075505/85/Pollen-morphological-variation-of-berberis-l-from-pakistan-and-its-systematic-importance-6-320.jpg)
Pollen morphological variation of berberis l. from pakistan and its systematic importance
- 1. R E S E A R C H A R T I C L E Pollen morphological variation of Berberis L. from Pakistan and its systematic importance Saeed ur Rahman1,2 | Shujaul Mulk Khan1 | Muhammad Zafar1 | Mushtaq Ahmad1 | Raees Khan1 | Sajad Hussain1 | Muhammad Khalid3 | Sadaf-Ilyas Kayani2 1 Department of Plant Sciences, Quaid-i-Azam University, Islamabad, Pakistan 2 Joint International Research Laboratory of Metabolic & Developmental Sciences, Key Laboratory of Urban Agriculture (South) Ministry of Agriculture, Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China 3 School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai, China Correspondence Shujaul Mulk Khan, Department of Plant Sciences, Quaid-i-Azam University, Islamabad 44000, Pakistan. Email: [email protected] Review Editor: Alberto Diaspro Abstract Due to overlapping and diverse morphological characters, Berberis is among the most taxonomically complex genera. Palynology is one of the taxonomic tools for delimitation and identification of complex species. In this study, pollens of 10 Berberis species were analyzed through light and scanning electron microscopy. Qualitative as well as quantita- tive features (pollen shape, size, presence or absence of colpi, colpi length and width, exine thickness, ornamentation, pollen class, aperture, and polar–equatorial ratio) were measured. Five species were observed to have colpate (pantocolpate) with elongated ends, radially symmetrical, isopolar, monads, and psilate-regulate pollens. In polar view, six pollen were spheroidal, two were ovoid, one spherical, and one oblate. Similarly, varia- tion in pollen length was prominent and the largest pollen on polar view was recorded for B. psodoumbellata 60–65 μm (62.4 ± 0.9), while the smallest one was observed for B. lycium 29–35 μm (32.2 ± 1). The observed variation in both quantitative and qualitative features were important in taxonomic identification. This shows that palynological char- acters are helpful in identification of Berberis genus at the species level. K E Y W O R D S Berberis genus, identification problems, Pakistan, pollen types 1 | INTRODUCTION With 500 species, Berberis is an important genus of the family Berberidaceae (Ahrendt, 1961; Frodin, 2004; Ghavipanje, Nasri, Farhangfar, & Modaresi, 2016; Mokhber-Dezfuli, Saeidnia, Gohari, & Kurepaz-Mahmoodabadi, 2014; Rao, Husain, Dutt, & Garg, 1998; Rounsaville & Ranney, 2010; Schneider, 1905). The genus consists of perennial shrubs, found in different countries of the world (Harber, 2012). From Pakistan, the genus is represented by 29 species (20 species, 6 subspecies, 2 varieties, and 1 forma) distributed mainly in the northern mountainous ranges (Khan, Ahmad, Ahmad, & Azam, 2014; Khan, Shah, et al., 2018). Different overlapping characters not only make the genus complex but also problematic taxonomically which makes species identifi- cation challenging (Sodagar et al., 2012). Environmental conditions also play an important role in species variation (Ahrendt, 1961; Chapman, 1936; Khan et al., 2014; Rao et al., 1998). Leaf texture and serrations show variation as the plant age increases (Rao et al., 1998; Tiwari, Rawat, & Adhikari, 2012). Various morphological characters have been extensively revised by taxonomists and concluded that the overlapping characters not only vary from population to population but within the population and even within the same plant (Rao et al., 1998). Various studies have been conducted on the importance of morphology (Ahrendt, 1961), palynology (Khan, Abidin, et al., 2017; Khan et al., 2018), floral anatomy, (Rao et al., 1998; Khan, Ahmad, et al., 2017; Khan, Abidin, et al., 2019), and DNA barcoding but still taxonomic problems exist in the genus (Roy et al., 2010). Pollen characters have long been used to resolve taxonomic problems in a number of plant families (Castro, Silveira, Navarro, Paiva, & Coutinho, 2009). Pollen characters have not only importance in taxonomic and phylogenetic study but also in paleobotany, aeropalynology, pollination biology, pollen-pistil interaction, potential Received: 26 December 2018 Revised: 30 April 2019 Accepted: 8 June 2019 DOI: 10.1002/jemt.23325 Microsc Res Tech. 2019;82:1593–1600. wileyonlinelibrary.com/journal/jemt © 2019 Wiley Periodicals, Inc. 1593
- 2. source of nectar, pollen allergy, and in the recognition of bee plants (Paul, Essien, Idachaba, Edegbo, & Tamenku, 2014; Saensouk, Chantaranothai, & Theerakulpisut, 2009; Yuanhui, 1988). At different taxonomic levels, pol- len morphology has not only a crucial role in identification but also in the interpretation of relationship among different species (Song, Park, & Song, 2012). Blackmore and Heath (1984) have been studied the pollen morphology of three species of the family Berberidaceae and observed 3-colpate, micro-reticulate pollen in Epimedium alpinum and syncolpate, psilate or punctate pollen in Mahonia aquifolium and Berberis vulgaris. Similarly, Perveen and Qaiser (2010) studied the pollen morphology of 12 species representing two genera of the family Berberidaceae from Pakistan using light microscope (LM) and scanning electron microscope (SEM). Their results show radially symmetrical, spheroidal, or subprolate pollen, which was isopolar or apolar, foveolate-fossulate or sub-psilate often regulate-reticulate. The taxonomy of Berberis is still contradictory and perhaps the morphological variation of barberries is responsible (par- tially) for the existing taxonomic confusion (Karlovic, Kremer, Liber, Šatovic, Vršek, 2009). SEM and LM studies are critically important for the taxonomic identification of complex species (Khan et al., 2018, Khan et al., 2019). Considering these facts, the main aim of the current work was to contribute to the knowledge of Berberis species identification, especially on the palynological level. TABLE 1 List of examined plants name its location and accession numbers S# Taxa Collection site Collected by Flowering period Accession No. 1 B. aitchisonii Ahrendt Chitral (Boni) Saeed ur Rahman May–June 129521 2 B. baluchistanica Ahrendt Balochistan Zain ul Abidin and Raees Khan June 129522 3 B. calliobotrys Bien Chitral (Ziarat) Saeed ur Rahman April–June 129523 4 B. chitria Buch. -Ham Murree Saeed ur Rahman, Benazir Abbasi June–July 129524 5 B. glaucocarpa Stapf Kashmir (Rawlakot) Saeed ur Rahman, Sajad Hussain April–May 129525 6 B. kunawurensis Royle Kashmir (Ponch) Saeed ur Rahman May–July 129526 7 B. lycium Royle Dir Upper, Dir Lower, Murree Saeed ur Rahman, Fazal Ullah April–June 129527 8 B. orthobotrys Bien Kashmir (Rawlakot) Saeed ur Rahman May–June 129528 9 B. parkeriana C.K. Schneid Dir Upper Saeed ur Rahman June 129529 10 B. psodoumbellata R.Parker Gilgit Saeed ur Rahman, Siraj Khan June–July 129530 FIGURE 1 Light microscope (LM) photos of the studied pollens. (a) B. aitchisonii, (b) B. baluchistanica, (c) B. chitria, (d) B. calliobotrys, (e) B. glaucocarpa, (f) B. kunawurensis, (g) B. lycium, (h) B. orthobotrys, (i) B. parkeriana, (j) B. psodoumbellata 1594 UR RAHMAN ET AL.
- 3. 2 | MATERIALS AND METHODS 2.1 | Taxon sampling, identification, preservation, and herbarium deposition Fresh plant specimens were collected between March and August 2016 from different areas of Pakistan. These areas were visited several times during the flowering season to ensure the collection of taxa with mature flowers. The collected plant species were identified with the help of taxon- omists from Quaid-i-Azam University through personal communication. These species were also confirmed using available literature. Flora of Pakistan (http://www.efloras.org) was also reviewed to ensure confirma- tion of exact species. Voucher specimens were deposited in the Herbar- ium of Pakistan, Quaid-i-Azam University, Islamabad, Pakistan (Table 1). 2.2 | Light microscopy For LM previously described method by Harley (1992) was used with minor modifications. Glacial acetic acid was used to dehydrate the sam- ple prior to acetolysis (nine parts acetic anhydride and one part sulphuric acid). The samples were treated again with glacial acetic acid in the test tube before washing. Anthers from mature flowers were separated care- fully with the help of forceps and a piercing needle. The anthers were then placed on microscope glass slides. After crushing with a small glass rod, impurities from the glass slides were removed by using a soft camel hair brush. A drop of glycerin jelly (safranine) was pipetted onto the glass slides for visibility of the pollen grains. The prepared slides (four to six for each species) were examined under Nikon microscope (E-40×/0.65). Photographs were taken through a LM (MT 4300H) connected with CCD digital camera (Model: HDCE-50B; Figure 1). 2.3 | Scanning electron microscopy For SEM, pollens were dehydrated with ethanol mounted on metallic stubs and sputter coated with gold at high vacuum (Ion-sputter Jeol JFC-1100). Pollens were clearly observed and photographs were taken at different magnifications with a JEOL-(Model-JSM5910) SEM at the Physics Department, University of Peshawar, Pakistan. The observed qualitative and quantitative characters were recorded as polar–equatorial diameter, the shape of pollen, number of colpi, length and width of colpi, P/E ratio, exine thickness, and so on (Table 2; Figures 3–7). Samples (five to seven) were analyzed for each species. The terminologies used were followed that of Erdtman (1986). 3 | RESULTS Pollen of 10 Berberis species were studied for different quantitative and qualitative palyno-morphological characteristics in which five species, that is, B. aitchisonii, B. chitria, B. glaucocarpa, B. parkeriana, and B. orthobotrys have been reported for the first time from Pakistan. For each pollen grain (four to eight samples), qualitative and quantitative features such as pol- len shape, size, presence or absence of colpi, colpi length and width, exine thickness, ornamentation, aperture, pollen class, and P/E ratio were inves- tigated (Table 2; Figures 3–7). Variation was seen in all noted characters and the studied pollen grains were of large size. The quantitative FIGURE 2 Scanning electron microscope (SEM) images of (a) B. aitchisonii, (b) B. baluchistanica, (c) B. chitria, (d) B. calliobotrys, (e) B. glaucocarpa, (f) B. kunawurensis, (g) B. lycium, (h) B. orthobotrys, (i) B. parkeriana, (j) B. psodoumbellata UR RAHMAN ET AL. 1595
- 4. characters are represented as a minimum–maximum (length and width) μm (mean ± SE). Both polar and equatorial measurements were recorded and largest pollen was observed in B. psodoumbellata (zonocolpate) 60–65 μm (62.4 ± 0.9) while the smallest one was observed in B. lycium 29–35 μm (32.2 ± 1) as shown in Table 2. Pollens were usually monad, radi- ally symmetrical, isopolar, and predominantly a psilate-regulate sculpture (Figure 2). Among the studied pollen grains five were colpate (pantocolpate). Colpus length ranges from 2 μm in B. calliobotrys to 3 μm in B. psodoumbellata. Colpus width was found to be variable and ranged from 1.2 μm in B. glaucocarpa to 2 μm in B. chitria. The shape index (P/E ratio) var- ies from 1 μm in B. chitria to 2.6 μm in B. lycium. Shapes of pollen grains vary from ovoid as observed in B. aitshisonii and B. chitria, spheroidal pollen shapes were common as observed in B. orthobotrys, B. baluchistanica, B. calliobotrys, B. kunawurensis, B. parkeriana, and B. psodoumbellata. Other pollen types were oblate in B. lycium and spherical in B. glaucocarpa. Psilate- regulate and foveolate-fossulate were the ornamentation observed in the studied species. B. aitshisonii, B. chitria, B. glaucocarpa, and B. lycium have psilate-regulate ornamentation and B. baluchistanica, B. calliobotrys, B. orthobotrys, B. parkeriana, and B. psodoumbellata with foveolate-fossulate ornamentation while B. kunawurensis was observed having sub-psilate orna- mentation. Five pollen were pantocolpate while five were observed as spiroaperturate with long spirals. Exine thickness ranges from 1 μm in B. chitria to 2.5 μm in B. glaucocarpa and B. kunawurensis. 3.1 | Taxonomic key to the Berberis species 1 + Pollen grain pantocolpate, psilate-regulate Berberis aitchisonii. − Pollen spiroaperturate 2 2 + Foveolate-fossulate, spheroidal Berberis baluchistanica − Psilate-regulate, pantocolpate 3 3 + Small colpi, ovoid, 40–45 μm in diameter Berberis chitria − Pollen grain pantocolpate, small colpi 4 4 + Foveolate-fossulate, spheroidal, 50–53 μm in diameter Berberis calliobotrys − Small copli, pantocolpate, spherical 5 5 + Psilate-regulate, exine thickness 2.5 μm Berberis glaucocarpa − Spiral long, sub-psilate 6 6 + Spiroaperturate, spheroidal, 41–48 μm in diameter Berberis kunawurensis − Oblate, psilate-regulate, colpi small 7 7 + Pantocolpate, exine thickness 1.9 μm Berberis lycium − Spiroaperturate, spheroidal, 35–40 μm in diameter 8 8 + Foveolate-fossulate, spheroidal, spiral long Berberis orthobotrys − Spheroidal, exine thickness 1.3 μm 9 9 + Foveolate-fossulate, spiroaperturate, 49–54 μm in diameter Berberis parkeriana − Pollen 60–65 μm in diameter, spheroidal 10 10 + spiroaperturate, foveolate-fossulate, P/E ratio 1.9 Berberis psodoumbellata TABLE2Qualitativeandquantitativecharactersofstudiedpollens S#SpeciesPolardiameter(μm)Equatorialdiameter(μm)PollenclassApertureOrnamentationShapeofpollenNo.ofcolpiL.C(μm)W.C(μm)P/EratioE.T(μm) 1B.aitshisonii44–50(46.9±1.1)32–35(33.3±0.5)PantocolpateSmallcolpiPsilate-regulateOvoid52.51.41.41.5 2B.baluchistanica37–41(39±1)31–34(32.5±0.7)SpiroaperturateSpiral,longFoveolate-fossulateSpheroidal0001.22 3B.chitria40–45(42.6±0.9)28–31(29.5±0.5)PantocolpateSmallcolpiPsilate-regulateOvoid52211 4B.calliobotrys50–53(51.6±0.5)30–35(32.3±0.8)PantocolpateSmallcolpiFoveolate-fossulateSpheroidal521.31.41.2 5B.glaucocarpa55–60(57.8±0.8)33–36(34.7±0.5)PantocolpateSmallcolpiPsilate-regulateSpherica52.51.21.72.5 6B.kunawurensis41–48(44.2±1.1)21–25(23±0.7)SpiroaperturateSpiral,longSub-psilateSpheroidal0001.92.5 7B.lycium29–35(32.2±1)10.5–13.5(12±0.5)PantocolpateSmallcolpiPsilate-regulateOblate521.22.61.9 8B.orthobotrys35–40(37±0.8)25–30(27±0.8)SpiroaperturateSpiral,longFoveolate-fossulateSpheroidal0002.31.3 9B.parkeriana49–54(51.4±0.9)19–23(21±0.7)SpiroaperturateSpiral,longFoveolate-fossulateSpheroidal0002.41.3 10B.psodoumbellata60–65(62.4±0.9)29–34(31.2±0.8)SpiroaperturateSpiral,longFoveolate-fossulateSpheroidal0001.92 Abbreviations:E.T,exinethickness;L.C,lengthofcolpus;P/Eratio,polar/equatorialratio;W.C,widthofcolpus. 1596 UR RAHMAN ET AL.
- 5. FIGURE 3 Comparative number of colpi in different pollen species [Color figure can be viewed at wileyonlinelibrary.com] FIGURE 4 Comparative length and width of colpi in different pollen species [Color figure can be viewed at wileyonlinelibrary.com] FIGURE 5 Comparative polar–equatorial length in different pollen species [Color figure can be viewed at wileyonlinelibrary.com] UR RAHMAN ET AL. 1597
- 6. 4 | DISCUSSION In this study, it was intended to assess Berberis pollen flora from Pakistan using LM and SEM for its taxonomic importance in Berberis species identification. Using different observed palyno-morphological features, a taxonomic key was proposed for fast and correct identifi- cation of collected Pakistani Berberis species. Because of species identification problems in the field, the pollen morphology of family Berberidaceae is still poorly understood. Our study sought to provide important and useful palynological information and is the first report of the characters studied in some species. In this part, we provide a discussion on various variable characters observed during our study which has implications in palynology. Price and Ayers (2008) stated that pollen shape can be influenced by various factors such as drying FIGURE 6 P/E ratio of different pollen species. P/E ratio, polar/ equatorial ratio [Color figure can be viewed at wileyonlinelibrary.com] FIGURE 7 Variation in exine thickness in different pollen species [Color figure can be viewed at wileyonlinelibrary.com] 1598 UR RAHMAN ET AL.
- 7. time, process, and preparation. Pollen of 10 species from the genus Berberis in the family Berberidaceae shows a reasonable variation in various aspects such as size, shape, pollen class, ornamentation, aper- ture, and presence or absence of colpi. For a systematic study of a genus, pollen characters such as shape, size, number of apertures, and exine ornamentation are not only important but also deciding factor for species identification (Myoung Yuon, 2012). Pollen varies in shape, size (both polar and equatorial diameter), ornamentation, and other aspects studied in the current study. These are the main findings and identification characteristics on the basis of which species may separate. The results show that LM and SEM palyno-morphological characters are important in the identification of Berberis species. In addition, LM and SEM studies can play an important role to identify complex pollen flora without knowing its species and particularly in the fields of geopalynology and aeropalynology. Perveen and Qaiser (2010) studied pollen of 12 Berberis species and stated that pollens were tri and pantocolpate. Our results are in accordance with them based on palyno-morphological investigations for some species. As pantocolpate pollen were observed in B. aitchisonii, B. chitria, B. calliobotrys, B. glaucocarpa, and B. lycium. Our results corroborate with information reported by Perveen and Qaiser (2010) regarding pollen shapes as they reported spheroidal. The current study shows that different palynological characters, studied through LM and SEM are helpful tools in the identification of Berberis species and will encourage palynologists to use LM and SEM for identification of different plant species. 5 | CONCLUSION AND FUTURE RECOMMENDATIONS The current study demonstrates the importance of LM and SEM studies for accurate and fast identification of Berberis flora using dif- ferent palyno-morphological characters. The very specific objective of the current study was simply to separate and/or identify Berberis species based on palyno-morphological features. Of course, there are some crucial limitations to this work, mainly, that we could not collect the maximum number of species because of seasonal varia- tion, that is, as flowering period varies from species to species. In-depth studies of palynological characters are suggested to the researchers to explore the remaining species which may lead to the identification of some new species. ACKNOWLEDGMENTS The authors are thankful to Prof. Dr. Mir Ajab Khan for guidance, encouragement, data analysis, and skilled technical support. Quaid-i- Azam University Islamabad Pakistan is also acknowledged for provid- ing facilities to complete this work. CONFLICT OF INTEREST The authors declare that there is no conflict of interest to claim. ORCID Raees Khan https://orcid.org/0000-0003-1388-7141 REFERENCES Ahrendt, L. W. A. (1961). Berberis and Mahonia: A taxonomic revision. Botanical Journal of the Linnean Society, 57(369), 1–410. Blackmore, S., Heath, G. (1984). Berberidaceae. Review of Palaeobotany and Palynology, 42(1–4), 7–21. Castro, S., Silveira, P., Navarro, L., Paiva, J., Coutinho, A. P. (2009). Pollen morphology of Chamaebuxus (DC.) Schb., Chodatia Paiva and Rhinotropis (Blake) Paiva (Polygala L., Polygalaceae). Grana, 48(3), 179–192. Chapman, M. (1936). Carpel anatomy of the Berberidaceae. American Jour- nal of Botany, 23, 340–348. Erdtman, G. (1986). Pollen morphology and plant taxonomy: Angiosperms (Vol. 1). Brill Archive, New York. Frodin, D. G. (2004). History and concepts of big plant genera. Taxon, 53 (3), 753–776. Ghavipanje, N., Nasri, M. F., Farhangfar, H., Modaresi, J. (2016). In situ, in vitro and in vivo nutritive value assessment of Barberry leaf as a roughage for goat feeding. Small Ruminant Research, 141, 94–98. Harber, J. (2012). Two new Berberis section Wallichianae from western China. Curtis's Botanical Magazine, 29(2), 112–121. Harley, M. (1992). The potential value of pollen morphology as an addi- tional taxonomic character in subtribe Ociminae (Ocimeae: Nepetoideae: Labiatae). Advances in Labiate Science. Kew: Royal Botanic Gardens, Kew, 5(1), 125–138. Karlovic, K., Kremer, D., Liber, Z., Šatovic, Z., Vršek, I. (2009). Intra-and interpopulation variability and taxonomic status of Berberis croatica Horvat. Plant Biosystems, 143(1), 40–46. Khan, I., Ahmad, H., Ahmad, B., Azam, S. (2014). Antiglycation and anti- oxidation properties of Berberis lyceum and Terminalia chebula: Possible role in curing diabetes and slowing aging. Pakistan Journal of Botany, 46, 1469–1471. Khan, R., Abidin, S. Z. U., Mumtaz, A. S., Jamsheed, S., Ullah, H. (2017). Comparative leaf and pollen micromorphology on some grasses taxa (Poaceae) distributed in Pakistan. International Journal of Nature and Life Sciences, 1(2), 72–82. Khan, R., Ahmad, M., Zafar, M., Ullah, A. (2017). Scanning electron and light microscopy of foliar epidermal characters: A tool for plant taxon- omists in the identification of grasses. Microscopy Research and Tech- nique, 80(10), 1123–1140. Khan, R., Ul Abidin, S. Z., Ahmad, M., Zafar, M., Liu, J., Amina, H. (2018). Palyno-morphological characteristics of gymnosperm flora of Pakistan and its taxonomic implications with LM and SEM methods. Microscopy Research and Technique, 81(1), 74–87. Khan, K. U., Shah, M., Ahmad, H., Khan, S. M., Rahman, I. U., Iqbal, Z., … Aldubise, A. (2018). Exploration and local utilization of medicinal vege- tation naturally grown in the Deusai plateau of Gilgit, Pakistan. Saudi Journal of Biological Sciences, 25(2), 326–331. Khan, R., Abidin, S. Z. U., Ahmad, M., Zafar, M., Liu, J., Jamshed, S., Kiliç, Ö. (2019). Taxonomic importance of SEM and LM foliar epidermal micro-morphology: A tool for robust identification of gymnosperms. Flora, 255, 42–68. Khan, R., Kiliç, Ö., Abidin, S. Z. U., Ullah, A., Ullah, H., Zafar, M., … Khan, S. M. (2019). Utilization of foliar cuticle morphology for the identification of weedy grasses. Microscopy Research and Technique, 82(6), 1–9. Mokhber-Dezfuli, N., Saeidnia, S., Gohari, A. R., Kurepaz-Mahmoodabadi, M. (2014). Phytochemistry and pharmacology of Berberis species. Pharmacog- nosy Reviews, 8(15), 8. Myoung, L. S., Yuon, L. H. (2012). Pollen morphology of the family Lamiaceae in Mongolia. Journal of Korean Nature, 5(2), 169–179. UR RAHMAN ET AL. 1599
- 8. Paul, E., Essien, B., Idachaba, S., Edegbo, E., Tamenku, M. (2014). Compara- tive study of pollen morphology of some members of Euphorbiaceae fam- ily, standard research. The Journal of Agricultural Science, 2(4), 054–058. Perveen, A., Qaiser, M. (2010). Pollen flora of Pakistan–LXV. Berberidaceae. Pakistan Journal of Botany, 42(1), 1–6. Price, K. J., Ayers, T. J. (2008). Pollen morphology in Lysipomia (Campanulaceae: Lobelioideae) and interpretation of shape artifacts. Brittonia, 60(3), 297–302. Rao, R., Husain, T., Dutt, B., Garg, A. (1998). Revision of the family berberidaceae of India-I. Rheedea, 8(1), 1–66. Rounsaville, T. J., Ranney, T. G. (2010). Ploidy levels and genome sizes of Berberis L. and Mahonia Nutt. Species, hybrids, and cultivars. Hor- tScience, 45(7), 1029–1033. Roy, S., Tyagi, A., Shukla, V., Kumar, A., Singh, U. M., Chaudhary, L. B., … Nair, N. K. (2010). Universal plant DNA barcode loci may not work in complex groups: A case study with Indian Berberis species. PLoS ONE, 5(10), e13674. Saensouk, P., Chantaranothai, P., Theerakulpisut, P. (2009). Pollen mor- phology of the genus Cornukaempferia (Zingiberaceae) in Thailand. Journal of Systematics and Evolution, 47(2), 139–143. Schneider, C. (1905). Die gattung Berberis (Euberberis): vorarbeiten für eine monographie. Bulletin de l'Herbier Boissier II, 5, 33–48. Sodagar, N., Bahrami, A. R., Memariani, F., Ejtehadi, H., Vaezi, J., Khosravi, A. R. (2012). Biosystematic study of the genus Berberis L. (Berberidaceae) in Khorassan, NE Iran. Plant Systematics and Evolution, 298(1), 193–203. Song, U., Park, J., Song, M. (2012). Pollen morphology of Pinus (Pinaceae) in Northeast China. Forest Science and Technology, 8(4), 179–186. Tiwari, U. L., Rawat, G. S., Adhikari, B. S. (2012). Rediscovery of two endemic species of Berberis from Uttarakhand, Western Himalaya, India. Biodiversity: Research and Conservation, 28, 19–24. Yuanhui, L. (1988). Pollen morphology of the family Zingiberaceae in China-pollen types and their significance in the taxonomy. Acta Phytotaxonomica Sinica (China)., 26, 265–281. How to cite this article: ur Rahman S, Khan SM, Zafar M, et al. Pollen morphological variation of Berberis L. from Pakistan and its systematic importance. Microsc Res Tech. 2019;82:1593–1600. https://doi.org/10.1002/jemt.23325 1600 UR RAHMAN ET AL.