Bioscience Discovery, 8(3): 574-581, July - 2017 © RUT Printer and Publisher Print & Online, Open Access, Research Journal Available on http://jbsd.in ISSN: 2229-3469 (Print); ISSN: 2231-024X (Online) Research Article Stomatal Studies in the Genus Chlorophytum (Asparagaceae) Nalawade A S* and R V Gurav Department of Botany, Shivaji University, Kolhapur (MS)-416004 *[email protected] Article Info Abstract Received: 08-06-2017, Genus Chlorophytum has been studied for various aspects such as taxonomy, Revised: 29-06-2017, anatomy, molecular biology and ethnobotany by various workers but in the case Accepted: 30-06-2017 of epidermal and cuticular studies, less attention has been given. Data regarding stomatal studies is either absent or obscure. In present investigation stomatal Keywords: studies has been carried out to know the size, structure and distribution of Asparagaceae, epidermal cells, stomata, stomatal density and stomatal index of genus Chlorophytum, Stomatal Chlorophytum. In all the studied species stomata were predominantly Density, Stomatal Index. hypostomatic and anomocytic. Maximum length (L) and width (W) is observed in C. indicum (50.7±0.9 µm & 42±0.8µm) and minimum in C. breviscapum (25.2±0.7µm & 18.6±0.4µm). Highest stomatal density was observed in C. gothanense (227±7.4/mm2) and lowest in C. indicum (25±7.5/mm2). The Stomatal index was highest in C. glaucoides (51.28) followed by C. gothanense (46.9) and lowest on C. indicum (29.41) followed by C. kolhapurense (33.33). A negative correlation was observed in respect to stomatal density and chromosome number. INTRODUCTION in Anthericaceae and now it is being treated under Stomatal characters are among the epidermal subfamily Agavoideae of family Asparagaceae characteristic, which has been very useful in (APG III). In India, the genus is represented by taxonomic delimitation apart from functional about 20 species and most of them occur in Western relevance of epidermal structures (Hari Babu and Ghats (Adsul, 2014). Most of the Chlorophytum Savithramma, 2014) Distribution, size and species are usually forest dwellers and ephemerals frequency of stomata have been reported to be making it difficult to find them in flower and fruit. specific for a genus or a species (Miller 1938). The (Chandore et al., 2012) The restricted distribution, leaf epidermal cells are of significant taxonomic remote habitat, seasonal flowering and fruiting importance; the length and width are regarded as period, most of the species have similar sized useful aids in distinguishing varieties with similar flowers and leaf pattern and identification key with flowering dates in perennial rye grass (Wilkins & overlapping characters have made the genus Sabanci, 1990). difficult to identify (Adsul et al., 2014). However The Chlorophytum is old world genus there is no detail study done on stomatal and related distributed in Africa, Australia and Asia and is anatomical characters on genus Chlorophytum till represented by about 214 species, 6 subspecies and date. In present study an attempt was made on 14 varieties (Govaerts et al., 2012). Formerly the cuticular and epidermal studies of genus genus was considered under family Liliaceae then Chlorophytum from India. http://jbsd.in 574 ISSN: 2229-3469 (Print) Nalawade and Gurav gothanense, and C. kolhapurense with lesser MATERIALS AND METHODS frequency and all others have hypostomatic leaves. All the 16 species of Chlorophytum were collected Stomata in all are anomocytic. Meidner & from various localities of India (Table 1) and Mansfield (1968) reported linked chain of germplasm is maintained in the Botanical Garden of epidermal cells with stomata at joints in family Department of Botany, Shivaji University, Liliaceae. This type of arrangement was commonly Kolhapur. The foliar epidermal peels were taken observed in all studied species. from the middle of both surfaces of mature leaves. Stomatal size: Minimum value for length and Leaves of Chlorophytum species were collected, width is observed in C. breviscapum (25.2±0.7µm washed and peeled off by tearing the leaf suddenly & 18.6±0.4µm) and maximum in C. indicum and with forcing the lower epidermis upward. A (50.7±0.9 µm & 42±0.8µm). thin membranous epidermis gets separated near the broken edges (Singha et al., 2013). These peels Stomatal density: Lowest stomatal density is were taken in the Petri plates containing distilled observed in C. indicum (25±7.5/mm2) and highest water for study. These uniformed peels were taken in C. gothanense (227±7.4/ mm2). on glass slides and a drop of 1% safranin stain was Stomatal index: Minimum value is obtained in C. added, kept for 1 minute, washed with water and indicum (29.41) while maximum in C. glaucoides then mounted in a drop of glycerine to prevent the (51.28). drying. A coverslip was placed and excess of water was blotted with the help of blotting paper. Slides Stomatal cell arc: Arc with angle, radius and were examined and photographed under Biological length in single guard cell of stomata is measured research microscope LM 52 1803 with CMOS which shows lowest value for all three parameters 15500 - 5.0MP Camera (figure 2). The stomata of of arc in C. breviscapum (133±11.90, 15.6±1.3µm, each species are illustrated line drawing (figure 3). 33.9±1.5µm) while highest value for angle is found In the stomatal studies, images were analysed by in C. laxum (189±5.10) and for radius and length is software TS View version 7.3.1.7 (figure 1) and found in C. indicum (25.9±0.5 µm & 69.5±2.1 µm). readings were noted in 10 randomly selected fields Plants or tissues with the smaller cell size are more and mean value was estimated. The values for the tolerant of low water potential (Ainsworth et al., stomatal index were calculated using Salisbury’s 2006; Muller et al., 2007; Weijschede et al., 2008) formula (Salisbury, 1927). but in present investigation it is found that C. indicum, C. laxum, C. borivlianum and C. kolhapurense have larger epidermal cell size though RESULTS AND DISCUSSION they were collected from moist or dry deciduous Data on epidermal cells, stomatal cells, stomatal arc forest area. Epidermal cell density is higher than the and their density and stomatal index are presented stomatal density in all taxa except C. glaucum. This in Table 2. character is useful in identification of respective species. Patil and Patil (1987) observed the Epidermal cell size: Lowest value for length is presence of stomata on adaxial surface of C. noted in C. breviscapum (71.4±17.9 µm) and tuberosum. During present study similar highest in C. indicum (332.4±44.8µm) and however observations were observed in the leaves of C. minimum width is noted in C. gothanense arundinaceum, C. breviscapum, C. gothanense, and (25.9±2.8µm) and maximum in C. heynei C. kolhapurense. In all studied Chlorophytum (70.5±4.1µm). On the adaxial surface of all taxa, species stomata are aperigenous i.e., without the leaf epidermal cells are either hexagonal or subsidiary cells. Present investigation supports rectangular. observations made by Stebbins & Khush (1961) that Epidermal cell density: Density value is lowest in plants without subsidiary cells are predominantly C. laxum (58±10mm2) while highest value is geophytes. Naik and Nirgude (1981) observed that observed in C. gothanense (257±16.2mm2). taxa certain anatomical features such as stomatal Stomatal distribution: Out of 16 species studied, studies indicates close correlation with chromosome amphistomatic leaves were observed in C. number. In present study taxa with chromosome tuberosum with the maximum frequency of stomata number 2n=16 show higher length and width in while in C. arundinaceum, C. breviscapum, C. epidermal cells. However, stomatal density showed a negative correlation with chromosome number. http://biosciencediscovery.com 575 ISSN: 2231-024X (Online) Bioscience Discovery, 8(3): 574-581, July - 2017 Figure 1. Measurement of dimension of A. Epidermal cell length and width (40X), B. Stomatal length and width (100X), C. Stomatal density (10X), D. Stomatal arc (100X) A negative correlation was also observed in APG III, 2009. An update of the Angiosperm stomatal index, stomatal density and stomatal size. Phylogeny Group classification for the orders and Stomatal density and index decreases as stomatal families of flowering plants: APG III. Botanical size increases. Journal of the Linnean Society, 161: 105-121. Chandore AN, Malpure NV, Adsul AA and Acknowledgment Yadav SR, 2012. Chlorophytum belgaumense, a Authors are thankful to the Head, Department of new species of Asparagaceae from the Western Botany, Shivaji University, Kolhapur for providing Ghats of India. Kew Bulletin, 67: 527-531. research facilities and the financial assistance from Govaerts R, Zonneveld BJM and Zona SA, 2012. UGC-BSR Fellowship, New Delhi is gratefully World Checklist of Asparagaceae. Royal Botanic acknowledged. Gardens, Kew. Available from: http://apps.kew.org/wcsp/qsearch.do. 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