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Cytomorphological Diversity in Some Members of Family Asteraceae from the Ecologically Disturbed Habitats of Solang Valley, Kullu District, Himachal Pradesh

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Cytomorphological Diversity in Some Members of Family Asteraceae from the Ecologically Disturbed Habitats of Solang Valley, Kullu District, Himachal Pradesh © 2015 The Japan Mendel Society Cytologia 80(2): 203–222 Cytomorphological Diversity in Some Members of Family Asteraceae from the Ecologically Disturbed Habitats of Solang Valley, Kullu District, Himachal Pradesh Maninder Kaur* and Vijay Kumar Singhal Department of Botany, Punjabi University, Patiala-147002, Punjab, India Received September 30, 2014; accepted January 5, 2015 Summary Present cytomorphological surveys include meiotic studies and chromosome counts in 42 species under 93 accessions collected from the various localities of Solang Valley in Kullu district, Himachal Pradesh at different altitudes ranging from 2,400 to 3,100 m. Artemisia salsaloides (n=10), Dandranthema boreale (n=36), Ligularia fischeri (n=30) and Tussilago farfara (n=12) are recorded as the first ever chromosomal counts from India. Additional/variable cytotypes are recorded in Anaphalis nepalensis (2n=6x=42), A. triplinervis (2n=6x=42), Artemisia nilagirica (2n=4x=36; 2n=6x=54), Artemisia salsoloides (n=10), Brachyactis pubescens (2n=4x=36), Dandranthema boreale (2n=8x=72), Taraxacum officinale (2n=8x=64) and Tussilago farfara (2n=2x=24). Intraspecific polyploid cytotypes are reported in Anaphalis nepalensis (4x, 6x), Artemisia nilagirica (4x, 6x) and Taraxacum officinale (2x, 3x, 4x, 5x, 6x) from the valley. Existence of B-chromosome has been reported in Cnicus wallichii (2n=34+0-1B) which is the first ever record for the species at world level. Key words Chromosome count, Male meiosis, Intraspecific cytotype, Asteraceae, Meiotic abnormality. Kullu district, situated in the central part of Himachal Pradesh with an altitudinal range of 1,100–4,300 m, is represented by three valleys̶Kullu Valley, Parvati Valley and Solang Valley (Map 1a, b). Solang Valley, named after Solang River, lies between 77°2′29″–77°12′2″E longitude and 32°18′25″–32°24′57″N latitude. The valley originates from Beas Kund Glacier, culminates into Beas River near Palchan and covers an area of 11,215.03 ha. The valley, which is situated at the border of Greater Himalaya and Trans-Himalaya, harbors a rich biodiversity of plants. Based on its tourism potential, the area is being developed to attract tourists and, in doing so, several construction activities are going on in the valley. Also, the area is being intruded by more and more number of people resulting in serious threat to its bio-resources. Besides, the valley which was once ecologically undisturbed, has now become fragile and there is every possibility of erosion of many of its bio-resources in the near future. Keeping in view the ongoing threats to the existing plant diversity, the studies have been undertaken by the authors to explore the cytomorphological diversity in the flowering plants of Solang Valley. Some of the findings in the form of an asynaptic mutant in the tetraploid cytotype (2n=4x=48) of Solanum nigrum L. (Singhal et al. 2012), reciprocal translocations in Achillea millefolium L. (Singhal et al. 2014) and cytomixis in Nepeta govaniana (Wall. ex Benth.) Benth. (Kaur and Singhal 2014) have already been published. The present communication includes the chromosomal counts, male meiotic studies and pollen fertility analysis in the members of family Asteraceae. * Corresponding author, e-mail: [email protected] DOI: 10.1508/cytologia.80.203 204 M. Kaur and V. K. Singhal Cytologia 80(2) Map 1. (a–c) a. Location of Himachal Pradesh in India. b. Location of Kullu district and Solang Valley in Himachal Pradesh. c. A broader view of Solang Valley. The family Asteraceae contains 1,600–1,700 genera and approximately 23,600 species distributed all around the globe except Antarctica (Funk et al. 2009). This group furnishes more than 10% of the total flowering plants and is cosmopolitan in distribution (Mabberley 1997). In India, the family contains 900 species under 167 genera (Hajra et al. 1995). Several species possess medicinal properties and many are known as ornamentals. Insecticides, oils, dyes and edible products are obtained from some members. The members are characterized by florets arranged on a receptacle in centripetally developing heads surrounded by bracts, anthers fused in a ring, with pollen pushed or brushed out by the style, and by the presence of achenes (cypselas) usually with a pappus (Funk et al. 2009). Extensive cytological information has been made available on the members of Asteraceae owing to the major cytological surveys conducted by Stebbins et al. (1953), Raven et al. (1960), Moore and Frankton (1962), Ornduff et al. (1963), Turner and Irwin (1960), Turner et al. (1961, 1962, 1967), Turner and King (1964), Turner and Lewis (1965), Solbrig et al. (1969), Strother (1976), Stuessy (1977), Turner and Powel (1977), Heywood et al. (1977), Razaq et al. (1988), Keil et al. (1988), Watanabe et al. (1995, 1997, 1999), Xiong et al. (1995), Carr et al. (1999), Valles et al. (2008) and Garcia et al. (2013). Cytologically, the members have also been studied by various workers from India̶Khoshoo and Sobti (1958), Sobti and Singh (1961), Koul (1964a, b), Mehra et al. (1965), Sharma and Sarkar (1967–1968), Subramanyam and Kamble (1967), Singh (1972), Mehra and Remanandan (1969, 1974, 1975, 1976), Shukur et al. (1977), Mathew and Mathew (1978, 1983, 1987, 1988), Gupta and Gill (1979, 1980, 1983, 1984a, b, 1989), Nirmala and Rao (1984), Kaul and Bakshi (1984), Gupta et al. (1989, 2010, 2013), Rajalakshmi and Jose (2011), Bala and Gupta (2013) and Usha et al. (2013). However, no such studies have ever been undertaken on the members of Asteraceae from this geographically isolated and ecologically disturbed area. Materials and methods The materials for cytological studies were collected from the wild plants growing at various altitudinal ranges of Solang Valley, Kullu district of Himachal Pradesh, India, during the months of April–September, 2010–2013 from the different localities (Map 1c). Cytologically investigated species were identified by consulting floras of different regions, such as the Flora of Himachal Pradesh (Chaudhary and Wadhwa 1984), Flora of Lahaul-Spiti (Aswal and Mehrotra 1994) and 2015 Cytomorphological Diversity in Some Members of Family Asteraceae 205 Flora of Kullu district (Dhaliwal and Sharma 1999). The plant specimens were also compared to the samples lying in the Herbaria, Department of Botany, Punjabi University, Patiala (PUN), Botanical Survey of India, Dehra Dun (BSI), and Forest Research Institute, Dehra Dun (FRI). Voucher specimens of the cytologically worked out accessions were deposited in the Herbarium, Department of Botany, Punjabi University, Patiala. For male meiotic studies, capitula of suitable sizes randomly collected from five individuals in each population, were fixed in Carnoy’s fixative (mixture of ethanol, chloroform and glacial acetic acid in a volume ratio of 6 : 3 : 1) for 24 h, transferred to 70% alcohol and stored in a refrigerator until analysis. Pollen mother cells (PMCs) were prepared by squash technique and stained with 1% acetocarmine. Freshly prepared slides were carefully examined to determine the chromosome number. PMCs were also analysed for detailed meiotic behaviour at different meiotic stages, diakinesis, metaphase I (MI), anaphases-I/II (AI/II), telophases-I/II (TI/II) and sporad stage. In accessions with a normal meiotic course, a total of 10–30 PMCs were examined for determining the chromosome counts, while in cytologically abnormal accessions, 30–50 slides prepared from different anthers/florets were analyzed. Pollen fertility was estimated through stainability tests by squashing the anthers from mature capitula in glyceroacetocarmine (1 : 1) mixture, and 1% aniline blue dye. Well-filled pollen grains with stained nuclei were taken as apparently fertile while shriveled and unstained ones were counted as sterile. Chromosome spreads were analyzed with an Olympus light microscope, and the best plates of chromosome counts, meiotic abnormalities, sporads and pollen grains (fertile, sterile) were photographed from the temporary mounts with a Nikon Eclipse 80i microscope. The previous chromosome counts in the worked out species were compiled from chromosome indexes, viz. Darlington and Wylie (1955), Ornduff (1968, 1969), Bolkovskikh et al. (1969), Moore (1970, 1971, 1972, 1973, 1974, 1977), Fedorov (1969), Löve and Löve (1980, 1982, 1986), Goldblatt (1981, 1984, 1985, 1988), Goldblatt and Johnson (1990, 1991, 1994, 1996, 1998, 2000, 2003, 2006), Kumar and Subramaniam (1986) and Khatoon and Ali (1993). Besides, IAPT/IOPB Chromosome Reports and various journals and the website of IPCN (http://www.tropicos.org/ Project/IPCN) were also consulted. Results and discussion Present cytomorphological surveys include the exact chromosome counts, detailed meiotic course and pollen fertility in 103 wild accessions falling under 8 tribes, 28 genera and 42 species of family Asteraceae collected from the geographically isolated and severely disturbed habitats of Solang Valley. The data regarding specific locality with altitude along with GPS coordinates, voucher specimen number (PUN), meiotic chromosome number, ploidy level, pollen fertility (%age) and previous chromosome counts are given in Table 1. Four species, Artemisia salsaloides (n=10), Dandranthema boreale (n=36), Ligularia fischeri (n=30) and Tussilago farfara (n=12) present the first ever chromosomal records from India. Additional/variable cytotypes are recorded in Anaphalis nepalensis (2n=6x=42), A. triplinervis (2n=6x=42), Artemisia nilagirica (2n=4x=36; 2n=6x=54), Artemisia salsaloides (n=10), Brachyactis
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