New Chromosome Number Reports in Some Polypetalous Species from District Kangra (Himachal Pradesh), India

New chromosome number reports in some polypetalous species from District Kangra (Himachal Pradesh), India

Savita Rani1, Santosh Kumari and Raghbir Chand Gupta

Department of Botany, Punjabi University Patiala 147 002, India

1Author for correspondence: ([email protected]) Received June 8, 2011; accepted August 3, 2011

ABSTRACT. As a part of our programme to explore and evaluate genetic diversity of fl owering plants from District Kangra (Himachal Pradesh) India, at present 150 species of Polypetalae have been meiotically worked out. It brings to light new/ varied chromosome number reports for 23 species. The species being cytologically worked out for the fi rst time on world-wide basis include Chaerophyllum capnoides (n = 11), Heracleum brunonis (n = 11), Berberis ceratophylla (n = 14), Sedum trifi dum (n = 18), Corydalis meifolia (n = 8), C. thyrsifl ora (n = 8), Indigofera hamiltonii (n = 8), Viola betonicifolia (n = 7), V. canescens (n = 6), and V. sempervirens (n = 6) along with new intraspecifi c cytoypes in case of six species as Astragalus ladakensis (n=8), Geranium pratense (n = 13), G. wallichianum (n = 28), Pelargonium hortorum (n = 18), Hypericum dyeri (n = 9) and Argemone mexicana (n = 7). The species being cytologically worked out for the fi rst time from India include Barbarea vulgaris (n = 8), Capsella bursa- pastoris (n = 8), Thlaspi alpestre (n = 7), Astragalus hamosus (n = 8), Caragana pygmaea (n = 8), Desmodium retusum (n = 11), Lespedeza cuneata (n = 10) and Oxalis corymbosa (n = 7). KEYWORDS. Chromosome number reports, Cytotypes, District Kangra, Polypetalae.

Kangra is the most popular district located on the were taken as apparently fertile, while shrivelled and un- Southwestern end of the Himachal Pradesh, a hilly state in stained pollen grains were counted as sterile. Photomicro- Western Himalayas. Its area extends from low hill sub- graphs of pollen mother cells and pollen grains were made tropical zone of altitude range (400-650 m) in Shiwaliks from freshly prepared slides using Nikon 80i eclipse Dig- to high hill wet sub-temperate zone (above 3000 m) in ital Imaging System. Voucher specimens were deposited Dhauladhar and Pir Punjal (Bara Bhangal and Chhota in the Herbarium, Department of Botany, Punjabi Univer- Bhangal) through mid hill sub-humid zone (651-1800 m) sity, Patiala (PUN). in Palampur and Dharamshala. The geographical area of District Kangra is 5,739 sq. km and lies in 31°45’0” to RESULTS AND DISCUSSION 32°28’05” N latitude and 75°35’34” to 77°04’46” E It is interesting to note that the present study has brought longitudes supports diverse species of Angiospermic fl ora. to light in one or other way, newer chromosome counts in In District Kangra, subclass Polypetalae is represented by 23 species belonging to 18 genera and 11 families from 52 families and 140 genera (Chowdhery and Wadhwa different localities with altitudinal range of 500-3,000 m 1984). It harbours enormous economically, medicinally from District Kangra. The data regarding locality with and ethnobotanically important Angiosperms (Sood et.al. altitude, accession number, present chromosome number, 1982; Kapur 1985; Sharma and Maheshwari 2005; Uniyal ploidy level, nature of meiotic course and pollen fertility et al. 2006). The perusual of literature shows that for of these species have been given in Table 1. The morpho- district Kangra, no attempts has been made for cytological logical fi eld notes are provided only for those species picture of the vast diverse fl ora except for a few sporadic which have been cytologically worked out for the fi rst reports by Bir and Kumari (1975, 1981) on 17 Legume time on world-wide basis. The previous chromosome taxa from Dharamshala. To study the genetic diversity at reports are based on chromosome number compilations intra- and interspecifi c levels and to further enrich the by Darlington and Wylie (1955), Fedorov (1974), Kumar chromosomal database of the lesser known Angiospermic and Subramaniam (1986), Index to Plant Chromosome species, the present meiotic studies have been carried out Numbers from 1968 onwards, various Journals, Proceeding from wide spread area covering up to 3,000 m altitude. volumes and internet. Further, in case of any plant species having same chromosome number reported by many MATERIALS AND METHODS authors, at the most only four latest references have been For meiotic studies, fl ower buds were collected from mentioned in the text. Meiotic abnormalities have been various localities of the Kangra district of Himachal observed in the form of cytomixis, chromosomal sticki- Pradesh (Table 1). Smears of appropriate sized fl ower ness, unoriented bivalents, inter-bivalent connections or buds were made after fi xing these in the Carnoy’s fi xative formation of laggards and bridges in the species marked (6 ethanol: 3 chloroform: 1 acetic acid v/v), using standard with abnormal meiotic course. Brief observations for each acetocarmine technique. Pollen fertility was estimated by species are discussed below. mounting mature pollen grains in glycero-acetocarmine (1:1) mixture. Well-fi lled pollen grains with stained nuclei Heracleum brunonis Benth. ex C. B.Clarke The species is 92 RANI ET AL.

Table 1. Location, altitude, accession number, present chromosome number, meiotic course, ploidy level and pollen fertility in different species from District Kangra. Taxon Location/Altitude (m) Accession Meiotic Meiotic Ploidy Pollen Number Chromosome Course* Level Fertility (PUN) (n) (x) (%age) Family: Apiaceae 1. Heracleum brunonis Patti, 32°13’N 76°10’E/ 55199 11 N 2x 95.50 (DC.) Benth. 900 Family: Berberidaceae 2. Berberis ceratophylla Chhota Bhangal, 55080 14 ABN 2x 70.56 G. Don 32°02’N 76°50’E/ 2,300 Family: Brassicaceae 3. Barbarea vulgaris R.Br. Chhota Bhangal, 52796 8 N 2x 93.70 32°02’N 76°50’E/ 2,300 4. Capsella bursa-pastoris. Chhota Bhangal, 52722 8 N 2x 98.00 (L.) Medik 32°02’N 76°50’E/ 2,300 5. Thlaspi alpestre L. Baragran, 32°02’N 76°50’E/ 52742 7 N 2x 92.30 3,000 Family: Crassulaceae 6. Sedum trifi dum Wall. Triund, 32°16’N 76°22’E/ 55122 18 N 4x 91.45 3,000 Family: Fumariaceae 7. Corydalis meifolia Wall. Lohadari, 32°02’N 76°50’E/ 55202 8 N 2x 96.00 1,600 8. Corydalis thyrsifl ora Prain Lohadari, 32°02’N 76°50’E/ 55128 8 N 2x 95.89 1,600 Family: Fabaceae 9. Astragalus hamosus L. Chhota Bhangal, 52737 8 N 2x 93.20 32°02’N 76°50’E/ 2,300 10. Astragalus ladakensis Chhota Bhangal, 52710 8 ABN 2x 73.50 N. P. Balakr. 32°02’N 76°50’E/ 2,300 11. Caragana pygmaea (L.) DC Triund, 32°16’N 76°22’E/ 55195 8 N 2x 90.00 3,000 12. Desmodium retusum Patti, 32°13’N 76°10’E/ 55201 11 N 2x 90.76 G. Don 900 13. Indigofera hamiltonii Chhota Bhangal, 55078 8 ABN 2x 60.78 Graham 32°02’N 76°50’E/ 2,300 14. Lespedeza cuneata G. Don Patti, 32°13’N 76°10’E/ 55200 10 N 2x 91.00 Family: Geraniaceae 900 Family: Geraniaceae 15. Geranium pratense L. Chhota Bhangal, 55094 13 ABN 2x 62.87 32°02’N 76°50’E/ 2,300 16. Geranium wallichianum Chhota Bhangal, 52734 28 ABN 4x 63.97 D. Don 32°02’N 76°50’E/ 2,300 17. Pelargonium hortorum Chhota Bhangal, 52739 18 N 4x 86.60 L. H. Bailey 32°02’N 76°50’E/ 2,300 Family: Hypericaceae 18. Hypericum dyeri Rehder Biching, 32°02’N 76°50’E/ 54300 9 N 2x 89.90 2,200 Family:oxalidaceae 19. Oxalis corymbosa DC. Tal-mata, 32°14’N 76°12’E/ 52738 7 ABN 2x 61.90 1,400 Family: Papavaraceae 20. Argemone mexicana L. Dehra, 31°52’N 76°12’E/ 52743 7 N 2x 90.00 530 Family: violaceae 21. Viola betonicifolia Sm. Baragran, 32°02’N 76°50’E/ 54303 7 N 2x 98.00 3,000 22. Viola canescens Tal-mata, 32°14’N 76°12’E/ 54303 6 N 2x 91.69 Wall. 1,400 23. Viola sempervirens Chhota Bhangal, 55096 6 N 2x 92.80 Greene 32°02’N 76°50’E/ 2,300m Meiotic course *N = Normal; *ABN = Abnormal. CHROMOSOME NUMBERS OF SOME POLYPETALOUS SPECIES IN KANGRA, INDIA 93 found on moist rocky slopes between the altitudinal range Corydalis thyrsifl ora Prain The species is found between of 500-1,500 m in District Kangra. It is characterized as the altitudinal range of 1,500-2,500 m, distributed on pubescent herb, fl owers greenish - white to purple and moist and shady places in forest. It is a short stemmed fruits glabrous browm. The fl owering and fruiting is seen herb with woody rootstocks and yellow fl owers with during the months of August-September. The species is purple tip. The fl owering and fruiting is seen during the cytologically worked out for the fi rst time on world-wide months of July-September. The species is cytologically basis showing chromosome count of n = 11 (Fig. 1A) and worked out for the fi rst time on world-wide basis showing which is in conformity with the earlier chromosomal haploid count of n = 8 (Fig. 1H) conforms to chromosomal reports of the allied species of the genus. reports of the allied species of the genus.

Berberis ceratophylla G. Don The species is found be- Astragalus hamosus L. The chromosome number n = 8 tween the altitudinal range of 2,000-3,000 m in District (Fig. 1I) is reported for the fi rst time from India and is in Kangra. It is charecterized as an undershrub, stem pale line with the earlier report of 2n =16 (Maassoumi 1986, yellowish, fl owers yellow and fruit dark red. The fl owering 1987) from outside India. and fruiting is seen during the months of May-August. The species is cytologically worked out for the fi rst time Astragalus ladakensis N. P. Balakr. The present chromo- on world-wide basis and the present haploid count of n = some count of n = 8 (Fig. 1J) adds a new diploid cytotype 14 (Fig. 1B) conforms to chromosomal reports of the for the species. The species is earlier known to exhibit allied species of the genus. only 2n = 32 (Ashraf and Gohil 1986; Wang et al. 1994; Gu and Sun 1996). Barbaraea vulgaris W. T. Aiton The gametic number n = 8 (Fig. 1C) for the species is reported for the fi rst time Caragana pygmaea (L.) DC. The species exhibits n = 8 from India and conforms to the earlier report of 2n = 16 (Fig. 1K) at various stages of meiosis, makes a new report (Harriman 1981; Hill 1989; Albers and Probsting 1998; from India and confi rms the earlier report of 2n =16 Koch et al. 1999) from outside India. (Frahm-Lelivela 1962; Mesicek and Sajak 1995) from outside India. Capsella bursa-pastoris (L.) Medik The gametic number n = 8 (Fig. 1D) for the species is reported for the fi rst time Desmodium retusum G. Don The chromosome number from India and conforms to the earlier report of 2n = 16 n =11 (Fig. 1L) is reported for the fi rst time from India and (Khatoon 1991; Neuffer and Eschner 1995; Aksoy et al. is in line with the earlier report of 2n = 22 (Manandhar 1999; Runemark 2000) from outside India. and Sakya 2003) from outside India.

Thlaspi alpestre L. The present chromosome count of n = Indigofera hamiltonii Graham. The species is found 7 (Fig. 1E) is reported for the fi rst time from India and is between the altitudinal range of 2,000-3,000 m in District in line with the previous chromosomal reports from Kangra. It is characterized as a tall shrub with dark red outside India by Manton (1932), Gadella and Kliphuis fl owers and pods glabrous and linear. The fl owering and (1966), Holmgren (1971) and Laane (1971). fruiting is seen during the months of April-July. The present chromosome count of n = 8 (Fig. 1M) is the fi rst Sedum trifi dum Wall. The species is found between the chromosome count for the species on world-wide basis altitudinal range of 2,500-3,500 m in District Kangra. It is and conforms to chromosomal reports of the allied species characterized as a perennial herb with glabrous stem, of the genus. leaves long, strap- shaped, tapering to a stalkless base and fl owers pink. The fl owering and fruiting is seen during the Lespedeza cuneata G. Don The chromosome number n = months of August-September. The n = 18 (Fig. 1F) is the 10 (Fig. 1N) is reported for the fi rst time from India and is fi rst chromosome count for the species on world-wide in line with the earlier report of 2n = 20 (Lee 1969; Clewell basis and is in conformity with the earlier chromosomal 1971; Liu et al. 1985; Hill 1989) from outside India. reports of the allied species of the genus. Geranium pratense L. The present chromosome count of Corydalis meifolia Wall. The species is found between n = 13 (Fig. 1O) for the species adds an anuploid cytotype the altitudinal range of 1,500-2,500 m in District Kangra. on world-wide basis. The species is already known to It is an erect herb with leafy stems and fl owers yellow have variable chromosome numbers as 2n = 24 (Tjebbes with purple tips. The fl owering and fruiting is seen during 1928; Tischler 1934) and 2n = 28 (Hollingsworth et al. the months of July-September. The species is cytologically 1992; Dobes et al. 1997; Albers and Probsting 1998; worked out for the fi rst time on world-wide basis and the Petrova and Stanimirova 2001). present haploid count of n = 8 (Fig. 1G) conforms to chromosomal reports of the allied species of the genus. G. wallichianum D. Don The haploid count of n = 28 94 RANI ET AL.

Fig. 1. New chromosome numbers in some polypetalous species from District Kangra (Himachal Pradesh), India. A. Heracleum brunonis, PMC at metaphase I, n=11. B. Berberis ceratophylla, PMC at anaphase I, n =14. C. Barbarea vulgaris, PMC at metaphase I, n=8. D. Capsella bursa-pastoris, PMC at metaphase I, n=8. E. Thlaspi alpestre, PMC at metaphase I, n=7. F. Sedum triﬁ dum, PMC at metaphase I, n=18. G. Corydalis meifolia, PMC at anaphase I, n=8. H. Corydalis thyrsiﬂ ora, PMC at metaphase I, n=8. I. Astragalus hamosus, PMC at anaphase I, n=8. J. Astragalus ladakensis, PMC at anaphase I, n=8. K. Caragana pygmaea, PMC at metaphase I, n=8. L. Desmodium retusum, PMC at anaphase I, n=11. M. Indigofera hamiltonii, PMC at anaphase I, n=8. N. Lespedeza cuneata, PMC at anaphase I, n=10. O. Geranium pratense, PMC at anaphase I, n=13; P. Geranium wallichianum, PMC at diakinesis, n=28. Q. Pelargonium hortorum, PMC at metaphase I, n=18. R. Hypericum dyeri, PMC at metaphase I, n=9; S. Oxalis corymbosa, PMC metaphase I, n=7. T. Argemone mexicana, PMC at metaphase I, n=7. U. Viola betonicifolia, PMC at metaphase I, n=7. V. Viola canescens, PMC at diakinesis, n=6; W. Viola sempervirens, PMC at anaphase I, n=6. Bar=10μm. CHROMOSOME NUMBERS OF SOME POLYPETALOUS SPECIES IN KANGRA, INDIA 95

(Fig. 1P) for the species adds a new tetraploid cytotype on basis showing chromosome count of n = 6 (Fig. 1W) and world-wide basis. Earlier the species is known to have 2n is in conformity with the chromosomal reports of the = 26 (Chaterjee and Sharma 1970; Roy et al. 1988) and 2n allied species of the genus. = 28 (Warburg 1938). ACKNOWLEDGEMENTS. The authors are grateful to the University Grants Commission, New Delhi for providing fi nancial assistance under Pelargonium hortorum L. H. Bailey The present chromo- the DRS SAP III of UGC as well as to Department of Science and some count of n = 18 (Fig. 1Q) adds a new tetraploid cy- Technology, New Delhi for FIST programmes. We are also highly thankful to the Director and other staff of Herbarium of Botanical Survey totype for the species on world-wide basis. 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