Cytological Diversity in Some Members of Tribe Agrostideae and Eragrosteae (Poaceae) from Haryana and Adjoining Shiwalik Hills

Raghbir Chand Gupta1, Akshita Gupta2 and Navjot Kaur1*

1 Department of Botany, Punjabi University, Patiala, 147002, Punjab, India 2 Department of Agriculture, Khalsa College, Patiala, 147001, Punjab, India

Received June 1, 2017; accepted August 12, 2017

Summary The present study deals with the male meiosis of 20 species covered under 12 genera and two tribes. Four species shows new chromosome count from world as well as India level. Agrostis pilosula (2n=28), Polypogon monspeliensis (2n=56) and Eragrostis viscosa (2n=32) are the ﬁrst chromosome reports in the world, whereas, Diplachne fusca (2n=20) is the ﬁrst report from India. The meiotic course of all the investigated plant species is normal with high pollen fertility.

Key words Agrostideae, Eragrosteae, Male meiosis, Pollen fertility.

The present work is undertaken from the area of Materials and methods Haryana and its adjoining hills. Haryana is a state lo- cated in north India with 44,212 km2 of area. Shiwaliks Wild plants were collected from various localities of are also known as subhimalayas or the foothills. The Haryana, Shivalik hills and Rajasthan. The young in- altitude ranges from 900–1,500 m and up to 2,000 m. florescences were fixed in Carnoy’s fixative (6 alcohol : 3 The information regarding chromosomal data is gath- chloroform : 1 acetic acid) for 24–48 h and then stored ered from Indexes to Plant Chromosome Number by in 70% alcohol until use. For cytological analysis, an- Darlington and Wylie (1955), Federov (1969), Moore thers were squashed in 1% acetocarmine. Chromosome (1973, 1974, 1977), Goldblatt (1981, 1984, 1985, 1988), counts were confirmed by observing a number of well Kumar and Subramanian (1987), Goldblatt and Johnson spread PMCs and meiotic abnormalities if any, were (1990, 1991, 1994, 1996, 1998, 2000, 2003), Khatoon analyzed. Pollen fertility was estimated by mounting and Ali (1993) and various internet sites. the mature pollen grains in 50% glycero-acetocarmine The research on cytology of monocots mainly grasses stain. Well-stained pollen grains were taken as fertile had been undertaken from Punjab, North West and and shriveled or unstained nuclei as sterile. Photomicro- Central India, Himachal Pradesh, Kashmir by vari- graphs were taken by using Nikon 80i eclipse and Leica ous workers from the Department of Botany, Punjabi Qwin Digital Imaging System at 100x. University, Patiala (Gupta and Gupta 2008, Kaur et al. 2010a, 2010b, 2011a, 2011b, 2011c, 2011d, 2015, 2016a, Results and discussion 2016b, 2017, Chauhan et al. 2010, Saggoo and Kumari 2013, Gupta et al. 2014, 2017, Singhal et al. 2014, Kaur During the present investigation, 20 species belonging and Gupta 2016a, 2016b, Kumari and Saggoo 2016a, to 12 genera of tribe Agrostideae and Eragrosteae have 2016b, Singhal and Kumari 2017). All these studies re- been worked out cytologically. The information regard- ported interesting findings in the form of intraspecific ing the meiotic chromosome numbers (PUN), pollen cytotypes, hybrids, apomicts, and triploids. Not much fertility, ploidy level, previous chromosome reports from has been done on the exploration and evaluation of cy- India, and meiotic course (either normal or abnormal) tological diversity on grasses has been undertaken from are given in Table 1. The results on species with new the study area. meiotic chromosome report from India or outside India The present work is in continuation with the work have been explained below. done by Gupta et al. (2017). Presently, the detailed meiosis and microsporogenesis of 20 species under 12 genera Tribe: Agrostideae and two tribes has been compiled. Agrostis pilosula Trin The present meiotic course of the species has been * Corresponding author, e-mail: [email protected] carried out on single population collected from Solan at DOI: 10.1508/cytologia.82.501 an altitude of 1,350 m. It reveals the presence of 14 : 14 502 R. C. Gupta et al. Cytologia 82(5)

Table 1. Information with voucher data, chromosome number (n), pollen fertility, ploidy level, previous chromosome reports, and meiotic behavior of the species studied presently.

Taxa Previous reports** Herbarium Meiotic S. with Pollen Ploidy Meiotic Localiy with Altitude Acc. No. Chr. No. base fertility level course PUN No. (n) World India number

Tribe: Agrostideae 1. Agrostis pilosula Trin. P-I Dharampur, Solan (1,350 m) 50575 14 75.6 4x 44, 56 42: Mehra and Sharma 1975, Koul and Gohil 1991, Gupta et al. New chromosome 2014 report from world 2. Alopecurus nepalensis Trin. P-I Ballabgarh, Faridabad (197 m) 50623 7 60.4 2x 14: Mehra and Sunder 1969, Bir and Sahni 1986 14, 28: Bir and N Sahni 1987 28: Kaur et al. 2011c 42: Kaur et al. 2011 3. Muhlenbergia himalayensis Hack. ex. Hook.f. P-I Millitary Hospital, Kasauli (1,927 m) 50648 20 90.3 4x 40: Mehra and Sharma 1975, Gupta et al. 2014 N 4. Polypogon fugax Nees ex Steud. P-I Jabli, Shiwaliks (900 m) 49792 21 57.4 6x 42 28: Mehra and Sunder 1969 28, 42: Singhal et al. 2014 42: N P-II Barog, Solan (1,350 m) 49794 21 51.7 6x Mehra and Sharma 1975, Mehra 1982, Bir and Sahni 1986, Koul N and Gohil 1991, Kaur et al. 2010b 42, 42+1B: Sharma and Sharma 1979 42+0-1B: Parkash 1979 42, 84: Gohil and Koul 1986 5. P. monspeliensis (L.) Desf. Cytotype-I (n=14) 14, 26, 28: Mehra and Sunder 1969, Parkash 1979, Koul and Gohil 1991, N P-I Mullana, Ambala (272 m) 49731 14 99.0 4x 28, Bir and Chauhan 1990 28, 42: Mehra et al. 1968, Bir and Sahni P-II Kalka, Shiwaliks (624 m) 49738 14 97.1 4x 28+0- 1986 35: Sindhe 1969, Sindhe and Narayan 1976 42: Koul and N P-III KUK, Kurukshetra (350 m) 49754 14 98.2 4x 1B Gohil 1991, Kumari and Saggoo 2016 N Cytotype-II (n=28) New chromosome P-IV Morni, Panchkula (365 m) 49802 28 65.4 8x report from world Tribe: Eragrosteae 6. Acrachne racemosa (Heyne) Ohwi P-I Indri, Karnal (252 m) 50654 9 75.6 2x 18 9: Bir and Sahni 1986 12: Kaur et al. 2011c 18: Mehra et al. N P-II Jyotisar, Kurukshetra (350 m) 50661 9 84.0 2x 1968 N P-III Samalkha, Panipat (218 m) 50847 9 85.3 2x N 7. Dactyloctenium aegyptium (L.) Willd. Cytotype-I (n=22) 18, 20, 18: Bir and Sahni 1988 18, 20: Bir and Sahni 1983 18, 20, 38: N P-I Guhla, Kaithal (219 m) 50287 22 98 4x 36, 38, Bir and Sahni 1985 18, 20, 38, 46: Bir and Sahni 1986 20, 40: Cytotype-II (n=23) 40, 44, Mehra et al. 1968 20, 22, 23, 27: Sachdeva and Kals 1981 20, N P-II Shahpur, Ambala (272 m) 50312 23 82.8 4x 46, 48 23, 27, 38, 40, 44, 46: Sharma and Salam 1980 22+: Bir Cytotype-III (n=24) et al. 1987 22, 23, 46: Bir et al. 1980 36: Mehra and Sharma N P-III KUK, Kurukshetra (350 m) 50290 24 92.1 6x 1975 38: Bir and Sahni 1983 1985 40: Sharma and Sharma 1979 40, 44, 46: Sharma and Salam 1984 42: Bhattacharya 1973 44, 46: Rao and Mwasumbi 1981 46: Bir and Sahni 1986 48: Gupta 1971, Christopher and Abraham 1974 52: Sharma et al. 1978 8. D. aristatum Link. P-I Mirpur, Rewari (242 m) 50335 10 93.2 2x 20,40: Sharma and Sharma 1979 20, 38, 40: Sharma and Salam N 1984 9. Desmostachya bipinnata (L.) Stapf. P-I KUK, Kurukshetra (350 m) 50267 10 98.1 2x 20: Mehra et al. 1968, Christopher and Abraham 1974, Bir et al. N 1987 40: Bir and Sahni 1984 10. Dinebra retroﬂexa (Vahl.) Penz. P-I CCS Univ. Hissar (216 m) 50729 10 96 2x 20 20: Krishnaswamy 1941 20, 40, 60: Bir and Sahni 1986 N 11. Diplachne fusca (L.) P. Beauv. P-I Sanauli, Panipat (218 m) 50745 10 98.2 2x 20, 38, 40: Bir and Sahni 1986 New chromosome 40 report from India 12. Eleusine indica (L.) Gaertn. Cytotype-I (n=9) 18, 36 18: Krishnaswamy 1941, Mehra et al. 1968, Gupta 1971, Bhat- N P-I Barog, Solan (1,350 m) 50353 9 95.2 2x tacharya 1973, Christopher and Abraham 1974, Mehra and P-II Kalka, Shiwaliks (624 m) 50368 9 98.0 2x Sharma 1975, Sharma et al. 1978, Bir and Sahni 1986, Koul N P-III Paonta Sahib, Sirmour (932 m) 50259 9 76.4 2x and Gohil 1991, Singhal et al. 2014, 18, 36: Bir and Chauhan N Cytotype-II (n=27) 1990 18, 36, 54, 60: Kalia 1978 18, 40: Mehra et al. 1968 28: P-IV Trilokpur Shiwaliks (932 m) 50360 27 98.3 6x N Kaur et al. 2011b 13. Eragrostis ciliaris (L.) R. Br. P-I Mirpur rewari, (242 m) 50817 20 92.9 4x 20, 40 20: Mehra et al. 1968 20: Rao and Mwasumi 1981 20, 40: Bir N and Sahni 1988 40: Bir and Sahni 1984, 1986 14. E. japonica (Thunb.) Trin P-I Nizampur, Mahendragarh (261 m) 50823 20 97.5 4x 20, 40 20: Mehra and Kalia 1976 40: Bir and Sahni 1988 60: Christo- N pher and Abraham 1974 15. E. minor Host. P-I KUK, Kurukshetra (350 m) 50825 20 84.9 4x 20, 30, 40, 60: Bir and Sahni 1986, 1988 60: Bir and Sahni 1986 N 40, 60, 80 16. E. pillosa (L.) P. Beauv. P-I Takeli, Gurgaon (225 m) 50854 18 80.2 4x 20, 30, 20, 36, 40: Bir and Sahni 1986, 1988 30: Bir and Sahni N 40, 60 1988 36: Bir and Sahni 1983 36+: Bir and Sahni 1985 40: Mehra et al. 1968 40: Christopher and Abraham 1974 17. E. tenella (L.) P. Beauv. Ex Roem & Schult. P-I Sanauli, Panipat (218 m) 50849 9 93.4 2x 20, 40, 20: Mehra et al. 1968 60: Christopher and Abraham 1974 N 60 18. E. viscosa (Retz.) Trin. P-I Devsar, Bhiwani (224 m) 50873 16 89.2 4x 40, 60 16: Bir and Chauhan 1990 First chromosome report from world 19. Leptochloa chinensis (L.) Nees P-I KUK, Kurukshetra (350 m) 50573 18 82.9 4x 40 20, 40: Bir and Sahni 1986 36: Mehra and Kalia 1976 40: N Mehra et al. 1968, Bir and Sahni 1986, Christopher and Abraham 1974 20. L. panacea (Retz.) Ohwi P-I Agroha, Hissar (216 m) 50570 20 100.0 4x 20 20: Mehra et al. 1968, Bir and Sahni 1985 20, 40: Bir and Sahni N 1986 2017 Cytological Diversity in Some Members of Tribe Agrostideae and Eragrosteae (Poaceae) from Haryana and Adjoining Shiwalik Hills 503

Figs. 1–24. (1) Agrostis pilosula, PMC at A-I with 14 : 14 chromosomes; (2) Alopecurus nepalensis, PMC showing 7II at diakinensis; (3) Muhlenbergia himalayensis, PMC with 20II at diakinensis; (4) Polypogon fugax, PMC with 21II at M-I; (5, 6) P. monspeliensis, (5) (Cyt-I): PMC with 14II at M-I, (6) (Cyt-II): PMC with 28II at M-I; (7) Acrachne racemosa, PMC with 9II at M-I; (8–10) Dactyloctenium aegyptium, (8) (Cyt-I) PMC with 22II at M-I, (9) (Cyt-II) PMC at M-I with 23II, (10) (Cyt-III) PMC with 24 : 24 chromosomes at A-I; (11) D. aristatum, PMC with 10II at M-I; (12) Desmostachya bipinnata, PMC with 10II at M-I, (13) Dinebra retroﬂexa, PMC with 10II at M-I, (14) Diplachne fusca, PMC with 10II at M-I, (15, 16) Eleusine indica, (15) (Cyt-I) PMC with 9II at M-I, (16) (Cyt-II) PMC with 27II at diakinensis; (17) Eragrostis ciliaris, PMC with 20 : 20 chromosomes at A-I; (18) E. japonica, PMC with 20 : 20

chromosomes at A-I; (19) E. minor, PMC with 20II at M-I; (20) E. pillosa, PMC with 18 : 18 chromosomes at A-I; (21) E. tenella, PMC with 9II at diakinensis; (22) E. viscosa, PMC with 16II at M-I; (23) Leptochloa chinensis, PMC with 18II at M-I; (24) L. panacea, PMC with 16II at M-I. 504 R. C. Gupta et al. Cytologia 82(5) chromosome at A-I (Fig. 1). Meiosis is found to be nor- (x=9, 10), Eleusine (x=9, 10), Eragrostis (x=9, 10), and mal with 75.6% pollen fertility. The present report of Leptochloa (x=9, 10) are dibasic. n=14 is the new chromosome count for the species from The present research is a first attempt to study the world level. Previously, 2n=42 by Mehra and Sharma chromosome numbers of the Tribe Agrostideae and (1975) from Nainital, Koul and Gohil (1991) from Kash- Eragrosteae from Haryana and its adjoining Shiwalik mir, and Gupta et al. (2014) from Lahaul Spiti, H.P. Out- hills. We compiled and analyzed these chromosome side India, 2n=44 by Narayan and Muniyamma (1972); number reports on a population basis and reconsidered 2n=56 by Gould and Soderstrom (1970) from Ceylon. the basic chromosome numbers of both the tribes up to Polypogon monspeliensis (L.) Desf date. Thus, the data belonging to both the tribes shoul Presently, two cytotypes, tetraploid (n=14) (Fig. 5) prove useful in further cytotaxonomic and phylogenetic and octaploid (n=28) (Fig. 6), are recorded. The octa- research. ploid cytotype is very common. The present report of tetraploid cytotype with n=14 is in conformity, whereas, Acknowledgements octaploid cytotype with n=28 is a new chromosome report in the world. Besides this, other chromosome The authors are thankful to BSR-Indira Gandhi numbers reported from India are 2n=35 and 2n=42 by Single Girl Child Scholarship (Award letter number: Koul and Gohil (1991) from Kashmir, and Kumari and F.7-152/2007 BSR) to Navjot Kaur and DBT-IPLS proj- Saggoo (2016b) from Kinnaur. Outside India, 2n=26 by ect (Project no. BT/PR-4548/INF/22/146/2012) sanc- Faruqi et al. (1987) from Pakistan. Thus the species has tioned to Punjabi University, Patiala for financial as- well developed intraspecific polyploidy series i.e., 2x, 4x, sistance. The authors are also thankful to the Head of 5x, 6x, and 8x. Department of Botany, Punjabi University, Patiala for providing us necessary laboratory facilities. Tribe: Eragrosteae Diplachne fusca (L.) P. Beauv References The present diploid (n=10) based on the basic chromosome number 10 is a new chromosome count from Bhattacharya, B. 1973. Chromosome analysis of some Indian mem- India. 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