Cytological Studies on the Monochlamydeae from North India

Cytological studies on the Monochlamydeae from North India

Santosh Bala1, Raghbir C. Gupta and Preet B. Attri

Department of Botany, Punjabi University, Patiala 147002, Punjab, India

1Author for correspondence: ([email protected]) Received April 7, 2011; accepted May 5, 2011

ABSTRACT. Chromosome numbers of 24 species (34 populations) of eight families under subclass Monochlamydeae of the dicots, collected from North India are presented. Seven new chromosome counts are recorded for six species: Aerva scandens (n=27), Croton bonplandianum (n=25), Euphorbia helioscopia (n=14), Euphorbia hirta (n=20), Boerhavia diffusa (n=26, 52) and Mirabilis jalapa (n=27). Indian populations of Polygonum multifl orum (n=11, 22) are counted for the fi rst time. Number of total species and their chromosome numbers of presently investigated genera and their families have been reviewed to fi nd out polyploidy percentage and probable base numbers in India.

KEYWORDS: Base number, Chromosome number, Monochlamydeae, North India, Polyploidy

Members of Monochlamydeae are characterized by the For meiotic studies, the appropriate sized buds were presence of perianth, which may be uniseriate or biseriate fi xed in Carnoy’s fi xative for 24 hrs and preserved in 70% or may be altogether absent. It comprises of eight series, alcohol. Smears of pollen mother cells (PMCs) were 36 families, 801 genera and 11784 species. It includes a subsequently prepared in 1% acetocarmine. Chromosome number of important medicinal plants such as Achyranthes numbers were confi rmed from large number of PMCs at aspera (Girach et al. 1992), Cannabis sativa (Shah 1997, different meiotic stages. Photomicrographs of desired Grigoriev 2002) as ethanomedicine; Chenopodium album PMCs were obtained using Leica-Qwin Digital Imaging (Yadav et al. 2007), Fagopyrum cymosum (Yao et al. system. 1989) and Euphorbia splendens (Lee 1982). Some of other examples are of Croton bonplandianum (Islam et al. RESULTS AND DISCUSSION 2010) as anti-tumour; Polygonum multifl orum (Ip et al. Presently, cytological studies have been carried out on 34 1997) as antioxidant, (But et al. 1996) as antihepatic; Eu- populations of 24 species under 17 genera covering eight phorbia splendens (Bani et al. 1997) as antiinfl ammatory; families of subclass Monochlamydeae of Dicot Angio- Euphorbia hirta (Lanhers et al. 1996) with neurophysio- sperms from North India. The haploid (n) chromosome logical effects; Mirabilis jalapa (Kusamba et al. 1991) as numbers in the investigated species along with specifi c antibacterial. Anti-HIV fl avonoids (Luo et al. 1995) and locality, accession numbers (PUN) and previous chromo- antiviral fl avonoids (Du et al. 2003) are reported from some reports are presented in the Table 1. The families, Morus alba. Most of the chromosome studies on these genera and species are arranged alphabetically under their families were carried out from South and Central India respective families. Only the interesting taxa with new with scattered reports from North India. There are reports reports are further discussed. Information on previous on chromosome numbers but no detailed cytogenetical chromosome counts of presently investigated species is investigations seem to have been carried out on these based on data collected from Fedorov (1969), Moore families from North India. Keeping in view the economic (1973, 1974, 1977), Goldblatt (1981, 1984, 1985, 1988), importance and chromosome diversity in members of Goldblatt and Johnson (1990, 1991, 1994, 1996, 1998, these families, the present attempt has been made. The 2000, 2003), Kumar and Subramanian (1986), IAPT/ present paper deals with the detailed cytological analysis IOPB Chromosome data (Marhold 2006-2010). on 34 populations of 24 species of subclass Monochla- mydeae. Aerva scandens Roxb. The species showed n=27 at diakinesis (Fig. 1) with MATERIALS AND METHODS normal meiosis and pollen fertility (87%). The present Plant materials were collected from wild and some chromosome number of n=27 is counted for the fi rst time cultivated plants throughout the year in 2006-2007 from as the previous reports are of n=36 (Sarkar et al. 1975) three different states: Himachal Pradesh, Punjab and and n=52 (Pal 1964). Uttrakhand of North India. Voucher specimens have been deposited in the Herbarium, Department of Botany, Croton bonplandianum Baill. Punjabi University, Patiala (PUN), India. The area of It is an important medicinal herb commonly called collection with exact location and altitude are mentioned ‘Dantibhed’ distributed in tropical and subtropical areas, in Table 1. and is naturalized throughout India. Meiotic studies made 46 BALA ET AL.

Table 1. Information on taxa, voucher data, accession number, chromosome number and previous chromosome reports of presently investigated 24 species including 34 populations of subclass Monochlamydeae

Accession Chromosome Previous chromosome Name of taxa Voucher data (PUN) number number reports (2n) (PUN) (n) India World Family: Amaranthaceae Achyranthes aspera L. Punjab; Hoshiarpur, 295 m 49512 21 42, 48, 84, 96 14, 42 *Aerva scandens Wall. Himachal Pradesh; Sakri, 527 m 53342 27 36, 52 36 Alternanthera sessilis R. Br. Punjab; Landran, 293 m 49515 30 34, 36, 40, 96 34 Amaranthus spinosus L. Himachal Pradesh; Palampur, 1219 m 53341 17 32, 34 34 Himachal Pradesh; Sakri, 527 m 53360 17 Amaranthus virdis L. Punjab; Moga, 216 m 49516 17 34 34 Himachal Pradesh; Sakri, 527 m 53340 17 Family: Cannabinaceae Cannabis sativa L. Punjab; Patiala, 250 m 49525 10 20 20, 40, 80 Himachal Pradesh; Sakri, 527 m 53343 10 Family: Chenopodiaceae Chenopodium album L. Punjab; Moga, 216 m 49527 18 18, 36, 54 18, 36, 54 Himachal Pradesh; Sakri, 527 m 53344 27 Chenopodium ambrosioides L. Punjab; Bathinda, 201 m 49528 16 16, 32 16, 32, 36, 64 Family: Euphorbiaceae *Croton bonplandianum Baill. Punjab; Patiala, 250 m 49536 10 20 20 Punjab; Nabha, 245 m 49537 25 *Euphorbia helioscopia L. Punjab; Bathinda, 201 m 49549 21 42 12, 42 Himachal Pradesh; Sakri, 527 m 53337 14 *Euphorbia hirta L. Punjab; Patiala, 250 m 49550 20 18, 20 12, 18, 20 Euphorbia pulcherrima Willd. Punjab; Dhuri, 235 m 49551 14 28, 42, 44 26, 28, 56 Euphorbia royaleana Boiss Punjab; Boghpur, 231 m 49552 60 30, 120 + 0-2B - Euphorbia splendens Boz. ex Hook Punjab; Bandhni, 224 m 49553 20 40 24, 36, 40 Phyllanthus niruri Auct. Punjab; Patiala, 250 m 49565 13 26 14, 26 Ricinus communis L. Punjab; Faridkot, 195 m 49571 10 20 20 Family: Nyctaginaceae *Boerhavia diffusa L. Punjab; Patiala, 250 m 49522 26 26 - Punjab; Nabha, 245 m 49523 52 *Mirabilis jalapa L. Himachal Pradesh; Dharamshala, 53336 27 58 54, 58, 1500 m ~116 Family: Polygonaceae Fagopyrum cymosum Meissn. Uttrakhand; Mussoorie, 2000 m 52176 16 24, 32 16, 32 *Polygonum multiﬂ orum Thunb. Punjab; Ropar, 259 m 49570 11 -22 Himachal Pradesh; Manali, 2200 m 49575 22 Rumex dentatus D. Don Punjab; Ropar, 259 m 49572 20 40 40 Himachal Pradesh; Bir, 1300 m 53339 20 Rumex hastatus L. Punjab; Barnala, 226 m 49573 9 18 18 Uttrakhand; Mussoorie, 2000 m 49074 9 Family: Ulmaceae Trema orientalis (L.) Blume Punjab; Patiala, 250 m 49077 10 20, 22, 40, 40+0-2B c. 120 Family: Urticaceae Debregeasia hypoleuca Wedd. Uttrakhand; Mussoorie, 1600 m 49078 14 28 - *New records CYTOLOGICAL STUDIES, MONOCHLAMYDEAE, NORTH INDIA 47

Figs. 1-9. Chromosomes of some plants of North India. Scale bar=10μm. 1. Aerva scandens, PMC at diakinesis showing 27 bivalents; 2. Croton bonplandianum, PMC at metaphase I with n=25; 3. Euphorbia helioscopia, PMC at diakinesis showing 14 bivalents; 4. Euphorbia hirta, PMC at metaphase I showing 20 bivalents; 5. Boerhavia diffusa, PMC at metaphase I with 26 bivalents; 6. PMC at diakinesis with 52 bivalents; 7. Mirabilis jalapa, PMC at diakinesis showing 27 bivalents; 8. Polygonum multiﬂ orum, PMC at metaphase I with 11 bivalents, 9. PMC at metaphase I with 5 IV + 12 II. on two populations from Patiala and Nabha revealed the and hexaploid levels based on x=7. The species seems to presence of n=10 and n=25 (Fig. 2), respectively. Meiotic be a balanced polyploid as there is no meiotic abnormality course is normal in both the populations with high observed during the present studies. Microsporogenesis is percentage of pollen fertility. The present report of n=10 normal with high pollen fertility in hexaploid (92%) and is in conformity with the previous reports (Sidhu and Bir tetraploid (82.15 %). The present report of n=21 is in line 1983; Soontornchainaksaeng and Chaiyasut 1999). The with the previous reports (Bhalla 1942; Wang et al. 1999; chromosome count of n=25 is a new chromosome report. Montgomery et al. 1997; Lovkvist and Hultgard 1999), but n=14 is a new chromosome report. Euphorbia helioscopia L. In India, it is abundant throughout the North India and Euphorbia hirta L. grows up to 2,400 m in the Himalayas as a common weed In India, it is found up to 3,000m as a weed of cultivated of cultivation during winter, and frequently found in waste ﬁ elds, lawns and waste places. Present studies made on places. Meiotic studies made on two populations of the the species reported the tetraploid cytotype with 20 species revealed the presence of two cytotypes showing bivalents (Fig. 4). Further meiotic course is normal with n=14 (Fig. 3) and 21 indicating them to be at tetraploid 87% pollen fertility. The present report of 2n=40 is 48 BALA ET AL.

Table 2. Data on total number of taxonomically and cytologically known species; level and frequency of polyploidy; frequency of different cytotypes and probable base number/s in the genera studied during present investigations, at India level Name of Number of Number of Number Number %age Frequency Number Number of Ploidy Probable genus Taxonomically Cytologically of of of of of spp. with levels Base known spp. worked out spp. Diploids Polyploids polyploidy cytotypes cytotypes intraspeciﬁ c number/s polyploidy (x) Achyranthes 100 2 - 2 100 2n=14(1), 21 (1), 9 2 2x, 3x, 7, Linn. 24(1), 36(2), 6x, 12x 12 42(2), 84(2)

Aerva Forssk. 10 4 1 3 75 2n=16(2), 26(1), 9 2 2x, 8, 36(2), 42(1), 4x 9, 44(1), 52(1), 11, 72(1). 13

Alternanthera 200 9 1 8 88.9 2n=28(1), 34(2), 13 2 2x, 14, Forssk. 36(1), 60(1), 4x, 17, 64(1), 68(4), 6x 18 96(2), 100(1) Amaranthus 60 25 21 4 16 2n=16(1), 32(4), 26 3 2x, 8, Linn. 34(14),49(3), 8x, 17 49-59(1), 64(1), 16x 81(1), 128(1)

Cannabis Linn. 1 1 1 - 0 2n=20(1) 1 - 2x 10

Chenopodium 150 8 2 6 75 2n=16(1), 18(6), 15 4 2x, 8, Linn. 32(1), 36(3), 4x, 9 42(10, 54(3) 6x

Croton Linn. 750 8 7 1 12.5 2n=16(1), 20(8), 11 1 2x, 4x, 8, 40(1), 50(1) 5x 10

Euphorbia 2000 32 13 19 59.37 2n=12(2), 16(4), 50 9 2x, 6, Linn. 18(6), 20(6), 4x, 7, 20+2B(1), 22(1), 6x, 8, 24(2), 28(6), 30(1), 32(2), 8x, 9, 38(1), 40(2), 9x, 10, 42(2), 44(1), 12x, 11, 54(1), 56(2), 18x, 12 60(3), >60(1), 70x 80(1), 90(2), 120(1), 180(1), 700(1)

Phyllanthus 600 11 7 4 36.36 2n=14(1), 24(2), 14 2 2x, 7, Linn. 26(7), 28(1), 4x 12, 13 52(3)

Ricinus Linn. 1 1 1 - 0 2n=20(1) 1 - 2x 10

Boerhavia 40 4 4 - 0 2n=26(1), 40(1), 5 - 2x 13, 20, Linn. 42(1), 94(1) 21, 47

Mirabilis Linn. 4 1 1 - 0 2n=58(1) 1 - 2x 29

Fagopyrum 15 2 1 1 50 2n=16(1), 32(1) 2 - 2x, 8 Mill. 4x

Polygonum 300 16 10 6 37.5 2n=20(7), 32 5 2x, 10, Linn. 22(10), 24(4), 4x, 11, 26(1), 32(1), 6x 12 40(5), 44(1), 60(2), 66(1) Rumex Linn. 200 7 2 5 71.4 2n=18(2), 20(1), 8 1 2x, 9, 40(3), 42(1), 4x, 10 120(1) 12x

Trema Lour. 30 3 1 2 66.67 2n=20(2), 5 2 2x, 10 20+1B(1), 40(1), 4x, 160(1) 16x

Debregeasia 6 2 2 - 0 2n=28(2) 2 - 2x 14 Gaudich. CYTOLOGICAL STUDIES, MONOCHLAMYDEAE, NORTH INDIA 49

Table 3. Data on total number of taxonomically and cytologically known species; level and frequency of polyploidy; frequency of different cytotypes and probable base number/s in the families studied during present investigations, at India level Name of Family Total number of Total number of Total number Total number %age **Frequency of Total Taxonomically Cytologically worked of of of cytotypes number of known spp. out spp. Diploids Polyploids polyploidy cytotypes Amaranthaceae 850 59 36 23 38.98 2n=12(1), 14(1), 16(5), 96 18(7), 20(1), 21(1), 24(1), 25(1), 26(3), 28(2), 32(6), 34(18), 35(1), 36(11), 40(2), 42(4), 48(1), 49(4), 50(1), 52(2), 60(1), 64(2), 68(4), 72(2), 81(1), 84(2), 96(3), 100(2), 102(1), 128(1)

Cannabinaceae 3 1 1 - 00 2n=20(1) 1

Chenopodiaceae 1400 31 16 15 48.38 2n=12(1), 16(1), 42 18(23), 32(1), 36(10), 42(1), 54(4), 72(1)

Euphorbiaceae 5000 150 79 71 47.3 2n=12(2), 14(2), 16(6), 216 18(8), 20(23), 22(33), 24(6), 26(7), 28(11), 30(1), 32(2), 34(5), 36(6), 38(3), 40(4), 42(4), 44(11), 48(5), 50(2), 52(14), 54(1), 56(4), 60(5), 64(2), 66(3), 72(2), 80(4), 88(3), 90(3), 96(3), 98(1), 100(2), 104(1), 108(2), 112(2), 116(1), 120(3), 124(1), 130(1), 140(1), 180(1), 196(1), 200(1), 700(1)

Nyctaginaceae 30 9 8 1 11.11 2n=26(1), 30(1), 34(1), 8 40(1), 42(1), 58(1), 94(1), 116(1)

Polygonaceae 800 27 13 14 51.85 2n=14(3), 16(2), 18(2), 49 20(8), 22(10), 24(4), 26(1), 28(2), 32(2), 40(8), 42(1), 44(1), 48(1), 60(2), 66(1), 120(1)

Ulmaceae 200 9 5 4 44.4 2n=20(6), 28(2), 40(2), 12 60(1), 160(1)

Urticaceae 550 33 20 13 39.39 2n=16(2), 20(3), 22(2), 44 24(9), 26(8), 28(7), 32(2), 38(1), 40(1), 50(1), 52(6), 56(2) **Figures in parenthesis represent number of species. reported for the fi rst time. Cytological studies made on the populations of B. diffusa revealed that the two morphotypes are actually Boerhavia diffusa L. two different cytotypes at different levels of polyploidy. It is indigenous to India and is found throughout the The one with pinkish fl owers is having n=26 and the warmer parts of the country up to an altitude of 2,000 m in purple colored morphotype has n=52. These two cytotypes the Himalayas. It is very common in waste places and in show the formation of 26 and 52 bivalents respectively, fi elds after the rainy season. Leaves are used against during meiosis I (Figs 5, 6). Further meiotic course is abdominal tumours and cancers (Asolkar et al. 1992). normal with high pollen fertility, tetraploid (88%) and Two morphotypes with purple and white fl owers have octaploid (86%). The present chromosome counts of been reported. n=26, 52 are new to India. 50 BALA ET AL.

Morphologically, the two cytotypes are differing from compared to taxonomically known species. Total numbers each other in having profusely branched habit of 45.3- of cytotypes are found to be maximum in Euphorbiaceae 65.5 cm height bearing broadly ovate with round based family. leaves of 2.2-6.0 x 1.3-1.35 cm size in tetraploids as compared to less profusely branched habit of 84.2-91.6 ACKNOWLEDGEMENTS. The authors are thankful to DRS SAP- II, ASIST Programme and Rajiv Gandhi National Fellowship Scheme cm height bearing sub-orbicular with sub-cordate based under UGC, New Delhi for fi nancial assistance. leaves of 1.4-3.8 x 0.9-1.7 cm size in octaploids. LITERATURE CITED Mirabilis jalapa L. Asolkar, L. V., Kakkar, K. K. and Chakre, O. J. 1992. Second Meiotic studies revealed 27 bivalents at diakinesis (Fig. 7) Supplement to Glossary of India. 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