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A Karyotype Analysis in Ledebouria Revoluta (Hyacinthaceae) with a New Cytotype

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A Karyotype Analysis in Ledebouria Revoluta (Hyacinthaceae) with a New Cytotype 8 KAWAKAMI ET AL Chromosome Botany (2019) 13 (2): 61-67 ©Copyright 2019 by the International Society of Chromosome Botany A Karyotype analysis in Ledebouria revoluta (Hyacinthaceae) with a new Cytotype Priyanka T. Giranje1, Mayur D. Nandikar2* and Shrirang R. Yadav3 1,2 Naoroji Godrej Centre for Plant Research, 431, Lawkim Campus, Shindewadi, Shirwal, Dist. Satara, Maharashtra, India 412 801. 3Professor and BSR Faculty Fellow, Department of Botany, Shivaji University, Kolhapur, Maharashtra, India 416 004. *Author for correspondence ([email protected]) Received November 21, 2018; accepted December 21, 2018 ABSTRACT: Ledebouria Roth is a complex genus and it shows a wide range of chromosome counts. Present paper reports chromosome number 2n (6X) = 90 in L. revoluta (L.f.) Jessop a number that is not known so far in the entire genus Ledebouria which exhibits highly asymmetrical (2C) karyotype. The karyotype symmetry of the chromosomes and extreme level of polyploidy indicates that the L. revoluta is actively evolving in varied climatic and geographical regions. KEYWORDS: Karyotype, Ledebouria, Hyacinthaceae, new cytotype, India The genus Ledebouria Roth (Hyacinthaceae) consists of Rahangdale 2016) were collected from the hill slopes of ca. 61 species and 2 infraspecific species in the world Junnar, Pune district, Maharashtra (Fig. 1A). The plants (WCSP 2018). Although, Ledebouria was described as for the cytological study were grown in Naoroji Godrej early as 1821 by Roth, it has been treated as a synonym of Centre for Plant Research (NGCPR) Botanical garden, Scilla in most Indian literature (Giranje and Nandikar Shirwal, Satara, Maharashtra. The voucher specimens 2016). The genus has very few qualitative characters of (M.D. Nandikar 1403, M.D. Nandikar 1418 and P.T. diagnostic value in determination of the species. Although, Giranje 1434) have been deposited at NGCPR. the characters such as blotches on the leaf, size, shape and orientation of the leaves (erect, sub–erect or prostrate), Sampling of karyotype: The bulbs were kept for rooting. inflorescence size, flower number, etc have been used for Water grown healthy root tips (0.5 cm long) were pre– identification of the species, however, these characters are treated with a saturated solution of para–dichlorobenzene varies greatly with reference to their climatic and (pDB) at 10 ±2°C for 3–4 hrs. Root tips were hydrolyzed geographical conditions. in 1 N HCL and then stained and squashed with 2 % In India the genus is represented by Ledebouria aceto–orcein. revoluta Roth and other three ill–defined species viz. L. The well separated somatic plates were photographed viridis S. Dutta and P. Harvey ex M. R. Almeida, L. by OPTSCOPES IS–300 camera under LEICA DM 500 karnatakensis Punekar and Lakshmin., and L. microscope at 100x magnification. Five well separated hyderabadensis M. V. Ramana, Prasanna and Venu. somatic chromosome plates were selected for karyotype Ledebouria revoluta is wide spread from Africa to analysis. Chromosomes were sorted into different peninsular India and shows great diversity both in categories on the basis of type of chromosome and arm morphological and chromosome number in different ratio by following Levan et al. (1964). Evaluation of habitats. Leaf tip bulbils is a predominant mode of Karyotype symmetry has been determined by using vegetative reproduction in majority of the populations categories of Stebbins (1971). which is subjected to cytological variations (Jessop 1973; Dixit et al. 1989; Jehan et al. 2014). The chromosome RESULTS counts reported so far in Ledebouria revoluta shows 10 The karyotype analysis of Ledebouria revoluta has been cytotypes (2n = 28, 30, 44, 45, 46, 58, 60, 64, 68, and 70: given in Table 2. Different karyotypic parameters such as Table 1) indicating active cytological differentiation. The a total chromosome length of haploid compliment (THCL), present report (2n= 90) confirms hexa–ploidy (6X) nature total chromosome length percent (TCL %), total from of populations, which is first time recorded not only in percent (TF %), symmetric index (SI), Gradient Index Ledebouria revoluta but also in entire genus Ledebouria. (GI) Coefficient of variation indices (CVcl and CVci) (Paszko 2006), and asymmetry indices (Ai, A1, A2) MATERIALS AND METHODS (Zarco 1986) were examined for L. revoluta, and are Plant material: Plant material of Ledebouria revoluta tabulated in Table 3. (= described as L. junnarensis by Rahangdale and 62 GIRANJE ET AL. Table 1. A detailed account of previous chromosome counts in Ledebouria revoluta (L.f.) Jessop Reported as taxa n (2n) Studied by Country/Provision Ledebouria revoluta (L. f.) Jessop 15 30 Haque and Ghosh (2015) India (West Bengal, Kolkata, Rahara) Ledebouria revoluta (L.f.) Jessop 64, 68 Sheeba and Vijayavalli (1998) (=Scilla indica (Wight) Baker) Ledebouria revoluta (L.f.) Jessop 22 Johnson and Brandham (1997) South Africa (=Scilla revoluta (L.) Baker) Ledebouria revoluta (L.f.) Jessop 28 Stedje (1996) East Africa, Kenya, Machakos, between Koma Rocks Ledebouria revoluta (L.f.) Jessop 30 Dixit et al. (1989) India (Maharashtra, Mumbai, Kanheri caves) (=Scilla hyacinthina (Roth) J. F. Macbr) 45 India (Maharashtra, Satara, Kas plateau) 60 India (Maharashtra, Pune, Dive ghat) Ledebouria revoluta (L.f.) Jessop 30 Jha and Sen (1980), India (Maharashtra, Satara, Mahabaleshwar) (=Scilla hyacinthina (Roth) J. F. Macbr.) 60 Nair (1989) India (Maharashtra, Mumbai) Ledebouria revoluta (L.f.) Jessop 45 Sheriff and Rao (1981) India (Tamil Nadu, Chennai, Adyar & Saidapet) (=Scilla indica Roxb.) Ledebouria revoluta (L.f.) Jessop 20,22, Subramanian (1981) (=Scilla indica Baker ) 44,45, 46,58 Ledebouria revoluta (L.f.) Jessop 10 Sarkar et al. (1980) India (Maharashtra, Uklapani) (=Scilla hyacinthina (Roth) J. F. Macbr.) Ledebouria revoluta (L.f.) Jessop 30 Sen (1973), Western Coastal belts of India (=Scilla indica (Wight) Baker ) Chakravarty and Sen (1992) Ledebouria revoluta (L.f.) Jessop 9 Jessop (1973) South Africa (Rhodes University garden; Henderson's 10 Farm; 34 miles west of Potgietersrust; 11 5 miles west of Colenso; Kroomie, Fort Beaufort; 2 miles 12 north-west of Grahamstown; Willowmore; 13 SWAZILAND;34 miles west of Potgietersrust). 15 16 17 22 A KARYOTYPE ANALYSIS IN LEDEBOURIA REVOLUTA (HYACINTHACEAE) WITH A NEW CYTOTYPE 63 Table 1. Continued Reported as taxa n (2n) Studied by Country/Provision Ledebouria revoluta (L.f.) Jessop 15 Jessop (1970) South Africa (Eastern Cape,Grahamstown 20 miles N. of Pretoria Rhodes Botany Dept. Garden) Ledebouria revoluta (L.f.) Jessop 30 Rao (1956) India (Maharashtra, Mumbai; Karnataka, Dharwad, (=Scilla indica (Wight) Baker ) 44 Mysore; Tamil Nadu, Tiruchirapalli, Chennai; 45 Andhra Pradesh, Krishna, Machilipatnam; Madhya 46 Pradesh, Sagar; Maharashtra, Nagpur) 58 60 Ledebouria revoluta (L.f.) Jessop 44, 45, Darlington and Wylie (1955) (= Scilla indica (Wight) Baker ) 46, 58 Ledebouria revoluta (L.f.) Jessop (=Scilla 30, 44, Rao (1953) indica (Wight) Baker ) 45, 46, 58 Ledebouria revoluta (L.f.) Jessop 30 Sheriff and Murthy (1946) India (Karnataka, Kolar hills) (=Scilla indica (Wight) Baker ) Ledebouria revoluta (L.f.) Jessop 44 Raghavan and Venkatasubban (1939) India (Tamil Nadu, Chennai, Adyar) (=Scilla indica Roxb.) 46 India (Tamil Nadu, Chennai, Saidapet) 45 India (Tamil Nadu, Chennai, Soundarya Nursery) 64 GIRANJE ET AL. Fig. 1: Ledebouria revoluta A. habitat, B. habit, C. chromosome count (2n =90), D. idiogram. A KARYOTYPE ANALYSIS IN LEDEBOURIA REVOLUTA (HYACINTHACEAE) WITH A NEW CYTOTYPE 65 Table 2. Karyotype details of Ledebouria revoluta Chromosome Length of Length of Total length d=l-s Arm ratio Centromeric Centromeric pairs long arm (l) short arm (s) c=l+s (µm) r=l/s index Position (µm) (µm) i=s/c*100 1 4.30 ±1.08 2.03 ±0.48 6.33±1.53 2.27 2.12 32.05 sm 2 3.96±0.87 1.83±0.50 5.79±1.32 2.13 2.16 31.63 sm 3 3.38±0.91 1.67±0.60 5.05±1.47 1.71 2.02 33.08 sm 4 2.61±0.61 1.55±0.47 4.16±1.05 1.06 1.68 37.28 sm 5 2.46±0.41 1.28±0.36 3.74±0.76 1.18 1.92 34.21 sm 6 1.91±0.56 1.19±0.06 3.10±0.58 0.72 1.60 38.46 m 7 1.53±0.43 1.20±0.17 2.73±0.60 0.33 1.27 44.03 m 8 1.45±0.31 0.99±0.14 2.44±0.38 0.46 1.47 40.51 m 9 1.42±0.25 0.96±0.11 2.38±0.34 0.45 1.47 40.47 m 10 1.32±0.21 0.95±0.14 2.27±0.32 0.37 1.38 41.95 m 11 1.22±0.22 0.95±0.07 2.17±0.28 0.28 1.29 43.62 m 12 1.12±0.14 0.96±0.14 2.07±0.27 0.16 1.16 46.22 m 13 1.26±0.18 0.81±0.13 2.07±0.24 0.46 1.57 38.93 m 14 1.18±0.18 0.77±0.16 1.96±0.26 0.41 1.53 39.49 m 15 1.11±0.20 0.80±0.12 1.91±0.25 0.31 1.39 41.90 m 16 1.22±0.24 0.65±0.20 1.87±0.22 0.57 1.87 34.87 sm 17 1.00±0.10 0.81±0.15 1.81±0.22 0.19 1.23 44.82 m 18 1.05±0.14 0.71±0.10 1.76±0.21 0.34 1.48 40.32 m 19 0.98±0.17 0.72±0.05 1.70±0.20 0.26 1.36 42.34 m 20 0.99±0.13 0.66±0.14 1.65±0.21 0.32 1.48 40.24 m 21 1.07±0.09 0.55±0.24 1.63±0.18 0.52 1.95 33.94 sm 22 0.96±0.16 0.66±0.07 1.62±0.18 0.30 1.46 40.71 m 23 1.06±0.06 0.60±0.19 1.66±0.18 0.46 1.76 36.17 sm 24 0.94±0.09 0.63±0.08 1.57±0.16 0.30 1.48 40.32 sm 25 0.91±0.08 0.63±0.21 1.54±0.15 0.29 1.46 40.73 sm 26 1.03±0.14 0.51±0.14 1.54±0.15 0.52 2.00 33.31 sm 27 0.98±0.18 0.55±0.15 1.52±0.16 0.43 1.79 35.82 sm 28 0.94±0.22 0.55±0.17 1.49±0.18 0.39 1.70 37.01 sm 29 0.89±0.08 0.60±0.20 1.49±0.18 0.29 1.48 40.39 m 30 0.82±0.07 0.63±0.11 1.45±0.17 0.19 1.31 43.35 m 31 1.08±0.18 0.37±0.25 1.44±0.16 0.71 2.92 25.51 sm 32 0.93±0.14 0.49±0.29 1.42±0.18 0.44 1.90 34.49 sm 33 0.90±0.15 0.44±0.25 1.35±0.30 0.46 2.04 32.84 sm 34 0.95±0.20 0.42±0.29 1.37±0.19 0.52 2.24 30.84 sm 35 0.98±0.26 0.36±0.23 1.34±0.18 0.63 2.74 26.72 sm 36 0.87±0.16 0.44±0.19 1.30±0.18 0.43 1.98 33.51 sm 37 0.99±0.21 0.30±0.33 1.29±0.17 0.68 3.25 23.54 st 38 0.89±0.20 0.38±0.25 1.27±0.15 0.52 2.37 29.66 sm 39 0.84±0.15 0.40±0.19 1.24±0.15 0.44 2.11 32.12 sm 40 0.81±0.21 0.38±0.14 1.19±0.15 0.42 2.10 32.21 sm 41 0.82±0.21 0.33±0.21 1.15±0.13 0.50 2.52 28.43 sm 42 0.95±0.23 0.14±0.21 1.08±0.14 0.81 6.88 12.69 st 43 0.74±0.17 0.20±0.12 0.94±0.10 0.54 3.69 21.30 st 44 0.90±0.22 0.07±0.16 0.97±0.10 0.83 12.87 20.21 st 45 0.82±0.08 0.04±0.08 0.86±0.11 0.78 21.91 14.37 st The karyotype analysis of Ledebouria revoluta can be µm.
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