Urginea Indica (Roxb) Kunth
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© 2016 The Japan Mendel Society Cytologia 81(3): 329–334 Karyotypic Variations in Different Accessions of Urginea indica (Roxb) Kunth. and Urginea wightii Lakshmin. Hyacinthaceae H. L. Geetha, M. N. Shivakameshwari* and R. Nijagunaiah Department of Botany, Jnana Bharathi Campus, Bangalore University, Bangalore 560056, India Received December 2, 2015; accepted June 22, 2016 Summary Urginea indica Kunth. Hyacinthaceae, commonly called Indian Squill, represents a species com- plex. Intraspecies variation is common. Karyological studies were made in six accessions of Urginea collected from six different places in India, of which four accessions of Urginea indica were collected from Kanakapura, Trichendur, Kashmir valley, and Kerala. Two accessions of Urginea wightii were from Gubbi and Bellary. Varia- tions in ploidy were noted. Among the Urginea indica accessions, diploids (2n=20) were found in Kanakapura, Trichendur, and Kashmir Valley; a pentaploid (2n=50) was noticed in the Kerala accession, whereas two dif- ferent ploidies were noticed in the Urginea wightii accessions of Gubbi and Bellary (2n=20, 2n=40) within the same plant. There is close homology between Kanakapura and Tiruchendur with subtelocentric being a major component. Key words Urginea, Karyotype, Accession, Diploid, Tetraploid, Pentaploid. Urginea genus belongs to the family Hyacinthaceae. placed Drimia in the Tribe Urgineae, Order Scillodeae, Urginea indica is also called Indian squill. Squill bulbs and Family Asparagaceae. Urginea is known for pheno- have long been used as a source of medicine with phar- typic and genotypic complexities, and variations in ac- maceutical and biocidal applications. There is mention cessions within species lead to the appearance of a wide of Indian squill in British and European pharmacopias. range of intermediate forms, which led to confusion Urginea indica accessions are unexplored from a cyto- taxonomically. taxonomic point of view. The species has been studied Several cytological studies have been conducted by by Shiva Kameshwari and Muniyamma (1999, 2001). Dixit and Yadav (1989), Deb and Dasgupta (1974), Jha Karyotype analysis of different accessions is very use- and Sen (1980, 1983a, b), and Shiva Kameshwari et al. ful in detecting the probable relationship, origin, and na- (2014) to solve these complexities. However, even to ture of chromosome variations. The present report deals this day, none of the studies have completely elucidated with karyomorphology of Urginea indica and Urginea the complexities. Thus, Urginea is a taxonomically, wightii. The karyotype differentiation between acces- genetically, and phenotypically complex genus. Further sions was elucidated. Urginea indica is an aggregate of detailed studies of Urginea are required, with a large diploid, tetraploid, aneuploid, pentaploid, and hexaploid collection of specimens from various localities to sup- cytotypes growing in different localities in India. Mor- port the research. In the present study, karyomorpho- phological and cytological differences have been detect- logical studies were made in U. indica (four accessions) ed within the accessions of U. indica (Shiva Kameshwari and U. wightii (two accessions) aiming to characterize et al. 2010). Urginea wightii is morphologically and these species cytologically and explore the pattern of cytologically distinct from Urginea indica. choromosomal differentiation and karyological varia- The plant is known for its cardiac glycosides (Mulhol- tions within and between the species. The basic number land et al. 2013). It is gaining global importance due to of Urginea according to Goldblatt et al. (2012) is X=10. its multiple properties as a medicine (Nath et al. 2014). According to Manning et al. 2004, U. indica and their Materials and methods accessions are controversial and need further resolution. The medicinal plant Urginea is separated from Bulbs of Urginea indica of four different accessions Urginea and included in Drimia (Jessop 1977, Stedje were collected from Kashmir, Kanakapura, Thiruchen- 1987). Molecular studies by Pfosser and Speta (1999) dur, and Kerala, and two accessions of Urginea wightii were collected from Gubbi and Bellary and grown in * Corresponding author, e-mail: [email protected] the department of Botany, Bangalore University, Banga- DOI: 10.1508/cytologia.81.329 lore, under uniform environmental conditions. Root tips 330 H. L. Geetha et al. Cytologia 81(3) Fig. 1. a–d. Somatic metaphase chromosomes and idiograms of accessions of U. wightii. a: Somatic metaphase chromosomes of Bellary accession showing 2n=40. b: Somatic metaphase chromosomes of Bellary accession showing 2n=20. c: Idiogram of Bellary accession 2n=40. d: Idiogram of Bellary accession 2n=20. Scale bar=10 µm. Table 1. Karyometric data in different accessions of Urginea indica Kunth. Total chromosome Sl. no. Place of collection Chromosome number Ploidy Karyotype formula Karyotype length in µm 1. Kanakapura 2n=20 Diploid St10 81.64 Asymmetrical 2. Trichendur 2n=20 Diploid St10 99.72 Asymmetrical 3. Kashmir 2n=20 Diploid Sm2+St7+T1 115.5 Asymmetrical 4. Kerala 2n=50 Pentaploid Sm2+St20+t3 301.51 Asymmetrical 5. Gubbi (U. wightii) 2n=20 Diploid St10 94.64 Asymmetrical 2n=40 Tetraploid Sm5+St15 222.45 6. Bellary (U. wightii) 2n=20 Diploid Sm5+St5 82.22 Asymmetrical 2n=40 Tetraploid Sm6+St14 189.08 were excised from potted plants between 9 : 30 a.m. to mosome numbers of all the accessions were recorded 11 : 30 a.m. and pretreated with 0.1% 8-hydroxy quino- (Table 1). Karyotypes were made for each accession line and kept at 4°C for 6 h and fixed in acetic alcohol following the methods employed by Levan et al. (1964). (3 : 1) for 24 h and stored in 70% alcohol. The roots were Symmetry and asymmetry have been determined ac- hydrolysed in 1 N HCl for 15 min and kept in feulgen cording to the system proposed by Stebbins (1958). stain for 1 h and squashed in 45% acetic acid. Temporary preparations for feulgen staining were made. The chro- 2016 Karyotypic Variations in Different Accessions of Urginea indica (Roxb) Kunth. and Urginea wightii Lakshmin. Hyacinthaceae 331 Fig. 2. a–d. Somatic metaphase chromosomes and idiograms of accessions of U. wightii. a: Somatic metaphase chromosomes of Gubbi accession showing 2n=40. b: Somatic metaphase chromosomes of Gubbi accession showing 2n=20. c: Idiogram of Gubbi accession 2n=40. d: Idiogram of Gubbi accession 2n=20. Scale bar=10 µm. Results and discussion Such inconsistency has been reported in over 240 species belonging to angiosperms by Nirmala and Rao The cytological variations obtained in the present (1996). Similar cases of polysomaty have been reported study provided a basis for recognizing them as distinct in growing shoot apex by Sharma (1956). cytotypes (Figs. 1–4). The four cytotypes of Urginea Karyotype asymmetry in different accessions of U. indica investigated revealed three diploids of 2n=20 and indica varies greatly due to different conditions of the one pentaploid of 2n=50. The karyotypes were asym- locality. Similar studies have been made by Wang et metrical in all the accessions, with subtelocentric be- al. (2013) in the ophiopogoneae tribe from Southwest ing the major component of the karyotype followed by China. submetacentric, terminal point, and finally telocentric Currently, we do not know how many more cytotypes chromosomes. These accessions are phenotypically dis- from the U. indica complex are present in South India. tinct types. The karyotype complementary shows that it Cytotaxonomic studies in the genus Urginea and karyo- should be diploid with the basic chromosome number of type variation in U. indica revealed by Oyewole (1987) X=10. Diploid and pentaploid accessions were detected have aided our understanding of karyotypic variations. in Urginea indica. The high frequency of diploid cyto- He reported that the difference in the chromatin material types is most common. is correlated with both morphological differences and In the Gubbi and Bellary accessions of Urginea ecological preferences. wightii, chromosomal variations were observed in the The main objective of this study is to identify the ploi- cells of the same root tip. Of the cells studied, 50% dy level of the Urginea accessions and to compare their showed 2n=20 chromosomes, while the remaining 50% morphological characteristics with plants collected from showed 2n=40 chromosomes. Such variations within the different localities of India with the same chromosome same tissue are termed chromosome mosaics. number or different cytotypes. The results obtained sug- 332 H. L. Geetha et al. Cytologia 81(3) Fig. 3. a–d. Somatic metaphase chromosomes and idiograms of accessions of U. indica. a: Somatic metaphase chromosomes of Kerala accession showing 2n=50. b: Somatic metaphase chromosomes of Kanakapura accession showing 2n=20. c: Idio- gram of Kerala accession 2n=50. d: Idiogram of Kanakapura accession 2n=20. Scale bar=10 µm. gest that the specific categories assigned to the bulbs col- terning might be involved in rearranging small gene lected from different localities should be autopolyploidy segments. Hence, U. indica seems to be a stable poly- and mixoploidy, which is reported in Gubbi and Bellary morphism in which the different forms have attained with 2n=20 and 2n=40 at equal frequency. reproductive isolation and genetic variability with each Each accession varies in their morphology and karyo- form retaining its morphological identity. According to type. The total length of chromatin material at meta- Oyewole