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In Vitro Androgenesis in Anther Cultures of Crateva nurvala Buch. Ham.
Neetika Walia1, Suman Sharma2, Sadhana Babbar3* and Shashi B. Babbar1*
1Department of Botany, University of Delhi, Delhi-110007, India
2Ramjas College, University of Delhi, Delhi-110007, India
3Swami Shraddhanand College, University of Delhi, Delhi-110036, India
ABSTRACT
The anthers of Crateva nurvala, at late uninucleate to early binucleate stages of microspore development, were cultured on Murashige and Skoog’s basal medium (BM) supplemented with different concentrations of sucrose, 6-Benzylaminopurine (BAP, 0.5 or 1.0 mg l-1) and/or 2,4-Dichlorophenoxyacetic acid (2,4-D, 0.5 mg l-1). Among the media tested, high frequency of callusing was observed from the anthers cultured on BM adjuvated with 2,4-D alone or along with BAP. The calli were repeatedly sub-cultured on BM adjuvated with 1-6 mg l-1 BAP at intervals of one month each. Three shoots differentiated from two of the calli during the fifth sub-culture on the medium adjuvated with 3 mg l-1 BAP and one during the sixth passage on the medium fortified with 4 mg l-1BAP. The regenerated shoots exhibited differences in their growth characteristics; especially the size of leaves; two being slow growing with smaller leaves. These shoots were individually multiplied using their nodal explants. Up to the fourth passage, cloned shoots maintained the original phenotype of the shoots from which they had originated. DNA content of the four clones was estimated by flow cytometry. Two of these clones had DNA content comparable to the diploid, while other two had near haploid DNA content, indicating the possible origin of these clones from microspores. This inference was further strengthened when mean guard cell size of the clones with near-haploid contents was found to be smaller than those of the mature tree and in vitro raised seedlings. Though, the ultimate aim of regenerating haploid plants of C. nurvala was not realized, flow cytometric and stomatal size analyses strongly suggested androgenic origin of two shoots.
Key words: Androgenesis, Anther culture, Crateva nurvala
*Authors for correspondence: Sadhana Babbar, sadhanababbar@gmail.com; SB Babbar, sbbabbar@gmail.com
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