Blepharis sindica-T. Anders is a vulnerable species whose natural population is decreasing due to unscientific exploitation, scanty rainfall and habitat destruction. It has been notified as threatened species in the report of UNDP -CCF II Project and given A2cd Vulnerable category as per IUCN red list. So, this plant needs urgent efforts for conservation including both in situ and ex situ. In ex situ conservation the seeds were utilized first as an explant to develop complete seedling plants. Seeds were excised from fruit capsule after removing coat and inoculated in Basal modified MS medium supplemented with charcoal and ms medium supplemented with plant growth regulators, NAA, 2,4-D. Complete seedling plants and callus was achieved from seeds in basal modified MS and ms supplemented with 2,4-D, combination of 0.5mg/l NAA, 1.0mg/lBAP supplemented MS medium. Callus initiation and callus multiplication achieved from inoculation of cotyledons and subcultured callus parts. Callus initiation achieved by inoculation of cotyledons in combination of 0.5mg/l NAA, 1.0mg/l BAP, with only 1.0 mg/l 2,4-D supplemented MS medium. Multiplication of callus achieved from subculturing of callus parts in MS medium supplemented with 1.0mg/l&2.0 mg/l BAP. Enormous rooting and shooting along with leaf development showed on basal modified MS media. Better responses of rooting and shooting along with leaf development showed on Basal modified MS media supplemented with charcoal as compare to basal modified MS medium devoid of charcoal from seeds. Browning was also observed from cultures due to which callus appeared with brown colour after few days of subculture.

Tissue culture, Blepharis sindica, MS medium, Growth regulators

Through this research work it is concluded that addition 2,4-D or NAA and BAP in nutrient medium(modified ms) are responsible for callus initiation from seed and cotyledonary part of seedling plant. Due to hard coat around seeds in Blepharis sindica, it is not easy to initiate response from seed, so a cut by scalpel is required in coat to remove it and seed isolation. Basal MS with little change is sufficient to initiate and develop complete seedling plants with roots, epicotyl, hyhpocotyl and leaves in whorl of four which is necessary for its conservation due to its vulnerable status and typical habitat. Callus multiplication can be achieved with low concentration of BAP in ms medium from subculturing of callus parts and cotyledonary parts.

Rajneesh Sharma and Shikha Roy (2023). In vitro propagation of Blepharis sindica ex T.Anders. Biological Forum – An International Journal, 15(12): 05-08.