Journal of Biotechnology & Biomaterials
Open Access

Abstract

Development of an efficient plant regeneration system is a prerequisite for any plant tissue culture approach for crop improvement. The work was undertaken to develop a regeneration system for B. juncea genotypes RH 406 and RH 555 using seedling explants. Hypocotyls and cotyledons from 5-day old seedlings were used as explants. Hypocotyl and cotyledon explants excised from in vitro grown 5-day old seedlings were cultured on MS medium supplemented with different concentrations and combinations of growth regulators. Highest percent shoot formation was observed on MS medium with 2.5 mg/l BAP (64.85�±1.42) from cotyledon explants in genotype RH 406. Highest shoots per explant were obtained on MS medium fortified with 2.5 mg/l BAP in genotype RH 406 and in genotype RH 555; it was obtained on MS medium supplemented with 1.0 mg/l BAP. Eight rooting media were tried for root formation in regenerated shoots. Maximum rooting response was obtained on MS medium supplemented with 0.2 mg/l NAA i.e., 93.44�±0.80 and 90.67�±1.45 in genotypes RH 406 and RH 555 respectively. The regenerated plants were transferred to a mixture of sand: Soil in 1:1 ratio in pots. Percent survival obtained was higher in genotype RH 406 i.e., 81.8% as compared to RH 555 where it was 67%. Transformation protocol was developed in genotype RH 406 as hypocotyls and cotyledons explants of this genotype showed GUS assay test positive. Cotyledon explants showed 75% while hypocotyl explants showed 80% GUS expression. Hence, regeneration and Agrobacterium mediated transformation conditions were developed.

Biography

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