International Research Journal of Biotechnology

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Study on the development of salt tolerant kenaf varieties through Agrobacterium-mediated genetic transformation

Abstract

S. K. Bhajan, N. Biswas, N. M. R. B. Rahman, M. M. Hasan, S. K. Saha, and A. Khatun

Cotyledons with attached petioles of two kenaf varieties were cultured on MS medium supplemented with 3.0 mg/l BAP and 0.5 mg/l IAA after co-cultivation with salt and drought tolerant Agrobacterium vector LBA-4404 (pBI121CIPKsense). Variety HC-2 showed higher percentage of shoot regeneration (80%) than variety HC-95. The regenerated shoots of both of the kenaf varieties showed resistance against kanamycin (50 mg/l). Non-transformed shoots became albino and died within 6 weeks. GUS histochemical assay was performed for the kanamycin resistant shoots. Maximum of 90.0% GUS positive explants were found from the variety HC-2 and variety HC-95 showed 80.0% GUS positive response. In both varieties, the stable GUS gene integration was proved by PCR amplified DNA band using GUS specific primers set. MS medium was supplemented with different concentrations of NaCl e.g. 25, 50, 75, 100 and 125 mM. Transformed shoots were able to survive upto 100 mM NaCl. Non transformed shoots died on salt containing medium. Salt tolerant transgenic shoots were rooted on hormone free MS medium and transferred to soil. After maturity these plants produced flowers and seeds.

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