Plant Tissue Cult. 12(2) : 155-165, 2002 (December)

In vitro Plantlet Regeneration and Agrobacterium- mediated Genetic Transformation of Wheat (Triticum aestivum L.)

R. H. Sarker and Ashapurno Biswas

Department of Botany, University of Dhaka, Dhaka-1000, Bangladesh

Key words: Regeneration, Wheat, Transformation, Agrobacterium

Abastract

Different explants such as mature and immature embryos, seeds, endosperms, leaves, shoot bases and root tips of four local wheat (Triticum aestivum L.) varieties, namely Sourav, Gourav, Kanchan and Protiva were evaluated for their in vitro ability to callus and produce regenerants as well as their response to Agrobacterium- mediated genetic transformation. Among explants immature embryos were found to be the best for callus induction and shoot regeneration. Profuse callus developed, when MS was supplemented with 5.5 mg/1 2,4-D. The maximum shoot regeneration was obtained on MS medium, containing 0.5 mg/1 BAP, 0.5 mg/1 Kn and 25 mg/1 tyrosine. The majority of regenerated shoots (90%) developed roots on half strength of MS without any hormonal supplement. Plantlets were successfully established in soil. No significant variation was observed among the wheat varieties, regarding their ability for callus induction and shoot regeneration. Two strains of Agrobacterium , namely, LBA4404 containing pBI121 and EHA105 having plasmid pCAMBIA1301 were used in transformation experiments. Among the two strains, EHA105 was found to be compatible to the aforesaid four local varieties of wheat. The maximum transformation efficiency, monitored by transient GUS histochemical assay, was obtained in callus, derived from immature embryos with a bacterial suspension of optical density 0.75 at 600 nm.

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