Plant Tissue Cult. 10(2) : 137-142, 2000 (December)

Agrobacterium-mediated Genetic Transformation of Peanut (Arachis hypogaea L.)

R. H. Sarker, M. Nurul Islam, Aparna Islam and Zeba I. Seraj¹

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

Key words: Genetic transformation, Peanut, GUS expression, Kanamycin resistance, PCR amplification

Abastract

A protocol for direct in vitro regeneration as well Agrobacterium-mediated genetic transformation was developed for peanut (Arachishypogea L.) cv. Tridana Badam. Best multiple shoot regeneration was achieved from leaflet explants on MS medium containing 5.0 mg/1 BAP and 0.5 mg/1 Kn. Half strength MS medium, supplemented with 0.2 mg/1 of NAA was found suitable for root induction from the excised shoots. Transformation experiments using the same explant were conducted using Agrobacterium tumefaciens strain LBA4404 containing the binary plasmid pBI121 with GUS (?-glucuronidase) and NPT II (neomycin phosphotransferase) genes. Optimal conditions of infections were determined by transient GUS assays. Shoot regeneration did not however occur under selection pressure of 50 mg/1 kanamycin. Infected explants were therefore transferred to regeneration medium without kanamycin. Four to eight week-old shoots were then transferred to rooting medium containing 50 mg/1 Kanamycin. Roots developed from 85% of the regenerated shoots. Histochemical assay revealed the presence of the NPT II gene in leaf DNA was confirmed through polymerase chain reaction (PCR) analysis.

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