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AGROBACTERIUM MEDIATED TRANSFORMATION
OF SOYBEAN (GLYCINE MAX L.): SOME
CONDITIONS STANDARDIZATION

Abstract

Present study was aimed to standardize some transformation conditions for soybean cultivar NARC-4 using Agrobacterium tumefaciens strain EHA 101 harboring pGUSintNPTII. Soybean half seed cotyledonary node method was opted. It was found that explant preparation in Agro-suspension culture resulted in highest transformation efficiency (48.3%) than in infection medium and water. One hour infection time was found optimum (55.9% transformation efficiency) in culture OD600 1.0. Co-cultivation of soybean half seed explants with Agrobacterium for five days showed better results as compared with three and four days. Two hr washing in washing medium containing 1g/L cefotaxime controlled prevalence of Agrobacterium in further steps. It was found that varying kanamycin concentration in selection medium resulted in high survival rate of transformed shoots. NARC-4 soybean cultivar showed better transformation efficiency than NARC-7 when genotype dependency was examined using Agrobacterium strain EHA101.

Introduction

Plant genetic engineering mainly depends upon Agrobacterium tumefaciens mediated transformation that accounts so far production of 80% transgenics (Wang & Fang, 1998). Soybean transformation has shown significant improvement and enabled public and private sector for production of commercial cultivars with transgenic traits.
Hinchee et al., (1988) first time reported soybean transformation with Agrobacterium strain. They successfully regenerated plants on media containing kanamycin or glyphosate but transformation efficiency was quite low (Hinchee et al., 1988) but modifications in regeneration protocol may produce high rate of transformants (Parrott et al., 1989; McKenzie & Cress, 1992). Although a number of factors that affect on transformation efficiency has been studied that includes sonication assisted Agrobacterium mediated


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