Assessment of the efficiency of hairy roots induction using soybean, sugar beet and tobacco explants

Document Type: Research Paper

Authors

Institute of Biotechnology, Shiraz University, Shiraz, Iran.

Abstract

Agrobacterium-mediated gene transfer method is one of the used methods for genetic transformation in the plant regeneration program. Transformation efficiency can be optimized depending on the strain of bacteria, the genotype of plant and conditions of growth. In this study, the gfp gene was transferred into sugar beet, tobacco, and soybean by Agrobacterium rhizogenes strain AR15834. The effects of bacterial concentrations, antibiotic concentrations and the types of explants and genotypes on the gene transfer efficiency and transgenic hairy roots production were investigated. The explants were inoculated with the bacteria at the adjusted concentrations and two days after the transformation, the explants were transferred to a solid MS medium containing different concentrations of kanamycin antibiotic. According to the results and the examined factors, the optimal conditions to achieving of the maximum production of transgenic hairy roots included bacterial concentration with OD600 = 0.2, cotyledon explant, 50 mg/L kanamycin concentration and Djakel genotype for soybean; bacterial concentration with OD600 = 0.2, leaf with petiole explant and SBSI004 genotype for sugar beet, and bacterial concentration with OD600nm = 0.8 and 100 mg/L kanamycin concentration for tobacco.

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Main Subjects


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