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A simple and efficient method for obtaining transgenic soybean callus tissues
- Liu, Dong, Liu, Sheng, Chang, Dasong, Wang, Lu, Wang, Dan, Wang, Ning Ning
- Acta physiologiae plantarum 2013 v.35 no.7 pp. 2113-2125
- 2,4-D, Agrobacterium, Glycine max, antioxidants, benzyladenine, callus, explants, genes, genotype, hypocotyls, phenotype, screening, soybeans
- In the present study, a simple and efficient method for obtaining transgenic callus tissues of soybean [Glycine max (L.) Merr.] was developed based on Agrobacterium-mediated transformation. Hypocotyl segments of soybean were used as the starting material. Several factors such as soybean genotype, Agrobacterium concentration, inoculation time, co-cultivation period and addition of antioxidants in co-cultivation medium affecting the transformation efficiency were examined. The explants were cultured on callus induction medium containing 0.5 mg L⁻¹6-benzylaminopurine and 2.0 mg L⁻¹, 2,4-Dichlorophenoxyacetic acid for callus induction. Callus tissues were induced at both the acropetal and basipetal ends. CaMV35S::GUS and CaMV35S::GFP transgenic callus tissues were obtained using the optimized protocol. The average transformation efficiency reached up to 87.7 % based on GUS detection. From inoculation with Agrobacterium to obtaining transgenic soybean callus will take about 3 weeks. In order to validate this method for gene function investigation, GVG::GmSARK transgenic soybean callus tissues were obtained and their senescence-associated phenotypes were assessed. To our knowledge, this is the first report using hypocotyl segments as starting materials to obtain transgenic callus, and this system provides a method for high-throughput screening of functional genes of interest in transformed soybean callus.