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Studies on Agrobacterium-mediated genetic transformation of embryogenic suspension cultures of sweet potato
Abstract
In this study, genetic transformation of embryogenic suspension cultures of sweet potato (Ipomoea batatas) cultivar Xu55-2 was conducted utilizing the Agrobacterium tumefaciens strain EHA105 that
contains the binary vector pBIN19/SBD2 with SBD2 (starch binding domain 2) gene and neomycin phosphotransferase (NPT II) gene. The presence of the SBD2 gene in the genomic DNA of transgenic
plants was verified by PCR amplification and confirmed by Southern blot analysis. Results suggested that cefotaxime (Cefo), at the concentration of 200 mg/L, was able to effectively suppress the growth of
Agrobacterium after co-cultivation. The optimal concentration for kanamycin (Kan) was 10 mg/L for selecting resistance calli, somatic embryo formation and plant regeneration. The highest frequency of
shoot induction (30.9%) was obtained on the MS medium containing 10 mg/L Kan, 200 mg/L Cefo, 1.0 mg/L abscisic acid (ABA) and 1.0 mg/L gibberellic acid (GA3).
contains the binary vector pBIN19/SBD2 with SBD2 (starch binding domain 2) gene and neomycin phosphotransferase (NPT II) gene. The presence of the SBD2 gene in the genomic DNA of transgenic
plants was verified by PCR amplification and confirmed by Southern blot analysis. Results suggested that cefotaxime (Cefo), at the concentration of 200 mg/L, was able to effectively suppress the growth of
Agrobacterium after co-cultivation. The optimal concentration for kanamycin (Kan) was 10 mg/L for selecting resistance calli, somatic embryo formation and plant regeneration. The highest frequency of
shoot induction (30.9%) was obtained on the MS medium containing 10 mg/L Kan, 200 mg/L Cefo, 1.0 mg/L abscisic acid (ABA) and 1.0 mg/L gibberellic acid (GA3).
