Genotype-Independent Transformation of Italian Ryegrass Using Agrobacterium tumefaciens

摘要

In the present study, genotypic variation of Agrobacterium-mediaied transformation of Korean Italian ryegrass has been evaluated. Mature seedderived calli of a total of seven cultivars were infected and co-cultured with Agrobacterium tumefaciens carrying the binary vector pCAMBIA1301, which contains a reporter gene (gus) and a plant selectable marker gene conferring resistance to hygromycin (hpt) in the T-DNA region. The effects of several factors such as callus type and callus age on transformation frequency and the expression of the GUS gene were investigated. The highest transformation frequency (6.7%) was obtained with the Hwasan 101 cultivar when 9-week-old calli (type-Ⅰ) were inoculated with Agrobacterium. The overall transformation rates of the examined cultivars ranged from 0.4% to 6.7%. GUS histochemical assays, PCR, and southern analysis of transgenic plants demonstrated that transgenes were successfully integrated into the genome of Italian ryegrass. Thus, optimization of transformation frequency and selection of a suitable cultivar of Italian ryegrass may improve molecular breeding of this species.