Studies On Shoot Regeneration From Leaf Pieces In Vitro And Phytase Genetic Transformation Of Raspberry

Taking six raspberry cultivars as tested materials, this paper studied the factors affecting the adventitious shoot regeneration from leaf pieces in vitro, resulting in the establishment of a stable and efficient adventitious shoot regeneration system of raspberry clone B.It was discovered through experiment that genotype was the major restrictive factor affecting shoot regeneration from leaf disk in vitro . Among the 6 genotypes tested, clone B had the strongest ability in shoot regeneration with shoot induction frequency of 90% and mean number 12.0 shoots per leaf. Phytohormone type, concentration and their combinations in medium were the most important factors influencing shoot regeneration from leaf disk of raspberry . The phytohormone combination BA 2.0 mg/L and IBA 0.1 mg/L had the best effect on shoot induction frequency , mean number of shoots per leaf and shoot quality in our experiment. The 21-30 days old leaf had the highest shoot induction frequency and the highest mean number of shoots per leaf. MS medium was the most suitable basal medium for shoot induction. Dark and 3000 Lux light intensity were harmful to regenerate shoots .The paper also studied the factors affecting the phytase genetic transformation -mediated by Agrobacterium tumefaciens , we obtained the antibiotic plant of raspberry clone B and tobacco clone A . Results of PCR of the tobacco clone A showed that phytase gene had integrated into the genome of the tobacco clone A ,and was expressed properly. But raspberry clone B was failed . We gave several reasons about the failure of phytase genetic transformation of raspberry clone B.