Establishment Of The Regeneration System And Agrobacterium-mediated Transformation With Antisense AcInv Gene

Reducing sugar in potato(Solanum tuberosum L.) tubers will accumulate during storage in low temperature , which leads to starch content decreased, and dark color of chips when fried in high temperature. It seriously affects appearance quality and edible quality of the potato products. Therefore, it is a very important object to cultivate new lines which resist to low-temperature sweetening. Introducing AcInv antisense gene into potato cultivars to restrain itself AcInv offspring for reducing sugar content is a very important approach. Studies on regeneration and agrobacterium-mediated transformation system were carried out with 6 potato varieties cultivated by Gansu academy of agricultural science.Through this system an antisence AcInv gene were introduced into potato. The main results were showed as following:1. Regeneration medium of 6 potato varieties was screening. The optimal medium for callus induction from stem was MS+6-BA 2.5mg/L+NAA 0.5mg/L+GA3 5mg/L+2,4-D 1mg/L for"Longshu No.3","Longshu No.4"and"L0031-17", MS+6-BA 2.5mg/L+NAA 0.5mg/L+GA3 2.5mg/L+2,4-D 0.5mg/L for"Longshu No.5"and"Longshu No.6"and MS+6-BA 2.5mg/L+NAA 1mg/L for"LK99"; The optimal medium for adventitious bud differentiation from stem-derived calluses was found to be MS+6-BA 3.5mg/L+NAA 1mg/L+GA3 10mg/L for"Longshu No.3","Longshu No.6"and"L0031-17", MS+6-BA 3.5mg/L+NAA 1mg/L+GA3 2.5mg/L for"Longshu No.4"and"Longshu No.5"and MS+6-BA 1.5mg/L+NAA 0.5mg/L+GA3 10mg/L for"LK99"; and the optimal regeneration medium for tube chip was MS+ZT 2mg/L+IAA 1mg/L. Among six varieties,"Longshu No.3","Longshu No.5"and"Longshu No.6"have strong ability of regeneration and"LK99"and"L0031-17"have weak ability of regeneration. Analyzing the effects of different recipetent material on regeneration, tube chip is better than stem cutting.2. Studies on agrobacterium-mediated transformations showed that the transformation frequence of stem cutting and microtube chip from 6 varieties by no-pre-culture was higher than that by pre-culture.Different explant types needed different time of infection and co-culture.For the stems explant,the optimal time of infention was...