Of. Ossnac1 Gene On Early Japonica Kongyu The 131 Genetic Transformation

Concentration of saline-alkaline have imoprotent effect on rice growth, today stress resistance has long been targeted in designing better crops. In this research use three type of vector: pCAMBIA1301s-SNAC1, p1301U-CIPK15 and pC2003H-SNAC1 were transformed into rice Kongyu131 (Oryza sativa L.) nuclear genome via the Agrobacterium tumefaciens-meidiated method. Exression level, saline-alkaline tolerence and copy number of T-DNA were analyzed by RT-PCR and Southern blotting respectively. The main results in this study were as follows:1. The vector pCAMBIA1301s-SNAC1, pC2003H-SNAC1 and p1301U-CIPK15 were confirmed in E.coli DH5αwith restriction enzyme digestion and PCR detection.2. 36 independent resistant calli were regenerated from obtainable which the rice Kongyu131 transformed with SNAC1 gene of the vector pCAMBIA1301s-SNAC1. 42 living transgenic plants were obtained. 12 independent resistant calli were regenerated from the transformation of vector pC2003H-SNAC1. 5 living transgenic plants were obtained. 15 independent resistant calli were regenerated form transformation with CIPK15 gene. 31 living transgenic plants were obtained.3. PCR detection indicated that the SNAC1 gene and CIPK15 gene were integrated into the genome of T0 generation of the rice variety Kongyu131.4. Evaluation for saline-alkalng tolerance through added 100mmol·L-1NaHCO3, the result showed that T1 generation of transgenic rice had higher ability of saline-alkalng tolerance.5. Analysis of Southern bloting and RT-PCR showed that SNAC1 gene and CIPK15 gene stable expression in T1 generation of the rice variety Kongyu131.