RPW2表达载体构建及对烟草的遗传转化  

CONSTRUCTION OF RPW2 EXPRESSION VECTOR AND ITS GENETIC TRANSFORMATION IN TOBACCO

作　　者：何道一[1] 段培树 陈丽萍[1] 

机构地区：[1]淮北煤炭师范学院生命科学学院,资源植物生物学安徽省重点实验室,安徽淮北235000

出　　处：《核农学报》2009年第4期602-605,共4页Journal of Nuclear Agricultural Sciences

基　　金：安徽省自然科学基金(070411008)

摘　　要：将RPW2基因插入表达载体pBI121的表达盒中,构建了RPW2组成型表达载体。通过农杆菌叶盘转化法转化烟草叶片,经卡那霉素筛选、PCR扩增和Southern杂交鉴定证明,RPW2基因已成功转入烟草细胞中,获得6个转基因株系。RT-PCR半定量分析表明,转基因烟草株系的叶片中有RPW2基因的表达。不同转基因株系的基因表达量不同,转基因株系T-R2和T-R4表达量较高。转基因植株叶片上人工接种烟草赤星病菌液检测转基因植株的抗病性,结果表明,与对照植株相比,转RPW2基因植株对烟草赤星病抗性显著增强,说明RPW2基因在烟草中也具有抗病能力。另外,转基因株系间RPW2基因表达量虽有差异,但抗病性与其他株系相比没有显著差异。RPW2 gene was cloned from a Triticum aestivum-Elytrigia elongatum aline substitution line,and the RPW2 expression vector was constructed by inserting RPW2 into the expression box of vector pBI121.Leaf disease of tobacco were transformed by Agrobacterium tumefacien and the transformed plants were selected by kanamycin.And then,6 transformed plants were identified by PCR and southern blot.RT-PCR semi-quantitative analysis results indicated that the RPW2 expressed in leaves of all transformed plants,however,the expression levels differed with different tansformed plants, high expression in the transformed T-R2 and T-R4. Resistance to Ahernaria alternata in the transformed plants of tobacco with RPW2 was significantly increased comparing with the check plants. The RPW2 transformed plants which were characterized by different expression levels of the foreign gene showed no difference in resistance to Ahernaria alternata of tobacco.

关 键 词：RPW2基因 转化 烟草 抗烟草赤星病 

分 类 号：S572[农业科学—烟草工业] S572.01[农业科学—作物学]