Pib基因编码区功能的转基因初步鉴定

Initial determination of function for Pib gene ORF region by rice transformation

将Pib结构基因克隆到双元载体,构建了3个植物表达载体pNAR501、pNAR502和pNAR503,这3个载体分别携带由35S驱动Pib结构基因的外显子2外显子3、5′端部分缺失外显子1和5′端部分缺失的外显子1外显子2外显子3的不同片段。应用农杆菌介导法获得水稻品种日本晴转基因植株30株,经PCR、Southern b lot和后代潮霉素抗性分离检测确定外源基因已整合进水稻基因组。T0代转基因植株分蘖期大苗离体叶片抗稻瘟病鉴定结果表明,含3个不同载体的转基因植株对稻瘟病E1、F1、G1生理小种的抗性都显著高于对照日本晴。T1代3至4叶期幼苗活体接种鉴定结果与T0代离体叶片鉴定结果不同,对E1、F1、G1生理小种的抗性低于对照日本晴。

Three plant binary expression vectors pNAR501, pNAR502 and pNAR503 were constructed which carried exon 2-exon 3, 5＇partial deletion exon 1 and 5＇partial deletion exon 1 -exon 2-exon 3 of Pib gene driven by 35S separately. These three vectors were transformed into Nipponbare by Agrobecterium mediated method and 30 transgenic rice plants were obtained and confirmed by PCR, Southern blot and hygromycin resistance segregation in their progeny. Rice blast resistance test for in vitro leaves of To transgenic plants in tillering stage showed higher resistance than that of control Nipponbare to E1, F1, G1 races. But the results of rice blast resistance test for Tl transgenic plants in 3 to 4 leaves stage were different. All T1 transgenic plants had lower level of resistance than that of control Nipponbare to E1, F1, G1 races of rice blast.