摘要
本研究对基因枪导入了pBAC128F/R质粒(含有玉米Adh1内含子1的CaMV35S启动子驱动的bar基因作为选择标记,以来自拟南芥菜逆境诱导表达类型———亲水蛋白rd29b基因的启动子驱动脱水应答转录因子DREB1B基因的逆境诱导表达类型质粒)的T4代转基因小麦进行了稳定表达研究。PCR、PCR-Southern和Southern杂交分析表明,外源转录因子DREB1B基因已稳定整合到转基因株系的基因组中。半定量RT-PCR结果表明,部分转基因株系的DREB1B基因的相对表达量有所增强。叶片除草剂抗性检测结果显示有8个转基因株系可抗到150mg/L。叶片脯氨酸含量测定结果表明,有4个株系(编号为1,18,30和76)的脯氨酸含量提高幅度较大,同一株系,干旱与未干旱处理相比,脯氨酸含量提高了3～5倍。干旱处理后的转基因植株与非转基因植株相比,脯氨酸含量高2～3倍。在干旱条件下,T4代田间小区产量统计数据分析结果表明,有4个转基因株系(编号为30,51,70和76)的产量与非转基因植株相比有显著增加。研究表明,利用逆境诱导型rd29b基因的启动子来增强外源DREB1B基因的表达,能显著改良小麦的抗旱性。
Transgenic wheat offspring, T4 generation, with plasmid pBACI28F/R （7.0 kb）, which contain rd29b （induced by drought stress） promoter driven DREB1B gene, and CaMV35S promoter plus maize （Zea rnays L.） Adhl first intron driven bar gene were studied on expression stability. Analysis ofPCR, PCR-Southem and Southern blot for transgenic lines of T4 generation proved that exogenous DREB1B has been integrated into genome. And se- mi-quantitative RT-PCR analysis indicated that DREB1B was well expressed in part of lines. Herbicide resistance of 8 lines were more than 150 mg/L. Content of proline in leaves of T4 lines was determined and analysed. Content ofproline of 4 transgenic lines （number 1, 18, 30 and 76） exhibited increasing 2-3 folds as compared to that ofnon- transgenic plants under drought stress. In addition, content ofproline after drought treatment has been increased by 3-5 times as compared with that of non-drought treatment for the same one among the 4 transgenic lines. In drought stress condition, field yield data analysis of T4 generation indicated that yield of 4 transgenic plants （number 30, 51, 70 and 76） were obviously higher than negative control. Our research suggested that enhancement expression of alien transcriptional factors DREB1B driven rd29b promoter into wheat was an effective way to improve its drought tolerance ability.
出处
《分子植物育种》
CAS
CSCD
2009年第3期437-443,共7页
Molecular Plant Breeding
基金
北京市重大转基因育种平台项目(D08070503010000)
国家转基因生物新品种培育重大专项课题(2008ZX08002-002)共同资助
