反义磷脂酶D γ基因转化小麦的研究

Transformation of Wheat with Anti-Phospholipase D γ Gene

目的:获得抗寒、耐盐等能力增强的小麦新种质或新品种。方法:利用穗茎注射法将含有反义磷脂酶Dγ基因(PLDγ)的根癌农杆菌菌液注入到春小麦富硒乌麦旗叶鞘内,T1代经田间表型选择筛选到10株变异株,通过特异PCR扩增和Southern杂交技术对获得的变异株进行分子检测,并对转基因植株进行耐盐性鉴定。结果:变异株均扩增出大小为430bp的目的片段,Southern印迹结果与其一致,证实反义PLDγ基因已经整合到小麦的核基因组中;6株转基因植株的抗寒力比对照显著增强,4株的熟期提早3～4d;在室内耐盐性筛选中,2株变异植株耐NaCl的能力比对照有不同程度的提高。结论:反义PLDγ基因在提高小麦的抗寒、耐盐性上具有很大的应用价值。

Objective： To enhance the abilities of tolerance adversity as salt and cold injury of wheat. Methods： Antisense phospholipase D γ（PLDγ） gene was introduced into wheat Fuxi Wumai mediated by earstem injecting. 10 individual plants were obtained from T1 generation by selecting phenotype, and variant plants were detected by PCR and Southern test. Then seeds of transgenic plants selected in MS containing NaCI. Results： A 430 bp fragment was amplified from variant plants by PCR analysis, which accorded with the result of Southern test, so it was primarily confirmed that anti- PLDy gene had been integrated into wheat genome. 6 transgenic plants became more tolerant freezing injury and the mature period of 4 plants were 3 to 4 days shorter than that of receptor. Salt-tolerance test demonstrated that 2 transgenic plants were enhanced to some extent. Conclusion： It was obvious that anti-PLDy had great value in enhancing cold resistance and salt tolerance of wheat.