外源λDNA导入普通小麦雄性不育变异的分子验证

MOLECULAR VERIFICATION ON MALE STERILE MUTANT AFTER INJECTED EXOGENOUS λDNA INTO WHEAT

将外源λDNA导入普通小麦品系 81 45 2 7,获得一细胞质雄性不育变异体 ,稳定成系 ,简称D型不育系 ,受体 81 45 2 7可作其保持系。用32 P标记的λDNA作为探针 ,对受体 81 45 2 7、D型不育系及其与鲁麦 1 4号的杂种F1 核DNA和叶绿体DNA进行点杂交。实验结果表明 :D型不育系及杂种F1 核DNA和叶绿体DNA点杂交均呈阳性反应 ,而受体无阳性反应。说明外源λDNA片段已经整合进了不育变异体核基因组和叶绿体基因组中 ,并且可以稳定遗传。

A cytoplasmic male sterile mutant and then a stable cms line named D type sterile line were obtained after injected exogenous λDNA into wheat line 814527,and line 814527 could be its maintainer line.By using λDNA labled with 32 P as probe,unlabled λDNA as positive check,dot blottings of nuclear DNA and chloroplast DNA of receptor 814527,D type sterile line and its hybrid F 1 with Lumai 14 were carried out.Positive dots appeared in nuclear DNA and chloroplast DNA of D type sterile line and its hybrid F 1,but did not appear in the receptor.It showed that fragments of exogenous λDNA existed in nuclear genome and chloroplast genome of D type sterile line,and could be inherited stably.All these results,on a molecular level,proved the reliability of exogenous DNA injection.