为培育转Bt基因抗虫水稻品种材料,以传统杂交的方法将3个Bt明恢63中的Bt基因cry1C*、cry2A*和cry1Ac/Ab,转育到水稻品种秀水123、湘晴、旱恢3号和沪旱1B中;以回交二代自交三代(BC2F3)水稻叶片为材料,利用分子标记辅助选择、田间除草剂筛选和试纸条检测方法,以获得新型转Bt基因株系。并在全生育期实行除不喷施农药外的正常田间管理的转基因植株的田间抗虫鉴定方法筛选出抗虫性良好、农艺性状优良的株系,为后续抗虫育种提供材料。 BC2 F3代PCR检测及试纸条检测的结果显示分别获得cry1C*和cry2A*基因的沪旱1B、秀水123和湘晴的纯合株系。同时,在田间自然诱发虫害条件下,转基因株系对稻纵卷叶螟虫表现出强抗性。
In order to breed trans-Bt gene rice materials with pest resistance,three Bt genes,cry1Ac/Ab, cry1C* and cry2A*,from the donor parents developed by Huazhong Agricultural University were introgressed into Xiushui123,Xiangqing,Hanhui No.3 and Huhan1B,namely Minghui63( Cry1C*) ,Minghui63( Cry2A*) and Minghui63 ( Cry1Ab/Ac) respectively using the method of traditional hybrid;then the new type lines of trans-Bt gene rice were by obtained means of maker-assisted selection ( MAS) ,screening of basta test in the field and test paper strip detection.In addition trans-gene rice lines of good pest resistance and fine agricultural traits were scteened by using the assays of insect-resistance in the field under normal field management but without spraying pesticide in the whole bearing stage of these trans-gene lines providing the materials for fur-ther anti-pest breeding.The homozygous lines with cry1C* and cry2A were obtained in the different back-grounds ( Xiushui123,Xiangqing and Huhan1B) respectively by the detection of PCR and test paper strip.At the same time,Bt lines showed excellent resistance to rice leaf roller in the field under the condition of its natu-ral occurrence.