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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。BC2F3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 transgenic Bt insect-resistant rice varieties, Bt genes cry1C *, cry2A * and cry1Ac / Ab in three Bt Minghui 63 were transferred to rice variety Xiushui 123, Xiangqing, 3 and Shanghai-Harbin 1B. Using the second generation BC2F3 backcrossing rice leaves as material, molecular marker-assisted selection, field herbicide screening and test strip detection were used to obtain new transgenic Bt lines. In the whole growth period, the field insect resistance identification methods of the transgenic plants except the non-sprayed pesticides except the pesticide were screened out for the lines with good insect resistance and agronomic traits to provide materials for subsequent insect-resistant breeding. BC2F3 generation of PCR detection and strip test results show that cry1C * and cry2A * gene were Shanghai drought 1B, Xiushui123 and Xiangqing homozygous lines. At the same time, the transgenic lines showed strong resistance to rice leafroller rice stem borers under natural insect-induced conditions in the field.