Ye Rongjian, Huang Haiqun, Yang Zhou, Chen Taiyu, Liu Li, Li Xianghua, Chen Hao, Lin Yongjun
National Key Laboratory of Crop Genetic Improvement and National Centre of Plant Gene Research, Huazhong Agricultural University, Wuhan 430070, PR China.
Pest Manag Sci. 2009 Sep;65(9):1015-20. doi: 10.1002/ps.1788.
Yellow stem borer (Tryporyza incertulas Walker), striped stem borer (Chilo suppressalis Walker) and leaf folder (Cnaphalocrocis medinalis Guenec) are three lepidopteran pests that cause severe damage to rice in many areas of the world. In this study, novel insect-resistant transgenic rice was developed in which Bt protein expression was nearly absent in the endosperm. The resistant gene, cry1C*, driven by the rice rbcS promoter (small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase), was introduced into Zhonghua 11 (Oryza sativa L. ssp. japonica) by Agrobacterium-mediated transformation.
A total of 83 independent transformants were obtained, 19 of which were characterised as single-copy foreign gene insertion. After preliminary screening of the T(1) families of these 19 transformants in the field, six highly insect-resistant homozygous lines were selected. These six homozygous transgenic lines were field tested for resistance to leaf folders and stem borers, and for their agronomic performance. The Cry1C* protein levels in leaves and endosperm were measured by ELISA. Subsequently, the elite transgenic line RJ5 was selected; this line not only possessed high resistance to leaf folders and stem borers, normal agronomic performance, but also Cry1C* expression was only 2.6 ng g(-1) in the endosperm.
These results indicated that RJ5 has the potential for widespread utility in rice production.
二化螟(Tryporyza incertulas Walker)、三化螟(Chilo suppressalis Walker)和稻纵卷叶螟(Cnaphalocrocis medinalis Guenec)是三种鳞翅目害虫,在世界许多地区对水稻造成严重危害。在本研究中,开发了一种新型抗虫转基因水稻,其胚乳中几乎不存在Bt蛋白表达。由水稻rbcS启动子(核酮糖-1,5-二磷酸羧化酶/加氧酶小亚基)驱动的抗性基因cry1C*,通过农杆菌介导的转化被导入到中华11(Oryza sativa L. ssp. japonica)中。
共获得83个独立转化体,其中19个被鉴定为单拷贝外源基因插入。在田间对这19个转化体的T(1)家系进行初步筛选后,选择了6个高抗虫纯合系。对这6个纯合转基因系进行了对稻纵卷叶螟和螟虫的抗性以及农艺性状的田间测试。通过ELISA测定叶片和胚乳中的Cry1C蛋白水平。随后,选择了优良转基因系RJ5;该品系不仅对稻纵卷叶螟和螟虫具有高抗性,农艺性状正常,而且胚乳中Cry1C表达量仅为2.6 ng g(-1)。
这些结果表明RJ5在水稻生产中具有广泛应用的潜力。