【作者】 成卓敏； 何小源； 吴茂森； 陈彩层； 贾力； 田苗英； 刘太国； 张文蔚； 肖红； 周广和；

【Author】 Cheng Zhuo-min He Xiao-yuan Wu Mao-sen Chen Cai-ceng Jia LiTian Miao-ying Liu Tai-guo Zhang Wen-wei Xiao Hong Zhou Guang-he(Institute of Plant Protection, Chinese Academy of Agricultural Sciences, State KeyLaboratory for Biology of Plant Diseases and Insect Pests, Beijing 100094)

【机构】 中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室；

【摘要】 GPV是大麦黄矮病毒(barley yellow dwarf virus,BYDV)中国分离物,引起的小麦黄矮病是小麦上最重要的病毒病害。根据BYDV GPV的序列,我们应用RT-PCR获得了外壳蛋白基因和复制酶基因的cDNA。将cDNA插入到不同的质粒构建了其表达载体。通过花粉管通道法,建立了一个可快速、高效地获得抗BYDV的转基因小麦的技术体系。更多还原

【Abstract】 Barley yellow dwarf virus (BYDV) is the type member of the luteovirus group, and is considered to be of economic importance in most cereal-producing countries. So far, no natural forms of resistance have been described in common wheat.The cDNAs of the coat protein gene or replicase genes of BYDV GPV was obtained by RT-PCR and were inserted into different plasmids to construct expression plasmids named pPPI1 to pPPI20 based on different promoter. The recombinant plasmids were identified by molecular analysis to confirm real GPV coat protein gene or replicase gene cDNA in plasmids.An efficient pollen tube pathway-mediated transformation protocol was developed for the generation of transgenic wheat plants that express coat protein (CP) of BYDV GPV. Molecular analysis of the CP gene in transgenic plants confirmed the stable integration of the CP gene into the wheat genome and inheritance of the gene to T1,T2, T3 and T4 generations. Upon inoculation with GPV, transgenic plants that expressed the CP gene exhibited a significant delay in symptom development and reduced virus accumulation compared with control plants.Resistance evaluation of the T2 generation of transgenic wheat plants carrying GPV replicase gene, were carried out in both field and greenhouse conditions. Compared with non-transgenic plants, two transgenic lines of Shan 160 showed high level of resistance with no or very weak symptom on the leaf apex after virus infection.We have developed an improved protocol for the rapid and efficient production of transgenic wheat with resistance to BYDV by using no selectable markers and the pollen tube pathway to transfer the BYDV replicase gene into the common wheat varieties, then sowing transgenic seeds into the field for resistance screening and molecular assay. Transgenic wheat plants resistant to BYDV GPV were obtained.更多还原