转cry1C~*基因抗虫超级粳稻田间目的基因表达及抗螟虫性研究

Target Gene Expression in the Insect-resistant Super Japonica Rice Transformed with cry1C~* and Stem Borer Resistance in Field

为了研究转基因抗虫超级粳稻田间目的基因表达及抗螟虫性,将10个来自于不同独立抗性愈伤组织的转cry1C*基因抗虫超级粳稻品系种植于田间,利用实时荧光定量PCR方法检测孕穗期叶片、茎鞘和幼穗等器官目的基因mRNA,利用酶联免疫吸附(ELISA)法检测孕穗期叶片、茎鞘和幼穗及收获后糙米的Cry1C蛋白,利用田间目测调查二化螟危害的白穗率。结果显示,转基因超级粳稻不同品系及不同器官cry1C*基因田间表达量明显不同,cry1C*基因mRNA表达量:叶片>茎鞘>幼穗,蛋白质表达量:叶片>茎鞘>幼穗>糙米。转基因超级粳稻田间目的基因表达,在mRNA水平和蛋白质水平,不同器官间存在正相关关系,各器官Cry1C蛋白质含量和糙米Bt蛋白质含量呈正相关。在本研究范围内,不论转基因粳稻植株Cry1C蛋白质含量高或低的品系,田间均表现为高抗二化螟。培育转基因抗虫粳稻品种时,应注意对目的基因适量表达的抗虫基因型的选择。

To study the target gene expression and stem borer resistance in transgenic insect-resistant super Japonica rice in field, 10 super rice strains with insect-resistant genes Japonica rice transformed with cry1C＊ from different independent resistant callus were grown in field. The mRNA of target gene in rice leaf, stemsheath and young panicle at booting stage were detected by real-time PCR. The Cry1C protein in rice leaf,stem sheath, young panicle at booting stage and in brown rice after harvest were detected by enzyme-linked immunosorbent assay（ELISA）. The white head plants damage of Chilo suppressalis was investigated by visual inspection in field. The results showed that the expression of cry1C＊ in different organs and different transgenic super Japonica rice strains was obviously different. The quantities of mRNA expression of cry1C＊ gene followed an order： leafstem sheathyoung panicle, and the quantities of cry1C＊ protein followed an order：leafstem sheathyoung panicle brown rice. The positive correlation of target gene expression in transgenic super Japonica rice existed in different organs at mRNA and protein levels. The positive correlation existed between Cry1C protein content in organs and Bt protein content in brown rice. The stains, no matter with highor low Cry1C protein contents, had high resistance to Chilo suppressalis in filed in our study. The insectresistant genotype which properly expressed target genes should be paid attention in breeding transgenic insect-resistant Japonica varieties.