水稻抗黑条矮缩病转基因株系的鉴定分析

发布时间：2018-08-04 18:22

【摘要】：水稻黑条矮缩病毒(Rice black streaked dwarf virus, RBSDV)是一种水稻病毒病,该病极具暴发性、间歇性和迁移性,对我国水稻安全生产造成了极大威胁。本研究以RNAi (RNA interference)技术为依托,针对RBSDV基因组构建RNAi载体,以高感RBSDV的水稻品种武陵粳1号为转化受体材料,通过对转化植株进行PCR和qRT-PCR的转基因阳性鉴定及表达量测定以及灰飞虱传毒表型鉴定,以期获得抗RBSDV的转基因株系,为水稻抗RBSDV育种提供抗性基因资源及种质。结果如下：1.对构建的RNAi载体转化得到的转基因T1代植株进行阳性鉴定与表达量分析。2.对pANDA-S8, pANDA-S9和pANDA-S8S9的转基因材料加代繁殖,获得了T3代纯系。3.通过qRT-PCR表达量分析,选取了高表达量的转基因阳性株系进行RBSDV传毒实验,并进行田间表型抗性分析。初步实验结果表明,沉默S8、S9以S8S9嵌合片段的转基因材料对RBSDV具有抗性,且嵌合片段的抗性更好。
[Abstract]:Rice black stripe dwarf virus (Rice black streaked dwarf virus, RBSDV) is a kind of rice virus disease, which is extremely sudden, intermittent and migratory, which poses a great threat to the safe production of rice in China. In this study, based on RNAi (RNA interference) technique, RNAi vector was constructed according to RBSDV genome, and Wulingjing 1, a rice variety with high susceptibility to RBSDV, was used as the transformant receptor material. In order to obtain the transgenic lines resistant to PCR and qRT-PCR, the transgenic gene resources and germplasm for RBSDV resistant breeding of rice were obtained through the identification of PCR and qRT-PCR transgenic positive expression and the identification of transfered phenotype of planthopper. The result is as follows: 1. Positive identification and expression analysis of T 1 transgenic plants transformed by RNAi vector. 2. 2. The transgenic materials of pANDA-S8, pANDA-S9 and pANDA-S8S9 were propagated and the T3 generation pure line. 3 was obtained. Through the analysis of qRT-PCR expression, transgenic positive lines with high expression were selected for RBSDV transmission experiment, and field phenotypic resistance analysis was carried out. The preliminary results showed that the silenced S8 S9 was resistant to RBSDV with S8S9 chimeric fragments, and the chimeric fragments were more resistant to RBSDV.
【学位授予单位】：湖南农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S435.111.4

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