【摘要】：水稻黑條矮縮病毒(Rice black streaked dwarf virus, RBSDV)是一種水稻病毒病,該病極具暴發(fā)性、間歇性和遷移性,對我國水稻安全生產(chǎn)造成了極大威脅。本研究以RNAi (RNA interference)技術(shù)為依托,針對RBSDV基因組構(gòu)建RNAi載體,以高感RBSDV的水稻品種武陵粳1號為轉(zhuǎn)化受體材料,通過對轉(zhuǎn)化植株進(jìn)行PCR和qRT-PCR的轉(zhuǎn)基因陽性鑒定及表達(dá)量測定以及灰飛虱傳毒表型鑒定,以期獲得抗RBSDV的轉(zhuǎn)基因株系,為水稻抗RBSDV育種提供抗性基因資源及種質(zhì)。結(jié)果如下：1.對構(gòu)建的RNAi載體轉(zhuǎn)化得到的轉(zhuǎn)基因T1代植株進(jìn)行陽性鑒定與表達(dá)量分析。2.對pANDA-S8, pANDA-S9和pANDA-S8S9的轉(zhuǎn)基因材料加代繁殖,獲得了T3代純系。3.通過qRT-PCR表達(dá)量分析,選取了高表達(dá)量的轉(zhuǎn)基因陽性株系進(jìn)行RBSDV傳毒實(shí)驗(yàn),并進(jìn)行田間表型抗性分析。初步實(shí)驗(yàn)結(jié)果表明,沉默S8、S9以S8S9嵌合片段的轉(zhuǎn)基因材料對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.
【學(xué)位授予單位】：湖南農(nóng)業(yè)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2016
【分類號】：S435.111.4

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