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本研究采集健康马铃薯块茎和被PVY侵染的马铃薯块茎为材料,选取马铃薯RCCR基因和LSD1基因为目标基因。首先提取总RNA后通过反转录PCR(RT-PCR)、琼脂糖凝胶电泳,证明RCCR基因和LSD1基因成功获得,然后进行荧光定量PCR(qPCR)测定在PVY侵染的马铃薯块茎中RCCR基因和LSD1基因的相对表达量,最后又进行了目标基因编码蛋白的结构分析。结果表明,这2个基因的相对表达量均有所下降,推测这2个基因与马铃薯块茎细胞坏死相关,为之后研究PVY引起马铃薯块茎细胞坏死的相关分子机制做了前期探索,为今后的分子育种和抗病药物筛选提供了可选的靶点。
In this study, we collected healthy potato tubers and potato tubers infected with PVY, and selected the potato RCCR gene and LSD1 gene as the target genes. The total RNA was first extracted and then verified by reverse transcription PCR (RT-PCR) and agarose gel electrophoresis to confirm that the RCCR gene and the LSD1 gene were successfully obtained. Then the quantitative PCR (qPCR) was used to determine the RCCR gene in PVY-infected potato tubers And LSD1 gene relative expression, and finally carried out the structural analysis of the target gene encoding protein. The results showed that the relative expression of these two genes decreased, speculated that these two genes are related to potato tuberous cell necrosis, which will make a preliminary exploration for the molecular mechanism of PVY induced potato tuber cell necrosis. Breeding and resistance drug screening provide an alternative target.