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根据抗病基因(Resistance genes,R基因)的功能保守域,利用生物信息分析方法在绒毛状烟草(Nicotiana tomentosiformis)全基因组上获得6个完整可靠的抗真菌病候选基因。在这6个基因的非保守区设计特异引物,以普通烟草品种红花大金元反转录cDNA为模板克隆抗真菌病相关基因的干扰片段,利用GatewayTM克隆技术构建上述候选基因的RNAi载体,经酶切和测序证明载体构建正确,为进一步鉴定这6个候选基因的具体抗病性,并为将其应用于烟草抗病新品种的选育工作奠定了基础。
According to the conserved domains of resistance genes (R genes), 6 complete and reliable antifungal gene candidates were obtained from the whole genome of Nicotiana tomentosiformis using bioinformatics analysis. Specific primers were designed in the non-conserved regions of these six genes. The anti-mycosis-related gene fragment was cloned by reverse transcription cDNA of Nicotiana tabacum, and the RNAi vector of the candidate gene was constructed by GatewayTM cloning technology. The result of restriction analysis and sequencing showed that the vector was constructed correctly, which laid the foundation for the further identification of these six candidate genes and their application in the breeding of new tobacco disease-resistant varieties.