| 摘要: |
| [目的]从中国野生华东葡萄白河35-1经白粉菌接种诱导后叶片所构建的cDNA文库中,筛选抗病相关基因.[方法]根据植物抗病基因保守序列NBS位点设计特异引物,对葡萄白粉病接种诱导后的环化cDNA文库进行菌落PCR筛选,从随机挑选的大量菌落中筛选出82个PCR阳性菌落,提取阳性菌落的质粒进一步进行PCR检测确认,对确认的36个阳性质粒插入片段进行了测序.[结果]测序结果和序列生物信息学分析结果均表明,有3个基因序列与Genbank中的已知序列有较高的同源性,其中1个基因序列与葡萄抗白粉病基因 (DT661587.1)序列同源性达99%,另外2个基因序列与水分胁迫下葡萄特异表达基因(DT031657.1)和葡萄抗皮尔斯病基因(CF202948.1)的同源性分别为89%和96%.[结论]初步推测获得的3个基因均为抗性相关基因. |
| 关键词: 中国野生葡萄 cDNA基因 文库筛选 抗病基因 |
| DOI： |
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| 基金项目:国家转基因专项(JY03-A-19-02)；国家自然科学基金项目(No.30571280) |
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| Resistance genes screening in Uncinular necator induced cDNA library of Chinese wild grape |
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| Abstract: |
| 【Objective】 This paper aimed to screen expressed resistance genes from Chinese wild grape cDNA library which was constructed from leaves induced by grape powdery mildew (Uncinular necator) inoculation.【Method】 Specific oligo nucleotide primers were designed according to conservative NBS sequence of resistance genes and the universal sequence primers on vector.Colony PCR was employed to screen the circled cDNA library.82 positive colonies were acquired by colony PCR,out of which 36 colonies were identified by plasmid PCR assay.【Result】 Positive plasmids were sequenced and the sequence analysis result showed that 3 expressed sequence tags were highly homologous to known sequences,one was 99% homology to a registered Chinese wild grape sequence DT661587.1,the other two were 89% and 96% homology to Vitis vinifera water deficit response gene DTO31657.1 and grape Pierce′s disease resistance gene CF202948.1 respectively.【Conclusion】 These genes are supposed to be related with grape powdery mildew resistance. |
| Key words: Chinese wild grape cDNA library library screening resistance gene |
