| 一株突破va基因型烟草抗性的PVY病毒分离物的鉴定 |
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| 引用本文： | 李若,李尊强,万秀清,乔婵,郭玉双,潘红杏,崔晓,陈荣平,郭兆奎.一株突破va基因型烟草抗性的PVY病毒分离物的鉴定[J].中国烟草学报,2020,26(4):72-77. |
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| 作者姓名： | 李若 李尊强 万秀清 乔婵 郭玉双 潘红杏 崔晓 陈荣平 郭兆奎 |
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| 作者单位： | 1.中国烟草总公司黑龙江省公司烟草科学研究所, 哈尔滨市哈药路17号 150076 |
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| 基金项目： | 黑龙江省烟草行业科技开发项目“应用基因组编辑技术改良黑龙江烟区烟草主栽品种的PVY抗性”HN201704中国烟草总公司重大专项项目“基于基因组编辑技术的烟草重要基因功能研究”110201501015(JY-02) |
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| 摘 要： | 背景和目的 近来，能突破va基因型抗性的马铃薯Y病毒(Potato virus Y，PVY)毒株在烟草上不断出现。2016年，我们分离到一株抗性突破的毒株PVY-CJ，本文对该病毒分离物进行了验证和鉴定。 方法 采用病毒重新接种、多重RT-PCR、RT-PCR分段扩增病毒全长基因组、PVY病毒分子进化分析及病毒蛋白VPg蛋白序列比对等方法对PVY-CJ毒株进行了验证和鉴定。 结果 三种重新接种了PVY-CJ的va基因型烟草品种均发病。多重RT-PCR结果将PVY-CJ鉴定为PVYNTN株系。序列分析表明，PVY-CJ全长基因组包含一个全长9180 nt的开放读框，其编码一个3060 AA的多聚蛋白。分子进化分析进一步验证了其归类于PVYNTN株系。相比其他PVYNTN株系毒株，PVY-CJ的VPg蛋白105位氨基酸出现了一个由赖氨酸到谷氨酸的替换，而这一氨基酸替换导致了其对va基因来源抗性的突破。 结论 我们首次报道并鉴定了国内一株能突破va基因型烟草抗性的PVY毒株—PVY-CJ。这为进一步研制PVY抗病烟草品种提供了材料。
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| 关 键 词： | 烟草 PVY va基因型 抗性突破 鉴定 PVY-CJ |
| 收稿时间： | 2019-12-10 |
Identification of a PVY virus isolate breaking down va-genotype tobacco resistance |
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| Affiliation: | 1.Heilongjiang Institute of Tobacco Science of CNTC, Harbin 150076, China2.Molecular Genetics Key Laboratory of China Tobacco, Guizhou Academy of Tobacco Science, Guiyang 550081, China |
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| Abstract: | Background Recently, the Potato virus Y (PVY) strains breaking down the resistance of va-genotype tobacco varieties have emerged on cultivated tobacco constantly. In 2016, we isolated a va-resistance breaking-down PVY strain, which was named as PVYCJ. This isolate was verified and identified in this study. Methods Re-inoculation on the va-genotype tobacco varieties, multiple RTPCR, segmented amplification of the whole PVY genome by RT-PCR, molecular phylogenetic analysis and the sequence alignment of VPg protein of PVY strains were adopted to confirm and identify PVY-CJ strain. Results Test data showed: 1) All the 3 va-genotype tobacco varieties showed symptoms of infection when they were inoculated with PVY-CJ inoculum; 2) PVY-CJ was classified as PVYNTN strain according to the result of multiple RT-PCR; 3) Sequence analysis showed that the whole genome of PVY-CJ contained an open reading frame of 9180 nt encoding a polyprotein of 3060 amino acids; 4) the result of molecular phylogenetic analysis further classified PVY-CJ as PVYNTN strain; 5) Compared with other PVYNTN strains, a substitution of lysine to glutamic acid happened at 105 amino acid of VPg protein of PVY-CJ, which led to a breaking down of va-geno source resistance. Conclusion We reported and identified a PVY strain which could break down the resistance of va-genotype tobacco varieties in the mainland of China for the first time. This study provides material for breeding new PVY-resistant tobacco varieties. |
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