| TRV介导的葡萄叶片VvANR基因瞬时表达分析 |
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| 引用本文： | 杨波,刘海霞,牛铁泉,张鹏飞,梁长梅,赵旗峰,温鹏飞.TRV介导的葡萄叶片VvANR基因瞬时表达分析[J].核农学报,2021,35(4):826-836. |
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| 作者姓名： | 杨波 刘海霞 牛铁泉 张鹏飞 梁长梅 赵旗峰 温鹏飞 |
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| 作者单位： | 山西农业大学园艺学院,山西太谷 030801;山西农业大学信息科学与工程院,山西太谷 030801;山西农业大学果树研究所,山西太谷 030801 |
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| 基金项目： | 山西省重点研发计划重点项目(201703D211001-04-02),山西省重点研发计划项目(201803D211016-4),山西省应用基础研究计划(201901D111222),山西省新兴产业领军人才资助计划 |
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| 摘 要： | 为建立葡萄叶片TRV-VIGS系统,分析验证VvANR基因功能,本研究以烟草脆裂病毒(Tobacco rattle virus,TRV)介导基因沉默表达载体pTRV2-ANR,采用真空侵染和主叶脉针孔注射法分别侵染葡萄幼嫩、成熟叶片,观察表型并测定原花青素含量,实时荧光定量PCR测定被侵染叶片中VvANR及其相关基因表达。结果表明,真空侵染的幼嫩、成熟叶片分别于第6、第7天出现漂白,侵染15 d后,幼嫩叶片几乎全部漂白,成熟叶片大面积漂白。主叶脉针孔注射侵染的幼嫩、成熟叶片分别于第17、第19天出现漂白;侵染25 d后,幼嫩和成熟叶片发生全叶漂白,且漂白均沿主叶脉逐渐扩展至整个叶面;对照均未发生漂白现象。pTRV2-ANR侵染后叶片中原花青素含量极显著降低,VvANR基因表达量极显著降低,且原花青素生物合成相关基因VvLAR1、VvLAR2、VvMYBPA1、VvMYBPA2、VvUFGT表达量显著降低,除VvUFGT表达量呈显著差异外,其余均呈极显著差异;而VvANS、VvDFR表达量轻微上调,且与对照差异不显著。本研究建立了葡萄叶片VIGS体系,为基因功能快速验证提供了新方法;同时,揭示了葡萄叶片中原花青素含量、VvANR表达及与相关基因的关系,本研究为完善葡萄原花青素积累机制奠定了一定的理论基础。
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| 关 键 词： | 葡萄(Vitis vinifera L.) 叶片 VvANR VIGS 表达分析 |
| 收稿时间： | 2020-05-27 |
Transient Expression Analysis of VvANR Gene in Grape Leaves Mediated by TRV |
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| YANG Bo,LIU Haixia,NIU Tiequan,ZHANG Pengfei,LIANG Changmei,ZHAO Qifeng,WEN Pengfei.Transient Expression Analysis of VvANR Gene in Grape Leaves Mediated by TRV[J].Acta Agriculturae Nucleatae Sinica,2021,35(4):826-836. |
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| Authors: | YANG Bo LIU Haixia NIU Tiequan ZHANG Pengfei LIANG Changmei ZHAO Qifeng WEN Pengfei |
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| Institution: | 1College of Horticulture, Shanxi Agricultural University, Taigu, Shanxi 030801; 2College of Information Science and Engineering, Shanxi Agricultural University, Taigu, Shanxi 030801; 3Pomology Institute, Shanxi Agricultural University, Taigu, Shanxi 030801 |
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| Abstract: | To analyse and verify the function of the VvANR gene, the Tobacco rattle virus (TRV) mediated gene silencing vector pTRV2-ANR was used to establish TRV-VIGS system of grape leaves. The young and mature leaves of grapes were infected through Vacuum infection and injection at the main vein with TRV, the phenotype of leaves, and the content of proanthocyanidins, relative expression of VvANR after infection were determined. The results indicated that the young and mature leaves were bleached on the 6th and 7th days infected through vacuum infection, respectively. 15 days later after infection, the young leaves were almost bleached and the mature leaves were bleached to a large extent. The young and mature leaves infected by injection at the main veinbecame bleached on the 17th and 19th days; 25 days later after infection, the young and mature leaves were all bleached and the bleaching gradually expanded, along the main vein to the whole leaves. While no bleaching occurred in the control group. After pTRV2-ANR infection, the proanthocyanidin content in the leaves and the VvANR gene expression were significantly reduced, the expression levels of proanthocyanidin biosynthesis related genes VvLAR1, VvLAR2, VvMYBPA1, and VvMYBPA2 were extremely significantly reduced, and the expression of VvUFGT significant reduced; While the expression of VvANS and VvDFR were slightly up-regulated, and the differences were not significant. In this study, VIGS system of grape leaves was established, which provided a new method for rapid verification of gene function; at the same time, the proanthocyanidin content, the expression of VvANR and their relationship with related genes in grape leaves were revealed, which enriched the study of proanthocyanidins metabolism in grape. |
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| Keywords: | grape(Vitis vinifera L ) leaf VvANR VIGS expression analysis |
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