首页|沙地葡萄茎痘相关病毒在'阳光玫瑰'葡萄树不同物候期和不同部位的变化规律

沙地葡萄茎痘相关病毒在'阳光玫瑰'葡萄树不同物候期和不同部位的变化规律

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[背景]沙地葡萄茎痘相关病毒(grapevine rupestris stem pitting associated virus,GRSPaV)是皱木复合病中最常见的一种病毒,主要通过无性繁殖传播,灵敏的检测技术和培育无病毒植株是预防GRSPaV危害的关键.[目的]建立 GRSPaV 检测体系,确定适宜样品采集时期及部位,为病毒检测和无病毒材料培育及样品采集提供参考.[方法]建立基于AugeGreen染料法和外壳蛋白(coat protein,CP)基因区域设计引物的RT-qPCR检测方法,对携带GRSPaV的'阳光玫瑰'葡萄树地上部不同物候期、不同部位的样品进行 GRSPaV 检出率及基因表达量分析.[结果]以 GRS q CP1 为引物的GRSPaV RT-qPCR检测方法灵敏度较RT-PCR高 10 倍.GRSPaV在萌芽期、新梢生长期和花期的检出率均为 100%,果实膨大期为 91.7%;成熟卷须为 94.4%,成龄叶片、成龄叶柄和幼嫩枝条均为 100%.检测为阴性的样品均为幼嫩部位.GRSPaV CP 基因表达量在花期幼嫩卷须中最高,其次为花期成龄叶片;成龄叶中表达量在果实膨大期至成熟期均为同物候期内表达量最高部位;当年冬芽的萌芽期及二次枝的新梢生长期表达量均较低.[结论]'阳光玫瑰'葡萄GRSPaV检出率和GRSPaV CP 基因表达量随物候期进程表现差异,检出率在萌芽期至花期最高,果实成熟期最低;GRSPaV CP 基因表达量在花期最高.综合检出率及表达量因素,花期成龄叶适合作为GRSPaV病毒检测样品;果实膨大期至成熟期当年冬芽及萌发的二次枝适合作为脱除病毒材料.
The Variation of GRSPaV in Different Parts of Shine Muscat Grapevines During Their Phenological Periods
[Background]Grapevine rupestris stem pitting associated virus(GRSPaV)is the most common virus causing rugose wood disease.It is mainly transmitted via asexual reproduction.Sensitive detection technology and cultivation of virus-free plants are essential for preventing GRSPaV.[Objective]The purpose of this study was to establish a GRSPaV detection system,and to determine the appropriate periods and sites of sample collection for virus detection,so as to provide a reference for virus detection and obtaining virus-free materials.[Method]An RT-qPCR detection method was established using the AugeGreen dye method and primers according to the coat protein gene(CP)region.The detection rate and gene expression of GRSPaV in samples from different parts of Shine Muscat grapevines during various phenological stages were examined.[Result]The RT-qPCR detection method for GRSPaV using GRS q CP1 as primer showed 10 times higher sensitivity than that of RT-PCR method.The detection rates of GRSPaV were 100%during the budbreak,shoot growing period,and florescence,91.7%during the fruit enlargement stage,94.4%for mature tendrils,and 100%for mature leaves,mature petioles,and tender shoots.Samples showing negative results were all from the young parts of the plants.The young tendrils at florescence showed the highest level of GRSPaV CP gene expression,followed by the mature leaves at florescence.The expression level in mature leaves was the highest among the parts in the same phenological period from berry expansion to maturity.The expression levels in secondary winter buds at the budbreak stage and the secondary laterals at the shoot growing period were both relatively low.[Conclusion]The detection rate of GRSPaV and the GRSPaV CP gene expression level of Shine Muscat grape varied with the progression of phenological periods.The detection rate was the highest from budbreak to florescence,while which was the lowest at berry maturity period.The GRSPaV CP gene expression level was the highest at florescence.Based on both the detection rate and the expression level factors,it could be concluded that the mature leaves at florescence were suitable as samples for GRSPaV virus detection,and the winter buds from the berry expansion to maturity period and the lateral shoots germinated from these buds was appropriate for obtaining virus-free materials.

Shine Muscat grapevinesgrapevine rupestris stem pitting associated virusreal-time fluorescence quantitative PCRdetection rate of virusesgene expression level

李美璇、张向昆、王莉、乔月莲、师校欣、杜国强

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河北农业大学园艺学院,河北保定 071000

'阳光玫瑰'葡萄 沙地葡萄茎痘相关病毒 实时荧光定量PCR 病毒检出率 基因表达量

河北省现代农业产业技术体系葡萄创新团队项目热杂果现代种业科技创新团队项目

HBCT202315020121326310D

2023

10.3864/j.issn.0578-1752.2023.21.008

中国农业科学
中国农业科学院

中国农业科学

CSTPCDCSCD北大核心
影响因子：1.899
ISSN：0578-1752
年,卷(期)：2023.56(21)
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