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

doi:10.3864/j.issn.0578-1752.2023.21.008

Abstract

Abstract:

【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.

Key words: Shine Muscat grapevines, grapevine rupestris stem pitting associated virus, real-time fluorescence quantitative PCR, detection rate of viruses, gene expression level

LI MeiXuan, ZHANG XiangKun, WANG Li, QIAO YueLian, SHI XiaoXin, DU GuoQiang. The Variation of GRSPaV in Different Parts of Shine Muscat Grapevines During Their Phenological Periods[J].Scientia Agricultura Sinica, 2023, 56(21): 4234-4244.

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References 30

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引物名称 Primer	基因区域 Gene region	引物序列 Primer sequence	产物长度 Product length (bp)
GRS q CP1	CP	F: TCTCACTGCTCTGATGTTGGTAG R: TAGATGGTGGTATCCCCGTC	184
GRS q CP2	CP	F: GGCTATGAAACACGAAACGGT R: CCTTAGGTCACAAAATGGACTCTT	205
GRS q TGB	TGB	F: TCGGACCCATACCAAAACAT R: CCAAACCGTCACTGCTAACC	130
GRS q RdRp1	RdRp	F: GTATCGGGAGGTGCGTTGT R: GATTTCCTTTATGCCTTGGGT	131
GRS q RdRp2	RdRp	F: GCTTCCTTATGGCTGTCCG R: TTCTTCGCCTCAAACTCCTG	184

物候期 Phenological period	休眠枝条 Dormant shoot	嫩芽 Bud	嫩叶片 Tender leaf	嫩叶柄 Tender petiole	成龄叶片 Mature leaf	成龄叶柄 Mature petiole	幼嫩卷须 Tender tendril	成熟卷须 Mature tendril	1—2片叶副梢 1-2 leaves lateral shoot	3—5片叶副梢 3-5 leaves lateral shoot	幼嫩新梢 Tender shoot	木质化新梢 Lignify shoot
休眠期 Dormant period	2/3
萌芽期 Budbreak stage		3/3
新梢生长期 Growing period			3/3	3/3			3/3	3/3	3/3	3/3
花期 Florescence			3/3	3/3	3/3	3/3	3/3	3/3
果实膨大期 Berry expansion period			1/3	3/3	3/3	3/3	3/3	3/3	3/3	3/3
果实转色期 Grape fruit veraison			1/3	1/3	3/3	3/3	1/3	2/3	0/3	1/3
果实成熟期 Maturity			0/3	0/3	3/3	3/3	0/3	3/3	1/3	0/3
落叶期 Defoliating period					3/3	3/3					3/3	2/3
当年冬芽萌芽期 Secondary winter bud embryonic stage		2/3
当年冬芽新梢生长期 Secondary winter bud growing period			1/3	3/3			3/3	3/3	0/3	1/3

The Variation of GRSPaV in Different Parts of Shine Muscat Grapevines During Their Phenological Periods

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