Study on establishment and optimization of rapid tissue culture propagation technology of three wine grape cultivars

doi:10.6048/j.issn.1001-4330.2024.01.012

Abstract

Abstract:

【Objective】 In order to improve the quality of wine grape seedlings and the breeding speed of improved varieties,the tissue culture virus-free and rapid propagation technology system of wine grape varieties Dunkelfelder T,Petit Verdot and Sauvignon Blanc was preliminarily established.Which has provided the basis for improving the quality of seedlings of small wine grape varieties and rapid propagation. 【Methods】 Three one-year axillary buds of wine grape were used as explants,and the orthogonal test design was used to explore the effects of different disinfection methods on the survival rate of explants,and the effects of different concentrations of 6-BA,IBA,NAA and different combinations of media on the germination,proliferation and rooting of axillary buds. 【Results】 The results showed that the survival rate of axillary buds could be improved and the pollution rate could be reduced by 10% sodium hypochlorite disinfection for 7.5min,75% ethanol disinfection for 30 s,and 0.1% HgCl₂ disinfection for 10min; When the sterilized explants were inoculated into MS medium supplemented with 1.0 mg/L 6-BA and 0.05 mg/L NAA,the germination rate was the highest. When the sterile new buds of the start-up culture were added into the 1/2MS medium supplemented with 1.0 mg/L 6-BA and 0.3 mg/L IBA,the clustered buds had the best proliferation effect. When the single buds after the proliferation culture were put into the 1/2MS rooting medium supplemented with 0.4 mg/L IBA and 0.4 mg/L NAA,the overall quality of rooting of tissue culture seedlings reached the best effect. 【Conclusion】 The technical system of tissue culture and rapid propagation and virus-free of three wine grape varieties has been preliminarily established. The heat treatment combined with shoot tip for virus-free of tissue culture seedlings was applied in the experiment,and the results showed that this method could remove most of grape viruses.

Key words: wine grape; tissue culture; detoxification

摘要：

【目的】研究提高酿酒葡萄苗木品质(质量)和良种繁育速度(效率)的技术,建立酿酒葡萄品种紫大夫、小味儿多和长相思等品种的组培脱毒快繁技术体系,为提高小众酿酒葡萄品种苗木的品质及快速繁殖提供依据。【方法】以3个酿酒葡萄1年生腋芽为外植体,采用正交试验设计方法,分析不同消毒方式对外植体成活率的影响,研究不同浓度6-BA、IBA、NAA和不同培养基组合对腋芽萌发、增殖及生根培养的影响。【结果】腋芽经10%次氯酸钠消毒7.5 min,75%乙醇消毒30 s,0.1%HgCl₂消毒10 min能提高腋芽的成活率,降低污染率;经消毒后的外植体接种到添加1.0 mg/L 6-BA和0.05 mg/L NAA的MS培养基中,其萌芽率最高;将启动培养的无菌新芽接种到添加1.0 mg/L 6-BA和0.3 mg/L IBA 的1/2MS培养基中,丛生芽增殖效果最佳,将增殖培养后的单芽切下接种到添加0.4 mg/L IBA和0.4 mg/L NAA的1/2MS生根培养基中,组培苗的生根综合质量最佳。【结论】建立了3个酿酒葡萄品种组培快繁脱毒技术体系。采用热处理结合茎尖进行组培苗的脱毒,能够脱除绝大部分的葡萄病毒。

关键词: 酿酒葡萄, 组织培养, 脱毒

CLC Number:

S663

YANG Bo, WANG Hao, CHEN Yongwei, HA Rong, JIN Wei, XU Can, ZHANG Min, YANG Guili, YANG Hongbo, MA Wenli. Study on establishment and optimization of rapid tissue culture propagation technology of three wine grape cultivars[J]. Xinjiang Agricultural Sciences, 2024, 61(1): 109-117.

杨波, 王昊, 陈永伟, 哈蓉, 靳韦, 徐灿, 张敏, 杨桂丽, 杨宏波, 马文礼. 3个酿酒葡萄品种组培快繁体系的建立及优化[J]. 新疆农业科学, 2024, 61(1): 109-117.

Figures/Tables 10

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处理编号 Process number	培养基类型 Type of culture medium	6-BA (mg/L)	NAA (mg/L)
D₁	MS	0.5	0
D₂	MS	1	0.05
D₃	MS	1.5	0.1
D₄	1/2MS	0.5	0.05
D₅	1/2MS	1	0.1
D₆	1/2MS	1.5	0
D₇	GS	0.5	0.1
D₈	GS	1	0
D₉	GS	1.5	0.05

处理编号 Process number	培养基类型 Type of culture medium	6-BA (mg/L)	NAA (mg/L)
D₁	MS	0.5	0
D₂	MS	1	0.05
D₃	MS	1.5	0.1
D₄	1/2MS	0.5	0.05
D₅	1/2MS	1	0.1
D₆	1/2MS	1.5	0
D₇	GS	0.5	0.1
D₈	GS	1	0
D₉	GS	1.5	0.05

处理编号 Process number	培养基类型 Type of culture medium	6-BA (mg/L)	IBA (mg/L)
E₁	MS	0.5	0.1
E₂	MS	1	0.2
E₃	MS	1.5	0.3
E₄	1/2MS	0.5	0.2
E₅	1/2MS	1	0.3
E₆	1/2MS	1.5	0.1
E₇	GS	0.5	0.3
E₈	GS	1	0.1
E₉	GS	1.5	0.2

处理编号 Process number	培养基类型 Type of culture medium	6-BA (mg/L)	IBA (mg/L)
E₁	MS	0.5	0.1
E₂	MS	1	0.2
E₃	MS	1.5	0.3
E₄	1/2MS	0.5	0.2
E₅	1/2MS	1	0.3
E₆	1/2MS	1.5	0.1
E₇	GS	0.5	0.3
E₈	GS	1	0.1
E₉	GS	1.5	0.2

处理编号 Process number	紫大夫Dunkelfelder T		长相思Sauvignon blanc		小味儿多Petit verdot
处理编号 Process number	污染率 Pollution rate(%)	成活率 Survival rate(%)	污染率 Pollution rate(%)	成活率 Survival rate(%)	污染率 Pollution rate(%)	成活率 Survival rate(%)
C₁	68.75±3.06^b	31.25±0.82^b	38.46±0.78^b	61.54±1.26^a	68.75±2.41^c	31.25±0.61^d
C₂	81.25±1.02^a	18.75±1.77^c	40.74±0.41^b	59.26±2.82^a	75.00±2.45^b	25.00±1.63^e
C₃	81.25±2.65^a	18.75±1.22^c	48.00±2.45^a	52.00±1.22^b	62.50±1.63^d	37.50±1.63^c
C₄	68.75±3.06^b	31.25±1.85^b	50.00±1.63^a	50.00±4.08^b	68.75±2.41^c	31.25±2.24^d
C₅	67.75±2.24^b	32.25±1.71^b	37.93±1.16^b	62.07±1.28^a	50.00±1.63^e	57.00±1.22^a
C₆	62.50±2.04^c	37.50±1.67^a	39.29±1.46^b	60.71±1.80^a	75.00±2.45^b	25.00±0.41^e
C₇	62.50±3.67^c	37.50±1.57^a	38.36±0.09^b	63.64±2.77^a	53.75±18.4^e	56.25±0.86^a
C₈	68.75±2.24^b	31.25±1.36^b	40.00±1.63^b	60.00±3.26^a	51.00±2.04^e	50.00±1.22^b
C₉	68.75±3.06^b	31.25±2.24^b	48.28±2.02^a	51.72±1.31^b	81.25±1.43^a	18.75±1.02^f