3个酿酒葡萄品种组培快繁体系的建立及优化

doi:10.6048/j.issn.1001-4330.2024.01.012

摘要/Abstract

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

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

中图分类号:

S663

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.

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.

图/表 10

参考文献 24

[1]	韦伟, 单守明. 葡萄脱毒与快繁技术研究进展[J]. 分子植物育种, 2022, 20(1):259-265.
	WEI Wei, SHAN Shouming. Research Progress on Virus-free Grape and Rapid Propagation Technology[J]. Molecular Plant Breeding, 2022, 20(1):259-265.
[2]	胡泽军, 张亚红, 张晓煜, 等. 宁夏贺兰山东麓酿酒葡萄不同子产区土壤肥力特征与评价[J]. 中国土壤与肥料, 2021,(6):35-41.
	HU Zejun, ZHANG Yahong, ZHANG Xiaoyu, et al. Characteristics and evaluation of soil fertility in different grape production areas at the eastern foot of Helan Mountain,Ningxia[J]. Soil and Fertilizer Sciences in China, 2021,(6):35-41.
[3]	朱峰, 杨微. 宁夏贺兰山东麓酿酒葡萄产业发展措施探讨[J]. 现代农业科技, 2018,(21):98,101.
	ZHU Feng, YANG Wei. Discussion on the development measures of wine grape industry at the eastern foot of Helan Mountain in Ningxia[J]. Modern Agricultural Science and Technology, 2018,(21):98,101.
[4]	刘世松. 进口葡萄酒对我国葡萄酒产业的影响及对策[J]. 中国市场, 2011,(45):157-163.
	LIU Shisong. Influence of imported wine on Chinese wine industry and Its Countermeasures[J]. China Market, 2011,(45):157-163.
[5]	齐向丽, 师校欣, 杜国强. ‘红国王’葡萄组织培养快速繁殖[J]. 分子植物育种, 2020, 18(3):982-987.
	QI Xiangli, SHI Xiaoxin, DU Guoqiang. Rapid Propagation of ‘Hongguowang’ Grape Tissue Culture in Vitro[J]. Molecular Plant Breeding, 2020, 18(3):982-987.
[6]	沈传进, 王利民, 张平, 等. 鲜食葡萄组织培养快速繁育系统的建立[J]. 河北农业科学, 2011, 15(7):16-21.
	SHEN Chuanjin, WANG Limin, ZHANG Ping, et al. Tissue culture rapid proliferation system establishment of table Grape[J]. Journal of Hebei Agricultural Sciences, 2011, 15(7):16-21.
[7]	林茜, 高营营, 覃换玲, 等. ‘阳光玫瑰’葡萄组培脱毒快繁技术研究[J]. 果树学报, 2021, 38(3):435-443.
	LIN Qian, GAO Yingying, QIN Huanling, et al. Study on the technology of virus-free and rapid propagation of ‘Shine Muscat’grape in tissue culture[J]. Journal of Fruit Science, 2021, 38(3):435-443.
[8]	闵首军, 李爱玲. 葡萄组织快繁技术研究[J]. 农学学报, 2012, 2(10):55-57,67.
	MIN Shoujun, LI Ailing. Study on Rapid Propagation Technology of Grape Tissues[J]. Journal of Agriculture, 2012, 2(10):55-57,67. DOI
[9]	王婷, 肖慧琳, 唐美玲, 等. ‘黑虎香’葡萄离体快繁技术研究[J]. 烟台果树, 2020,(1):17-18.
	WANG Ting, XIAO Huiling, TANG Meiling, et al. Study on in vitro Rapid Propagation of ‘Heihuxiang’ grape[J]. Yantai Fruits, 2020,(1):17-18.
[10]	蔡文博, 段虹, 王军, 等. 4个鲜食葡萄品种组培快繁体系的建立[J]. 核农学报, 2019, 33(2):248-254. DOI
	CAI Wenbo, DUAN Hong, WANG Jun, et al. Establishment of in vitro rapid micropropagation of four table grape cultivars[J]. Journal of Nuclear Agricultural Sciences, 2019, 33(2):248-254. DOI
[11]	冯文华, 代红军. ‘赤霞珠’葡萄组培快繁体系研究[J]. 北方园艺, 2016, 40(13):104-106.
	FENG Wenhua, DAI Hongjun. Study on tissue culture and rapid propagation system of ‘Cabernet Sauvignon’ grape[J]. Northern Horticulture, 2016, 40(13):104-106.
[12]	刘娜, 许轲, 张文, 等. 四个酿酒葡萄品种组培快繁体系的初建[J]. 植物生理学报, 2013, 49(10):1071-1076.
	LIU Na, XU Ke, ZHANG Wen, et al. Preliminary establishment of in vitro rapid micropropagation of four grape-wine(Vitis vinifera L.) cultivars[J]. Plant Physiology Journal, 2013, 49(10):1071-1076.
[13]	徐美隆, 刘静, 王巧珍, 等. 酿酒葡萄‘赤霞珠’组织培养快繁技术优化[J]. 北方园艺, 2016,(22):117-120.
	XU Meilong, LIU Jing, WANG Qiaozhen. Optimization of Tissue Culture and Rapid Propagation of ‘Cabernet Sauvignon’ Grape[J]. Northern Horticulture, 2016,(22):117-120.
[14]	王婷, 卢建声, 唐美玲, 等. 蛇龙珠组培繁殖试验初报[J]. 烟台果树, 2019,(1):9-10.
	WANG Ting, LU Jiansheng, TANG Meiling, et al. Preliminary report on tissue culture and propagation of Rosa laevigata[J]. Yantai Fruits, 2019,(1):9-10.
[15]	李琴, 舒木林, 甘祖燕, 等. 大花美人蕉种子高效无菌萌发及离体快繁体系的建立[J]. 植物生理学报, 2022, 58(4):667-676.
	LI Qin, SHU Mulin, GAN Zuyan, et al. Establishment of efficient seed germination and in vitro rapid propagation system of Canna generalis[J]. Plant Physiology Journal, 2022, 58(4):667-676.
[16]	叶伟, 刘凉琴, 史学正, 等. 基质中添加多孔改良剂对南方高丛蓝浆果品种‘蓝美1号’组培苗茎段瓶外生根的影响[J]. 植物资源与环境学报, 2020, 29(3):69-71.
	YE Wei, LIU Liangqin, SHI Xuezheng, et al. Effects of substrate complemented with porous soil amendment on ex vitro rooting of stem segments of tissue culture seedlings of Vaccinium corymbosum‘Lanmei 1’[J]. Journal of Plant Resources and Environment, 2020, 29(3):69-71.
[17]	陈卫平, 尚红莺, 周军, 等. 贺兰山东麓酿酒葡萄的生态适应性[J]. 西北植物学报, 2007, 27(9):1855-1860.
	CHEN Weiping, SHANG Hongying, ZHOU Jun, et al. Ecological Adaptability of Wine Grape at the Eastern Foot of Helan Mountain[J]. Acta Botanica Boreali-Occidentalia Sinica, 2007, 27(9):1855-1860.
[18]	胡文斌, 张少飞, 孙娜. 陇南红提葡萄茎段组织培养研究[J]. 陕西林业科技, 2018, 46(1):6-9.
	HU Wenbin, ZHANG Shaofei, SUN Na. Stem Tissue Culture of Red Grape in Longnan Area[J]. Shaanxi Forest Science and Technology, 2018, 46(1):6-9.
[19]	陈玉霞, 邱建辉, 谷峰. 葡萄新品种组织培养快繁技术研究[J]. 安徽农业科学, 2022, 50(4):106-109.
	CHEN Yuxia, QIU Jianjun, GU Feng. Study on Tissue Culture Rapid Propagation Technology of New Grape Varieties[J]. Journal of Anhui Agricultural Sciences, 2022, 50(4):106-109.
[20]	刘晓芹, 冯美, 王振平, 等. 葡萄砧木组培快繁研究[J]. 安徽农业科学, 2010, 38(17):8866-8868,8876.
	LIU Xiaoqin, FENG Mei, WANG Zhenping, et al. Study on Tissue Culture and Rapid Propagation of Grape Rootstock[J]. Journal of Anhui Agricultural Sciences, 2010, 38(17):8866-8868,8876.
[21]	Miransari M, Smith D L. Plant hormones and seed germination[J]. Environmental and Experimental Botany, 2014, 99:110-121. DOI URL
[22]	苏玲, 宫磊, 尹向田, 等. 贵妃玫瑰葡萄组培技术探究[J]. 安徽农业科学, 2020, 48(8):51-53,65.
	SU Ling, GONG Lei, YIN Xiangtian, et al. Study on Tissue Culture Technology of the Guifeimeigui Grape[J]. Journal of Anhui Agricultural Sciences, 2020, 48(8):51-53,65.
[23]	齐永顺, 张志华, 王同坤, 等. 同源四倍体玫瑰香葡萄嫩枝扦插不定根发生过程中内源激素的变化[J]. 园艺学报, 2009, 36(4):565-570.
	QI Yongshun, ZHANG Zhihua, WANG Tongkun, et al. Changes of Endogenous Hormones in Adventitious Root Formation in Softwood Cuttings of Autotetraploid Grape Muscat Hamburg[J]. Acta Horticulturae Sinica, 2009, 36(4):565-570.
[24]	张尊平, 范旭东, 胡国君, 等. 葡萄试管苗热处理脱毒技术研究[J]. 中国果树, 2013,(1):39-41.
	ZHANG Zunping, FAN Xudong, HU Guojun, et al. Study on virus-free treatment technology of grape testtube seedlings[J]. China Fruits, 2013,(1):39-41.

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