摘要
Nursery plant propagation by grafting has been widely used in modern viticulture to minimize the damage caused by biotic and abiotic stresses.In grapevine(Vitis spp.),an effective way to control disease damage is to provide producers and growers with pathogen-free stock plants.In this study,five grapevine rootstock varieties,‘SO4’,‘101-14’,‘5BB’,‘110R’and‘1103P’,were selected as explants to establish an in vitro culture protocol,and three species of grapevine viruses were tested by real-time RT-PCR.The results showed that MS medium supplemented with 0.2 mg/L IBA,1.0 mg/L 6-BA,0.5 mg/L KT,4.0 mg/L adenine for culture initiation,and WPM supplemented with 0.2 mg/L IBA for subculture were suitable for all five rootstock varieties,with multiplication coefficients ranging from 1.6 to 4.4.Virus testing showed that single RT-PCR was more effective for detecting the three viruses compared to double or triple RT-PCR.Only plantlets free from the aforementioned viruses were retained for subculture.Plantlets were hardened at room temperature under natural lighting in Hoagland solution for 2 weeks and transplanted to pots filled with mixed media in a greenhouse.This protocol is applicable for rapid propagation of the five grapevine rootstock varieties and can be used for commercial production of virus-free grapevine stocks.
组培、嫁接快繁体系的建立被广泛应用于现代葡萄栽培生产中,以减少生物和非生物胁迫造成的危害。为葡萄种植产业提供无病毒砧木是控制病害的有效途径。本研究中,以“SO4”、“101-14”、“5BB”、“110R”和“1103P”五种常见的葡萄砧木品种为外植体,通过建立组培快繁体系快速获得无病毒葡萄砧木。快繁体系建立结果显示,快繁体系中适合不同砧木启动培养的培养基为MS基本培养基添加0.2 mg/L IBA、1.0 mg/L 6-BA、0.5 mg/L KT、4.0 mg/L腺嘌呤;适合不同砧木继代的培养基为WPM基本培养基添加0.2 mg/L IBA,且增殖系数在1.6~4.4之间。病毒检测结果显示,单次RT-PCR检测三种病毒比双重和三重RT-PCR更有效。通过不断继代获得无GLRaV-3,GFKV和GRSPa病毒组培苗进行大棚移栽驯化。幼苗不同霍格兰溶液中室温自然光照下驯化2周,移栽到温室中混合培养基质中,缩短驯化周期。该方案适用于5个葡萄藤砧木品种的快速繁殖,可用于无病毒葡萄砧木的商业化生产。
基金
现代农业产业技术体系葡萄种苗扩繁与生产技术(CARS-30)
北京无病毒葡萄种苗繁育基地建设及推广示范(D171100002317001)。
