摘要
【目的】明确川西高原酿酒葡萄根癌病的致病菌种类,评价不同药剂对病原菌的抑制及对根癌病的防治效果。【方法】采集川西高原酿酒葡萄根瘤组织进行病原菌的分离纯化,通过形态观察、生理生化鉴定、PCR检测及16s rDNA测序分析确定分离菌株种类,并对分离菌株进行系统进化分析(极大似然法maximum-likelihood)、生长特性分析及致病性检测。利用皿内抑菌圈法初步研究不同药剂对病原菌的抑制作用,并通过葡萄苗接种试验明确药剂对根癌病的防治效果。【结果】成功由葡萄根瘤组织中分离得到9株菌株,经鉴定为生物Ⅲ型葡萄土壤杆菌(Agrobacterium vitis),进化分析表明,9株菌株聚类在同一分支。虽然9株菌株生长特性略有差异,但均能使胡萝卜及葡萄形成瘿瘤,有致病性。进一步研究11种商品化药剂对分离菌株的抑制效果,发现中生菌素、福美双、多菌灵、春雷霉素、枯草芽孢杆菌和氢氧化铜对分离菌株抑制效果微弱或无效果,对分离菌株抑制效果较好的依次为四霉素、百菌清、喹啉铜、解淀粉芽孢杆菌KN-527及乙蒜素。进一步验证这5种效果较好的药剂对葡萄根癌病的防治效果,其中乙蒜素对葡萄根癌病的防治效果最好,发病率仅为6%,防治效果高达48.05%;其次,喹啉铜及四霉素对葡萄根癌病有防治效果;在所试药剂中,百菌清及解淀粉芽孢杆菌KN-527的防治效果较差。【结论】川西高原酿酒葡萄根癌病的致病菌为葡萄土壤杆菌,对酿酒葡萄根癌病防治效果最好的为乙蒜素,然后依次为喹啉铜、四霉素、百菌清及解淀粉芽孢杆菌KN-527。其中乙蒜素在根癌病田间防治可能具有较广阔的应用前景,生防菌株解淀粉芽孢杆菌KN-527对早期葡萄根癌病的发生有抑制作用。
【Objective】The Western Sichuan Plateau in China is considered as a high-quality wine grape producing area in China,due to its climatic characteristics such as sufficient sunlight,large temperature difference between day and night,little rainfall and large evaporation.However,the crown gall disease of grape frequently occurs in this region,directly threatening plant health and grape production and causing huge economic losses.Therefore,the premier objective of this study is to identify the pathogen species that cause grape crown gall disease specifically in the Western Sichuan Plateau for better clarifying the disease-controlling agents.Recent studies on grape crown gall disease are mainly carried out in the northern China,and the causes and pathogens causing grape crown gall disease in the Western Sichuan Plateau region have barely been investigated.Moreover,the limited control measures and unclear efficacy of commercial microbicides on grape crown gall disease have impeded the prevention and control in the field.Therefore,this study also aimed to find out the suitable disease managements by evaluating the controlling effect of 11 types of commercial microbicides on pathogenic bacteria.【Methods】Fresh grape crown galls were collected from the Western Sichuan Plateau for tissue grinding with the pathogen isolation by using MW selection medium,followed by yeast extract mannitol broth(YEB)medium.The isolated strains were then identified by morphological observation.The 16s rDNA was amplified with 27F(5'-AGAGTTTGATCCTGGCTCAG-3')/1492R(5'-AAGGAGGTGATCCAGCCGCA-3')for sequencing and phylogenetic analysis(Maximum-Likelihood)for each isolates.To better character the isolates and define their opine type,genes in T-DNA of Agrobacterium producing arginine,including octopine type and nopaline type,were amplified with primers AP-F/AP-R,OP-F/OPR and NP-F/NP-R,respectively.The growth curve of the isolates was drawn by measuring optical density at 600 nm(OD600nm).The biovar types of the isolates were applied to conduct phy
作者
葛青
许寰铎
王家乐
余楚维
刘伟
刘旭
GE Qing;XU Huanduo;WANG Jiale;YU Chuwei;LIU Wei;LIU Xu(College of Enology,Northwest A&F University/Shaanxi Provincial Key Laboratory of Viti-Viniculture,Yangling 712100,Shaanxi,China;Horticulture Research Institute,Sichuan Academy of Agricultural Sciences,Chengdu 610066,Sichuan,China)
出处
《果树学报》
CAS
CSCD
北大核心
2024年第8期1636-1648,共13页
Journal of Fruit Science
基金
陕西省重点研发计划项目(2022NY-118、2023-ZDLNY-21)
陕西省自然科学基础研究计划项目(023-JC-QN-0225)
秦创原引用高层次创新创业人才项目(QCYRCXM-2022-309)
中央高校基本科研业务费专项资金(2452022013)。
关键词
葡萄
根癌病
葡萄土壤杆菌
杀菌剂
病害防治
Grapevine
Crown gall
Agrobacterium vitis
Microbicide
Disease control
