茶炭疽病拮抗链霉菌的筛选鉴定与拮抗能力测定  

Screening,Identification and Determination of Antagonistic Actinomycetes Strain against Tea Anthracnose

作　　者：张玉丹 谭琳 任佐华[3] 杨艺帅[3] 杨学宇 胡秋龙 ZHANG Yudan;TAN Lin;REN Zuohua;YANG Yishuai;YANG Xueyu;HU Qiulong(Key Laboratory of Tea Science of Ministry of Education,Hunan Agricultural University,Changsha 410128,China;College of Horticulture,Hunan Agricultural University,Changsha 410128,China;College of Plant Protection,Hunan Agricultural University,Changsha 410128,China)

机构地区：[1]湖南农业大学茶学教育部重点实验室,长沙410128 [2]湖南农业大学园艺学院,长沙410128 [3]湖南农业大学植物保护学院,长沙410128

出　　处：《中国生物防治学报》2023年第3期646-656,共11页Chinese Journal of Biological Control

基　　金：2021年度湖南省重点研发计划项目(2022NK2051);2021年度湖南省科技创新重大项目(2021NK1020)。

摘　　要：茶炭疽病是茶树的一种重要叶部病害,发生时严重影响茶叶产量和品质,给茶叶生产带来经济损失。为获得对茶炭疽病菌刺盘孢菌Colletotrichum camelliae具有良好拮抗效果的链霉菌,采用稀释涂布平板法从茶园根际土壤中分离放线菌菌株,利用平板对峙法筛选具有较高拮抗活性的菌株;结合其形态特征、培养特征、生理生化特性和分子生物学对其进行种属鉴定;采用扫描电镜观察菌株JS4-F对茶炭疽病菌的生长抑制作用,离体叶片法和盆栽防效测定其发酵液对茶炭疽病的室内防治效果,平板对峙法测定其抗菌谱。结果表明,分离筛选获得一株对茶炭疽病菌具有较好拮抗作用的菌株JS4-F,根据该菌的形态特征、生理生化特性初步将其鉴定为链霉菌属,通过联合16S rRNA、gyrB、ropB、recA、trpB基因测序结果构建系统发育树,JS4-F菌株被鉴定为藤黄生孢链霉菌Streptomyces luteosporeus。菌株JS4-F对茶炭疽病菌的拮抗效果达75.59%,扫描电镜表明其能使茶炭疽菌菌丝畸形、皱褶、菌丝之间相互缠绕;氮源利用试验表明,菌株JS4-F能利用L-精氨酸、L-甘氨酸、L-丙氨酸、L-色氨酸,能在6%NaCl浓度下生长,pH的耐受范围为5~12。菌株JS4-F发酵液原液室内离体叶片防治效果达52.54%,稀释10倍后防治效果为39.94%,稀释100倍后防治效果为30.56%;盆栽防效试验表明,菌株JS4-F发酵液原液对茶炭疽病的防治效果达44.59%;拮抗菌株JS4-F对其它8种植物病原菌均具有较好的拮抗作用,抑制效果均大于60%。综上,菌株JS4-F对茶炭疽病有较好的防治潜力,能给茶炭疽病的生物防治提供参考资源和技术依据。Tea anthracnose is an important foliar disease of tea plants,which seriously affects the yield and quality of tea when it occurs and brings economic losses to tea production.In order to obtain actinomycetes with good antagonistic effect against tea anthracnose caused by Colletotrichum camelliae,dilution coated plate method was used to isolate the actinomycetes from the rhizosphere soil of tea plantations,and strains with high antagonistic activity were screened using the plate confrontation method;their morphological characteristics,culture characteristics,physiological and biochemical properties and molecular biology were combined to identify their species.The growth inhibition of JS4-F against tea anthracnose was observed by scanning electron microscopy,the indoor control effect of its fermentation solution against tea anthracnose was determined by in vitro leaf method and pot experiment,and its antibacterial spectrum was determined by plate standoff method.The results showed that strain JS4-F with good antagonistic effect against tea anthracnose was isolated and screened,and it was initially identified as Streptomyces spp.based on its morphological characteristics and physiological and biochemical properties,and a phylogenetic tree was constructed by combining the sequencing results of 16SrRNA,gyrB,ropB,recA and trpB genes,and the strain JS4-F was identified as Streptomyces luteospore.The antagonistic effect of strain JS4-F against tea anthracnose reached 75.59%,and scanning electron microscopy showed that strain JS4-F could deform,wrinkle and intertwine the mycelium of tea anthracnose.The nitrogen source utilization test showed that strain JS4-F could utilize L-arginine,L-glycine,L-alanine,and L-tryptophan,could grow at 6%NaCl concentration and pH tolerance range was 5-12.The control effect of strain JS4-F fermentation broth was 52.54%on isolated leaves indoors,39.94%after 10 times dilution and 30.56%after 100 times dilution.The pot control test showed that the control effect of strain JS4-F fermentation brot

关 键 词：茶炭疽病 链霉菌 拮抗作用 生物防治 

分 类 号：S476[农业科学—农业昆虫与害虫防治]