云南白菜根肿病拮抗菌的筛选及鉴定  

Screening and identification of antagonistic bacteria against clubroot of Brassica rapa var.glabra Regel in Yunnan province

作　　者：张怡宁 刘秋艳 朱美竹 曾民[2,3,4] 苏源 梁家灿 刘庆 姚永兴 周晓罡 ZHANG Yi-ning;LIU Qiu-yan;ZHU Mei-zhu;ZENG Min;SU Yuan;LIANG Jia-can;LIU Qing;YAO Yong-xing;ZHOU Xiao-gang(College of Agriculture and Life Sciences,Kunming University,Kunming 650214,China;Biotechnology and Germplasm Resources In-stitute,Yunnan Academy of Agricultural Sciences,Kunming 650223,China;Yunnan Provincial Key Laboratory of Agricultural Biotech-nology,Kunming 650223,China;Key Laboratory of Southwestern Crop Gene Resources and Germplasm Innovation,Ministry of Agricul-ture,Kunming 650223,China;Yunan Haomeng Agricultural Technology Co.,Ltd.,Chuxiong,Yunnan 651604,China)

机构地区：[1]昆明学院农学与生命科学学院,昆明650214 [2]云南省农业科学院生物技术与种质资源研究所,昆明650223 [3]云南省农业生物技术重点实验室,昆明650223 [4]农业部西南作物基因资源与种质创制重点实验室,昆明650223 [5]云南昊盟农业科技有限公司,云南楚雄651604

出　　处：《西南农业学报》2024年第1期137-144,共8页Southwest China Journal of Agricultural Sciences

基　　金：云南省人力资源与社会保障厅专家基层工作站项目(云人社通[2022]17号);云南省牟定县生态畜牧产业科技特派团(202304BI090019);云南省自然科学基金重点项目(2016FA016)。

摘　　要：【目的】获得对云南白菜根肿病具有高效拮抗作用的微生物菌株,为云南白菜根肿病的生物防治提供潜在的菌种资源。【方法】从云南省曲靖市麒麟区三宝乡大棚白菜根肿病发病严重的地块,采集正常生长或发病较轻的白菜植株及其根际土壤,分离筛选抑菌效果较好的菌株,并对其进行形态特征鉴定、生理生化检测及分子序列分析。【结果】经初步筛选共获得46株细菌。土壤样品中共分离出30株细菌,其中抑制效果最好的是TB⁃7菌株,该菌株对白菜根肿病菌休眠孢子萌发抑制率为76.88%;白菜植株内共分离出16株内生细菌,其中抑制效果最好的是EB⁃15菌株,该菌株对白菜根肿病菌休眠孢子萌发抑制率为67.38%。通过对比分析选择抑菌效果最好的TB⁃7菌株作为目标菌株,经鉴定,TB⁃7菌株为革兰氏阳性芽孢杆菌,菌体菌落较小,颜色淡黄色,表面光滑且扁平,菌体湿润,不透明,边缘规则;生理生化指标检测结果显示,TB⁃7菌株是好氧菌,V⁃P、淀粉水解、pH 5.7,生长3项检测指标均呈阳性,柠檬酸盐、丙酸盐、D⁃木糖、L⁃阿拉伯糖、D⁃甘露醇、明胶液化、7%NaCl生长、硝酸盐还原8项检测指标均呈阴性;16S rDNA扩增产物约为1500 bp,测序结果表明,TB⁃7菌株与坚强芽孢杆菌(Cytobacillus firmus)同源性达到100%,鉴定该菌株为芽孢杆菌属的坚强芽孢杆菌。【结论】通过初步筛选及鉴定,获得1株对白菜根肿病菌休眠孢子抑制率达到76.88%的拮抗菌株,该菌株的获得可为解决云南本地日益严重的白菜根肿病的生物防治奠定基础。【Objective】The microbial strains with effective antagonistic effect against Yunnan cabbage clubroot were obtained through screen⁃ing,which provided potential strain resources for the biological control of Yunnan cabbage clubroot.【Method】Chinese cabbage plants with normal growth or mild disease and their rhizosphere soil were selected from the plot of the greenhouse in Sanbao town,Qilin district,Qujing city,Yunnan province.The strains with good bacteriostatic effect were isolated and screened,and their morphological characteristics,physiological and biochemical detection and molecular sequence analysis were performed.【Result】A total of 46 bacterial isolates were obtained through preliminary screening.Thirty bacterial strains were isolated from soil samples,of which TB⁃7 had the best inhibitory effect,and the inhibition rate of dormant spore germination was 76.88%;Sixteen strains of endophytic bacteria were isolated from Chinese cabbage,of which EB⁃15 had the best inhibitory effect and the germination inhibition rate of dormant spores was 67.38%.Through comparative analy⁃sis,strain TB⁃7 with the best antibacterial effect was selected as the target strain,and identified as a gram⁃positive Bacillus,with small colo⁃nies,pale yellow color,smooth and flat surface,and moist cells,opaque,with regular edges.The results of physiological and biochemical indexes showed that TB⁃7 was aerobic,and the test indicators of V⁃P,starch hydrolysis and growth at pH 5.7 were positive,citrate,propio⁃nate,d⁃xylose,L⁃arabinose,D⁃mannitol,gelatin liquefaction,7%NaCl growth and nitrate reduction were negative;The amplified product of 16S rDNA was about 1500 bp.The sequencing results showed that the homology between TB⁃7 strain and Cytobacillus firmus of the genus Bacillus reached 100%.【Conclusion】Through preliminary screening and identification,an antagonistic strain was screened with an inhibi⁃tion rate of 76.88%to dormant spores of Chinese cabbage clubroot,which would lay a foundation for the b

关 键 词：根肿病 生防菌 休眠孢子 菌种鉴定 

分 类 号：S634[农业科学—蔬菜学]