甘肃省黄芪细菌性叶斑病病原的分离和鉴定  

Isolation and Identification of Pathogen Causing Bacterial Leaf Spot of Astragalus membranaceus in Gansu Province

作　　者：吕娇娇 晋玲 王艳 张婷 高国香 LYU Jiao-jiao;JIN Ling;WANG Yan;ZHANG Ting;GAO Guo-xiang(School of Pharmacy,Gansu University of Chinese Medicine,Lanzhou 730000,China;State Key Laboratory of Dao-di Herbs,Beijing 100700,China)

机构地区：[1]甘肃中医药大学药学院,甘肃兰州730000 [2]道地药材国家重点实验室,北京100700

出　　处：《中国现代中药》2023年第3期568-573,共6页Modern Chinese Medicine

基　　金：国家自然科学基金项目(31960004);中央本级重大增减支项目(2060302);中国科学院“西部之光”人才培养引进计划项目;甘肃省创新基地和人才计划项目;西北中藏药省部共建协同创新中心开放基金项目(Xbzzy-2022-02)。

摘　　要：目的:明确甘肃省黄芪细菌性叶斑病的病原。方法:通过植物病原细菌形态学、生理生化指标测定,以及16S rDNA、rpoB和rpoD基因位点联合构建系统发育树,对黄芪细菌性叶斑病病原菌进行鉴定。结果:病原菌在营养琼脂(NA)培养基上培养24 h后菌落直径为1~3 mm,圆形或近圆形,乳白色、光滑凸起、半透明状、边缘整齐;革兰染色反应为阴性,好氧型,在10~40℃均能生长,在pH为5.7、6.8的环境中生长良好,在2%、5%NaCl环境下可生长;可利用葡萄糖、木糖、甘露糖、柠檬酸盐及硝酸盐;在代谢活动中可产生酪氨酸酶、过氧化氢酶、苯丙氨酸脱氨酶;分解色氨酸,液化明胶;不产生卵磷脂酶、淀粉水解酶及乙酰甲基甲醇。致病性测定初步验证了黄芪细菌性叶斑病的致病菌;16S rDNA、rpoB和rpoD基因序列分析进一步确定引起黄芪细菌性叶斑病的病原菌为绿黄假单胞菌Pseudomonas viridiflava(Burkholder)Dowson。结论:明确了甘肃省黄芪细菌性叶斑病的病原,为黄芪细菌性叶斑病的有效防治提供参考。Objective:To identify the pathogen causing bacterial leaf spot of Astragalus membranaceus in Gansu province and lay a foundation for the prevention and control of this disease.Methods:The pathogen was identified based on the morphological,physiological,and biochemical characteristics and the phylogenetic tree constructed based on 16S rDNA,rpoB,and rpoD.Results:The pathogen colony on NA medium appeared round or nearly round,milky white,smooth,convex,and translucent,with entire margin and the diameter of 1-3 mm.It is an aerobic Gram-negative bacterium which can grow at 10-40℃,pH 5.7 and 6.8.The pathogen can tolerate 2%and 5%NaCl,while it cannot grow in the medium with 7%or 10%NaCl.It can utilize glucose,xylose,mannose,citrate,and nitrate and produce tyrosinase,catalase,and phenylalanine deaminase.It can decompose tryptophan,liquefy gelatin,and cannot produce lecithinase,starch hydrolase,or acetylmethylcarbinol.The pathogenicity test preliminarily verified the isolated bacterium was the pathogen causing bacterial leaf spot of A.membranaceus.The sequence analysis of 16S rDNA,rpoB,and rpoD further identified the pathogen as Pseudomonas viridiflava(Burkholder)Dowson.Conclusion:This study provides a theoretical basis for the prevention and treatment of bacterial leaf spot of A.membranaceus.

关 键 词：黄芪 细菌性叶斑病 病原鉴定 

分 类 号：R282.2[医药卫生—中药学]