菘蓝内生细菌的分离、筛选和鉴定  

Isolation,Screening and Identification of Endophytic Bacteria from Isatis indigotica

作　　者：王霞 薛林贵[1] 张晓华[2] 何小燕 范桃桃 尚海 WANG Xia;XUE Lin-gui;ZHANG Xiao-hua;HE Xiao-yan;FAN Tao-tao;SHANG Hai(College of Chemical and Biological Engineering, Lanzhou Jiaotong University, Key Laboratory of Extreme Environmental Microbial Resources and Engineering of Gansu, Lanzhou 730070;College of Environmental and Municipal Engineering, Lanzhou Jiaotong University, Lanzhou 730070)

机构地区：[1]兰州交通大学化学与生物工程学院甘肃省极端环境微生物资源与工程重点实验室,兰州730070 [2]兰州交通大学环境与市政工程学院,兰州730070

出　　处：《生物技术通报》2018年第3期163-169,共7页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31260135);甘肃省科技支撑计划项目(1504FKCA015);兰州市科技计划项目(2015-3-68)

摘　　要：筛选和鉴定药用植物菘蓝中对河西走廊常见植物病原菌禾谷镰刀菌、链格孢霉、大斑凸脐蠕孢有拮抗活性的内生细菌。利用常规分离法对菘蓝根、茎、叶、叶柄、花进行内生细菌的分离,采用同步培养法对分离的内生细菌进行活性菌株筛选,并对筛选的活性菌株进行形态观察,生理生化指标测定以及16S r RNA全序列鉴定。结果显示,共分离得到19株内生细菌,其中10株细菌对禾谷镰刀菌有不同程度的抑制作用,占分离菌总数的52.6%,其中菌株G2对禾谷镰刀菌的抑菌效果最强,抑菌率为94.63%,19株细菌均对链格孢霉,大斑凸脐蠕孢有不同程度的抑制作用,占分离菌总数的100%,其中菌株G2、J1、Y5及B2对这两种病原菌的抑菌率最大,均接近100%,经形态学观察,生理生化检测及16S r RNA序列分析,菌株G2为玫瑰色库克菌(Kocuriarosea);菌株J1,Y5为黄杆菌(Microbacterium maritypicum);菌株B2为人参短状杆菌(Brachybacterium ginsengisoli)。从菘蓝中分离筛选到了对河西走廊常见植物病原菌有强抑菌活性的内生细菌,尤其菌株G2、J1、Y5和B2的抑菌性较强,其抑菌活性值得进一步研究。This work is to screen and identify the endophytic bacteria from medicinal Isatis indigotica in the Hexi Corridor of GansuProvince against Fusarium moniliforme,Alternaria alternate,and Exserohilum turcicum. Endophytic bacteria were isolated from the roots,stems,leaves,petioles and flowers of I. indigotica using conventional separation method,from which the antagonistic bacteria were screenedby the synchronous culture method,and the screened active endophytic bacteria were identified based on phenotypical and biochemicalproperties as well as its 16 S r RNA gene sequence. As results,a total of 19 endophytes were isolated,and 10 strains of them had variedinhibition effects on F. moniliforme,accounting for the 52.6% of total isolates;the inhibition effect of strain G2 on F. moniliforme wasthe strongest with inhibition rates up to 94.63%. All 19 strains of the isolates had varied inhibition effects on A. alternata and E. turcicum,accounting for 100% of the total number of isolates. Among these strains,G2,J1,Y5 and B2 were selected for further study,and theirinhibitory rates nearly reached 100%. Combined with the physiological and biochemical characteristics,colony characteristics,cellmorphological characteristics and 16 S r RNA sequencing results,the strains J1 and Y5 were identified as Microbacterium maritypicum,and thestrain G2 and B2 as Kocuria rosea and Brachybacterium ginsengisoli,respectively. In summary,antagonistic bacteria on the plant pathogenswere isolated from I. indigotica,especially the strain G2,J1,Y5 and B2 presented the strongest inhibitory effects,and thus their inhibitoryactivities will be worthwhile for further studies.

关 键 词：菘蓝 内生菌 植物病原菌 抑菌活性 16S RRNA 

分 类 号：S476[农业科学—农业昆虫与害虫防治]