生物技术通报 ›› 2018, Vol. 34 ›› Issue (3): 163-169.doi: 10.13560/j.cnki.biotech.bull.1985.2017-0855

• 研究报告 • 上一篇    下一篇

菘蓝内生细菌的分离、筛选和鉴定

王霞1, 薛林贵1, 张晓华2, 何小燕1, 范桃桃1, 尚海1   

  1. 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
  • 收稿日期:2017-10-15 出版日期:2018-03-20 发布日期:2018-04-10
  • 作者简介:王霞,女,硕士,研究方向环境微生物、微生物生理及微生物工程;E-mail295101203@qq.com
  • 基金资助:
    国家自然科学(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)

Isolation,Screening and Identification of Endophytic Bacteria from Isatis indigotica

WANG Xia1, XUE Lin-gui1, ZHANG Xiao-hua2, HE Xiao-yan1, FAN Tao-tao1, SHANG Hai1,   

  1. 1. College of Chemical and Biological Engineering,Lanzhou Jiaotong University,Key Laboratory of Extreme Environmental Microbial Resources and Engineering of Gansu,Lanzhou 730070;
    2. College of Environmental and Municipal Engineering,Lanzhou Jiaotong University,Lanzhou 730070
  • Received:2017-10-15 Published:2018-03-20 Online:2018-04-10

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 rRNA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 rRNA序列分析,菌株G2为玫瑰色库克菌(Kocuria rosea);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。

关键词: 菘蓝, 内生菌, 植物病原菌, 抑菌活性, 16S rRNA

Abstract: This work is to screen and identify the endophytic bacteria from medicinal Isatis indigotica in the Hexi Corridor of Gansu Province 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 screened by the synchronous culture method,and the screened active endophytic bacteria were identified based on phenotypical and biochemical properties as well as its 16S rRNA gene sequence. As results,a total of 19 endophytes were isolated,and 10 strains of them had varied inhibition effects on F. moniliforme,accounting for the 52.6% of total isolates;the inhibition effect of strain G2 on F. moniliforme was the 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 their inhibitory rates nearly reached 100%. Combined with the physiological and biochemical characteristics,colony characteristics,cell morphological characteristics and 16S rRNA sequencing results,the strains J1 and Y5 were identified as Microbacterium maritypicum,and the strain G2 and B2 as Kocuria rosea and Brachybacterium ginsengisoli,respectively. In summary,antagonistic bacteria on the plant pathogens were isolated from I. indigotica,especially the strain G2,J1,Y5 and B2 presented the strongest inhibitory effects,and thus their inhibitory activities will be worthwhile for further studies.

Key words: Isatis indigotica, endophytic bacteria, phytopathogen, antifungal activity, 16S rRNA