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烟草赤星病菌拮抗性大豆根瘤内生菌的筛选及抑制作用
引用本文:赵龙飞,徐亚军,侯怡婷,邹艳慧,李亚楠,杨志华,李小雨,张梦瑶.烟草赤星病菌拮抗性大豆根瘤内生菌的筛选及抑制作用[J].生态学杂志,2016,27(5):1560-1568.
作者姓名:赵龙飞  徐亚军  侯怡婷  邹艳慧  李亚楠  杨志华  李小雨  张梦瑶
作者单位:河南省高校植物与微生物互作重点实验室/商丘师范学院生命科学学院, 河南商丘 476000
基金项目:本文由国家自然科学基金项目(U1204301)和河南省高校青年骨干教师项目(2012GGJS166)资助
摘    要:以烟草赤星病菌为供试病原菌,采用对峙法、代谢液培养法对分离自大豆根瘤的内生细菌进行抑菌筛选,对筛选菌株作用的病原菌菌丝进行显微观察,研究筛选菌株细胞培养特征及生理生化特性、16S rDNA测序序列、系统发育,以及进行温室接种防病试验.结果表明: 大豆根瘤内生菌经初筛、复筛,对烟草赤星病菌抑菌率42%以上的有7株菌,分别属于芽孢杆菌属、假单胞菌属、中华根瘤菌属和寡养单胞菌属.显微观察表明,在拮抗性内生菌作用下病原菌菌丝末端出现明显畸形,呈珊瑚状分枝、球状膨大等;代谢液试验表明,内生菌对病原菌的抑制作用主要是内生菌代谢产生的胞外物质在发挥作用;对峙试验表明,芽孢杆菌可迅速形成生物薄膜,有效阻碍病原菌丝生长和延伸.盆栽防病试验表明,拮抗性内生菌处理下烟草病情指数显著低于对照,表明筛选菌株可作为烟草赤星病的生防菌株资源.

关 键 词:烟草赤星病菌  根瘤内生菌  16SrDNA测序  系统发育  生物防治
收稿时间:2015-09-17

Screening and inhibition of antagonistic endophytic bacteria isolated from soybean (Glycine max)nodules against Alternaria longipes.
ZHAO Long-fei,XU Ya-jun,HOU Yi-ting,ZOU Yan-hui,LI Ya-nan,YANG Zhi-hua,LI Xiao-yu,ZHANG Meng-yao.Screening and inhibition of antagonistic endophytic bacteria isolated from soybean (Glycine max)nodules against Alternaria longipes.[J].Chinese Journal of Ecology,2016,27(5):1560-1568.
Authors:ZHAO Long-fei  XU Ya-jun  HOU Yi-ting  ZOU Yan-hui  LI Ya-nan  YANG Zhi-hua  LI Xiao-yu  ZHANG Meng-yao
Institution:(Henan Province Key Laboratory of Plant-Microbe Interactions/College of Life Sciences, Shangqiu Normal University, Shangqiu 476000, Henan, China).
Abstract:Using Alternaria longipes as tested phytopathogen, endophytic bacteria isolated from soybean nodules were selected to study antagonistic effects by confrontation and metabolic liquid culture methods. The inhibited hyphae were determined by microscopic observation, and the screened strains were characterized by cell culture, physiological and biochemical tests, 16S rDNA sequencing, phylogenetic analysis and inoculation trials in greenhouse. The results indicated that the seven of the endophytes exerted more than 42% inhibitory effects after the first and the second screening. These strains belonged to genus Bacillus, Pseudomonas, Sinorhizobium and Stenotrophomonas, respectively. Microscopic observation showed that the affected hyphae ends of A. longipes appear deformity of coralline branch, spherical expansions and so on. Antagonistic experiments with metabolites showed that the inhibition of endophytic bacteria against pathogenic fungus played an effective role mainly by bacteria producing extracellular substances. Confrontation tests suggested that endophytic Bacillus rapidly produced biofilm to effectively hinder the growth and extension of pathogen hyphae. Inoculation experiments showed that the disease index of treatment group was significantly lowered compared with the control, suggesting it could be utilized as a biological control resource inhibiting tobacco brown spot.
Keywords:Alternaria longipes  endophytic bacteria of root nodules  16S rDNA sequencing  phylogenetic analysis  biocontrol  
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