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烟草青枯病拮抗内生细菌的分离、鉴定及其田间防效

易有金1;刘如石2;尹华群3;罗宽1;刘二明1;刘学端3   

  1. 1湖南农业大学生物安全科技学院, 长沙 410128;
    2湖南师范大学基因功能与调控研究室, 长沙 410081;
    3中南大学资源加工与生物工程学院, 长沙 410083
  • 收稿日期:2006-09-16 修回日期:2006-11-26 出版日期:2007-03-18 发布日期:2007-03-18

Isolation, identification and field control efficacy of an endophytic strain against tobacco bacterial wilt (Ralstonia solanacarum)

YI You-jin1; LIU Ru-shi2; YIN Hua-qun3; LUO Kuan1; LIU Er-ming1; LIU Xue-duan3   

  1. 1College of Bio-Safety Science and Technology, Hunan Agricultural University, Changsha 410128, China;
    2Laboratory of Gene Function and Genome Engineering, Hunan Normal University, Changsha 410081, China;
    3School of Minerals Processing & Bioengineering, Central South University, Changsha 410083, China
  • Received:2006-09-16 Revised:2006-11-26 Online:2007-03-18 Published:2007-03-18

摘要: 从病区健康烟草植株茎杆内分离到一株对烟草青枯罗尔氏菌(Ralstonia solanacarum)有强拮抗作用的内生细菌,命名为B-001菌株.拮抗性研究表明,B-001菌株对多种革兰氏阳性细菌、革兰氏阴性细菌以及病原真菌均有较强的抑制作用.形态和生理生化特征初步表明菌株B-001为芽孢杆菌属(Bacillus)细菌.经扩增、测序得到B-001的16S rDNA序列,GenBank接收号为DQ444283.用ClustalX进行多重序列对比,并通过MEGA3方法构建16S rDNA系统发育树,表明:菌株B-001与Bacillus subtilis (DQ415893)的相似性为99.2%,并处于同一分支;结合形态和生理生化指标,将其鉴定为枯草芽孢杆菌(B. subtilis).2005和2006年在湖南省桂阳县、宁乡县进行了田间试验,防效在40.03%~78.14%,防治效果良好,且明显优于农用链霉素.

关键词: 九顶山, 红桦种群, 空间格局, 距离方法

Abstract: In this paper, an endophytic strain B-001 against tobacco bacterial wilt (Ralstonia solanacarum) was isolated from the stem of healthy tobacco in R. solanacarum-infected fields, which had a stronger inhibitory effect on some kinds of gram-positive bacteria, gram-negative bacteria, and pathogenic fungi. This strain belonged to Bacillus, and its 16S rDNA after PCR and sequencing had an accession of GenBank being DQ444283. The 16S rDNA phylogenetic tree was constructed with MEGA3, and compared with the published 16S rDNA sequences of relative bacteria species. B-001 had a 99.2% sequence similarity with Bacillus subtilis (DQ415893). According to the mor-phological, physiological and biochemical characteristics, and based on phylogenetic analysis, B-001 was identified as a strain of B. subtilis. Field experiments in Guiyang and Ningxiang counties of Hunan Province showed that in 2005 and 2006, the control efficacy of B-001 on R. solanacarum ranged from 40.03% to 78.14%, better than that of Streptomycini.

Key words: Jiuding Mountain, Betula albo-sinensis population, Spatial pattern, T-square