Liu C H, Chen X, Liu T T, Lian B, Gu Yucheng, Caer V, Xue Y R, Wang B T
State Key Laboratory of Pharmaceutical Biotechnology, School of Life Science, Nanjing University, 22# Hankou Road, Nanjing, 210093, People's Republic of China.
Appl Microbiol Biotechnol. 2007 Aug;76(2):459-66. doi: 10.1007/s00253-007-1010-0. Epub 2007 May 30.
An Acinetobacter strain, given the code name LCH001 and having the potential to be an endophytic antagonist, has been isolated from healthy stems of the plant Cinnamomum camphora (L.) Presl, guided by an in vitro screening technique. The bacterium inhibited the growth of several phytopathogenic fungi such as Cryphonectria parasitica, Glomerella glycines, Phytophthora capsici, Fusarium graminearum, Botrytis cinerea, and Rhizoctonia solani. Biochemical, physiological, and 16S rDNA sequence analysis proved that it is Acinetobacter baumannii. When the filtrate from the fermentation broth of strain LCH001 was tested in vitro and in vivo, it showed strong growth inhibition against several phytopathogens including P. capsici, F. graminearum, and R. solani, indicating that suppression of the growth of the fungi was due to the presence of antifungal compounds in the culture broth. Moreover, the antifungal activity of the culture filtrate was significantly correlated with the cell growth of strain LCH001. The active metabolites in the filtrate were relatively thermally stable, but were sensitive to acidic conditions. Three antifungal compounds were isolated from the culture broth by absorption onto macropore resin, ethanol extraction, chromatography on silica gel or LH-20 columns, and crystallization. The structures of the bioactive compounds were identified by spectroscopic methods as isomers of iturin A, namely, iturin A2, iturin A3, and iturin A6. The characterization of an unusual endophytic bacterial strain LCH001 and its bioactive components may provide an alternative resource for the biocontrol of plant diseases.
通过体外筛选技术,从健康的樟树(Cinnamomum camphora (L.) Presl)茎中分离出一株不动杆菌菌株,其代号为LCH001,具有作为内生拮抗物的潜力。该细菌抑制了几种植物病原真菌的生长,如寄生隐孢壳菌(Cryphonectria parasitica)、大豆炭疽菌(Glomerella glycines)、辣椒疫霉(Phytophthora capsici)、禾谷镰刀菌(Fusarium graminearum)、灰葡萄孢(Botrytis cinerea)和立枯丝核菌(Rhizoctonia solani)。生化、生理和16S rDNA序列分析证明它是鲍曼不动杆菌(Acinetobacter baumannii)。当对菌株LCH001发酵液的滤液进行体外和体内测试时,它对包括辣椒疫霉、禾谷镰刀菌和立枯丝核菌在内的几种植物病原体表现出强烈的生长抑制作用,这表明真菌生长受到抑制是由于培养液中存在抗真菌化合物。此外,培养液滤液的抗真菌活性与菌株LCH001的细胞生长显著相关。滤液中的活性代谢物相对热稳定,但对酸性条件敏感。通过大孔树脂吸附、乙醇萃取、硅胶或LH - 20柱色谱和结晶,从培养液中分离出三种抗真菌化合物。通过光谱方法将生物活性化合物的结构鉴定为伊枯草菌素A(iturin A)的异构体,即伊枯草菌素A2、伊枯草菌素A3和伊枯草菌素A6。对一种不寻常的内生细菌菌株LCH001及其生物活性成分的表征可能为植物病害的生物防治提供一种替代资源。