Isolation and identification of the antagonistic strain DM-54 of <italic>Bacillus amyloliquefacien</italic> against <italic>Verticillium dahliae</italic>, and optimization of antifungal protein producing conditions

doi:10.1007/s11703-009-0005-2

Front Agric Chin

2009, Vol. 3

Issue (1) : 16-23 https://doi.org/10.1007/s11703-009-0005-2

RESEARCH ARTICLE

Isolation and identification of the antagonistic strain DM-54 of Bacillus amyloliquefacien against Verticillium dahliae, and optimization of antifungal protein producing conditions

Ying ZHU¹(

), Shuna LI², Hongshui YUAN², Xiaojun GUO², Baocheng ZHU^1,2(

)

1. College of Life Science, Hebei University, Baoding 071002, China; 2. College of Life Science, Agricultural University of Hebei, Baoding 071001, China

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Abstract

The strains capable of resistance against Verticillium dahliae Kleb were isolated and screened from the soils of cotton fields from several different provinces in China. A strain, coded DM-54, with a rather high antagonistic activity was obtained. Its morphological characteristics, physiological and biochemical properties and a 16 S rDNA sequence of this strain were further studied. The DM-54 strain was finally identified as a kind of Bacillusamyloliquefacien. Through a single factor experiment and an orthogonal experiment, the optimal shaking flask fermentation condition of strain DM-54 was found to be: media composed of 5% dextrin, 3% soy peptone, 0.02% MgSO₄, 0.01% CaCl₂, initial pH 7.0 and 10% inoculum volume, media volume 30/250 (mL/mL), fermentation temperature at 32oC, rotating speed 200 r?min^-1, fermentation time of 48 h. Its antagonistic activity was distinguished to be elevated, at about 39.9%. Our research offers an effective means for the massive production of antagonistic proteins.

Keywords cotton verticilliumwilt antagonistic bacterium isolation and identification Bacillus amyloliquefacien fermentation

Corresponding Author(s): ZHU Ying,Email:aileenzy@126.com; ZHU Baocheng,Email:zhu2222@126.com

Issue Date: 05 March 2009

Cite this article:

Ying ZHU,Shuna LI,Hongshui YUAN, et al. Isolation and identification of the antagonistic strain DM-54 of Bacillus amyloliquefacien against Verticillium dahliae, and optimization of antifungal protein producing conditions[J]. Front Agric Chin, 2009, 3(1): 16-23.

URL:

https://academic.hep.com.cn/fag/EN/10.1007/s11703-009-0005-2
https://academic.hep.com.cn/fag/EN/Y2009/V3/I1/16

Fig.1 Colonial morphology of stain DM-54

Tab.1 Physiological and biochemical characteristics of strain DM-54

Fig.2 Phylogenetic tree of the 16 S rDNA sequence of strain DM-54 and related strains

Tab.2 Comparison of similarities between several norm stains and strain DM-54

Tab.3 Effect of carbon source on antagonistic protein production of strain DM-54

Tab.4 Effect of nitrogen source on antagonistic protein production of strain DM-54

Tab.5 Effect of ions on antagonistic protein production of strain DM-54

Tab.6 Orthogonal experimental design and results of culture media

Tab.7 Orthogonal experimental design and results of culture conditions

Fig.3 Effect of rotating speed on antagonistic protein activity of strain DM-54

Fig.4 Effect of temperature on antagonistic protein activity of strain DM-54

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