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Frontiers of Agriculture in China

ISSN 1673-7334

ISSN 1673-744X(Online)

CN 11-5729/S

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Front Agric Chin    2011, Vol. 5 Issue (4) : 581-587    https://doi.org/10.1007/s11703-011-1111-5
RESEARCH ARTICLE
Screening and Identification of the antagonistic strain DL-59 of B. velezensis against A. brassicae and biocontrol efficiency
Shuna LI(), Shutao DU, Chaoyu LI
College of Life Science, Agricultural University of Hebei, Baoding 071001, China
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Abstract

The aim of this study was to screen the antagonistic spore-forming bacteria of Alternaria brassicae and to assess their control effect. The test microorganism was A. brassicae in this experiment. One spore-forming strain named DL-59 with a rather stronger antagonistic activity than ever reported was obtained by using an improved agar plate diffusion method. Morphological, physiological and biochemical tests combined with 16S rDNA sequence analysis were carried out to identify DL-59. Its control effect was studied in the basin culture and the plot trials. Two hundred strains were isolated form soil and 20 antagonistic bacteria strains were obtained through preliminary screen. After the secondary screening, a strain named DL-59 with a rather stronger antagonistic activity was obtained and identified as B. velezensis with the largest diameter of inhibition zone of 23 mm. The control efficacy was 79.07%. The plot trials showed that DL-59 had a visible preventive effect on cabbage black spot disease. This can be applied for biological control of plant disease. The strain can produce spores, which is important for the storage of its preparation.
Keywords antagonistic bacterium      screening and identification      16S rDNA      control effect     
Corresponding Author(s): LI Shuna,Email:bdlishuna@126.com   
Issue Date: 05 December 2011
 Cite this article:   
Chaoyu LI,Shutao DU,Shuna LI. Screening and Identification of the antagonistic strain DL-59 of B. velezensis against A. brassicae and biocontrol efficiency[J]. Front Agric Chin, 2011, 5(4): 581-587.
 URL:  
https://academic.hep.com.cn/fag/EN/10.1007/s11703-011-1111-5
https://academic.hep.com.cn/fag/EN/Y2011/V5/I4/581
Disease levelDegreeNumeric value
1No symptoms0
2Leaves with small brown spots, l-31
3Leaves with brown spots, 4-82
4Leaves with brown spots, 9 or more3
Tab.1  Cabbage black spot disease classification standard
Fig.1  Antagonistic effect of isolated bacteria strains against the pathogen.
Fig.2  Antagonistic effect of isolated bacteria strains against the pathogen by the secondary screening.
Fig.3  Colony morphology of DL-59 on NA.
Fig.4  Morphological shapes of thallus and spore of the antagonistic bacteria strain DL-59.
ItemResultItemResult
Growth temperature and heat resistance 65°C+Sugar and spirits fermentation+
Growth temperature and heat resistance 45°C+Methyl red-
Growth temperature and heat resistance 37°C+V.P determination-
Growth temperature and heat resistance 4°C-Starch hydrolysis+
Nitrogen sources utilization(NH4)2HPO4+Production of dextrine crystallization-
Nitrogen sources utilization KNO3-Cellulose decomposition+
Carbon sources utilization mannose+3-ketone group lactose+
Carbon sources utilization maltose+NNitrate reduction-
Carbon sources utilization sucrose-Nitrite reduction+
Carbon sources utilization soluble starch+Denitrification-
Fluorochrome-Ammonia production+
Production of pyocyanin-Urease-
Malonic acid utilization-Phenylalanine deaminase+
Citrate utilization-Tryptophan deaminase+
Tartrate utilization-Lipase test+
Catalase+Hydrogen sulfide+
Oxidation and ferment of glucosefermentativeMilk decompositionpeptonization
Indole+
Tab.2  The results of physiological and biochemical identification
Fig.5  Phylogenetic tree of 16S rDNA sequence of DL-59 strain and related strains.
Sequence numberSpeciesStrain-numberSimilarity (%)
AJ496807Amphibacillus xylanusDSM 662687.90%
AJ276351Bacillus subtilisDSM1099.42%
EF433406Bacillus amyloliquefaciensBCRC 1160199.57%
AY603658Bacillus velezensisCR-50299.78%
AB245422Bacillus velezensisLMG2247899.78%
AB021191Bacillus mojavensisIFO1571899.28%
DQ993671Bacillus axarquiensisLMG 2247699.28%
DQ993673Bacillus malacitensisLMG 2247799.28%
AF547209Bacillus acidicola105-293.87%
DQ207729Bacillus cereusCCM 201093.19%
Tab.3  Comparison of similarity between antagonistic strain DL-59 and several norm stains
TreatmentTreatment patternP = 0.05Total numberDiseased plantDisease indexControl efficacy (%)
DL-59spraya601316.8576.37
Carbendazol wettable powderspraya601517.8275.00
Watersprayb605571.30-
Tab.4  The basin culture trials control effect of DL-59 against cabbage black spot disease
TreatmentTreatment patternP = 0.05Total numberDiseased plantDisease indexControl efficacy (%)
DL-59spraya12097.5079.07
Carbendazol wettable powderspraya1201428.1378.51
Watersprayb1204335.83-
Tab.5  The plot trials control effect of DL-59 against cabbage black spot disease
Fig.6  Diseased plant after artificial inoculation of
Fig.7  The healthy plant sprayed with the fermented liquid of DL-59 after artificial inoculation of
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