Biocontrol Efficiency of Bacillus subtilis SL-13 and Characterization of an Antifungal Chitinase

Abstract

Abstract: The seed germination and tomato seedling tests showed that Bacillus subtilis SL-13 could promote the sprouting and seedling growth of tomato.The fresh and dry weight of tomato seedlings increased 42.86%and 18.75%,respectively.The control efficacies of the SL-13 to tomato Rhizoctonia rot were 20.65% and 35.23% in the greenhouse and field,respectively.The growth of the plant-pathogenic fungus Rhizoctonia solani was considerably inhibited in the presence of the strain SL-13 culture supernatant.The main antifungal protein was detected to be chitinase through vitro assay.The chitinase was purified with DEAE-Sepharose fast flow ion exchange column chromatography and Sephadex G-75 gel filtration for further characterization.The optimal pH and temperature for the chitinase activity were 7.0 and 50℃,respectively.It was demonstrated that the enzyme was stable at pH 5-9 and 40-60℃.70% of the enzyme activity was retained when incubated at 121℃ and 0.11 MPa for 20 min,and the enzyme was not sensitive to protease K and ultraviolet radiation.Thus it is suitable for effective biological control in relatively unstable environment.

Key words: Bacillus subtilis, growth-promoting, diseases control, antifungal chitinase, purification, Rhizoctonia solani

摘要： The seed germination and tomato seedling tests showed that Bacillus subtilis SL-13 could promote the sprouting and seedling growth of tomato.The fresh and dry weight of tomato seedlings increased 42.86%and 18.75%,respectively.The control efficacies of the SL-13 to tomato Rhizoctonia rot were 20.65% and 35.23% in the greenhouse and field,respectively.The growth of the plant-pathogenic fungus Rhizoctonia solani was considerably inhibited in the presence of the strain SL-13 culture supernatant.The main antifungal protein was detected to be chitinase through vitro assay.The chitinase was purified with DEAE-Sepharose fast flow ion exchange column chromatography and Sephadex G-75 gel filtration for further characterization.The optimal pH and temperature for the chitinase activity were 7.0 and 50℃,respectively.It was demonstrated that the enzyme was stable at pH 5-9 and 40-60℃.70% of the enzyme activity was retained when incubated at 121℃ and 0.11 MPa for 20 min,and the enzyme was not sensitive to protease K and ultraviolet radiation.Thus it is suitable for effective biological control in relatively unstable environment.

关键词: Bacillus subtilis, growth-promoting, diseases control, antifungal chitinase, purification, Rhizoctonia solani

LIU Yan, TAO Jing, YAN Yujun, LI Bin, LI Hui, LI Chun. Biocontrol Efficiency of Bacillus subtilis SL-13 and Characterization of an Antifungal Chitinase[J]. , 2011, 19(1): 128-134.

刘燕, 陶晶, 阎豫君, 李彬, 李晖, 李春. Biocontrol Efficiency of Bacillus subtilis SL-13 and Characterization of an Antifungal Chitinase[J]. , 2011, 19(1): 128-134.

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