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Enhancement of antifungal activity of Burkholderia pyrrocinia JK ‐ SH 007 genetically modified with Bacillus subtilis Chi113 gene
Author(s) -
He L.M.,
Ye J.R.,
Ren J.H.,
Huang L.,
Wu X.Q.
Publication year - 2016
Publication title -
forest pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.535
H-Index - 49
eISSN - 1439-0329
pISSN - 1437-4781
DOI - 10.1111/efp.12283
Subject(s) - biology , bacillus subtilis , microbiology and biotechnology , chitinase , plasmid , reporter gene , transformation (genetics) , gene , gene expression , bacteria , biochemistry , genetics
Summary Previous studies showed that Burkholderia pyrrocinia JK ‐ SH 007 is a potential biocontrol agent of poplar canker disease. In this report, B. pyrrocinia JK ‐ SH 007 was genetically modified by introducing the Bacillus subtilis Chi113 gene to enhance its antifungal activity. The green fluorescence of B. pyrrocinia JK ‐ SH 007E1 can be detected because of its transformation with plasmid pHKT 2‐ Chi113 containing a gfp reporter gene. Real‐time quantitative PCR ( qPCR ) showed that the B. subtilis Chi113 gene was highly expressed in B. pyrrocinia JK ‐ SH 007E1 at mRNA level. The chitinase activity and the antifungal activity of B. pyrrocinia JK ‐ SH 007E1 were significantly increased, and the clear halo was visible on the colloidal chitin agar plate. In addition, the constructed recombinant plasmid pHKT 2‐ Chi113 was stably maintained for at least 100 generations in the absence of antibiotic selection in B. pyrrocinia JK ‐ SH 007E1. In western blot analysis, the Chi113‐ GFP fusion protein was detected in B. pyrrocinia JK ‐ SH 007E1 using the GFP polyclonal antibody. In conclusion, the B. subtilis Chi113 gene was successfully transformed into B. pyrrocinia JK ‐ SH 007 and was expressed both at mRNA level and protein level. Moreover, the antifungal activity of B. pyrrocinia JK ‐ SH 007 was improved through this genetic modification.