Enhancement of antifungal activity of Burkholderia pyrrocinia JK-SH007 genetically modified with Bacillus subtilis Chi113 gene
Enhancement of antifungal activity of Burkholderia pyrrocinia JK-SH007 genetically modified with Bacillus subtilis Chi113 gene
复制标题
枯草芽孢杆菌 Chi113 基因修饰的吡咯伯克霍尔德氏菌 JK-SH007 抗真菌活性增强
DOI：
10.1111/efp.12283
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发表时间：
2016-12-01
期刊：
影响因子：
1.4
通讯作者：
Wu, X. -Q.
中科院分区：
文献类型：
--
作者：
He, L. -M.;Ye, J. -R.;Wu, X. -Q.
前期研究表明，Burkholderia pyrrocinia JK-SH 007是一种潜在的白杨溃疡病生防菌。在本报告中，B.通过引入枯草芽孢杆菌Chi 113基因对pyrrocinia JK-SH 007进行遗传修饰以增强其抗真菌活性。B的绿色荧光。pyrrocinia JK-SH 007 E1可以被检测到，因为它被含有gfp报告基因的质粒pHKT 2-Chi 113转化。实时荧光定量PCR（qPCR）结果显示，该B.枯草芽孢杆菌Chi 113基因在B. pyrrocinia JK-SH 007 E1基因的mRNA水平。测定了B. pyrrocinia JK-SH 007 E1的表达量显著增加，在几丁质琼脂平板上可见清晰的晕圈。此外，构建的重组质粒pHKT 2-Chi 113在B中不进行抗生素筛选的情况下稳定地保持了至少100代。核盘菌JK-SH 007 E1。Western blot分析显示，在B细胞中检测到Chi 113-GFP融合蛋白。pyrrocinia JK-SH 007 E1的多克隆抗体。总之，B.将Chi 113基因成功转化到B. pyrrocinia JK-SH 007的表达，并在mRNA水平和蛋白水平表达。此外，B. pyrrocinia JK-SH 007通过这种遗传修饰进行了改良。
Previous studies showed that Burkholderia pyrrocinia JK-SH007 is a potential biocontrol agent of poplar canker disease. In this report, B. pyrrocinia JK-SH007 was genetically modified by introducing the Bacillus subtilis Chi113 gene to enhance its antifungal activity. The green fluorescence of B. pyrrocinia JK-SH007E1 can be detected because of its transformation with plasmid pHKT2-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-SH007E1 at mRNA level. The chitinase activity and the antifungal activity of B. pyrrocinia JK-SH007E1 were significantly increased, and the clear halo was visible on the colloidal chitin agar plate. In addition, the constructed recombinant plasmid pHKT2-Chi113 was stably maintained for at least 100 generations in the absence of antibiotic selection in B. pyrrocinia JK-SH007E1. In western blot analysis, the Chi113-GFP fusion protein was detected in B. pyrrocinia JK-SH007E1 using the GFP polyclonal antibody. In conclusion, the B. subtilis Chi113 gene was successfully transformed into B. pyrrocinia JK-SH007 and was expressed both at mRNA level and protein level. Moreover, the antifungal activity of B. pyrrocinia JK-SH007 was improved through this genetic modification.