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从质粒pUC1965得到含有几丁质酶基因的6.5 kb DNA片断,将该基因与大肠杆菌-荧光假单胞杆菌穿梭质粒pDSK519连接,获得重组质粒pDSK51965。重组质粒转入荧光假单胞杆菌(Pseudomonasfluorescence)P5,获得工程菌株P.fluorescence P5-1。与野生菌株相比,平板拮抗实验表明, 工程菌株对小麦全蚀病(Gaeumannomycesgraminisvar. tricici)、水稻纹枯病(Rhizoctoniasalani)和棉花立枯病(R.salani )抑菌效果明显提高;盆栽防病实验表明,工程菌株显著地提高了对水稻纹枯病及棉花立枯病的防效,并提高了对小麦全蚀病的防效。
The 6.5 kb DNA fragment containing the chitinase gene was obtained from the plasmid pUC1965, and the gene was ligated with the E. coli-Pseudomonas fluorescens shuttle plasmid pDSK519 to obtain the recombinant plasmid pDSK51965. The recombinant plasmid was transformed into Pseudomonas fluorescein P5 to obtain the engineering strain P. fluorescein P5-1. Compared with the wild strains, the antagonistic experiments showed that the antibacterial effect of the engineering strains on Gaeumannomyces graminisvar. Tricici, Rhizoctonia sinensis and R.salani significantly increased. Experiments show that the engineering strain significantly improves the control effect on rice sheath blight and cotton blight and improves the control effect on wheat total eclipse.