2002-08-012024-05-18https://scholars.lib.ntu.edu.tw/handle/123456789/712520摘要：本計畫目的在利用不同類型的幾丁質分解酵素基因，加強螢光假單胞根圈細菌的抗菌能力，增進生物防治的效果。首先將從可以分解幾丁質的Bacillus拮抗細菌中選殖其不同類型的幾丁質分解酵素基因、完成DNA定序及進行幾丁質分解酵素純化及抗菌活性分析。其次利用不完整的幾丁質分解酵素基因片段插入Bacillus拮抗細菌染色體DNA中，破壞幾丁質分解酵素基因的完整性，觀察幾丁質分解能力減弱之變異菌株的抗菌及生物防治能力，以闡明幾丁質分解酵素在Bacillus拮抗細菌抗菌作用中參與的角色。並建構可以分泌幾丁質分解酵素的螢光假單胞根圈細菌，使遺傳工程菌株可產生抗生素外，還可產生幾丁質分解酵素。進一步組合應用不同類型的幾丁質分解酵素及其基因，希望可以促進螢光假單胞根圈細菌抗菌及生物防治的效果。<br> Abstract: The goal of this project is to utilize different types of chitinase to improve the antifungal activity of fluorescent Pseudomonas rhizobacteria and enhance the efficacy of biological control. Firstly, the genes of different types of chitinases in a chitinolytic Bacillus antagonist will be cloned and sequenced. The chitinase will be purified and analyzed on its antifungal activity. Secondly, the chitinase gene on the Bacillus antagonist will be disrupted by gene replacement. The effect of attenuated chitinolytic activity on the antifungal and biocontrol efficacy of mutants will be investigated to elucidate the role of chitinase in the antifungal activity of Bacillus antagonist. Then, the chitinase gene will be reconstructed for the secretion of chitinase out of the fluorescent Pseudomonas rhizobacteria. The bacteria were genetically engineered to produce chitinase beside of antibiotics. Furthermore, the combination of different types of chitnases and its genes will be applied to enhance the antifungal and biocontrol efficacy of fluorescent Pseudomonas rhizobacteria.拮抗細菌幾丁質分解酵素基因工程抗菌作用生物防治antagonistic bacteriachitinasegenetic engineeringantifungal activitybiological control