高产申嗪霉素和吩嗪-1-酰胺的水稻根际铜绿假单胞菌PA1201分离、鉴定与应用潜力

Isolation，identification and characterization of rice rhizobacterium Pseudomonas aeruginosa PA1201 producing high level of biopesticide “Shenqinmycin”and phenazine-1-carboxamide

摘要:

摘要:【目的】从水稻根际筛选能高效抑制水稻病原菌的细菌，分析和鉴定其形态和生化特征，为开发新型绿色农药奠定基础。【方法】从水稻根际分离能以1-氨基环丙烷-1-羧酸(ACC)为唯一碳源的菌株，根据菌株形态和生化特性、16S rDNA序列比对和磷脂脂肪酸图谱，对该菌株进行鉴定。通过氯仿萃取抽提、高效液相色谱分析，确定菌株PA1201在PPM培养基和黄豆粉培养基中申嗪霉素和吩嗪-1-酰胺的产量。【结果】菌株PA1201能有效抑制水稻纹枯病菌和水稻白叶枯病菌的生长，属于铜绿假单胞菌( Pseudomonas aeruginosa sp. PA1201);PA1201产生两种抑菌代谢产物申嗪霉素和吩嗪-1-酰胺，在PPM和黄豆粉培养基中申嗪霉素的产量可达81.7 mg/L和926.9 mg/L，吩嗪-1-酰胺的产量亦可达18.1 mg/L和489.5 mg/L;PA1201产生大量胞外蛋白酶，对人肺腺癌细胞系A549和黑腹果蝇具有一定毒性。【结论】PA1201的申嗪霉素产量比现有 生产菌株的出发菌株M18高3－4倍，还能产生另一种抑菌活性更高的衍生物吩嗪-1-酰胺，具有进一步开发的潜力。

关键词:关键词:假单胞菌，申嗪霉素，吩嗪-1-酰胺，绿色农药，应用潜力

Abstract:

Abstract:［Objective］To identify bacterial strains with the inhibitory activity to rice pathogens，and to evaluate their potentials for the development of new biopesticides.［Methods］Ｒice rhizosphere Pseudomonas strains were isolated using 1-aminocyclopropane-1-carboxylic acid as the sole carbon source． Strain PA1201 was further identified through morphological analysis，biochemical characterization，16S rDNA sequence analysis and phospholipid fatty acid profiling．Qualitative and quantitative analysis of the production of the green pesticide Shenqinmycin as well as phenazine-1-carboxamide produced by PA1201 was done by HPLC．Cytotoxicity of PA1201 was evaluated using human alveolar epithelial cell line A549 and Drosophila melanogaster as hosts．［Results］Strain PA1201 inhibited Rhizotonia solani Kühn and Xanthomonas oryzae pv．oryzae，the causal agents of rice sheath blight and bacterial blight，respectively．It was further identified as Pseudomonas aeruginosa PA1201，which produces shenqinmycin and phenazine-1-carboxamide．The fermentation titer of shenqinmycin and phenazine-1-carboxamide in the PPM medium was 81. 7 mg/L and 18.1 mg/L，respectively．In the medium supplemented with soybean meal and corn steep liquor，the level of shenqinmycin and phenazine-1-carboxamide reached 926.9 mg/L and 489.5 mg/L．PA1201 also produced high level of extracellular protease and was toxic to human cell line and fruit fly．［Conclusion］Strain PA1201 could be engineered for higher yield of Shenqinmycin or for a new biopesticide．

Key words:Keywords: Pseudomonas aeruginosa，Shenqinmycin; phenazine-1-carboxamide，biopesticide