五种假单胞菌的分离鉴定及其生物活性

Isolation，identification and characterization of five Pseudomonas strains

摘要:

摘要:【目的】从湖南长沙市采集到的土样中分离假单胞菌并进行归类，研究各菌株抑菌和抗肿瘤生物活性，以丰富假单胞菌菌种资源并为微生物次级代谢物的挖掘奠定基础。【方法】采用大蜡螟诱集法诱集分离假单胞菌，结合形态观察、生理生化特征和16S rRNA基因序列同源性分析，鉴定并归类各细菌，通过平板扩散法、对峙培养法和肿瘤细胞毒性试验分别研究各菌株抑制细菌、拮抗真菌和抗肿瘤细胞等生物活性。【结果】从湖南长沙市郊区菜地、林地中分离得到5 株假单胞菌，归类并命名为Pseudomonas protegens CY01、绿针假单胞菌CY02 (Pseudomonas chlororaphis CY02)、栖稻假单胞菌CY04(Pseudomonas oryzihabitans CY04)、Pseudomonas sp. CY05和恶臭假单胞菌CY06(Pseudomonas putida CY06)。P.protegens CY01 和P.chlororaphis CY02对枯草芽胞杆菌(Bacillus subtillis)和金黄色葡萄球菌(Staphylococcus aureus)具有较好的抑菌效果，P．chlororaphis CY02对水稻稻瘟病菌(Pyricularia oryzae)具有良好的拮抗作用，对小鼠黑色素瘤细胞B16 具有较强的细胞毒性。【结论】分离得到的P．chlororaphis CY02，在抑制病原细菌、拮抗水稻稻瘟病菌和抗肿瘤细胞等方面具有显著效果。

Abstract:

Abstract:［Objective］ We isolated Pseudomonas strains from soil samples collected from Changsha，Hunan province，catalogued them and studied the antimicrobial and antitumor activity.［Methods］We isolated Pseudomonas strains from soil samples through Galleria bait method，identified and catalogued the isolated strains according to morphological observation，physiological and biochemical characteristics and the homologous analysis of 16S rRNA sequences of nucleotides．Antimicrobial，antagonistic fungi and anti-tumor activities were studied by diffusion plate assay，dual-culture assay and cytotoxicity test respectively.［Results］We isolated 5 Pseudomonas strains from vegetable field and forest land of Changsha suburb，catalogued and named them as Pseudomonas protegens CY01，Pseudomonas chlororaphis CY02，Pseudomonas oryzihabitans CY04，Pseudomonas sp. CY05 and Pseudomonas putida CY06 respectively. P. protegens CY01 and P. chlororaphis CY02 have antibacterial activity against Bacillus subtillis and Staphylococcus aureus. P. chlororaphis CY02 has antagonistic activity against Pyricularia oryzae and anti-tumor cell activity against mouse melanoma B16 cells.［Conclusion］The isolated P. chlororaphis CY02 has a significant effect on pathogenic bacteria，Pyricularia oryzae and tumor cells.