一种新的黄瓜细菌性叶斑病拮抗细菌的筛选鉴定及其发酵条件的初步研究
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摘要
【目的】旨在从海南、安徽、浙江等地的土壤中分离、筛选出对黄瓜叶斑病新的病原菌菊苣假单胞(Pseudomonas cichorii)具有拮抗作用的细菌,并对其发酵条件进行初步研究。【方法】采用抑菌圈法筛选黄瓜叶斑病病原菌菊苣假单胞的拮抗菌,采用形态学观察、生理生化特性测定、16S rDNA同源性序列分析以及gyrA基因序列分析的方法对分离的拮抗菌进行鉴定。采用单因子变量法对拮抗菌的最佳发酵条件进行研究。【结果】从海南、安徽、浙江等地共分离出650株细菌,筛选出10株拮抗菌。其中编号为JNL的菌株拮抗效果最好。细菌鉴定结果表明,该菌为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)。最佳发酵条件研究结果表明,该菌在30℃、pH 7、碳源为半乳糖、氮源为硝酸钾、转速为150 r/min的条件下生长效果最好。【结论】JNL菌株对黄瓜细菌性叶斑病新的病原菌菊苣假单胞有较好的防治效果,具有开发成生物农药或生物肥料的潜力。
【Objective】The aim is to isolate, identificate the antagonism of a new pathogen(Pseudomonas cichorii)from the soils of Hainan,Anhui,Zhejiang,etc.and explore its optimal fermentation condition.【Method】The antagonistic bacteria was isolated by the inhibition zone method. The isolated antagonistic bacteria were identified by morphological characters,physiological and biochemical tests,16 S rDNA homology sequence analysis and gyrA gene sequence analysis. The optimal fermentation conditions of antagonistic bacteria were studied by single factor variable method.【Result】The results showed that 650 strains of bacteria were isolated from Hainan,Anhui,Zhejiang,ect.10 strains of antagonistic bacteria were screened. Among them, the strain numbered JNL had the best antagonistic effect. Bacterial identification results showed that the strain was Bacillus amyloliquefaciens.The optimal fermentation conditions showed that the strain had the best growth at 30 ℃, pH 7,carbon source galactose,nitrogen source of potassium nitrate and rotation speed of 150 r/min.【Conclusion】The strain JNL has a good control effect on the new pathogen of leaf spot of cucumber and the potential to be developed into biological pesticide or biological fertilizer.
【Objective】The aim is to isolate, identificate the antagonism of a new pathogen(Pseudomonas cichorii)from the soils of Hainan,Anhui,Zhejiang,etc.and explore its optimal fermentation condition.【Method】The antagonistic bacteria was isolated by the inhibition zone method. The isolated antagonistic bacteria were identified by morphological characters,physiological and biochemical tests,16 S rDNA homology sequence analysis and gyrA gene sequence analysis. The optimal fermentation conditions of antagonistic bacteria were studied by single factor variable method.【Result】The results showed that 650 strains of bacteria were isolated from Hainan,Anhui,Zhejiang,ect.10 strains of antagonistic bacteria were screened. Among them, the strain numbered JNL had the best antagonistic effect. Bacterial identification results showed that the strain was Bacillus amyloliquefaciens.The optimal fermentation conditions showed that the strain had the best growth at 30 ℃, pH 7,carbon source galactose,nitrogen source of potassium nitrate and rotation speed of 150 r/min.【Conclusion】The strain JNL has a good control effect on the new pathogen of leaf spot of cucumber and the potential to be developed into biological pesticide or biological fertilizer.
引文
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[9]卢娟,夏启玉,顾文亮,等.拮抗香蕉枯萎病菌的解淀粉芽孢杆菌LX1菌株的鉴定及其抗菌蛋白基因的克隆[J].热带作物学报,2013,34(1):117-124.
[10]Saitou N N M,Nei M C.Saitou N,Nei M.The Neighbor-Joining Method-a New Method for Reconstructing Phylogenetic Trees[J].Mol Biol Evol,1987,4(4):406-425.
[11]汪静杰,赵东洋,刘永贵,等.解淀粉芽孢杆菌SWB16菌株脂肽类代谢产物对球孢白僵菌的拮抗作用[J].微生物学报,2014,54(7):778-785.
[12]曹凤明,杨小红,马鸣超,等.枯草芽孢杆菌近缘种群鉴定方法研究进展[J].微生物学通报,2014,41(5):968-974.
[13]林海.环境工程微生物学.第2版[M].北京:冶金工业出版社,2014.
[14]王继华,徐世强,张木清.解淀粉芽孢杆菌的研究进展[J].亚热带农业研究,2017,13(3):191-195.
[2]孙福在,何礼远.黄瓜细菌性角斑病病原菌与寄主范围鉴定[J].植物病理学报,1988,18(1):23-28.
[3]李永镐.黄瓜细菌性叶斑病病原细菌鉴定[J].植物病理学报,1990,20(2):7-10.
[4]权春善,王军华,徐洪涛,等.一株抗真菌解淀粉芽孢杆菌的分离鉴定及其发酵条件的初步研究[J].微生物学报,2006,46(1):7-12.
[5]郭龙涛,邱思鑫,蔡学清,等.内生解淀粉芽孢杆菌TB2液体发酵条件的研究[J].热带作物学报,2010,31(2):259-264.
[6]范秀容,李广武,沈萍.微生物学实验(第二版)[M].北京:高等教育出版社,1989.
[7]王寅寒.芽孢杆菌防治丹尼斯凤梨细菌性软腐的初步研究[D].海口:海南大学,2013.
[8]东秀珠,蔡妙英.常见细菌系统鉴定手册[M].北京:科学出版社,2001.
[9]卢娟,夏启玉,顾文亮,等.拮抗香蕉枯萎病菌的解淀粉芽孢杆菌LX1菌株的鉴定及其抗菌蛋白基因的克隆[J].热带作物学报,2013,34(1):117-124.
[10]Saitou N N M,Nei M C.Saitou N,Nei M.The Neighbor-Joining Method-a New Method for Reconstructing Phylogenetic Trees[J].Mol Biol Evol,1987,4(4):406-425.
[11]汪静杰,赵东洋,刘永贵,等.解淀粉芽孢杆菌SWB16菌株脂肽类代谢产物对球孢白僵菌的拮抗作用[J].微生物学报,2014,54(7):778-785.
[12]曹凤明,杨小红,马鸣超,等.枯草芽孢杆菌近缘种群鉴定方法研究进展[J].微生物学通报,2014,41(5):968-974.
[13]林海.环境工程微生物学.第2版[M].北京:冶金工业出版社,2014.
[14]王继华,徐世强,张木清.解淀粉芽孢杆菌的研究进展[J].亚热带农业研究,2017,13(3):191-195.