拮抗放线菌ZZ-9菌株发酵液的抑菌谱及稳定性测定

拮抗放线菌ZZ-9菌株发酵液的抑菌谱及稳定性测定

范万泽;薛应钰;张树武;徐秉良

【摘要】为探究拮抗放线菌ZZ-9(Streptomyces rochei)菌株发酵液的抑菌谱,并明确传代培养、温度、光照、pH及紫外线等理化因素对发酵液稳定性的影响.以15种植物病原真菌为供试菌,采用菌丝生长速率法测定发酵液的抑菌谱,并以苹果树腐烂病菌(Cytospora sp.)为指示菌,测定不同条件下发酵液的稳定性.抑菌谱结果表明:ZZ-9菌株发酵液对供试的15种病原真菌均具有不同程度的抑制作用,其中对苹果树腐烂病菌和立枯丝核菌抑制效果最好,生长抑制率可达96％以上,且效果持久;而对苹果轮纹病菌、蚕豆褐斑病菌、月季叶霉病菌等抑制效果较差,抑制率在27％以下.稳定性试验结果表明:菌株传接6代时,活性稳定,从第7代开始菌株活性下降显著;发酵液耐高温,对强酸强碱处理、可见光及紫外线照射均有较好的稳定性;且在4℃及室温(20～25℃)下可贮藏至少30 d而保持其抑菌活性几乎不变.因此,拮抗放线菌ZZ-9菌株次生代谢产物具有较广谱的抑菌活性和较强的稳定性,具有开发成微生物农药的潜力.%To study bacteriostatic spectrum of fermentation broth from actinomycetes strain ZZ-9 identified as Streptomyces rochei,and to verify the effect of it on stability of fermentation broth under biochemical and physical factors such as subculture,temperature,illumination,pH and ultraviolet conditions.As the tested phytopathogen,method of growth rate was used to determine antimicrobial spectrum of fermentation broth with 15 kinds of plant pathogenic fungus.With Cytospora sp.as indicator phytopathogen,the stability of fermentation broth under different conditions was measured.The bacteriostatic spectrum results showed that the fermentation broth of ZZ-9 strain had inhibitory effects at different

degres on 15 kinds of pathogenic fungus.With growth inhibition rate of more than 96％ and the lasting effect,the fermentation broth of ZZ-9 strain had the best effect on Cytospora sp.and Rhizoctonia solani,and it had worse effect on Botryosphaeria piricola,Cladosporium cladosporioidesv and Ascochyta fabae,the inhibition rate was under 27 ％.The stability test showed that the activity of strain was stable until the 6th generation,it decreased significantly from the 7th generation,and the fermentation broth had high temperature resistance,good stability under strong acid and alkali,visible light and ultraviolet irradiation treatments.Moreover,the fermentation broth could be stored for at least 30 days at 4 ℃ and room temperature (20-25 ℃),the activity remained nearly

unchanged.Therefore,the secondary metabolites of antagonistic actinomycetes ZZ-9 strain had broad-spectrum antibacterial activity and strong stability,and the potential could be developed into the microbial pesticide.

【期刊名称】《西北农业学报》

【年(卷),期】2017(026)003

【总页数】8页(P463-470)

【关键词】拮抗放线菌;发酵液;抑菌谱;稳定性

【作者】范万泽;薛应钰;张树武;徐秉良

【作者单位】甘肃农业大学植物保护学院/甘肃省农作物病虫害生物防治工程实验室,兰州730070;甘肃农业大学植物保护学院/甘肃省农作物病虫害生物防治工程实

验室,兰州730070;甘肃农业大学植物保护学院/甘肃省农作物病虫害生物防治工程实验室,兰州730070;甘肃农业大学植物保护学院/甘肃省农作物病虫害生物防治工程实验室,兰州730070

【正文语种】中文

【中图分类】S482.2+92

中国是世界上农药和化肥污染最严重的国家之一，化学农药的滥用不仅对生态环境、人类健康产生影响，而且易造成病虫害抗性等问题。开发新型生物农药、减少和替代高毒高残化学农药是发展绿色、安全农产品的必要条件[1]。微生物农药因污染小、广谱性，与环境相容等特点符合绿色农业的发展要求而被深入研究[2]。放线

菌作为一种重要的微生物农药资源而倍受关注，其次生代谢产物在低浓度时被用来防治病、虫、草害或调节植物生长发育[3]。目前，已有多个商品化的产品被应用

于生产。例如，被用来杀虫的有阿维菌素、多杀菌素和南昌霉素；被用来杀菌的有放线菌酮、春雷霉素、万隆霉素、多效霉素、井冈霉素、农抗120以及多抗霉素、中生菌素、宁南霉素等；被用来除草的有双丙氨膦等[4-7]。因此，从放线菌及其

次生代谢产物中筛选广谱、稳定及低毒的农用抗生素具有十分广阔的前景。

由于放线菌产生的活性物质在自然环境中大多不稳定，从而影响放线菌的开发与应用。农用抗生素在施用到作物表面，或者进入到土壤中，都要受到光照、温度、土壤酸碱性等的影响。因此，农用抗生素在各种环境条件下的稳定性是评价其是否具有开发价值的一个重要指标[8]。甘肃农业大学植物病原实验室从甘肃陇东地区采

集的土壤中分离获得一株拮抗放线菌，菌株编号ZZ-9，前期研究结果表明，放线

菌ZZ-9对苹果树腐烂病菌(Cytospora sp.)具有较高抑菌活性，不同稀释倍数的

ZZ-9发酵滤液对腐烂病菌分生孢子萌发及菌丝生长均有明显的抑制作用，其中50倍发酵滤液对孢子萌发抑制率和菌丝生长抑制率均达80%以上，且受抑制菌丝颜