直丝紫链霉菌A8发酵条件优化、活性成分初步分析及对水稻纹枯病的防效研究  

Optimization of Fermentation Conditions,Preliminary Analysis of Active Ingredients of Streptomyces rectiviolaceus A8 and Its Control Effect on Rice Sheath Blight

作　　者：王明环 孙爱丽 赵联晶 刘列 赵添琦 董文汉 罗琼[1] 黄惠川 李成云[1] 杨根华[1] WANG Minghuan;SUN Aili;ZHAO Lianjing;LIU Lie;ZHAO Tianqi;DONG Wenhan;LUO Qiong;HUANG Huichuan;LI Chengyun;YANG Genhua(State Key Laboratory for Conversation and Utilization of Bio-resources in Yunnan/Yunnan Agricultural University,Kunming 650201,China;Science and Technology Bureau of Yunnan Agricultural University,Kunming 650201,China)

机构地区：[1]云南农业大学/省部共建云南生物资源保护与利用国家重点实验室,昆明650201 [2]云南农业大学科技处,昆明650201

出　　处：《中国生物防治学报》2021年第4期805-817,共13页Chinese Journal of Biological Control

基　　金：国家重点研发计划资助(2017YFD0200400);云南省高校科技创新团队支持计划资助;国家自然科学基金(31960525,31160352,31360423)。

摘　　要：为了获得直丝紫链霉菌Streptomyces rectiviolaceus A8最佳发酵水平,并初步确定其代谢生物活性物质以及对水稻纹枯病的防控效果。本研究通过碳、氮单因素试验,正交试验和发酵条件的优化,确定其最佳的发酵条件;通过气相色谱-质谱技术(GC-MS)对菌株A8产生的活性物质进行初步检测,将鉴定出的不同组分以立枯丝核菌Rhizoctonia solani和蜡状芽胞杆菌Bacillus cereus作为指示菌进行生物活性测定试验;同时在大田开展菌株A8对水稻纹枯病的生防效果研究。结果显示菌株A8的最佳发酵配方为:可溶性淀粉3%,黄豆粉0.6%,酵母膏0.4%,NaCl 0.05%,K_(2)HPO_(4)·3H_(2)O 0.05%,MgSO_(4)·7H_(2)O 0.05%,FeSO_(4)·7H_(2)O 0.001%;最佳发酵条件:装液量100-200 mL/500 mL,接种量3%-7%,培养温度28℃-31℃,初始pH 7.0-8.0;通过气相色谱-质谱联用技术对菌株A8发酵液粗提物进行了分离和鉴定,共鉴定出26个化合物,其中3-甲基-1,2-环戊二酮、1,2,4,5-四甲基苯和二氢-3-亚甲基-2,5-呋喃二酮对立枯丝核菌的抑菌率达到70%以上,二氢-3-亚甲基-2,5-呋喃二酮、2-呋喃羧酸,酰肼、邻苯二酚和3-甲基-1,2-环戊二酮对蜡状芽胞杆菌的抑菌作用较强,该放线菌具有潜在的同时抑制真菌和细菌的作用,对开发广谱生防制剂提供很好的材料。田间防效试验结果表明,菌株A8对水稻纹枯病有一定的防效,防治效果与枯草芽胞杆菌BT205相当。The aim of this research was to obtain the optimal fermentation conditions of Streptomyces rectiviolaceus A8,and to preliminarily determine its metabolic bioactive substances and control effect of rice sheath blight.The fermentation conditions were optimized by carbon and nitrogen single factor experiment and orthogonal experiment.The active substances produced by strain A8 were preliminarily detected by gas chromatography-mass spectrometry(GC-MS).Taken Rhizoctonia solani and Baccillus cereus as target bacteria,activities of the identified components were determined,and the control effect of strain A8 on rice sheath blight was investigated in the field test.The results showed that the optimal formula for strain A8 was starch soluble 3%,soybean mealy 0.6%,east extract 0.4%,NaCl 0.05%,K_(2)HPO_(4)·3H_(2)O 0.05%,MgSO_(4)·7H_(2)O 0.05%and FeSO_(4)·7H_(2)O 0.001%The optimal fermentation conditions was loaded liquid 100-200 mL/500 mL,inoculation amount 3%-7%,fermentation temperature 28℃-31℃and initial pH value 7.0-8.0.Twenty-six compounds were separated and identified from the crude extract.The biological activity test showed that the bacteriostasis rate of 3-methyl-1,2-cyclopentanedione,1,2,4,5-tetramethyl-benzene and dihydro-3-methylene-2,5-furandione against Rhizoctonia solani was over 70%,and dihydro-3-methylene-2,5-furandione,2-furancarboxylic acid,hydrazide,catechol and 3-methyl-1,2-cyclopentanedione had strong antibacterial effect on Bacillus cereus.The actinomycete has the potential to simultaneously inhibit fungi and bacteria,and provides good materials for the development of broad-spectrum biocontrol preparations.Field trials demonstrated that strain A8 had control effect on rice sheath blight,equal to that of Bacillus subtilis BT205.

关 键 词：直丝紫链霉菌A8 发酵条件 代谢生物活性物质 气相色谱-质谱技术 水稻纹枯病 

分 类 号：S476[农业科学—农业昆虫与害虫防治]