烟草马铃薯Y病毒拮抗细菌ZF-10的筛选鉴定及发酵条件优化  

Screening,identification and fermentation condition optimization of an antagonistic biocontrol bacterium ZF-10 for tobacco Potato Virus Y

作　　者：张诺妮 谢云波 李斌 张宗锦[3] 杨章明[3] 闫芳芳[3] 杨鹏[3] 陈寿明[3] 安德荣[1] ZHANG Nuoni;XIE Yunbo;LI Bin;ZHANG Zongjin;YANG Zhangming;YAN Fangfang;YANG Peng;CHEN Shouming;AN Derong(College of Plant Protection,Northwest A&F University,Xianyang 712100,Shaanxi,China;Sichuan Provincial Tobacco Company of China National Tobacco Corporation,Chengdu 610000,China;Panzhihua Branch of Sichuan Provincial Tobacco Company,Panzhihua 617000,Sichuan,China)

机构地区：[1]西北农林科技大学植物保护学院,陕西省咸阳市712100 [2]中国烟草总公司四川省公司,成都市610000 [3]四川省烟草公司攀枝花市公司,四川省攀枝花市617000

出　　处：《烟草科技》2024年第5期64-74,共11页Tobacco Science & Technology

基　　金：中国烟草总公司四川省公司重点科技项目“微生物菌剂筛选鉴定及其对烟株作用机理研究与产品开发应用”(SCYC202009);中国烟草总公司四川省公司科技项目“烟草绿色植保关键技术及体系集成研发与应用”(SCYC202115)。

摘　　要：为获得对烟草马铃薯Y病毒具有抑制作用的拮抗菌,从烟草根际土壤中筛选和测定了马铃薯Y病毒高效拮抗细菌的抑菌广谱性及生物学功能,并利用单因素试验对菌株进行发酵条件优化,同时验证了菌株对烟草马铃薯Y病毒病的盆栽防效。结果表明,共分离纯化出216株拮抗细菌。其中,有4株细菌对PVY具有显著拮抗活性,而ZF-10的拮抗效果最佳,拮抗率为90.20%。经形态学观察、生理生化特性检测及分子生物学分析鉴定该菌株为贝莱斯芽孢杆菌(Bacillus velezensis)。ZF-10对多种病原真菌具有抑制作用,并具有产氨、产纤维素酶、产蛋白酶、溶解无机磷和产铁载体能力,但无解钾能力。ZF-10的最优发酵培养基为可溶性淀粉15.0 g/L、酵母浸粉25.0 g/L、硫酸镁7.5 g/L,最优发酵条件为培养温度32℃、接种量1%、250 mL摇瓶装液量60 mL、初始pH值7.0、发酵时间36 h。盆栽试验结果显示,ZF-10发酵液对烟草马铃薯Y病毒病的相对防效为61.85%,表现出较好的生防潜力。In order to obtain antagonistic bacteria with inhibitory effects on tobacco Potato Virus Y(PVY),broad-spectrum antibacterial activities and biological functions of highly effective antagonistic bacteria against PVY from tobacco rhizosphere soil were screened and characterized.The fermentation conditions for the strain were optimized by a single-factor test,and the pot control effect of the strain on tobacco PVY was verified.The results showed that 216 strains of antagonistic bacteria were isolated and purified,of which 4 strains had a significant antagonistic activity against PVY,and ZF-10 had the best antagonistic effect with an antagonistic rate of 90.20%.The strain was identified as Bacillus velezensis by morphological observation,detection of physiological and biochemical characteristics,and molecular biological analysis.ZF-10 showed inhibitory effects on various pathogenic fungi,and had the ability to produce ammonia,cellulase production,protease production,dissolve inorganic phosphorus,and siderophore production,but no ability to release potassium.The optimal fermentation medium for ZF-10 was soluble starch at 15.0 g/L,yeast extract at 25.0 g/L,magnesium sulfate at 7.5 g/L,and the optimal fermentation conditions were culture temperature at 32°C,inoculum at 1%,250 mL flask volume 60 mL,initial pH at 7.0,fermentation time at 36 h.The results of the pot experiment showed that the relative control effect of ZF-10 fermentation broth on tobacco PVY disease was 61.85%,indicating a satisfactory biocontrol potential.

关 键 词：烟草马铃薯Y病毒 拮抗细菌 贝莱斯芽孢杆菌 抑菌谱 发酵 

分 类 号：S43[农业科学—农业昆虫与害虫防治]