抗香蕉枯萎病甲基营养型芽孢杆菌的鉴定及定殖  

Identification and Colonization of Bacillus methylotrophicus Against Banana Fusarium Wilt Disease

作　　者：周登博[1] 陈宇丰[1] 井涛[2] 起登凤[1] 张妙宜 冯仁军[1] 王飞[1] 谢江辉[1] 

机构地区：[1]中国热带农业科学院热带生物技术研究所,海口571101 [2]中国热带农业科学院海口实验站,海口570102

出　　处：《中国农学通报》2017年第36期145-151,共7页Chinese Agricultural Science Bulletin

基　　金：现代农业产业技术体系"国家香蕉产业技术体系"(CARS-31);国家重点研发计划"热带果树化肥农药减施增效技术集成研究与示范"(2017YFD0202105);中国热带农业科学院基本科研业务费专项资金"复合拮抗菌发酵液提高香蕉抗枯萎病能力的作用机理研究"(1630052016005);中国热带农业科学院基本科研业务费专项资金"肥料结构调整对香蕉生长及品质形成的影响"(1630012017007)

摘　　要：为系统评价分离自天然饼肥发酵液4-L-16菌株的抗真菌活性及定殖规律。以尖饱镰刀菌Race 4(Fusarium oxysporum f.sp.cubense Race 4)等9种病原菌为靶标菌株,采用平板对峙、含药介质法检测菌株抗真菌活性,并通过革兰氏染色、形态观察、生理生化特征和16S rDNA序列分析对其进行鉴定。结果表明:4-L-16菌株对9种植物病原菌具有广谱抗性,菌丝生长抑制率可达28.05%~76.07%,孢子萌发抑制率达24.44%~70.00%,其中对香蕉枯萎病菌的菌丝生长和孢子萌发抑制率分别为76.07%和70%,经鉴定该菌株为甲基营养型芽孢杆菌Bacillus methylotrophicus。经抗生素标记,获得抗600μg/mL利福平的标记菌株,盆栽接种试验表明标记菌在香蕉根际土壤和植株体内具有良好定殖能力,接种第10天土壤中菌量达到最大值为6.67×10~6CFU/g,随时间延长,定殖数量逐渐降低,25天后数量趋于稳定,为2.05×10~6CFU/g。香蕉植株内定殖以根系最多,为4.38×10~3CFU/g。甲基营养型芽孢杆菌4-L-16菌株广谱抗真菌活性、良好的定殖能力,已成为香蕉枯萎病生物防治新的菌种资源。Bacteria 4-L-16 was isolated from natural cake fertilizer fermentation liquid, in order to evaluate its anti-fungi activity and colonizing regulation systematically. Nine kinds of pathogen including Fusarium oxyspum f. sp.cubense Race 4 were taken as indicative strains to detect the anti-fungi activity of antagonisticbacteria by the methods of plate confrontation and containing medium. Bacteria 4-L-16 was also identified byGram staining, morphologic observation, physiological and biochemical characteristics analysis and 16 S rDNAsequence analysis. The results showed that the strain 4-L-16 had broad-spectrum resistance to the nine kindsof pathogen, the inhibition rates of mycelium growth were 28.05%-76.07%, and the inhibition rates of sporegermination were 24.44%-70.00%. For banana Fusarium wilt, the inhibition rates of mycelium growth andspore germination were 76.07% and 70%, respectively, and this strain was identified as Bacillus methylotrophicus. By the method of antibiotic marker, anti 600 μg/m L rifampicin marked strain of 4-L-16 wasobtained. The pot test showed that the strain 4-L-16 could colonize in banana rhizosphere soil and bananaplant, after inoculating 10 d, the soil colonization reached a maximum quantity of 6.67×10~6 CFU/g. But with the extension of days, the number of colonization decreased gradually, the quantity tended to be stable of 2.05×10~6 CFU/g after inoculating 25 d. The colonization mostly took place in the root of banana, was 4.38×103 CFU/g.Bacillus methylotrophicus 4-L-16 was taken as a new microbial resource for biological control of Banana Fusarium wilt disease for its broad-spectrum resistance activities and good colonization abilities.

关 键 词：香蕉枯萎病 甲基营养型芽孢杆菌 菌种鉴定 定殖 抗菌活性 

分 类 号：S432.1[农业科学—植物病理学]