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摘要:【目的】研究生防菌解淀粉芽孢種菌XP1对香蕉根际土壤细菌群落多样性的影响,揭示生防菌防治香蕉枯萎病的土壤微生物学机制,为利用生防菌防控香蕉枯萎病提供理论参考。【方法】试验设3个处理:CK,健康蕉园+每株0.5 L灭菌PDA培养基灌根+每株0.5 L灭菌LB培养基灌根;DI,健康蕉园+香蕉枯萎病菌(菌液浓度1×109个/mL,接种量0.5 L/株)+每株0.5 L灭菌LB培养基;TR,健康蕉园+香蕉枯萎病菌(菌液浓度1×109个/mL,接种量0.5 L/株)+生防菌(菌液浓度1×109个/mL,接种量0.5 L/株)。利用Illumina Hiseq 2500高通量测序平台,对3个处理香蕉植株的根际土壤分别提取总DNA并构建16S rDNA基因文库,进而对微生物群落多样性进行分析。【结果】生防菌XP1对香蕉枯萎病的防效试验结果显示,CK处理香蕉植株无发病现象;DI处理在接种香蕉枯萎病菌后蕉园的发病率达95.0%,病情指数为68.3;TR处理接种生防菌后蕉园的发病率为13.3%,病情指数降低至10.8,防治效果达84.2%。共测序获得886890个有效序列,在97%的相似水平下聚类后获得61660个OTUs,归属于40个细菌门。3个处理的OTUs数量表现为CK>DI>TR。变形菌门(Proteobacteria)、酸杆菌门(Acidobacteria)、浮霉菌门(Planctomycetes)、绿弯菌门(Chloroflexi)、芽单胞菌门(Gemmatimonadetes)、OD1、硝化螺旋菌门(Nitrospirae)、厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)、放线菌门(Actinobacteria)和疣微菌门(Verrucomicrobia)是3个处理共有的菌门,其中变形菌门是CK、DI和TR处理的优势菌门,分别占各自相对丰度≥1%物种的32.21%、28.94%和53.78%。TR处理的Shannon指数和Simpson指数均显著低于CK和DI处理(P<0.05,下同),Dominance指数显著大于CK和DI处理,Chao1指数和Observed species指数大于DI处理、小于CK处理;接种生防菌后,TR处理的Chao1指数和Observed species指数较DI处理分别提高7.3%和0.7%。【结论】接种生防菌XP1能有效防控大田香蕉枯萎病的发生,提高香蕉根际土壤细菌物种丰富度和有益菌比例,促进根际土壤细菌群落结构特征向健康土壤细菌群落结构特征的方向演替,具有一定的应用潜力。
关键词: 香蕉枯萎病;根际土壤;生防菌;高通量测序;微生物多样性
中国分类号: S436.681;S476 文献标志码: A 文章编号:2095-1191(2019)09-1981-09
Abstract:【Objective】The study was conducted to reveal the effects of Bacillus amyloliquefaciens XP1 on the diversity of bacterial communities in rhizosphere soil of banana tree, show the soil microbiological mechanism of biocontrol agents and provide theoretical basis for biological control of banana fusarium wilt. 【Method】Three treatments were set up in this study:CK, healthy banana orchard+0.5 L sterilized PDA medium irrigating root per plant+0.5 L sterilized LB me-dium irrigating root per plant;DI, healthy banana orchard+banana fusarium wilt strain(bacterial solution concentration was 1×109/mL, inoculation amount was 0.5 L/plant)+0.5 L per plant sterilized LB medium;TR, healthy banana orchard+banana fusarium wilt strain(bacterial solution concentration was 1×109 /mL, inoculation amount was 0.5 L/plant) and biocontrol agent(bacterial solution concentration was 1×109 /mL, inoculation amount was 0.5 L/plant). Using Illumina Hiseq 2500 high-throughput sequencing platform, total DNA was extracted from the rhizosphere soil of three banana plants treatments and 16S rDNA gene library was constructed to analyze the diversity of microbial communities. 【Result】The control test of XP1 on banana fusarium wilt indicated that there was no diseased banana plants in CK treatment, the incidence of banana orchard in DI treatment after inoculation of banana fusarium wilt fungus was 95.0% with disease index of 68.3. The incidence of banana orchard in TR treatment was 13.3% with disease index of 10.8, and the control effect was 84.2%. A total of 886890 valid sequences were obtained by sequencing, and 61660 OTUs(operational taxonomic unit) belonging to 40 phyla were obtained by clustering at 97% similarity level. The number of OTUs of the three treatments were in the following order:CK>DI>TR. Among them, Proteobacteria, Acidobacteria, Planctomycetes, Chloroflexi, Gemmatimonadetes, OD1, Nitrospirae, Firmicutes, Bacteroidetes, Actinobacteria and Verrucomrobia were the shared bacteria in three treatments. Among them, the dominant bacteria in CK, DI and TR were Proteobacteria, accounting for 32.21%, 28.94% and 53.78% of the species with relative abundance irrigating root per plant 1%. The Shannon index and Simpson index of TR treatment were significantly lower than those of CK and DI treatments(P<0.05, the same below), dominance index was significantly higher than that of CK and DI treatment, Chao1 index and Observed species index of TR treatment were larger than those of DI treatment and less than those of CK treatment. After biocontrol agent inoculation, the Chao 1 index and the Observed species index of TR treatment increased by 7.3% and 0.7%, respectively compared with DI treatment. 【Conclusion】The inoculation of XP1 can effectively prevent the occurrence of banana fusarium wilt in the field, improve the bacterial species richness in rhizosphere soil and proportion of beneficial bacteria, and promote the rhizosphere soil bacterial community succession towards direction of that in healthy soil. Therefore, it has potential in application.
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