引用本文:桂 莎,刘 芳,张立丹,樊小林.复合菌剂防控香蕉枯萎病的效果及其微生物学机制[J].土壤学报,2020,57(4):995-1007. DOI:10.11766/trxb201904180111
GUI Sha,LIU Fang,ZHANG Lidan,FAN Xiaolin.Effects of Complex Anti-Fungal Agents Biocontrolling Fusarium Wilt on Banana and Its Microbiological Mechanism[J].Acta Pedologica Sinica,2020,57(4):995-1007. DOI:10.11766/trxb201904180111
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复合菌剂防控香蕉枯萎病的效果及其微生物学机制
桂 莎, 刘 芳, 张立丹, 樊小林
广东高校环境友好型肥料工程技术研究中心,华南农业大学资源与环境学院
摘要:
由病原菌尖孢镰刀菌古巴专化型(Fusarium oxysporum f. sp. cubense (Foc))侵染引起的香蕉枯萎病对全世界香蕉产业带来了毁灭性的影响,且目前尚无广泛采用的防治方法。研究复合生防真菌制剂对香蕉枯萎病的防治效果,以期为大田香蕉枯萎病的防治提供依据。设置3组不同的菌剂处理,分别为对照组CK、复合菌剂NFP、复合菌剂NFPT,通过两季的盆栽试验,研究复合菌剂对香蕉枯萎病的防治效果及其对土壤微生物多样性的影响;利用 Illumina Miseq 高通量测序平台对细菌16S rRNA基因和真菌ITS区域进行测序分析,采用实时荧光qPCR定量分析各处理病原菌的数量。结果表明:(1)复合菌剂处理(NFP和NFPT)对香蕉枯萎病有较好的防治效果,其防效分别为43%和48%。(2)施用复合菌剂增加了细菌和真菌群落丰富度和多样性。基于Bray-curtis距离矩阵的主坐标分析(PCoA)结果表明NFP和NFPT改变了细菌和真菌群落结构。NFP和NFPT处理增加了潜在有益微生物中与香蕉枯萎病病情指数呈显著负相关的大理石雕菌属Marmoricola、类诺卡氏菌属Nocardioides、野野村式菌属Nonomuraea norank_c__AcidobacteriaDS-100属和norank_f__Blastocatellaceae__Subgroup_4属的相对丰度,显著减少了病原菌尖孢镰刀菌的数量,重塑了土壤微生物结构和功能,增强其抗病性。
关键词:  复合真菌制剂  香蕉枯萎病  Illumina Miseq 高通量测序  土壤微生物多样性
基金项目:国家重点研发计划项目(2018YFD0201100),广东省省级重大科研项目(2016KZDXM029)和国家现代农业产业技术体系建设专项(CARS-31-06)资助
Effects of Complex Anti-Fungal Agents Biocontrolling Fusarium Wilt on Banana and Its Microbiological Mechanism
GUI Sha, LIU Fang, ZHANG Lidan, FAN Xiaolin
College of Natural Resource and Environment Science, South China Agricultural University, R&D Center of Environment Friendly Fertilizer Science and Technology of Guangdong Provincial University
Abstract:
【Objective】Worldwidely, banana production is severely hindered by banana Fusarium wilt, a devastating soil-borne disease caused by Fusarium oxysporum f. sp. cubense (Foc). With no widely adopted effective methods available to control or prevent the disease, it causes serious economic losses every year. In this study, complex biocontrol fungal agents were introduced and effects of their application preventing banana Fusarium wilt and potential mechanisms were explored, in an attempt to provide certain references for controlling disease on the large field scale. 【Method】 A pot experiment, lasting for 2 seasons were conducted and designed to have three groups of pots, namely, CK (no controlling agent applied), NFP (NFP for application of a complex anti-fungal agent prepared by combining non-pathogenic Fusarium oxysporum.sp and Paecilomyces.sp in 1:1 ratio), and NFPT (application of a complex anti-fungal agents prepared by combining non-pathogenic Fusarium oxysporum sp., Paecilomyce sp. and Trichoderma sp. in 9:9:4 ratio) for comparison between the pots for effects of the applications controlling banana Fusarium wilt and effects on soil microbial diversity. The Illumina Miseq high-throughput sequencing platform was used to analyze bacterial 16SrRNA gene and fungal ITS regions, the real-time fluorescence quantification PCR (RT-qPCR) was to determine number of pathogens in the soil. 【Result】Applications of the complex fungal agents (NFP and NFPT) have good effects of controlling banana Fusarium wilt disease, with control efficiency being 43% and 48%, respectively, and improve richness and diversity of bacteria and fungi. Principal coordinate analysis (PCoA) based on Bray-curtis distance matrix shows that significant differences in composition of the bacterial and fungal communities exist between the pots applied with the complex fungal agents and the pots in CK. The first principal component (PC1) explains 29.45% and 43.14% of the variability in the bacterial and fungal communities, respectively, and differs sharply between the treatment pots and the CK pots in composition of the overall bacterial and fungal communities. The microbes (non-pathogenic Fusarium oxysporum sp., Paecilomyce sp. and Trichoderma sp.) introduced into the soil are found quite limited in survivability in this study, and their abundance has only a marginal direct effect on the number of F. oxysporum and disease severity of the Fusarium wilt disease. However, they suppress the disease by altering composition of the soil microbiome. In particular, application of the complex fungal agents (NFP and NFPT) increases relative abundances of the beneficial indigenous microbial groups, such asMarmoricola, Nocardioides, Nonomuraea, norank_c__Acidobacteria, DS-100 and norank_f__Blastocatellaceae__Subgroup_4. Their relative abundances are good indicators of the disease suppression effect and may play a keystone role in the process of the complex fungal agents suppressing banana Fusarium wilt disease. 【Conclusion】 In a word, application of the complex fungal agents (NFP and NFPT) significantly reduces the banana Fusarium wilt disease severity index. All the findings presented above show that relative abundance of the introduced non-pathogenic Fusarium oxysporum sp., Paecilomyce sp. and Trichoderma sp. has only a marginal effect on F. oxysporum. In contrast, the changes in abundance and community structures of the bacteria and fungi after application of the agents are the key factors suppressing the disease. Application of the agents stimulates the potential beneficial indigenous microbial groups that are significantly and negatively related to banana Fusarium wilt disease severity index. Thus, the effect of the complex fungal agents suppressing the disease seemed to be a joint one of the actual antagonism of the introduced microbes with the pathogens and their promoting growth of beneficial indigenous microbial groups.
Key words:  Complex biocontrol fungal agents  Banana Fusarium wilt  Illumina Miseq High-throughput sequencing  Soil microbial diversity