摘要
【目的】探究感染根腐病番荔枝根际土壤微生物群落的多样性和分布规律,为筛选鉴定番荔枝根腐病病原菌提供参考依据。【方法】采集感染根腐病的番荔枝根际土壤(感病根际土壤)和健康番荔枝根际土壤(健康根际土壤)样品,采用高通量测序技术,以ITS1和ITS4为引物,对其真菌ITS1区和细菌V3+V4区进行扩增测序,在门、纲、目、属和种水平上分析其微生物群落的结构和多样性。【结果】感病根际土壤的真菌数量明显增多,细菌数量明显减少。感病根际土壤中的真菌隶属于2界5门16纲36目48科81属96种,健康根际土壤中的真菌隶属于3界7门17纲43目73科113属160种。感病根际土壤微生物的群落丰富度、多样性指数均较健康根际土壤下降,真菌种类较少但均匀度上升。在PDA培养基上培养4 d后真菌类菌株形成中等量、呈放射状的气生菌丝;菌落直径4.5 cm,菌落平铺,全缘,呈圆形,白色,气生菌丝绒状。采用供试引物对根际土壤病原菌菌株进行扩增,显示菌株的ITS序列与Fusarium acutatum(登录号AY569567.1)同源性达99.45%,说明扩增出的ITS序列(RRD-1)为F.acutatum的基因片段。【结论】感染根腐病番荔枝根际土壤微生物的群落丰富度、多样性指数均较健康荔枝根际土壤明显下降,而真菌种类较少,均匀度上升。经病原菌形态学观察和生物比对鉴定,初步确定引起广西地区番荔枝根腐病的病原菌可能为真菌镰刀属的F.acutatum。
【Objective】The present paper aimed to explore the diversity and distribution rules of soil microbial community of Annona squamosa Lour.after the infection of root rot,and provide a reference basis for the screening and identification with A.squamosa pathogens of root rot.【Method】Samples of A.squamosa rhizosphere soil infected with root rot(infected rhizosphere soil)and healthy rhizosphere soil(healthy rhizosphere soil)were collected.Using high throughput sequencing technology and designing primers of ITS1 and ITS4,sequenced the fungal ITS1 and bacterial V3+V4 regions,and analyzed the structure and diversity of the phylum,class,order,genus,and species level.【Result】After infection with root rot,the number of fungi in its rhizosphere soil increased greatly,and the number of bacteria decreased.The classification of fungi in rhizosphere soil belonged to 2 kingdoms,5 phylum,16 classes,36 orders,48 families,81 genera and 96 species.The classification of fungi of healthy rhizome soil belonged to 3 kingdoms,7 phylum,17 classes,43 orders,73 families,113 genera and 160 species.The community richness and diversity index of the rhizosphere soil microorganisms decreased compared with healthy rhizosphere soil,along with fewer types of soil fungi,and uniform distribution of soil fungi.After 4 days,medium and radioactive strains were formed on PDA medium;the strains formed medium,radioactive gas bacteria;colony diameter 4.5 cm,bacteria flat spread,full edge,round,white,gas bacteria velvet.Rhizosphere soil pathogen strains were amplified using trial primers,the results showed that the displayed strain ITS sequence and Fusarium acutatum(accession number:AY569567.1)had 99.45% homology,indicating the amplified ITS segment sequence(RRD-1)was parts of F.acutatum sequences.【Conclusion】The community richness and diversity index of root soil microorganisms decreased greatly compared with that of healthy soil,and the types of soil fungi in pathogenic plants were less,and the uniformity increased.After pathogen morphological observation and biological comparison,the pathogen that caused root rot of A.squamosa in Guangxi might be F.acutatum.
作者
黄伟雄
安振宇
方仁
尧金燕
龙兴
HUANG Wei-xiong;AN Zhen-yu;FANG Ren;YAO Jin-yan;LONG Xing(Horticultural Research Institute,Guangxi Academy of Agricultural Sciences/Nanning Investigation Station of South Subtropical Fruit Trees,Ministry of Agriculture and Rural Affairs,Guangxi Nanning 530007,China;Guangxi Crop Genetic Improvement and Biotechnology Lab,Guangxi Nanning 530007,China)
出处
《西南农业学报》
CSCD
北大核心
2021年第8期1623-1629,共7页
Southwest China Journal of Agricultural Sciences
基金
广西自然科学基金项目(2020GXNSFAA297073)
广西重点研发计划项目(桂科AB19245004)
国家现代农业产业技术体系广西创新团队项目(nycytxgxcxtd-17-15)
广西农业科学院基本科研业务专项(桂农科2021YT049)。
关键词
番荔枝
根腐病
根际土壤
高通量测序
Annona squamosa Lour.
Root rot
Rhizosphere soil
High-throughput sequencing
