基于高通量测序分析天麻种植土壤的真菌变化

Analysis of fungal changes in Gastrodia elata planting soil based on high throughput sequencing

【目的】揭示天麻种植土壤真菌群落组成结构、天麻种植前后土壤真菌群落结构的变化及天麻种方竹[Chimonobambusa quadrangularis(Fenzi)Makino]种植结束后土壤真菌群落结构的变化,为研究天麻根际土壤微生态提供理论依据并为解决天麻连作障碍提供技术支持。【方法】基于高通量测序方法对采集的3组土样(原壤charlle001、一麻壤charlle002、方竹壤charlle006)进行基因组DNA提取,PCR扩增,产物检测、纯化和定量,建库测序,数据分析,物种组成多样性分析及差异分析。【结果】3组土样中真菌主要类群为Ascomycota、Basidiomycota和Mortierellomycota 3门真菌,其中Ascomycota真菌丰度普遍最大;3组样本特有真菌OTU:一麻壤>方竹壤>原壤;种水平下分析原壤charlle001中主要优势真菌为Clitocybe sp.、Solicoccozyma sp.和Spirosphaera sp.,一麻壤charlle002中主要优势真菌为Clavulinopsis sp.和Rosellinia sp.,方竹壤charlle006中主要优势真菌为Trichoderma sp.;Clavuli⁃nopsis和Rosellinia在一麻壤charllle002中丰度极大,在charllle001和charllle006中丰度极小;charlle001和charlle002组间真菌Mor⁃tierella相对丰度差异显著,charlle001和charlle006组间真菌Penicillium相对丰度差异显著;charlle002和charlle006组间真菌Peni⁃cillium相对丰度差异显著。种植收获一茬天麻后土壤中真菌种类数量递增,群落结构发生变化,一麻壤中真菌ASV、特有真菌数目明显增大;一麻壤种植方竹后木霉菌显著增多;Clavulinopsis和Rosellinia部分真菌易引起天麻病害,而方壤中Trichoderma多用于病原菌生物防治。【结论】天麻不同种植土壤的真菌群落结构不同,种植一年天麻后土壤中病原菌增多,再种植方竹呈现修复作用,但具体修复机制还有待后续深入研究。

【Objective】The study aimed to reveal the changes of soil fungal community structure of Gastrodia elata,the changes of soil fungi community before and after G.elata planting,and the changes of soil fungi after G.elata planting,so as to provide the theoretical basis for studying the rhizosphere microecology of G.elata,and the technical support for solving the obstacle of continuous cropping of G.elata in the future.【Method】Genomic DNA extraction,the PCR amplification,product detection,purification and quantification and the species compo⁃sition diversity and difference analyses were performed on three groups of the soil samples(the original soil charlle001,a cropping of soil charlle002,and restoration soil charlle006)based on the high⁃throughput sequencing method.【Result】The fungi in the three soil samples mainly belonged to phyla Ascomycota,Basidiomycota and Mortierellomycota,and Ascomycota had the largest fungal abundance.Specific fungus OTUs of 3 groups of samples showed:a croping soil>bamboo soil>original soil.The main dominant fungi were Clitocybe sp.and Solicoccozyma sp.in charlle001,and Spirosphaera sp.and Clavulinopsis sp.were in charlle002,and Trichoderma sp.was in charlle006.Clavulinopsis and Rosellinia had maximum abundance in charlle002 and minimal abundance in charlle001 and charlle006.The relative abun⁃dance of Mortierella was significantly different between charlle001 and charlle002,and Penicillium was significantly different between char⁃lle001 and charlle006,and Penicillium was significantly different between charlle002 and charlle006.There was a significant increase of Trichoderma in bamboo soil.Some fungi of Clavulinopsis and Rosellinia caused gastrodia diseases,while Trichoderma was used for biological control of pathogens in soil.【Conclusion】The fungal community structure of different soils planted with G.elata is different,and planting bamboo has a restorative effect,which pushes the process of the specific repaired mechanism of G.elata for further research.