白肉灵芝土传病原微生物的分离与鉴定  

Isolation and identification of soil-borne pathogenic microorganisms in Ganoderma leucocontextum

作　　者：姚春馨[1] 梁明泰[2] 马渊浩 刘家迅[2] 张绍松[1] 陈霞[2] 田果廷[1] YAO Chun-xin;LIANG Ming-tai;MA Yuan-hao;LIU Jia-xun;ZHANG Shao-song;CHEN Xia;TIAN Guo-ting(Biotechnology and Germplasm Resources Institute,Yunnan Academy of Agricultural Sciences,Kunming 650205,China;Horticultural Research Institute,Yunnan Academy of Agricultural Sciences,Kunming 650205,China)

机构地区：[1]云南省农业科学院生物技术与种质资源研究所,昆明650205 [2]云南省农业科学院园艺作物研究所,昆明650205

出　　处：《西南农业学报》2022年第12期2811-2818,共8页Southwest China Journal of Agricultural Sciences

基　　金：云南省重大科技专项计划(202102AE090051-2-02);云南省乡村振兴科技专项(202204BP090023)。

摘　　要：【目的】作物连作障碍与土壤微生物密切相关,分离与鉴定土传病原微生物,为解决白肉灵芝段木埋土栽培的连作障碍提供理论依据。【方法】针对大棚内段木埋土栽培的白肉灵芝,分别采集连作0年、1年、3年、5年不同栽培年限的覆土,以稀释平板法调查土壤可培养微生物类群的变化,分离白肉灵芝连作土中优势微生物;采用平板对峙试验筛选白肉灵芝连作障碍潜在的土传病原菌,结合形态、生理等微宏观特征与分子生物学技术鉴定各菌株,并进行物种分类。【结果】随着白肉灵芝连作年限的增加,覆土中可培养微生物的数量和种群发生变化,细菌、放线菌的数量下降趋势明显,真菌数量逐年增加,三大主要微生物种群中细菌占比从25.21%增加至95.29%,真菌占比从0.69%逐年增加至4.71%,放线菌占比则下降;连作土中分离到14个优势微生物,NJ聚类结果将其分为两大类群,结合DNA序列比对与形态分类结果,最终鉴定5个真菌菌株和9个细菌菌株;平板对峙培养试验结果显示,真菌N7及细菌N11、N9、N12、N13、N14共6个菌株对白肉灵芝菌丝生长的抑菌率高达80%以上。【结论】白肉灵芝的连作栽培显著影响了覆土层的土壤微生物区系,不同栽培年限其三大主要微生物种群差异显著;微生物种群中的芽孢杆菌属(Bacillus)、篮状菌属(Talaromyces)、红球菌属(Rhodococcus)和部分链霉菌属(Streptomyces)菌株可能是白肉灵芝段木埋土栽培连作障碍中潜在的土传病原菌。【Objective】Continuous cropping obstacle is considered to have a close connection with the soil microbial community.The present study aimed to isolate and identify soil-borne pathogenic microorganisms in Ganoderma leucocontextum and provide a theoretical basis for continuous cropping obstacles encountered in cut-log cultivation.【Method】Overburden soil used to cultivate G.leucocontextum in the greenhouse for various periods(0,1,3,5 years)were collected.Changes in culturable microbial community were investigated,and predominant microbes were isolated.Soil-borne pathogenic microorganisms of G.leucocontextum grown in continuously cropped soil were identified by plate confrontation culture.The species were further determined based on morphological and physiological features and molecular techniques.【Result】With the increase of G.leucocontextum continuous cropping years,the number of culturable microorganisms in the overburden soil and population changed,the number of bacteria and actinomyces declined significantly,while the fungi number increased.Specifically,bacteria accounted increased from 25.21%to 95.29%,fungi increased from 0.69%to 4.71%.By contrast,the proportion of actinomycetes decreased.Five dominant fungal species and nine dominant bacterial species were isolated and identified from continuously cropped soil.The antagonistic test showed that the inhibition rates on the growth of G.leucocontextum mycelium of six strains reached above 80%,namely,N7,N11,N9,N12,N13 and N14.【Conclusion】The continuous cultivation of G.leucocontextum significantly affected the soil microflora in the overburden soil,and the three main microbial populations had significant differences in different cultivation years.Bacillus,Talaromyces,Rhodococcus and some Streptomyces species might be potential soil-borne pathogenic microorganisms in the continuous cropping of G.leucocontextum,leading to obstacles encountered in cut-log cultivation.

关 键 词：白肉灵芝 土壤微生物 对峙培养 病原菌 

分 类 号：S646[农业科学—蔬菜学]