对三种苹果病原菌具抑制作用的新疆野苹果内生真菌的分离与鉴定  

Isolation and identification of endophytic fungi resistant to three apple pathogens from the branches of Malus sieversii

作　　者：王亚红 李永丽[1] 常乐 余海如 周洲[1] 曲良建[2] WANG Yahong;LI Yongli;CHANG Le;YU Hairu;ZHOU Zhou;QU Liangjian(Forestry College,Henan University of Science and Technology,Luoyang 471023,Henan,China;Research Institute of Forest Ecology,Environment and Protection,Chinese Academy of Forestry/Key Laboratory of Forest Protection,National Forestry and Grassland Administration,Beijing 100091,China)

机构地区：[1]河南科技大学林学院,河南洛阳471023 [2]中国林业科学研究院森林生态环境与森林保护研究所·国家林业和草原局森林保护重点实验室,北京100091

出　　处：《果树学报》2020年第3期390-396,共7页Journal of Fruit Science

基　　金：十三五国家重点研发计划课题(2016YFC0501503);国家自然科学基金(31600519,31870638)。

摘　　要：【目的】通过分离鉴定对苹果枝干病害有抑制作用的新疆野苹果枝干内生真菌,为苹果病害生物防治提供新型菌种资源。【方法】采用组织分离法从新疆野苹果枝干中分离内生真菌,平板对峙试验筛选对苹果树3种致病菌均具有显著抑菌作用的菌株,结合形态学、分子生物学分析,确定菌株的分类地位。【结果】共分离获得内生真菌75株,从其中筛选出对苹果树3种病原菌均具较强抑菌作用的菌株5株(E-m6、S-p5、S-m2、S-p6、E-p5)。对葡萄座腔菌Botryosphaeria dothidea的最高抑菌率为73.39%,对胶孢炭疽菌Colletotrichum gloeosporioides的最高抑菌率为73.52%,对苹果拟茎点霉Phomopsis mali的最高抑菌率为82.19%。结合形态学和ITS序列和TUB基因序列分析,菌株E-m6和S-p5为黑曲霉Aspergillus niger,S-m2为黄曲霉Aspergillus flavus,S-p6为Rosellinia sp.,E-p5为球毛壳菌Chaetomium globosum。【结论】从新疆野苹果枝条中筛选出具生防潜力的内生真菌菌株5株,为苹果枝干病害的生物防治提供了新的微生物资源,同时也为新疆野苹果内生菌研究奠定了基础。【Objective】In order to provide new fungi resources for the biological control of apple diseases,the endophytic fungi from healthy branches of Malus sieversii were isolated and identified.【Methods】Endophytic fungi were isolated from branches of M.sieversii by tissue separation method.Strains were screened out with significant antagonism activity against three pathogens(Botryosphaeria dothidea,Colletotrichum gloeosporioides and Phomopsis mali)of apple trees.The morphology and molecular biology were applied to determine the classification of the strains.【Results】A total of 75 endophytic fungi were isolated,among which 56 strains were from the phloem and 19 strains from the xylem.Five strains(E-m6,S-p5,S-m2,S-p6 and E-p5)were screened out with strong inhibitive effects on mycelia growth of three apple tree pathogens.The highest inhibitory rates of the selected strains against B.dothidea,C.gloeosporioides and P.mali were 73.39%,73.52%and 82.19%,respectively.The lengths of ITS regions of E-m6,S-p5,S-m2,S-p6 and E-p5 were 593 bp,554 bp,585 bp,430 bp and 575 bp,respectively;and the length of TUB gene of S-p6 was 420 bp.Combined with morphological,ITS and TUB phylogenetic trees analysis,the strains E-m6 and S-p5 were identified as Aspergillus niger,S-m2 was identified as Aspergillus flavus,S-p6 was identified as Rosellinia sp.,and E-p5 was identified as Chaetomium globosum【.Conclusion】In this study,five strains of endophytic fungi with biocontrol potential were screened out from M.sieversii branches,which could provide not only new microbial resources for the biological control of apple stem diseases,but also a basis for the study on endophytes of M.sieversii in the future.

关 键 词：新疆野苹果 内生真菌 抑菌作用 葡萄座腔菌 胶孢炭疽菌 苹果拟茎点霉 

分 类 号：S661.1[农业科学—果树学]