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林业科学 ›› 2020, Vol. 56 ›› Issue (5): 97-104.doi: 10.11707/j.1001-7488.20200511

• 论文与研究报告 • 上一篇    下一篇

新疆野苹果树内生细菌分离与鉴定及其对3种苹果病原菌的抑制作用

李永丽1,王亚红1,常乐1,余海如1,周洲1,赵文霞2,曲良建2,*   

  1. 1. 河南科技大学林学院 洛阳 471003
    2. 中国林业科学研究院森林生态环境与保护研究所 国家林业和草原局森林保护学重点实验室 北京 100091
  • 收稿日期:2018-12-05 出版日期:2020-05-25 发布日期:2020-06-13
  • 通讯作者: 曲良建
  • 基金资助:
    中国林业科学研究院基本科研业务费专项(CAFYBB2017SZ003);"十三五"国家重点研发计划课题(2016YFC0501503);国家自然科学基金项目(31600519);国家自然科学基金项目(31870638)

Isolation and Identification of Endophytic Bacteria from Xinjiang Wild Apple (Malus sieversii) and Their Antagonism against Three Pathogens of Apple Trees

Yongli Li1,Yahong Wang1,Le Chang1,Hairu Yu1,Zhou Zhou1,Wenxia Zhao2,Liangjian Qu2,*   

  1. 1. College of Forestry, Henan University of Science and Technology Luoyang 471003
    2. Research Institute of Forest Ecology, Environment and Protection, Chinese Academy of Forestry Key Laboratory of Forest Protection of National Forestry and Grassland Administration Beijing 100091
  • Received:2018-12-05 Online:2020-05-25 Published:2020-06-13
  • Contact: Liangjian Qu

摘要:

目的: 从我国优良的种质资源新疆野苹果树中分离和筛选用于苹果病害防治的内生细菌,以期为苹果病害生物防治提供新的菌种资源。方法: 采用稀释分离法和组织分离法从新疆野苹果枝干中分离内生细菌,通过平板对峙试验筛选出对葡萄座腔菌、胶孢炭疽菌、苹果拟茎点霉3种苹果树的致病菌均具有显著抑菌效果的菌株,结合形态学、生理生化特征及16S rDNA序列分析,明确分离和筛选出内生细菌的分类地位,经离体接种试验证实其是否具有致病性。结果: 共分离获得内生细菌217株;筛选出对苹果3种病原菌均具有较强抑菌作用的菌株7株(M-m7、M-m12、M-r12、P-m9、P-m11、L-m12、P-r21),对葡萄座腔菌的抑菌率分别为77.00%、72.60%、76.60%、60.64%、65.96%、65.81%、71.79%,对胶孢炭疽菌的抑菌率分别为69.85%、64.95%、68.87%、68.37%、56.12%、53.54%、65.66%,对苹果拟茎点霉的抑菌率分别为77.63%、79.22%、72.60%、69.37%、70.27%、69.30%、71.05%。经鉴定,M-m7、M-m12、P-m11、L-m12为枯草芽孢杆菌,M-r12、P-m9为贝莱斯芽孢杆菌,P-r21为解淀粉芽孢杆菌,接种试验证明这些生防菌株对苹果枝条不具致病性。结论: 从新疆野苹果枝条中筛选并鉴定出对苹果3种病原菌具有抑制作用的内生细菌菌株7株,对苹果树不具致病性。

关键词: 新疆野苹果, 内生细菌, 抑菌作用, 芽孢杆菌属

Abstract:

Objective: In this study,endophytic bacteria were isolated and screened from a wild apple (Malus sieversii) in Xinjiang,which is an excellent germplasm resource in China,in order to obtain new bacterial resources for biological control of apple diseases. Method: Endophytic bacteria were isolated from the branches of M. sieversii by the methods of dilution plating and tissue isolation. Seven endophytic bacteria strains,with the strong inhibitory effect on three pathogenic fungi (Botryosphaeria dothidea,Colletotrichum gloeosporioides and Phomopsis mali),were selected by the method of plate confrontation. The 7 strains were identified based on morphological,physiological and biochemical characteristics,as well as 16S rDNA sequence analysis. Their pathogenicity was determined by using in vitro inoculation tests. Result: In this study,a total of 217 strains of endophytic bacteria were isolated and purified,among which seven strains (M-m7,M-m12,M-r12,P-m9,P-m11,L-m12 and P-r21) with strong inhibitory effects on mycelium growth of three pathogenic fungi of B. dothidea,C. gloeosporioides and P. mali were screened out. Their inhibition rates were 77.00%,72.60%,76.60%,60.64%,65.96%,65.81% and 71.79% on B. dothidea; 69.85%,64.95%,68.87%,68.37%,56.12%,53.54% and 65.66% on C. gloeosporioides; 77.63%,79.22%,72.60%,69.37%,70.27%,69.30% and 71.05% on P. mali,respectively. Strains M-m7,M-m12,P-m11 and L-m12 were identified as Bacillus subtilis,strains M-r12 and P-m9 were identified as B. elezensis and strain P-r21 was identified as B. amyloliquefaciens. Inoculation tests showed the 7 strains had no pathogenicity to apple tree itself. Conclusion: Seven strains of endophytic bacteria in M. sieversii can effectively suppress mycelium growth of B. dothidea,C. gloeosporioides and P. mali,and have no pathogenicity to apple tree.

Key words: Malus sieversii, endophytic bacteria, inhibition activity, Bacillus

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