摘要
筛选获得对灰霉病菌具有抑制活性的芽孢杆菌菌株,明确其胞外抑菌物质的活性及稳定性,为灰霉病提供安全、高效、易行的生物防治方式,并为其后续生防制剂的研发提供科学依据。采集我国云南、黑龙江、新疆、河北等地蔬菜种植土壤样品129个,采用稀释涂平板法分离获得芽孢杆菌4 200株,采用平板对峙法获得191株具有灰葡萄孢菌拮抗性能的菌株,其中菌株KA4的抑菌能力最强,对灰霉病菌的抑菌率为65.34%。采用形态学、生理生化特征以及16S rDNA和gyrB基因序列分析相结合的鉴定方法进行了KA4菌株的鉴定,将其鉴定为解淀粉芽孢杆菌,命名为BA-KA4。BA-KA4的无菌发酵液具有较强的抑菌活性,稀释10,20,40倍的无菌发酵液对灰葡萄孢菌的抑菌率分别为78.83%,65.19%,45.38%。通过单因素法测定了不同处理温度、pH值和紫外照射时间对BA-KA4无菌发酵液抑菌活性的影响,该菌株的无菌发酵液具有良好的温度、pH和紫外线稳定性。综上,解淀粉芽孢杆菌BA-KA4能够抑制灰霉病菌生长、胞外抑菌物质活性稳定,可作为灰霉病生物防治的优良微生物资源,具有良好的开发前景。
In order to provide a safe,efficient and easy way of biological control against Botrytis cinerea,Bacillus strains with inhibitory activity were screened and the activity and stability of its extracellular antimicrobial components were determined.The research provided scientific basis for the research and development of biocontrol agents.4200 strains of Bacillus spp.were isolated by dilution plate method from 129 soil samples of vegetable planting soil from Yunnan,Heilongjiang,Xinjiang and Hebei Province.191 strains with antagonistic ability against Botrytis cinerea were obtained by dural culture method.Among them,strain KA4 had the strongest inhibition ability and the inhibition rate against Botrytis cinerea was 65.34%.The strain KA4 was identified as Bacillus amyloliquefaciens and named BA-KA4 by morphological,physiological and biochemical characteristics,16S rDNA and gyrB gene sequence analysis.The sterile fermentation broth of this strain had strong antimicrobial activity.The inhibition rates of BA-KA4 sterile fermentation broth with 10-fold,20-fold and 40-fold dilution were 78.83%,65.19%,45.38%,respectively.The single factor method was used to determine the effect of different treatment temperature,pH value and UV irradiation time on the antibacterial activity of BA-KA4 sterile fermentation broth.The sterile fermentation broth of this strain had good temperature,pH and UV stability.Comprehensively,B.amyloliquefaciens BA-KA4 could inhibit the growth of Botrytis cinerea and the activity of extracellular antimicrobial substances was stable,which could be used as a good microbial resource for biological control of Botrytis cinerea and had a good development prospect.
作者
刘昆昂
黄亚丽
贾振华
马宏
宋水山
LIU Kunang;HUANG Yali;JIA Zhenhua;MA Hong;SONG Shuishan(Institute of Biology,Hebei Academy of Sciences,Shijiazhuang 050081,China;Hebei University of Science and Technology,Shijiazhuang 050018,China;Hebei Engineering and Technology Center of Microbiological Control on Main Crop Disease,Shijiazhuang 050081,China)
出处
《华北农学报》
CSCD
北大核心
2020年第3期200-207,共8页
Acta Agriculturae Boreali-Sinica
基金
国家“十二五”水污染治理重大专项(2015ZX07204-007)
国家重点研发计划(2017YFD0300905)。
关键词
灰霉病
灰葡萄孢菌
解淀粉芽孢杆菌
生物防治
胞外抑菌成分
Gray mould
Botrytis cinerea
Bacillus amyloliquefaciens
Biocontrol
Extracellular bacteriostatic components
