摘要
【目的】从金银花叶片中筛选获得对金银花叶斑病病原菌具有良好拮抗效果的生防菌株,旨在为金银花叶斑病的生物防治提供合适的菌种资源,为金银花产业的高质量发展打下基础。【方法】以金银花叶片为材料,利用梯度稀释涂板法分离并纯化内生芽孢杆菌;采用牛津杯平板对峙试验筛选对金银花叶斑病病原菌具有拮抗作用的菌株;通过离心管温浴试验评估拮抗菌株对叶斑病菌形态的影响;通过牛津杯平板对峙试验检测拮抗菌株脂肽类抗生素粗提液对叶斑病菌菌丝生长的抑制作用;通过温室盆栽试验评估拮抗菌株对金银花叶斑病的防治效果;结合形态学、生理生化和分子生物学特征对拮抗菌株进行鉴定;利用平板对峙法测定拮抗菌株的抑菌谱。【结果】从金银花叶片中共分离得到120株内生芽孢杆菌,通过平板对峙法筛选获得4株对金银花叶斑病病原菌具有较好抑制效果的拮抗菌株,4株菌株对金银花叶斑病菌的拮抗效果无显著差异(P>0.05),随机选取菌株HC-14开展后续研究。平板对峙试验结果,菌株HC-14对金银花叶斑病菌具有良好的拮抗效果,其抑菌率为49.38%。离心管温浴试验结果,菌株HC-14的上清液对金银花叶斑病菌具有较强的抑制效果。牛津杯平板对峙试验结果,菌株HC-14的脂肽类抗生素粗提液对金银花叶斑病菌具有一定的抑制效果,其抑菌率为11.44%。盆栽试验结果,喷施接种菌株HC-14发酵液后第21 d,预防组、治疗组和保护组的病情指数分别仅为6.94、6.35和4.21,防治效果依次为76.62%、78.59%和85.79%。结合菌落形态、生理生化及分子生物学鉴定结果,将菌株HC-14鉴定为枯草芽孢杆菌(Bacillus subtilis)。菌株HC-14抑菌谱测定结果,菌株HC-14除对青枯劳尔氏菌无抑菌效果外,对10种常见的致病菌均有不同程度的抑制作用。【结论】枯草芽孢杆菌菌株HC-14的抑菌谱较广,具有开发
【Objective】The biocontrol strains with good antagonistic effect against the pathogens of honeysuckle leaf spot were selected from the leaves of honeysuckle,in order to provide suitable strain resources for the biological control of honeysuckle leaf spot and lay a foundation for the high-quality development of honeysuckle industry.【Method】In this study,honeysuckle leaves were used as materials to isolate and purify endophytic Bacillus strains using the dilution separa-tion method.The strains with antagonistic effects on Alternaria alternata were screened using the Oxford cup plate con-frontation method.The effects of antagonistic bacteria on the morphology of A.alternata were evaluated by a centrifuge tube co-culture test.The inhibitory effect of the lipopeptide crude extracts of antagonistic bacteria on the mycelial growth of A.alternata were detected using Oxford cup plate confrontation test.The biocontrol effect of antagonistic bacteria on honeysuckle leaf spot in pot assay were evaluated under greenhouse conditions.Antagonistic bacteria were identified based on morphological,physiological,biochemical,and molecular biological characteristics.Moreover,the antimicrobial spectrum of antagonistic bacteria were determined by plate confrontation method.【Result】A total of 120 endophytic Bacil-lus strains were isolated from honeysuckle leaves.Through the plate confrontation method,four strains were selected that exhibited effective antagonistic properties against the leaf spot pathogen of honeysuckle.The antagonistic effects of these four strains on the leaf spot pathogen were not significantly different(P>0.05).Strain HC-14 was randomly chosen for further study.The results of the plate confrontation test showed that strain HC-14 had a good antagonistic effect against the leaf spot pathogen with an inhibition rate of 49.38%.The results of the centrifuge tube temperature bath showed that the supernatant of strain HC-14 had a strong inhibitory effect on A.alternate.The results of the Oxford cup plate test sho
作者
吴小云
何朋杰
罗喜燕
崔文艳
WU Xiao-yun;HE Peng-jie;LUO Xi-yan;CUI Wen-yan(School of Basic Medicine,Guizhou University of Traditional Chinese Medicine,Guiyang,Guizhou 550025,China;Key Laboratory of Green Pesticide and Agricultural Bioengineering,Ministry of Education,Guizhou University,Guiyang,Guizhou 550025,China)
出处
《南方农业学报》
CAS
CSCD
北大核心
2023年第10期2958-2968,共11页
Journal of Southern Agriculture
基金
贵州省基础研究计划项目(黔科合基础-ZK〔2021〕一般146)
贵州中医药大学大学生创新创业训练计划项目(贵中医大创合字〔2022〕23号)
贵州省教育厅高等学校科学研究项目(青年项目)(黔教技〔2022〕208号)。
关键词
金银花
叶斑病
链格孢菌
枯草芽孢杆菌
共培养
生物防治
honeysuckle
leaf spot
Alternaria alternate
Bacillus subtilis
co-culture
biocontrol control
