茶轮斑病拮抗细菌kc-16的抑菌性能及田间防效  

Antimicrobial activity and field control effect of an antagonistic bacterial strain kc-16 against tea gray blight

作　　者：杨学宇 谭琳[3] 沈程文[1,2] 邓玉莲 李桂花 黄虹 胡秋龙 YANG Xueyu;TAN Lin;SHEN Chengwen;DENG Yulian;LI Guihua;HUANG Hong;HU Qiulong(Key Laboratory of Tea Science of Ministry of Education,Hunan Agricultural University,Changsha 410128,Hunan,China;College of Horticulture,Hunan Agricultural University,Changsha 410128,Hunan,China;College of Plant Protection,Hunan Agricultural University,Changsha 410128,Hunan,China)

机构地区：[1]湖南农业大学茶学教育部重点实验室,湖南长沙410128 [2]湖南农业大学园艺学院,湖南长沙410128 [3]湖南农业大学植物保护学院,湖南长沙410128

出　　处：《微生物学通报》2024年第11期4560-4573,共14页Microbiology China

基　　金：国家重点研发计划(2022YFD16000803-6);湖南省重点研发计划(2022NK2051,2023NK2013);湖南省科技创新重大项目(2021NK1020)。

摘　　要：【背景】茶轮斑病是茶树的主要病害,目前仍以化学防治为主,而化学防治引起的农残与抗药性问题亟待解决,发掘和利用生防微生物资源是保障茶叶安全生产和提高产量的重要举措。【目的】筛选对茶轮斑病菌(Pseudopestalotiopsis theae)具有拮抗能力的芽孢杆菌并初步明确该拮抗菌的抑菌性能及其对茶树的田间防病促生效果,为茶轮斑病菌生防技术开发提供支持。【方法】利用平板对峙法筛选对茶轮斑病菌具有拮抗效果的细菌;通过形态特征、生理生化特征结合16S rRNA基因测序对筛选所得拮抗菌进行种属鉴定;利用不同检测平板分析产真菌细胞壁裂解酶的特性,通过平板对峙试验测定菌株kc-16不同类型代谢产物对茶轮斑病菌的抑制能力,同时测定菌株kc-16对其他9种常见植物病原真菌的拮抗率以确定其广谱抑菌能力,最后进行田间试验评价菌株kc-16对茶轮斑病的防治和对茶树的增产效果。【结果】筛选到一株对茶轮斑病菌有较好抑制效果的菌株kc-16,抑菌率为(79.62±0.82)%。经生理生化特征比对与16S rRNA基因序列同源性分析,将菌株kc-16鉴定为枯草芽孢杆菌沙漠亚种(Bacillus subtilis subsp.inaquosorum);该菌具有固氮、产铁载体与溶磷能力,但不具有解钾能力;能够分泌蛋白酶、纤维素酶、β-1,3葡聚糖酶和几丁质酶。此外,其对9种病原真菌均有不同程度的抑制效果,抑制率为46.67%-81.00%。同时,其胞外抑菌物质均对茶轮斑病菌有抑制作用,菌株kc-16的无菌滤液、蛋白类代谢物、挥发性代谢产物和非挥发性代谢产物的平板抑制率分别为69.00%、58.00%、47.50%和78.71%;田间试验表明枯草芽孢杆菌沙漠亚种对茶树有良好的防病促生效果,在显著降低茶轮斑病发病率的同时,使茶叶鲜重(1芽1叶)相较空白对照提升了25.0%。【结论】枯草芽孢杆菌沙漠亚种对茶轮斑病菌有较好的抑制效果,同时对�[Background]Tea gray blight is a major disease attacking tea plants.Chemical methods are mainly used to control this disease,and the chemical residues and pathogen resistance caused by chemical control need to be solved urgently.Exploring biocontrol microorganisms is an important measure to ensure the safety and increase the yield of tea production.[Objective]To screen the Bacillus strains with inhibitory effects on tea gray blight and clarify the inhibitory activity and field control effect of the strain screened out,providing support for the development of biocontrol methods for tea gray blight.[Methods]A plate confrontation experiment was conducted to screen out the strain with antagonistic effect on Pseudopestalotiopsis theae,the pathogen causing tea gray blight.The strain was identified based on morphological,physiological,and biochemical characteristics and sequence analysis of the 16S rRNA.Various plates were utilized to examine the production of enzymes lysing fungal cell walls by the strain.Plate confrontation tests were conducted to determine the inhibitory activities of different metabolites produced by the strain on Ps.theae.The inhibition rates against nine common species of plant pathogenic fungi were measured to assess the antifungal spectrum of the strain.Finally,field experiments were conducted to evaluate the effects of the strain on tea gray blight and yield of tea plants.[Results]A strain kc-16 with antagonistic effect on Ps.theae was screened out,with the inhibition rate of(79.62±0.82)%,and it was identified as Bacillus subtilis subsp.inaquosorum by morphological,physiological,and biochemical characteristics and 16S rRNA gene sequence analysis.Strain kc-16 demonstrated nitrogen-fixing,iron carriers producing and phosphorus-solubilizing abilities but lacked the potassium-solubilizing ability,and it could secrete protease,cellulase,β-1,3-glucanase,and chitinase.In addition,strain kc-16 inhibited nine species of pathogenic fungi,with inhibition rates ranging from 46.67%to 81.00%.The extracellu

关 键 词：茶轮斑病 枯草芽孢杆菌沙漠亚种 生物防治 促生 

分 类 号：S476[农业科学—农业昆虫与害虫防治] S435.711[农业科学—植物保护]