烟草赤星病多功能生防菌株的筛选及菌群构建优化  

Screening,Construction and Optimization of Microbial Communities for Multifunctional Biocontrol of Tobacco Brown Spot

作　　者：吴旭艳 李行善 崔传斌 王平平 黄丽丽 王娜娜[1,2] WU Xu-Yan;LI Xing-Shan;CUI Chuan-Bin;WANG Ping-Ping;HUANG Li-Li;WANG Na-Na(College of Life Sciences,Northwest A&F University,Yangling 712100,China;National Key Laboratory of Crop Stress Resistance and Efficient Production,Yangling 712100,China;College of Plant Protection,Northwest A&F University,Yangling 712100,China;Shaanxi Provincial Company,China National Tobacco Corporation,Xi'an 710000,China)

机构地区：[1]西北农林科技大学生命科学学院,杨凌712100 [2]作物抗逆与高效生产全国重点实验室,杨凌712100 [3]西北农林科技大学植物保护学院,杨凌712100 [4]中国烟草总公司陕西省公司,西安710000

出　　处：《农业生物技术学报》2025年第3期628-641,共14页Journal of Agricultural Biotechnology

基　　金：中国烟草总公司重大科技项目(110202101056LS-16);中国烟草总公司陕西省公司科技项目(KJ-2021-02)。

摘　　要：烟草赤星病是危害烟草产业的主要真菌病害,生物防治是目前防治植物病害的重要手段。而合成菌群因其在农业生产中表现出的良好的促生抗逆等功能一直是生物防治领域的研究热点。本研究以陕西陕南烟区烟草赤星病重病烟田健康烟草植株(Nicotiana tabacum)的根际土壤为材料,采用稀释分离、平板对峙法和菌丝生长速率法分离筛选对烟草赤星病菌(Alternaria alternate)具有显著拮抗作用的菌株;通过测定细菌产吲哚乙酸(indole acetic acid, IAA)能力和促生盆栽实验筛选促生菌株;通过测定烟草叶片苯丙氨酸解氨酶(phenylalaninammo-nialyase, PAL)、多酚氧化酶(polyphenol oxidase, PPO)和过氧化物酶(peroxidase, POD)防御反应酶活筛选诱抗菌株,将筛选出的菌株进行形态学、生理生化特征测定和分子生物学方法鉴定,最终使用Box-Benhnken响应面法优化菌群配比,从而构建多功能生防菌群。筛选出的拮抗菌株Xh628A (GenBank No. PQ269269)和促生菌株X5616A (GenBank No. PQ269268)经鉴定均为贝莱斯芽孢杆菌(Bacillus velezensis),诱抗菌株Xh649A (GenBank No. PQ269267)经鉴定为皮特不动杆菌(Acinetobacter pittii);优化后菌群的最佳添加比例确定为Xh628A∶Xh649A∶X5616A=5.0∶1.0∶4.5,经验证该菌群的平均菌丝抑制率为77.24%,并相比单一菌株其促生和诱抗能力均有提升。本研究为后续开发高效、稳定和环境友好型多功能生物农药提供基础资料,并为多功能生物菌群的构建和配比优化提供科学方法。Tobacco brown spot is the main fungal disease that endangers the tobacco industry.Biological control is an important means to control plant diseases.Synthetic community has been a research hotspot in the field of biological control because of its good growth-promoting and stress-resistant functions in agricultural production.The rhizosphere soil of healthy tobacco(Nicotiana tabacum)plants in the tobacco field with severe brown spot disease in southern Shaanxi was used as the material,and the strains with significant antagonistic effect on Alternaria alternate were isolated and screened by dilution separation,plate confrontation method and mycelial growth rate method.The growth-promoting strains were screened by measuring the ability of bacteria to produce indole acetic acid(IAA)and growth-promoting pot experiments.The antibacterial strains were screened by measuring the activities of phenylalaninammo-nialyase(PAL),polyphenol oxidase(PPO)and peroxidase(POD)defense enzymes in tobacco leaves,and then the selected strains were identified by morphological,physiological and biochemical characteristics and molecular biological methods.Then,the Box-Benhnken response surface method was used to optimize the ratio of bacteria to finally construct a multi-functional biocontrol bacteria group.The screened antagonistic strain Xh628A(GenBank No.PQ269269)and growth-promoting strain X5616A(GenBank No.PQ269268)were identified as Bacillus velezensis,and the induced antibacterial strain Xh649A(GenBank No.PQ269267)was identified as Acinetobacter pittii.The optimal addition ratio of the optimized flora was determined to be Xh628A∶Xh649A∶X5616A=5.0∶1.0∶4.5,and the average mycelial inhibition rate of the flora was 77.24%,compared with the single strain,its growth-promoting and resistance-inducing ability has been improved.This study provides a research basis for the subsequent development of efficient,stable and environmentally friendly multifunctional biological pesticides,and provides a scientific method for the construction a

关 键 词：烟草赤星病菌 拮抗 促生 诱抗 细菌菌群 响应面法 

分 类 号：S435.72[农业科学—农业昆虫与害虫防治]