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中国生物防治学报 ›› 2024, Vol. 40 ›› Issue (3): 630-640.DOI: 10.16409/j.cnki.2095-039x.2024.02.020

• 研究论文 • 上一篇    

抗烟草PVY生防细菌D3-1的筛选鉴定及活性物质分析

李玲玲1, 陈奕冰1, 滕凯2, 肖志鹏2, 周向平2, 肖艳松2, 蔡海林2, 董鹏1, 梅斯钇1, 刘索妮1, 伍绍龙2, 刘天波2, 唐前君1   

  1. 1. 湖南农业大学植物保护学院, 长沙 410128;
    2. 中国烟草总公司湖南省公司, 长沙 410004
  • 收稿日期:2023-09-20 发布日期:2024-06-07
  • 通讯作者: 唐前君,教授,E-mail:Tangqianjun@126.com;刘天波,高级农艺师,E-mail:tianboliu@126.com。
  • 作者简介:李玲玲,硕士研究生,E-mail:2643476710@qq.com

Screening, Identification and Active Substance Analysis of Biocontrol Bacterium D3-1 against Tobacco PVY

LI Lingling1, CHEN Yibing1, TENG Kai2, XIAO Zhipeng2, ZHOU Xiangping2, XIAO Yansong2, CAI Hailin2, DONG Peng1, MEI Siyi1, LIU Suoni1, WU Shaolong2, LIU Tianbo2, TANG Qianjun1   

  1. 1. College of Plant Protection, Hunan Agricultural University, Changsha 410128, China;
    2. Hunan Provincial Tobacco Company, Changsha 410004, China
  • Received:2023-09-20 Published:2024-06-07
  • Contact: 湖南省重点领域研发计划(2023NK2019);中国烟草总公司项目(110202101050(LS-10),110202201019(LS-03));湖南省烟草公司衡阳市公司项目(HYYC2023KJ32)
  • Supported by:
    10.16409/j.cnki.2095-039x.2024.02.020

摘要: 由马铃薯Y病毒(Potato virus Y,PVY)引起的病毒病在烟草种植地区大面积发生,对烟草品质和产量产生重大影响。采用半叶枯斑法筛选出对烟草PVY防治效果优良的生防细菌D3-1,对PVY的枯斑抑制率达99.71%,经形态学、生理生化特性以及16S rDNA序列鉴定其为解淀粉芽胞杆菌Bacillus amyloliquefaciens。菌株D3-1具有广谱的抑菌能力,可抑制病原真菌和细菌生长,菌株D3-1发酵液(1×109 CFU/mL)对烟草PVY的田间防治效果为63.67%。全基因组测序分析发现菌株D3-1基因组含有表面活性素surfactin、丰原素fengycin、嗜铁素bacillibactin等12种具有抑菌作用的次生代谢产物合成基因簇,经HPLC-MS定性分析确定了表面活性素surfactin和丰原素fengycin等32个脂肽类提取物。菌株D3-1对PVY有良好防治效果,具有作为防治烟草PVY生防菌剂的潜力。

关键词: PVY, Bacillus amyloliquefaciens, 全基因组测序, 次生代谢产物合成基因簇, 活性物质

Abstract: Viral diseases caused by potato virus Y (PVY) occur extensively in tobacco planting areas, significantly impacting tobacco quality and yield. Through half-leaf necrosis spot method, the biocontrol bacterium D3-1 with excellent efficacy against PVY in tobacco was screened, exhibiting an inhibition rate of 99.71%. Based on morphological, physiological, biochemical characteristics and 16S rDNA sequence analysis, D3-1 was identified to be Bacillus amyloliquefaciens. D3-1 strain displayed broad-spectrum anti antimicrobial activity, inhibiting the growth of pathogenic fungi and bacteria. Field trials with D3-1 fermentation broth (1×109 CFU/mL) resulted in an efficacy of 63.67% against PVY in tobacco. Whole-genome sequencing revealed that strain D3-1 harbored 12 gene clusters responsible for the synthesis of secondary metabolites, including surfactin, fengycin, and bacillibactin, known for their antimicrobial properties. Qualitative analysis using HPLC-MS confirmed the presence of 32 peptide extracts, including surfactin and fengycin. Strain D3-1 exhibited promising efficacy against PVY, demonstrating its potential as a biocontrol agent for managing PVY in tobacco.

Key words: PVY, Bacillus amyloliquefaciens, whole-genome sequencing, gene cluster of secondary metabolite synthesis, active substance

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