蜡状芽孢杆菌的基因组和代谢特征，通过合成次生代谢物抑制核盘菌的生长。

Genomic and metabolic features of Bacillus cereus, inhibiting the growth of Sclerotinia sclerotiorum by synthesizing secondary metabolites.

机构信息

College of Horticulture and Plant Protection, Inner Mongolia Agricultural University, Hohhot, 010020, Inner Mongolia, China.

Inner Mongolia Cold and Arid Region Crop Protection Engineering Technology Center, Hohhot, 010020, Inner Mongolia, China.

出版信息

Arch Microbiol. 2022 Dec 1;205(1):8. doi: 10.1007/s00203-022-03351-5.

DOI:10.1007/s00203-022-03351-5

PMID:36454319

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9715469/

Abstract

We investigated the biocontrol mechanism of Bacillus cereus CF4-51 to find powerful microbes that effectively control Sclerotinia sclerotiorum. To assess its inhibitory effect on fungal growth, the plant pathogen (S. sclerotiorum) was co-cultured with Bacillus cereus. Scanning electron microscope (SEM) was used to study the morphology of S. sclerotiorum treated with CF4-51 biofumigant. The expression of sclerotium formation-related genes was analyzed by qRT-PCR. We performed whole genome sequencing of CF4-51 by PacBio Sequel platform. Lipopeptides were extracted from strain CF4-51 according to the method of hydrochloric acid precipitation and methanol dissolution. The volatiles CF4-51 were identified using gas chromatography-mass spectrometry (GC-MS). We found that the volatile organic compounds (VOCs) released by CF4-51 damaged the S. sclerotiorum hyphae and inhibited the formation of sclerotia. The qRT-PCR data revealed the down-regulated expression of the genes involved in sclerotial formation. Moreover, we analyzed the B. cereus CF4-51 genome and metabolites. The genome consisted of 5.35 Mb, with a GC content of 35.74%. An examination of the genome revealed the presence of several gene clusters for the biosynthesis of antibiotics, siderophores, and various other bioactive compounds, including those belonging to the NRPS-like, LAP, RIPP-like, NRPS, betalactone, CDPS, terpene, ladderane, ranthipeptide, and lanthipeptide (class II) categories. A gas chromatography-tandem mass spectrometry analysis identified 45 VOCs produced by strain CF4-51. Among these, technical grade formulations of five were chosen for further study: 2-Pentadecanone, 6,10,14-trimethyl-,1,2-Benzenedicarboxylic acid, bis(2-methylpropyl) ester, Dibutyl phthalate, Cyclododecane, Heptadecane. the five major constituents play important roles in the antifungal activity of the VOCs CF4-51 on the growth of S. sclerotiorum. The secondary metabolites produced by strain CF4-51are critical for the inhibition of S. sclerotiorum hyphal growth and sclerotial formation.

摘要

我们研究了蜡状芽孢杆菌 CF4-51 的生防机制，以寻找有效控制核盘菌的有力微生物。为了评估其对真菌生长的抑制作用，将植物病原菌（核盘菌）与蜡状芽孢杆菌共培养。用扫描电子显微镜（SEM）研究了 CF4-51 生物熏蒸剂处理后的核盘菌形态。通过 qRT-PCR 分析了与菌核形成相关基因的表达。我们使用 PacBio Sequel 平台对 CF4-51 进行了全基因组测序。根据盐酸沉淀和甲醇溶解的方法从菌株 CF4-51 中提取脂肽。使用气相色谱-质谱联用（GC-MS）鉴定 CF4-51 的挥发性物质。我们发现 CF4-51 释放的挥发性有机化合物（VOCs）破坏了核盘菌菌丝，抑制了菌核的形成。qRT-PCR 数据显示，与菌核形成相关的基因表达下调。此外，我们还分析了蜡状芽孢杆菌 CF4-51 的基因组和代谢物。基因组由 5.35 Mb 组成，GC 含量为 35.74%。对基因组的分析表明，存在几个抗生素、铁载体和各种其他生物活性化合物的生物合成基因簇，包括属于 NRPS 样、LAP、RIPP 样、NRPS、β-内酰胺、CDPS、萜烯、 ladderane、ranthipeptide 和 lanthipeptide（II 类）类别的化合物。气相色谱-串联质谱分析鉴定了 CF4-51 产生的 45 种 VOCs。其中，选择了技术级配方的五种进行进一步研究：2-十五烷酮、6,10,14-三甲基-1,2-苯二甲酸、双（2-甲基丙基）酯、邻苯二甲酸二丁酯、环十二烷、十七烷。这五种主要成分在 VOCs CF4-51 对核盘菌生长的抗真菌活性中发挥重要作用。CF4-51 产生的次生代谢物对抑制核盘菌菌丝生长和菌核形成至关重要。