基于全基因组测序的多粘类芽孢杆菌NBmelon-1生防机制探究及验证
作者:
作者单位:

1.宁波市农业科学研究院/宁波市特色园艺作物品质调控和抗性育种重点实验室, 宁波 315040;2.浙江万里学院, 宁波 315199

作者简介:

董文杰,E-mail:dongwenjie032020@163.com

通讯作者:

王毓洪,E-mail:yhwangsc@163.com

中图分类号:

Q933;S476

基金项目:

宁波市科技局重点研发计划暨“揭榜挂帅”项目(2023Z115);浙江省西甜瓜良种育繁推科技创新平台项目(ZJ2019-80);国家西甜瓜产业技术体系建设专项(CARS-25)


Exploration and verification of biocontrol mechanisms of Paenibacillus polymyxa NBmelon-1 based on whole genome sequencing and analysis
Author:
Affiliation:

1.Ningbo Academy of Agricultural Sciences/Ningbo Key Laboratory for Quality Control and Resistance Breeding of Characteristic Horticultural Crops, Ningbo 315040, China;2.Zhejiang Wanli University, Ningbo 315199, China

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    摘要:

    为研究多粘类芽孢杆菌NBmelon-1的抑菌促生机制,采用三代PacBio测序技术进行全基因组测序,分析潜在的功能基因以及预测次级代谢产物合成基因簇等;检测抑菌促生相关基因的表达情况;利用粗提物拮抗试验明确抑菌物质的主要成分,并验证该菌促生潜力,进一步明确其生防机制。结果显示,多粘类芽孢杆菌NBmelon-1基因组大小为5.7 Mb,GC含量为45.68%,含有4 984个编码基因;基因组中存在大量与诱导植物抗病性、水解酶和促进植物生长相关的基因,共预测到5个脂肽类次级代谢产物合成基因簇;采用酸沉淀法从NBmelon-1培养物中分离获得的粗提物对多种甜瓜病原真菌具有明显的抑菌活性,说明其中含有抑菌物质。促生潜力试验表明该菌株能够合成生长激素IAA、具有产铁载体能力,并为植物提供氮元素。以上结果表明,菌株NBmelon-1是一株多功能生防菌,具有广阔的开发应用前景。

    Abstract:

    Paenibacillus polymyxa NBmelon-1 is an efficient biocontrol bacterium isolated from the roots of sweet melons. The third-generation of PacBio sequencing technique was used to sequence the whole genome of strain NBmelon-1 to study the biocontrol mechanisms of P. polymyxa NBmelon-1. The potential functional genes were analyzed and clusters of genes involved in the synthesis of secondary metabolites were predicted. The expression of genes related with antibacterial effect and promoting the growth of plants was detected. The antagonism test of crude extracts was used to identify the main components of antibacterial substances and validate the potential of the bacteria to promote the growth of plants, further clarifying its biocontrol mechanism. The results showed that the genome of P. polymyxa NBmelon-1 was 5.7 Mb, with a GC content of 45.68% and 4 984 coding genes. There were a large number of genes involved in inducing plant disease resistance, hydrolytic enzymes, and promoting plant the growth of plants in the genome. A total of five clusters of genes involved in the synthesis of secondary metabolites of lipopeptides were predicted. The crude extracts isolated from NBmelon-1 culture with acid precipitation method had significant antibacterial activity against a variety of pathogenic fungi from melon, indicating the presence of antibacterial substances. The results of growth potential test showed that the strain was capable of synthesizing growth hormone IAA, producing iron carriers, and providing nitrogen elements for plants. It is indicated that P. polymyxa NBmelon-1 is a multifunctional biocontrol bacterium with broad prospects for development and application in agricultural production.

    图1 P. polymyxa NBmelon-1的基因组图Fig.1 Genome map of P. polymyxa NBmelon-1
    图2 P. polymyxa NBmelon-1基因组COG注释Fig.2 COG annotated map of the genome of P. polymyxa NBmelon-1
    图3 P. polymyxa NBmelon-1基因组GO注释Fig.3 GO annotation of the genome of P. polymyxa NBmelon-1
    图4 P. polymyxa NBmelon-1基因组KEGG注释Fig.4 KEGG annotation of the genome of P. polymyxa NBmelon-1
    图5 P. polymyxa NBmelon-1基因组CAZymesFig.5 CAZymes in the genome of P. polymyxa NBmelon-1
    图6 P. polymyxa NBmelon-1基因组中次级代谢产物基因簇Fig.6 Gene cluster of secondary metabolites in the genome of P. polymyxa NBmelon-1
    图7 P. polymyxa NBmelon-1抑菌促生相关基因分子验证结果Fig.7 Molecular verification results of genes related to inhibition and growth promotion of P. polymyxa NBmelon-1
    图8 P. polymyxa NBmelon-1抑菌物质拮抗试验Fig.8 Antagonism test of antimicrobial substances of P. polymyxa NBmelon-1
    图9 P. polymyxa NBmelon-1促进植物生长及产纤维素酶能力测定Fig.9 Determination of the plant growth promotion and cellulase production ability of P. polymyxa NBmelon-1
    表 1 PCR引物序列Table 1 Primer sequences used in this study
    表 2 菌株NBmelon-1基因组的基本特征Table 2 Basic properties of the genome of P. polymyxa NBmellon-1
    表 3 P. polymyxa NB melon-1毒力因子等级分类Table 3 Classification of virulence factors of P. polymyxa NB melon-1
    表 4 P. polymyxa NBmelon-1次级代谢产物合成基因簇Table 4 Synthesis of gene cluster by P. polymyxa NBmelon-1 secondary metabolites
    表 5 2种粗提物抑菌效果对比Table 5 Comparison of antibacterial effects between two crude extracts
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董文杰,郝芳敏,臧全宇,马二磊,丁伟红,王毓洪.基于全基因组测序的多粘类芽孢杆菌NBmelon-1生防机制探究及验证[J].华中农业大学学报,2025,44(2):200-211

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  • 收稿日期:2024-11-12
  • 在线发布日期: 2025-04-02
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