首頁資訊一株西瓜枯萎病生防菌Bacillus methylotrophicus的篩選、鑒定及抑菌物質(zhì)分析

一株西瓜枯萎病生防菌Bacillus methylotrophicus的篩選、鑒定及抑菌物質(zhì)分析

來源：泰然健康網(wǎng) 時(shí)間：2025年07月17日 03:17

作者:

余紅鳳

齊齊哈爾大學(xué)生命科學(xué)與農(nóng)林學(xué)院, 黑龍江齊齊哈爾 161006;黑龍江省農(nóng)業(yè)微生物制劑產(chǎn)業(yè)化技術(shù)創(chuàng)新中心, 黑龍江齊齊哈爾 161006;黑龍江省農(nóng)用生物制劑產(chǎn)業(yè)化協(xié)同創(chuàng)新中心, 黑龍江齊齊哈爾 161006
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張琳

畢鈺

王志剛

徐偉慧

黃欣冉

郭嘉儀

基金項(xiàng)目:

齊齊哈爾市食品產(chǎn)業(yè)研究院項(xiàng)目（SXSP-2021004）；黑龍江省高校基本科研業(yè)務(wù)費(fèi)項(xiàng)目（145209808）；齊齊哈爾大學(xué)研究生創(chuàng)新項(xiàng)目（YJSCX2022020）

Screening, identification, and antifungal substances of an antagonistic strain Bacillus methylotrophicus against Fusarium oxysporum f. sp. niveum

Author:

YU Hongfeng

College of Life Science and Agroforestry, Qiqihar University, Qiqihar 161006, Heilongjiang, China;Heilongjiang Provincial Technology Innovation Center of Agromicrobial Preparation Industrialization, Qiqihar 161006, Heilongjiang, China;Heilongjiang Provincial Collaborative Innovation Center of Agrobiological Preparation Industrialization, Qiqihar 161006, Heilongjiang, China
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ZHANG Lin

BI Yu

WANG Zhigang

XU Weihui

HUANG Xinran

GUO Jiayi

摘要 | | 訪問統(tǒng)計(jì) | 參考文獻(xiàn) [46] | | || 文章評論

摘要:

【背景】西瓜?；图怄哏牭毒‵usarium oxysporum f.sp.niveum，F(xiàn)on）是西瓜枯萎病的致病菌，嚴(yán)重威脅連作西瓜的生產(chǎn)?！灸康摹亢Y選拮抗西瓜枯萎病菌的菌株并推測其無菌上清液（cell-free supernatant，CFS）的抑菌成分，為其在農(nóng)業(yè)生產(chǎn)中的應(yīng)用提供理論依據(jù)。【方法】利用平板對峙法篩選拮抗Fon的菌株，通過盆栽試驗(yàn)驗(yàn)證其抑制西瓜枯萎病的能力，并通過掃描電鏡、共聚焦顯微鏡觀察和抗氧化酶活性測定等方法研究Fon暴露在拮抗菌株次級代謝物下孢子形態(tài)的變化、膜的完整性和造成的氧化損傷，通過二代Illumina Hiseq聯(lián)合三代PacBio Sequel測序平臺進(jìn)行全基因組測序、LC-MS非靶代謝組學(xué)、超高液相色譜系統(tǒng)和高分辨率質(zhì)譜（UHPLC-ESI-MS/MS），明確拮抗菌株次級代謝物中的抑菌功能物質(zhì)?！窘Y(jié)果】從健康西瓜根際土篩選到拮抗Fon的一株優(yōu)良菌株J4，盆栽試驗(yàn)表明其對西瓜枯萎病的防治效果為72.6%。經(jīng)全基因組學(xué)鑒定為Bacillus methylotrophicus J4，該菌株可產(chǎn)蛋白酶、淀粉酶、葡聚糖酶和纖維素酶并有分泌鐵載體的能力，其衰亡期無菌上清液對Fon菌絲生長抑制率為71.87%，菌株J4的CFS使Fon孢子表面凹陷，破壞其細(xì)胞膜的完整性，造成細(xì)胞死亡。菌株J4誘導(dǎo)Fon菌絲體活性氧（reactive oxygen species，ROS）的積累；菌株J4的CFS引起Fon菌絲超氧化物歧化酶（superoxide dismutase，SOD）、過氧化氫酶（catalase，CAT）、過氧化物酶（peroxidase，POD）、谷胱甘肽還原酶（glutathione reductase，GR）活性，以及可溶性蛋白和丙二醛（malondialdehyde，MDA）含量的顯著升高。該菌株基因組中含有13個(gè)抑菌功能的次級代謝產(chǎn)物合成基因簇，菌株J4的CFS中含有3種多肽化合物，并且這3種脂肽具有抑制Fon菌絲生長的能力，推測菌株J4上清液中的抑菌功能物質(zhì)是伊枯草菌素A （iturin A）、豐原素（fengycin）和表面活性素（surfactin）?！窘Y(jié)論】菌株J4對西瓜枯萎病有較強(qiáng)的防效，在西瓜枯萎病生物防治中具有廣泛的應(yīng)用前景。

Abstract:

[Background] Fusarium oxysporum f. sp. niveum (Fon) is the causal agent of watermelon wilt, seriously threatening the production of watermelon in continuous cropping. [Objective] To screen out the antagonistic strain against Fon and identify the antifungal components in the cell-free supernatant (CFS), providing a theoretical basis for application of the strain in agricultural production. [Methods] Plate confrontation was employed to screen the antagonistic strain against Fon. Pot experiments were conducted to verify the control effect of the antagonistic strain on watermelon wilt. Scanning electron microscopy and confocal microscopy were employed to observe the conidial morphology and membrane integrity of Fon. The activities of antioxidant enzymes were determined to evaluate the oxidative damage. Whole genome sequencing through second-generation Illumina Hiseq combined with third-generation PacBio Sequel sequencing platform, LC-MS-based non-target metabolomics, and ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UHPLC-ESI-MS/MS) were employed to identify the antifungal substances in the secondary metabolites of the strain. [Results] An antagonistic strain J4 was screened out from the rhizosphere soil of healthy watermelon. The pot experiments showed that its control effect against watermelon wilt was 72.6%. The strain was identified as Bacillus methylotrophicus J4 by whole genome sequencing, and it could produce protease, amylase, glucanase, cellulose, and siderophores. The CFS of J4 in the decline stage presented the inhibition rate of 71.87% on the growth of Fon. Moreover, it led to the collapsing of Fon spore surface, destroyed the integrity of the cell membrane, and caused cell death. Strain J4 induced the accumulation of reactive oxygen species (ROS) in the mycelia of Fon. The CFS of strain J4 significantly increased the activities of superoxide dismutase, catalase, peroxidase, and glutathione reductase and the content of soluble protein and malondialdehyde in the mycelia of Fon. The genome of strain J4 carried 13 gene clusters for the synthesis of secondary metabolites with antimicrobial functions. The CFS of strain J4 contained three peptides, which were proved to inhibit the growth of Fon. It was hypothesized that the antifungal functional substances in the CFS of strain J4 were iturin A, fengycin, and surfactin. [Conclusion] Strain J4 had a strong control effect on watermelon wilt and could be used to control watermelon wilt in the future.

引用本文

余紅鳳,張琳,畢鈺,王志剛,徐偉慧,黃欣冉,郭嘉儀. 一株西瓜枯萎病生防菌Bacillus methylotrophicus的篩選、鑒定及抑菌物質(zhì)分析[J]. 微生物學(xué)通報(bào), 2024, 51(2): 534-553

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一株西瓜枯萎病生防菌Bacillus methylotrophicus的篩選、鑒定及抑菌物質(zhì)分析

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一株西瓜枯萎病生防菌Bacillus methylotrophicus的篩選、鑒定及抑菌物質(zhì)分析

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一株西瓜枯萎病生防菌Bacillus methylotrophicus的篩選、鑒定及抑菌物質(zhì)分析

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