Article(id=1316714628260938674, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1316714454184739470, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20260197, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1773072000000, receivedDateStr=2026-03-10, revisedDate=null, revisedDateStr=null, acceptedDate=1775577600000, acceptedDateStr=2026-04-08, onlineDate=1791858830751, onlineDateStr=2026-10-13, pubDate=1791043200000, pubDateStr=2026-10-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1791858830751, onlineIssueDateStr=2026-10-13, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1791858830751, creator=13701087609, updateTime=1791858830751, updator=13701087609, issue=Issue{id=1316714454184739470, tenantId=1146029695717560320, journalId=1192105938417971205, year='2026', volume='66', issue='10', pageStart='4661', pageEnd='5310', issueExtLink='null', onlineDate='null', pubDate='1791043200000', pubDateStr='2026-10-04', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1791858789249, creator='13701087609', updateTime=1791885400527, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1316826070066774161, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1316714454184739470, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1316826070066774162, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1316714454184739470, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=5025, endPage=5037, ext={EN=ArticleExt(id=1316714629439538099, articleId=1316714628260938674, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Screening, identification, and control efficacy evaluation of biocontrol strains against maize stalk rot caused by Fusarium graminearum, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

Maize, an important food crop in China, directly affects national food security. In recent years, due to climate change and shifts in farming practices, maize stalk rot has become a major threat to maize production in China. [Objective] To screen and identify biocontrol bacterial strains against maize stalk rot, determine their optimal fermentation process, and develop effective bacterial agents, thereby providing theoretical support and a material basis for constructing a novel green prevention and control technology system. [Methods] Antagonistic bacterial strains against Fusarium graminearum were screened through a combination of dilution coating and plate confrontation methods. Candidate strains were identified through molecular biological techniques combined with physiological and biochemical characterization. Fermentation conditions were optimized via single-factor experiments, and seed coating agents were developed with the selected biocontrol bacteria. Indoor pot experiments and outdoor field plot trials were carried out for the biocontrol bacteria to systematically evaluate their maize growth-promoting effects and control efficacy against maize stalk rot. [Results] Two biocontrol strains, HS104 and HS107, with inhibition rates of approximately 70.00% against F. graminearum, were successfully screened out and identified as Bacillus velezensis. Both strains exhibited broad-spectrum antagonistic activity against major maize pathogens, including F. verticillioides, F. oxysporum, and Bipolaris maydis, with inhibition rates of around 60.00%. After optimization of fermentation conditions, indoor pot experiments showed that HS104 and HS107 not only significantly promoted maize growth but also achieved the control efficacy of 75.00% and 71.00% against maize stalk rot, respectively. The seed coating agents developed with the selected biocontrol bacteria performed well in field trials, significantly promoting maize growth and achieving the control efficacy of 63.93% and 77.33% against maize stalk rot, respectively. Moreover, they increased the yield of susceptible plants by 8.20% and 9.20%, respectively, compared with the control. [Conclusion] Strains HS104 and HS107 possess multiple excellent traits, including broad-spectrum antifungal activity, strong plant growth-promoting effects, and high disease control efficacy, demonstrating broad application prospects in the biocontrol of maize stalk rot and the enhancement of crop yields.

, authors=Zichen WANG1, 2, Liming WANG2, Shiyu LI2, Kang ZHANG1, 2, Hongzhe CAO2, Jihong XING1, 2, *, Jingao DONG1, 2, *, authorsList=Zichen WANG, Liming WANG, Shiyu LI, Kang ZHANG, Hongzhe CAO, Jihong XING, Jingao DONG, authorCompany=null, correspAuthors=Jihong XING, Jingao DONG, authorNote=

These authors contributed equally to this work.

, correspAuthorsNote=
*E-mail: XING Jihong,
DONG Jingao,
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玉米作为我国重要的粮食作物,其产量直接关系国家粮食安全。近年来,受气候变化和耕作模式改变的影响,玉米茎腐病已经成为我国玉米生产的主要威胁。 【目的】 筛选鉴定抗玉米茎腐病的生防菌株,明确其最佳发酵工艺,并开发为高效菌剂,为构建新型绿色防控技术体系提供理论支撑与物质基础。 【方法】 结合稀释涂布法与平板对峙法筛选禾谷镰孢(Fusarium graminearum)的拮抗菌株;结合生理生化特征与分子生物学手段对目标菌株进行物种鉴定;通过单因素试验优化菌株发酵条件,并研制生防菌株种衣剂;利用室内盆栽与室外田间小区试验,系统评价生防菌株种衣剂对玉米生长发育的促进作用及对玉米茎腐病的防治效果。 【结果】 成功筛选出2株对禾谷镰孢抑制率达70.00%左右的生防菌株HS104和HS107,经鉴定2株菌均为贝莱斯芽孢杆菌(Bacillus velezensis)。2株菌对多种玉米主要病原菌[拟轮枝镰孢(Fusarium verticillioides)、尖孢镰孢(Fusarium oxysporum)及玉蜀黍平脐蠕孢(Bipolaris maydis)]表现出拮抗活性,抑菌率均在60.00%左右。经发酵条件优化后,室内盆栽试验表明HS104和HS107不仅显著促进玉米生长,且对茎腐病的防效分别达到75.00%和71.00%。基于此研发的生防种衣剂在田间试验中表现优异,不仅显著促进玉米生长,对茎腐病的防效分别达到63.93%和77.33%;且感病植株的产量较对照分别提高8.20%和9.20%。 【结论】 菌株HS104和HS107兼具广谱抑菌、强促生及高效防病等多重优良特性,在玉米茎腐病生物防治及增产提质方面具有广阔的应用前景。

, authors=王紫宸1, 2, 王黎明2, 李时宇2, 张康1, 2, 曹宏哲2, 邢继红1, 2, *, 董金皋1, 2, *, authorsList=王紫宸, 王黎明, 李时宇, 张康, 曹宏哲, 邢继红, 董金皋, authorCompany=null, correspAuthors=邢继红, 董金皋, authorNote=

作者贡献声明

王紫宸:试验操作、收集数据、数据分析及文章撰写;王黎明:数据分析及文章撰写;李时宇:部分试验操作及数据分析;张康、曹宏哲:管理、协调研究活动的策划和执行;邢继红、董金皋:项目负责人,提供经费支持,对研究活动进行指导。

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A: Antagonistic effects of biocontrol strains HS104 and HS107 against four pathogenic fungi of maize; B: Inhibition rates of biocontrol strains HS104 and HS107 against four pathogenic fungi of maize., figureFileSmall=TLbJuWQUXwF3DaeYy09lQQ==, figureFileBig=Q3VjYMXiNAtBVG+rBdgCvQ==, tableContent=null), ArticleFig(id=1316829866582987641, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714628260938674, language=CN, label=图1, caption=HS104和HS107对禾谷镰孢及其他病原菌的平板抑制效果及抑菌率, figureFileSmall=TLbJuWQUXwF3DaeYy09lQQ==, figureFileBig=Q3VjYMXiNAtBVG+rBdgCvQ==, tableContent=null), ArticleFig(id=1316829866780119930, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714628260938674, language=EN, label=Figure 2, caption=Identification of strains HS104 and HS107. A: Colony and microscopic morphology of biocontrol strain HS104 (bar=2 μm); B: Colony and microscopic morphology of biocontrol strain HS107 (bar=2 μm); C: Phylogenetic tree constructed based on 16S rRNA; D: Phylogenetic tree constructed based on rpoB gene sequences. Numbers in parentheses are GenBank accession numbers; The bootstrap values are shown at the node; The scale bar indicates 0.010 and 0.050 substitutions per nucleotide position., figureFileSmall=CDQ9ydV8mRnb+ogXHd2ouw==, figureFileBig=Tg9YdW8bPwuAqrVUO/Os4Q==, tableContent=null), ArticleFig(id=1316829866897560443, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714628260938674, language=CN, label=图2, caption=菌株HS104和HS107的鉴定, figureFileSmall=CDQ9ydV8mRnb+ogXHd2ouw==, figureFileBig=Tg9YdW8bPwuAqrVUO/Os4Q==, tableContent=null), ArticleFig(id=1316829866981446524, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714628260938674, language=EN, label=Figure 3, caption=Optimization of fermentation conditions for strains HS104 and HS107. 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A: Pot experiment phenotypes of HS104 and HS107 in the laboratory (bar=30 cm) and statistical charts of plant height, stem diameter and fresh weight; B: Stem lesion phenotypes (bar=5 cm) and statistical chart of lesion area. * means significant difference at P<0.05, ** means significant difference at P<0.01, *** means significant difference at P<0.001, and ns means no significant difference., figureFileSmall=a8xKUvdxSU2DbuLs3DmK/A==, figureFileBig=Xoqij1ZHVxQIJ0lfaXqqMw==, tableContent=null), ArticleFig(id=1316829867245687679, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714628260938674, language=CN, label=图4, caption=室内试验中菌株HS104和HS107对玉米的影响, figureFileSmall=a8xKUvdxSU2DbuLs3DmK/A==, figureFileBig=Xoqij1ZHVxQIJ0lfaXqqMw==, tableContent=null), ArticleFig(id=1316829867321185152, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714628260938674, language=EN, label=Figure 5, caption=Screening and preparation of seed coating agents from strains HS104 and HS107. A: Screening of components for seed coating adjuvant; B: Comparison of the finished seed coating agents of HS104 and HS107 with commercially available seed coating agents., figureFileSmall=z8kGHxX/t+2T/li7n/2Z5w==, figureFileBig=llPq5h7Or1M9/RAQv1siGA==, tableContent=null), ArticleFig(id=1316829867400876929, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714628260938674, language=CN, label=图5, caption=菌株HS104和HS107种衣剂的筛选与制备, figureFileSmall=z8kGHxX/t+2T/li7n/2Z5w==, figureFileBig=llPq5h7Or1M9/RAQv1siGA==, tableContent=null), ArticleFig(id=1316829867589620610, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714628260938674, language=EN, label=Figure 6, caption=Effects of strains HS104 and HS107 on maize in field trials. A: Disease spot phenotypes of field-grown maize (bar=5 cm); B: Statistical chart of disease spot area; C: Plant phenotypes of HS104 and HS107 in field-grown maize (bar=30 cm); D: Statistical chart of plant height and stem diameter of field-grown maize; E: Ear phenotypes of HS104 and HS107 in field-grown maize (bar=5 cm); F: Statistical chart of kernel number per ear, 100-kernel weight, ear number per mu and yield of field-grown maize. * means significant difference at P<0.05 level, ** means significant difference at P<0.01 level, *** means significant difference at P<0.001 level and ns means no significant difference., figureFileSmall=W/IShohOnpmOyCs7djaxeQ==, figureFileBig=AWTr6oJ15ZBp+HwCfn+Uvw==, tableContent=null), ArticleFig(id=1316829867652535171, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714628260938674, language=CN, label=图6, caption=大田试验中菌株HS104和HS107包衣对玉米的影响, figureFileSmall=W/IShohOnpmOyCs7djaxeQ==, figureFileBig=AWTr6oJ15ZBp+HwCfn+Uvw==, tableContent=null), ArticleFig(id=1316829867715449733, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714628260938674, language=EN, label=Table 1, caption=

Physiological and biochemical characteristics of strains HS104 and HS107

, figureFileSmall=null, figureFileBig=null, tableContent=
Determination indexCharacteristics of strain HS104Characteristics of strain HS107
Methyl red determination++
VP assay--
Indole test-+
Utilization of citrate++
Nitrate reduction++
Starch hydrolysis+-
Hydrogen sulfide production--
Glucose catabolism++
Lactose catabolism+-
), ArticleFig(id=1316829868021633926, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1316714628260938674, language=CN, label=表1, caption=

菌株HS104和HS107的生理生化特性

, figureFileSmall=null, figureFileBig=null, tableContent=
Determination indexCharacteristics of strain HS104Characteristics of strain HS107
Methyl red determination++
VP assay--
Indole test-+
Utilization of citrate++
Nitrate reduction++
Starch hydrolysis+-
Hydrogen sulfide production--
Glucose catabolism++
Lactose catabolism+-
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玉米禾谷镰孢茎腐病生防菌株的筛选鉴定及其防治效果
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王紫宸 1, 2 , 王黎明 2 , 李时宇 2 , 张康 1, 2 , 曹宏哲 2 , 邢继红 1, 2, * , 董金皋 1, 2, *
微生物学报 | 研究报告 2026,66(10): 5025-5037
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微生物学报 |研究报告 2026 , 66 (10) : 5025 -5037
玉米禾谷镰孢茎腐病生防菌株的筛选鉴定及其防治效果
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王紫宸1, 2, 王黎明2, 李时宇2, 张康1, 2, 曹宏哲2, 邢继红1, 2, * , 董金皋1, 2, *
作者信息
  • 1.河北农业大学,华北作物改良与调控国家重点实验室,河北 保定
  • 2.河北省植物生理与分子病理学重点实验室,河北 保定
通讯作者:
邢继红, 董金皋
作者简介:

作者贡献声明

王紫宸:试验操作、收集数据、数据分析及文章撰写;王黎明:数据分析及文章撰写;李时宇:部分试验操作及数据分析;张康、曹宏哲:管理、协调研究活动的策划和执行;邢继红、董金皋:项目负责人,提供经费支持,对研究活动进行指导。

Screening, identification, and control efficacy evaluation of biocontrol strains against maize stalk rot caused by Fusarium graminearum
Zichen WANG1, 2, Liming WANG2, Shiyu LI2, Kang ZHANG1, 2, Hongzhe CAO2, Jihong XING1, 2, * , Jingao DONG1, 2, *
Affiliations
  • 1.State Key Laboratory of North China Crop Improvement and Regulation, Hebei Agricultural University, Baoding, Hebei, China
  • 2.Key Laboratory of Plant Physiology and Molecular Pathology, Baoding, Hebei, China
  • Corresponding Author:
    *E-mail: XING Jihong,
    DONG Jingao,
出版时间: 2026-10-04 doi: 10.13343/j.cnki.wsxb.20260197
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玉米作为我国重要的粮食作物,其产量直接关系国家粮食安全。近年来,受气候变化和耕作模式改变的影响,玉米茎腐病已经成为我国玉米生产的主要威胁。 【目的】 筛选鉴定抗玉米茎腐病的生防菌株,明确其最佳发酵工艺,并开发为高效菌剂,为构建新型绿色防控技术体系提供理论支撑与物质基础。 【方法】 结合稀释涂布法与平板对峙法筛选禾谷镰孢(Fusarium graminearum)的拮抗菌株;结合生理生化特征与分子生物学手段对目标菌株进行物种鉴定;通过单因素试验优化菌株发酵条件,并研制生防菌株种衣剂;利用室内盆栽与室外田间小区试验,系统评价生防菌株种衣剂对玉米生长发育的促进作用及对玉米茎腐病的防治效果。 【结果】 成功筛选出2株对禾谷镰孢抑制率达70.00%左右的生防菌株HS104和HS107,经鉴定2株菌均为贝莱斯芽孢杆菌(Bacillus velezensis)。2株菌对多种玉米主要病原菌[拟轮枝镰孢(Fusarium verticillioides)、尖孢镰孢(Fusarium oxysporum)及玉蜀黍平脐蠕孢(Bipolaris maydis)]表现出拮抗活性,抑菌率均在60.00%左右。经发酵条件优化后,室内盆栽试验表明HS104和HS107不仅显著促进玉米生长,且对茎腐病的防效分别达到75.00%和71.00%。基于此研发的生防种衣剂在田间试验中表现优异,不仅显著促进玉米生长,对茎腐病的防效分别达到63.93%和77.33%;且感病植株的产量较对照分别提高8.20%和9.20%。 【结论】 菌株HS104和HS107兼具广谱抑菌、强促生及高效防病等多重优良特性,在玉米茎腐病生物防治及增产提质方面具有广阔的应用前景。

玉米茎腐病  /  禾谷镰孢  /  贝莱斯芽孢杆菌  /  种衣剂  /  生物防治

Maize, an important food crop in China, directly affects national food security. In recent years, due to climate change and shifts in farming practices, maize stalk rot has become a major threat to maize production in China. [Objective] To screen and identify biocontrol bacterial strains against maize stalk rot, determine their optimal fermentation process, and develop effective bacterial agents, thereby providing theoretical support and a material basis for constructing a novel green prevention and control technology system. [Methods] Antagonistic bacterial strains against Fusarium graminearum were screened through a combination of dilution coating and plate confrontation methods. Candidate strains were identified through molecular biological techniques combined with physiological and biochemical characterization. Fermentation conditions were optimized via single-factor experiments, and seed coating agents were developed with the selected biocontrol bacteria. Indoor pot experiments and outdoor field plot trials were carried out for the biocontrol bacteria to systematically evaluate their maize growth-promoting effects and control efficacy against maize stalk rot. [Results] Two biocontrol strains, HS104 and HS107, with inhibition rates of approximately 70.00% against F. graminearum, were successfully screened out and identified as Bacillus velezensis. Both strains exhibited broad-spectrum antagonistic activity against major maize pathogens, including F. verticillioides, F. oxysporum, and Bipolaris maydis, with inhibition rates of around 60.00%. After optimization of fermentation conditions, indoor pot experiments showed that HS104 and HS107 not only significantly promoted maize growth but also achieved the control efficacy of 75.00% and 71.00% against maize stalk rot, respectively. The seed coating agents developed with the selected biocontrol bacteria performed well in field trials, significantly promoting maize growth and achieving the control efficacy of 63.93% and 77.33% against maize stalk rot, respectively. Moreover, they increased the yield of susceptible plants by 8.20% and 9.20%, respectively, compared with the control. [Conclusion] Strains HS104 and HS107 possess multiple excellent traits, including broad-spectrum antifungal activity, strong plant growth-promoting effects, and high disease control efficacy, demonstrating broad application prospects in the biocontrol of maize stalk rot and the enhancement of crop yields.

maize stalk rot  /  Fusarium graminearum  /  Bacillus velezensis  /  seed coating agent  /  biocontrol
王紫宸, 王黎明, 李时宇, 张康, 曹宏哲, 邢继红, 董金皋. 玉米禾谷镰孢茎腐病生防菌株的筛选鉴定及其防治效果. 微生物学报, 2026 , 66 (10) : 5025 -5037 . DOI: 10.13343/j.cnki.wsxb.20260197
Zichen WANG, Liming WANG, Shiyu LI, Kang ZHANG, Hongzhe CAO, Jihong XING, Jingao DONG. Screening, identification, and control efficacy evaluation of biocontrol strains against maize stalk rot caused by Fusarium graminearum[J]. Acta Microbiologica Sinica, 2026 , 66 (10) : 5025 -5037 . DOI: 10.13343/j.cnki.wsxb.20260197
玉米作为我国第一大粮食作物,其产量占全国粮食总产量的40%以上[1],国家统计局数据显示,截至2025年,我国玉米种植面积已经连续3年增长,产量达到了30 123.5万t[2],创历史新高,对保障粮食安全和农业可持续发展具有重要战略地位。然而,玉米茎腐病作为全球玉米主产区普遍发生的重大土传病害,已成为制约玉米稳产高产的关键生物胁迫因素。自20世纪70年代以来,该病害在我国西南、华南、华东及西北等产区广泛流行,发病率通常为5%-20%,重病田块可达40%-60%,严重威胁玉米产量与经济效益。
该病害由多种病原真菌复合侵染引发,其中禾谷镰孢(Fusarium graminearum)是我国最主要的致病菌株[3]。该病原菌不仅导致植株倒伏、籽粒灌浆受阻[4],其产生的脱氧雪腐镰刀菌烯醇(deoxynivalenol, DON)等真菌毒素还会在谷物及青贮饲料中持续累积[5],通过食物链威胁人畜健康,构成“从农田到餐桌”的全链条安全风险。
当前,玉米茎腐病的防控面临多重挑战。其中,抗病品种选育存在周期长、抗性易丧失等问题[6];农业防治(如轮作、深耕)虽能降低病原菌基数,但成本高昂且难以根治;化学防治虽见效快,但长期过量使用已导致环境污染、病原菌抗药性增强及农产品毒素超标等新问题[7]。在此背景下,生物防治凭借其生态友好、作用机制多元、不易产生抗药性等优势成为病害绿色防控的重要发展方向[8]。
芽孢杆菌(Bacillus spp.)因繁殖速度快、抗逆性强、代谢产物丰富(如脂肽类抗生素、酶类、挥发性物质等),已成为生防微生物研发的热点[9]。近年来,贝莱斯芽孢杆菌(Bacillus velezensis)[10]、枯草芽孢杆菌(Bacillus subtilis)[11]及解淀粉芽孢杆菌(Bacillus amyloliquefaciens)[12]等在防治土传病害、促进植物生长及诱导系统抗性等方面展现出巨大潜力。例如,贝莱斯芽孢杆菌(Bacillusvelezensis) Y6和F7可分泌伊枯草菌素、丰原素和表面活性素,实现对青枯雷尔氏菌(Ralstonia solanacearum)和尖孢镰孢(Fusarium oxysporum)的双重防控;病原菌的存在可诱导脂肽合成,其不仅直接抑制病原菌,还通过促进生物膜形成和运动性增强根际定殖能力[13]。有研究表明,芽孢杆菌是抗性玉米品种抵御茎腐病的核心微生物类群,其通过生态位竞争、分泌抗菌物质、诱导宿主合成小檗碱等异喹啉生物碱3种机制协同防御;甚至无直接抑菌活性的芽孢杆菌也可通过上调宿主酪氨酸脱羧酶1 (tyrosine decarboxylase 1, TYDC1)基因表达,激活代谢防御通路[14]。
本研究以禾谷镰孢为靶标,从玉米根际土壤中筛选高拮抗活性的细菌菌株,采用形态学观察与分子生物学技术(16S rRNA基因测序、rpoB基因系统发育分析)对其进行鉴定。通过发酵条件优化提升菌株活性物质产量,研制新型生防种衣剂,并经室内盆栽与大田试验评价其对玉米的促生作用(株高、茎粗、生物量)及对茎腐病的防控效果。本研究旨在为玉米茎腐病绿色防控提供新的菌种资源,为基于微生物组的病害综合治理策略开发提供关键技术支撑,对推动农业可持续发展具有重要意义。
‘郑单958’玉米品种,购自河南秋乐种业科技股份有限公司。
禾谷镰孢(Fusarium graminearum)、拟轮枝镰孢(Fusarium verticillioides)、尖孢镰孢(Fusarium oxysporum)、玉蜀黍平脐蠕孢(Busarium maydis),均由河北省植物生理与分子病理学重点实验室保存并提供。
试验土壤采集自河北省衡水市玉米种植区,随机选取3株健康玉米植株,并按照Wang等[15]的方法收集根际土壤样品,于4 ℃保存。
采用稀释涂布平板法[16]从玉米根际土壤中分离纯化获得单菌落。以禾谷镰孢为靶标病原菌进行平板对峙试验。将直径5.0 mm的活化病原菌菌饼接种于PDA平板中央,候选细菌分别接种于距离病原菌2.5 cm的上下左右4个位置。以仅接种病原菌的平板为对照,设置3个重复,置于27 ℃培养箱中培养。待对照平板菌落长满时测量对照组和处理组中病原真菌的菌落直径,抑制率计算如公式(1)所示。
$抑制率=(对照组病原真菌菌落直径-试验组病原真菌菌落直径)/对照组病原真菌菌落直径\times 100\%$
根据抑制率结果筛选高效菌株进行复筛,并进一步以拟轮枝镰孢、尖孢镰孢、玉蜀黍平脐蠕孢为病原真菌,测定抑菌谱。
依据《常见细菌系统鉴定手册》[17],对生防菌株进行菌落形态观察(如菌落颜色、质地、边缘特征)与生理生化特性分析(如碳源利用),并进行初步分类。同时,采用16S rRNA基因和rpoB基因的特异性引物对菌株进行分子生物学鉴定。以菌株菌液为模板,分别使用16S rRNA基因的通用引物27F (5′-AGAGTTTGATCCTG GCTCAG-3′)和1492R (5′-GGTTACCTTGTTAC GACTT-3′),以及rpoB基因特异性引物2292F (5′-GACGTGGGATGGCTACAACT-3′)和3354R (5′-ATTGTCGCCTTTAACGATGG-3′)进行PCR扩增,反应体系及程序参照李志普[18]的优化方案。扩增产物经1%琼脂糖凝胶电泳检测后,送北京擎科生物科技股份有限公司完成测序。序列通过NCBI数据库BLAST比对确定近缘种,下载相关同源序列后,利用MEGA 7.0软件进行多序列比对并构建系统发育树。
对生防菌发酵培养基的成分进行优化,微生物发酵培养基以1.0%葡萄糖(碳源)、1.0%胰蛋白胨(氮源)和0.5%氯化钠(无机盐)为基础。优化试验中,分别用蔗糖、麦芽糖、玉米淀粉和可溶性淀粉作为替代碳源,用酵母浸粉、氯化铵、蛋白胨、黄豆饼粉和牛肉膏作为替代氮源,用硫酸铜、氯化钙、硫酸镁和氯化钾作为替代无机盐,通过不同组合筛选最优发酵培养基成分。
在此基础上,进一步对发酵条件进行优化。优化条件包括装液量(30、60、90、120、150 mL)、接种量(1%、2%、3%、4%、5%种子液)、pH (5.0、6.0、7.0、8.0、9.0)、温度(31、34、37、40、43 ℃)、转速(140、160、180、200、220 r/min)、发酵时间(24、36、48、72 h)。每个条件均设置各梯度的单因素试验。
最后,基于单因素试验结果,选取培养基关键成分(如最优碳源、氮源、无机盐)作为三因素,采用三水平正交试验设计进行配比优化,通过极差分析与方差分析确定各成分的最佳配比。
选取‘郑单958’玉米种子(饱满均一),经5% NaClO消毒后播种,置于25 ℃、相对湿度30%-45%、光照12 h的培养室中。试验设生防菌处理组与无菌培养基对照组,每处理6-10株重复。
待玉米出苗后,处理组分2次灌根施用1×109 CFU/mL的生防菌发酵液(50 mL/株),对照组施等量无菌培养基。玉米8叶期时,测量株高、茎粗及鲜重以评估促生作用。10叶期时,通过针刺法在茎基部接种50 μL禾谷镰孢分生孢子悬液(1×105 CFU/mL)。接种7 d后,剖茎调查发病情况,利用ImageJ软件测量病斑面积并统计防效,防效计算如公式(2)所示。
$\begin{array}{c} \text { 防效 }= \\ \frac{(\text { 对照组病斑面积 }- \text { 试验组病斑面积 })}{\text { 对照组病斑面积 }} \times 100 \% \end{array}$
制备生防菌种衣剂需要对紫外保护剂和分散剂进行筛选。紫外保护剂在葡萄糖、蔗糖和山梨醇中筛选,分散剂以聚乙烯醇、多聚磷酸钠、CMC-Na、PEG 1000为备选材料。将1×109 CFU/mL的生防菌悬液吸取100 μL涂布在含有不同成分的平板上,每个处理重复3次。平板置于37 ℃培养箱培养24 h后,对各处理单菌落数量进行统计,选取菌落数最多的组合,分别确定为最佳紫外保护剂和最佳分散剂。随后,将筛选出来的分散剂、紫外保护剂按3.0%、1.5%的比例与1×109 CFU/mL生防菌悬液及载体混匀,然后与25.0%的成膜剂混合,2.5%的鲜红101色浆为警戒色,以药种比1:50的比例与玉米籽粒混匀包衣,即完成种衣剂的制备。
试验在河北农业大学河北省植物生理与分子病理学重点实验室试验田开展。以市售包衣‘郑单958’种子为对照(CK),生防菌包衣种子为处理组。玉米8叶期时,各处理选取20株测定株高与茎粗。10叶期时,选取30株/处理进行针刺接种禾谷镰孢分生孢子悬液(1×105 CFU/mL),伤口处覆膜保湿。接种28 d后,剖茎调查发病情况,利用ImageJ软件测量病斑面积并计算防治效果。
试验数据采用Microsoft Excel和IBM SPSS Statistics 22进行数据整理和统计分析,图表使用GraphPad Prism 8.0绘制。
通过平板稀释涂布法,从玉米根际土壤中共分离获得340株候选细菌菌株。经平板对峙试验验证,菌株HS104和HS107对禾谷镰孢(Fusarium graminearum)的拮抗效果最为突出,抑制率分别达69.85%和70.05%。广谱抑菌活性检测发现,2株菌对多种玉米重要病原真菌均表现出显著抑制作用:对拟轮枝镰孢(Fusarium verticillioides)抑菌率分别为61.15%和60.45%,对尖孢镰孢(Fusarium oxysporum)分别为62.59%和62.88%,对玉蜀黍平脐蠕孢(Bipolaris maydis)分别为60.97%和60.41% (图1A、1B)。上述数据表明,HS104和HS107不仅对禾谷镰孢具有强效抑制能力,其抑菌谱还覆盖了玉米生产中常见的多种土传及叶部病害病原菌,展现出优异的广谱抗真菌活性,具备作为多功能生防菌株的开发潜力。
形态观察显示,HS104和HS107菌落呈近圆形、乳白色、不透明(图2A、2B)。革兰氏染色均呈阳性,菌体呈杆状。培养特征表现为初期光滑隆起,后期塌陷褶皱并伴有气味释放。生理生化分析(表1)显示,HS104对甲基红试验、柠檬酸盐、硝酸盐、淀粉水解、葡萄糖利用及乳糖利用呈阳性,VP试验、吲哚试验和硫化氢产生试验为阴性;HS107则表现为甲基红试验、吲哚试验、柠檬酸盐利用、硝酸盐还原和葡萄糖分解为阳性,但VP试验、淀粉水解、硫化氢产生和乳糖利用为阴性。分子系统发育分析基于16S rRNA和rpoB基因序列构建,结果表明HS104和HS107与贝莱斯芽孢杆菌(Bacillus velezensis)聚为一支,具有极高的序列相似性(图2C、2D)。综合菌落形态、生理生化特性及分子系统发育证据,最终将HS104和HS107鉴定为贝莱斯芽孢杆菌(Bacillus velezensis)。
生防菌株发酵培养基成分优化的单因素试验显示,HS104的最适碳源为葡萄糖、氮源为酵母浸粉、无机盐为氯化钙;HS107的最适碳源为蔗糖、氮源为酵母浸粉、无机盐为氯化钙。进一步通过三因素三水平正交试验验证,确定HS104培养基的最佳配比为葡萄糖1.0%、酵母浸粉1.0%、氯化钙0.1%,HS107的最佳配比为蔗糖5.0%、酵母浸粉0.5%、氯化钙0.1% (图3A)。
最佳发酵条件筛选结果显示,2株菌均在37 ℃、pH 7.0条件下表现出最佳发酵性能,发酵周期均为24 h。HS104的最佳发酵转速为180 r/min、装液量150 mL (250 mL三角瓶)、接种量4%;HS107的最适转速为200 r/min、装液量120 mL (250 mL三角瓶)、接种量5% (图3B)。
在温室盆栽试验中,生防菌株HS104和HS107展现出显著的促生与防病双重效果。经菌液处理后,玉米植株的株高、茎粗和鲜重均显著增加。与对照组相比,HS104处理组株高提升14.35%,茎粗增加73.43%,地下部鲜重增长46.84%,地上部鲜重提高36.95%;HS107处理组则表现为株高提升16.06%,茎粗增加79.72%,地下部鲜重激增98.51%,地上部鲜重提升70.28% (图4A)。在病害防控方面,2菌株处理显著抑制茎腐病发生,病斑面积明显缩小,防效分别达75.00% (HS104)和71.00% (HS107) (图4B)。
对生防菌株HS104和HS107进行种衣助剂的筛选,结果显示,山梨醇(3.0%)作为紫外保护剂可有效减少菌体紫外线损伤,PEG 1000 (1.5%)作为分散剂能确保菌体均匀分布(图5A)。将上述助剂与1.00×109 CFU/mL的生防菌悬液充分混匀后,添加25.0%成膜剂(形成保护膜提升持效期)和2.5%鲜红101色浆(便于识别包衣效果),按药种比1:50的比例完成玉米籽粒包衣(图5B)。种衣剂质量检测显示,HS104种衣剂的有效活菌落数为8.41×108 CFU/mL,pH 6.3;HS107种衣剂有效活菌数9.13×108 CFU/mL,pH 6.4,均符合微生物种衣剂质量标准。
在田间试验中,生防菌株HS104和HS107种衣剂展现了显著的病害防控效果与促生能力。经包衣处理的玉米植株在接种禾谷镰孢孢子悬液28 d后,病斑面积显著减小,防效分别达63.93% (HS104)和77.33% (HS107),验证了其高效的病害抑制能力(图6A、6B)。处理组的生长指标也显著提升,与对照组相比,HS104处理组株高提高5.94%,茎粗增加20.34%;HS107处理组株高提升4.49%,茎粗增加18.64% (图6C、6D)。
产量评估显示,经种衣剂处理的植株较未处理对照产量显著提高。HS104处理组增产8.20%,HS107处理组增产9.20% (图6E、6F)。
本研究通过室内盆栽与田间试验系统证实了贝莱斯芽孢杆菌HS104和HS107具有显著的“促生-防病”双效功能。在促生方面,发酵液处理使玉米株高、茎粗及生物量分别提升了14.35%-16.06%、73.43%-79.72%及36.95%-70.28%;种衣剂处理也使玉米株高和茎粗显著增加(4.49%-5.94%和18.64%-20.34%)。这种促生作用可能源于多重途径:一方面,菌株分泌吲哚乙酸、细胞分裂素等植物激素,直接刺激细胞分裂与伸长[19];另一方面,通过固氮、解磷等生物转化功能,将土壤中难以利用的矿质元素转化为有效态,持续供给作物养分[20-22]。后续研究将重点解析其代谢组学特征,明确关键促生因子的分子调控网络。
HS104和HS107对玉米茎腐病表现出优异的防控效果(盆栽防效75.00%、71.00%,田间防效63.93%、77.33%)。其生防机制主要包括“直接拮抗”与“间接诱导”双重策略[23]。其中,直接拮抗作用是主要的防控途径。生防菌通过分泌次生代谢产物(如环脂肽、表面活性素等抗生素类物质)直接抑制病原菌[24]。例如,刘雪娇等[25]发现贝莱斯芽孢杆菌(B. velezensis) 3A3-15产生的抑菌物质C14-C15 surfactin A可使尖孢镰孢的菌丝畸形、断裂,分生孢子萌发抑制率达到93.20%;贝莱斯芽孢杆菌(B. velezensis) B-3分泌的伊枯草菌素A和表面活性素能破坏禾谷镰孢的细胞膜完整性,抑制其生长与代谢[26]。其次,生防菌通过快速繁殖占据根际生态位,与病原菌竞争营养与空间,实现竞争性排斥。同时,生防菌还能重塑根际微生态,促进有益菌增殖并抑制病原菌。Bacon等[27]证实枯草芽孢杆菌(B. subtilis)可通过根际定殖降低串珠镰孢(Fusarium moniliforme)的侵染;贝莱斯芽孢杆菌(B. velezensis) ZN-S10则通过增加根际微生物的α多样性降低烟草疫霉的丰度[28]。诱导植物系统抗性是间接防控途径,生防菌作为激发子,可激活植物防御信号通路(如茉莉酸JA和水杨酸SA途径),促进植保素、病程相关蛋白(PR蛋白)合成,并上调抗性基因表达,从而增强宿主免疫[29-30]。
从土壤微生态视角看,长期单一种植模式导致本土微生物群落丰富度下降及作物抗病能力减弱[31]。本研究筛选的HS104和HS107不仅能直接防病,更有望作为“微生物修复剂”,恢复失衡的根际微生态,构建健康的抗病微环境。这种基于“菌-植-土”协同的绿色防控体系符合农业可持续发展的需求。尽管2株菌表现出良好的应用潜力,但其具体的抑菌分子机理尚需深入解析。未来研究将聚焦于分离鉴定关键次生代谢产物,阐明其抑菌分子机制;同时,开展菌群复配研究,利用合成生物学理念构建多功能复合菌群,以增加大田环境下的定殖能力与防效稳定性,为玉米茎腐病的生物防治提供更有效的制剂产品。
本研究筛选并鉴定出2株贝莱斯芽孢杆菌HS104和HS107,其对禾谷镰孢的抑菌率分别为69.85%和70.05%。经培养基配方与发酵参数的系统优化,确定了2菌株的最适发酵工艺:HS104在葡萄糖1.0%、酵母浸粉1.0%、氯化钙0.1%、180 r/min条件下生长最佳;HS107在蔗糖5.0%、酵母浸粉0.5%、氯化钙0.1%、200 r/min条件下性能最优,二者均以pH 7.0、37 ℃为最佳发酵条件,发酵周期均为24 h。室内与田间试验表明,2株菌发酵液及种衣剂能显著促进玉米生长并高效防控茎腐病(田间防效最高达77.33%),增产幅度达8.20%-9.20%。本研究构建了完整的生防菌研发与应用技术体系,为玉米茎腐病的生物防治提供了高效菌株资源与新型种衣剂产品,具有广阔的农业应用前景。
  • 河北省高等学校科学技术研究项目(CXZX2026057)
  • 保定市科技计划(2494N032)
  • 国家玉米产业技术体系(CARS-02-30)
  • 河北省省级科技计划(24466301D)
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doi: 10.13343/j.cnki.wsxb.20260197
  • 接收时间:2026-03-10
  • 首发时间:2026-10-13
  • 出版时间:2026-10-04
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  • 收稿日期:2026-03-10
  • 录用日期:2026-04-08
基金
Science and Technology Research Project of Colleges and Universities in Hebei Province(CXZX2026057)
河北省高等学校科学技术研究项目(CXZX2026057)
Baoding Science and Technology Planning(2494N032)
保定市科技计划(2494N032)
Earmarked Fund for CARS(CARS-02-30)
国家玉米产业技术体系(CARS-02-30)
Science and Technology Program of Hebei Province(24466301D)
河北省省级科技计划(24466301D)
作者信息
    1.河北农业大学,华北作物改良与调控国家重点实验室,河北 保定
    2.河北省植物生理与分子病理学重点实验室,河北 保定

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2种不同金属材料的力学参数
科
Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)
属
Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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