Article(id=1280817655670084423, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1280817479555462000, articleNumber=null, orderNo=null, doi=10.13343/j.cnki.wsxb.20251011, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1767110400000, receivedDateStr=2025-12-31, revisedDate=null, revisedDateStr=null, acceptedDate=1773849600000, acceptedDateStr=2026-03-19, onlineDate=1783300325553, onlineDateStr=2026-07-06, pubDate=1783094400000, pubDateStr=2026-07-04, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1783300325553, onlineIssueDateStr=2026-07-06, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1783300325552, creator=13701087609, updateTime=1783300325552, updator=13701087609, issue=Issue{id=1280817479555462000, tenantId=1146029695717560320, journalId=1192105938417971205, year='2026', volume='66', issue='7', pageStart='3121', pageEnd='3677', issueExtLink='null', onlineDate='null', pubDate='1783094400000', pubDateStr='2026-07-04', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1783300283564, creator='13701087609', updateTime=1783326087324, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1280925708813832745, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1280817479555462000, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1280925708813832746, tenantId=1146029695717560320, journalId=1192105938417971205, issueId=1280817479555462000, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=3544, endPage=3557, ext={EN=ArticleExt(id=1280817656034988872, articleId=1280817655670084423, tenantId=1146029695717560320, journalId=1192105938417971205, language=EN, title=Preparation of chlamydospore wettable powder of Cladophialophoraguangxiense HX2 and pot experiment evaluation of its efficacy, columnId=1192149543992045670, journalTitle=Acta Microbiologica Sinica, columnName=Research Article, runingTitle=null, highlight=null, articleAbstract=

Objective To prepare the chlamydospore wettable powder with significant control efficacy against tomato bacterial wilt from Cladophialophora guangxiense HX2, a dark septate endophyte (DSE). Methods Single-factor experiments were carried out to screen the types and dosages of carriers, wetting agents, dispersants, and ultraviolet protectants for the wettable powder. Pot experiments were conducted to evaluate the effects of soaking tomato seeds with four concentrations (T1: 1×108 CFU/mL; T2: 1×107 CFU/mL; T3: 1×106 CFU/mL; T4: 1×105 CFU/mL) for 30 min on tomato plant growth, tomato bacterial wilt, and activities of five defense enzymes—peroxidase (POD), polyphenol oxidase (PPO), phenylalanine ammonialyase (PAL), catalase (CAT), and superoxide dismutase (SOD). Results The formulation was optimized as follows: 25% chlamydospore suspension, 8% dispersant polyethylene glycol (PEG8000) and wetting agent Tween-60 at a mass ratio of 1:3, 0.5%-1.5% UV protectant ascorbic acid (VC), and white carbon black as the carrier to make up the remaining proportion to 100%. The wettable powder prepared according to this formulation had the chlamydospore content of 2.35×108 CFU/g, the wetting time of 24.25 s, a suspension rate of 73.8%, pH 5.71, the moisture content of 16.67%, and the fineness of 98.81%. All indicators met the requirements of the national standard GB 20287—2006 Microbial Inoculants in Agriculture. The results of pot experiments indicated that the T2 treatment exhibited a significant plant growth-promoting effect, increasing the root length, plant height, stem diameter, fresh weight, and dry weight by 47.39%, 31.82%, 24.64%, 89.45%, and 90.97%, respectively, compared with the control group. On day 30 after pathogen inoculation, the control efficacy of this treatment against tomato bacterial wilt reached 50.9%, which was significantly higher than that of the Trichoderma harzianum treatment. Moreover, the T2 treatment significantly enhanced the activities of the five defense enzymes. Conclusion The HX2 chlamydospore wettable powder prepared in this study has good control efficacy against tomato bacterial wilt. This study provides a technical basis for the large-scale popularization and application of this agent.

, authors=Xinghao WANG1, 2, Fenghua ZENG1, Yanyan LONG1, Hongquan LIU2, Yanxin YU1, Jihui YU1, Ling XIE1, Yan ZHANG1, authorsList=Xinghao WANG, Fenghua ZENG, Yanyan LONG, Hongquan LIU, Yanxin YU, Jihui YU, Ling XIE, Yan ZHANG, authorCompany=null, correspAuthors=Ling XIE, Yan ZHANG, authorNote=

These authors contributed equally to this work.

, correspAuthorsNote=
E-mail: XIE Ling,
ZHANG Yan,
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目的 以深色有隔内生真菌(dark septate endophyte, DSE)广西枝孢瓶霉(Cladophialophora guangxiense) HX2为研究对象,制备对番茄青枯病具有显著防治效果的HX2厚垣孢子可湿性粉剂。 方法 通过单因素试验筛选可湿性粉剂的载体、润湿剂、分散剂及紫外保护剂种类与用量,采用盆栽试验评价了4种浓度(T1:1×108 CFU/mL;T2:1×107 CFU/mL;T3:1×106 CFU/mL;T4:1×105 CFU/mL)浸种番茄种子30 min后,对番茄植株生长、番茄青枯病防治效果以及过氧化物酶(peroxidase, POD)、多酚氧化酶(polyphenol oxidase, PPO)、苯丙氨酸解氨酶(phenylalanine ammonialyase, PAL)、过氧化氢酶(catalase, CAT)和超氧化物歧化酶(superoxide dismutase, SOD) 5种防御酶活性的影响。 结果 单因素试验结果表明,最佳配方为厚垣孢子浆25%,分散剂聚乙二醇(polyethylene glycol, PEG8000)和润湿剂吐温-60 (质量比为1:3)总用量8%,紫外保护剂抗坏血酸(ascorbic acid, VC) 0.5%-1.5%,并以白炭黑为载体配齐100%。按此配方制备的可湿性粉剂中厚垣孢子含量为2.35×108 CFU/g,润湿时间为24.25 s,悬浮率为73.8%,pH为5.71,含水量为16.67%,细度为98.81%,各项指标均符合《农用微生物菌剂》国家标准(GB 20287—2006)。盆栽试验结果显示,T2浓度处理促生效果显著,根长、株高、茎粗、鲜重、干重分别较对照增加47.39%、31.82%、24.64%、89.45%、90.97%;接种病原30 d后对番茄青枯病防效达50.9%,显著高于哈茨木霉处理,并能显著提升5种防御酶活性。 结论 本研究成功制备的HX2厚垣孢子可湿性粉剂对番茄青枯病具有显著防效,可为该制剂的规模化推广应用提供技术支撑。

, authors=王兴浩1, 2, 曾凤花1, 龙艳艳1, 刘红全2, 于延鑫1, 喻吉会1, 谢玲1, 张艳1, authorsList=王兴浩, 曾凤花, 龙艳艳, 刘红全, 于延鑫, 喻吉会, 谢玲, 张艳, authorCompany=null, correspAuthors=谢玲, 张艳, authorNote=

作者贡献声明

王兴浩:实验设计、数据分析、初稿撰写、论文修改;曾凤花:数据收集;龙艳艳:监督管理;刘红全:实验指导;于延鑫:数据整理;喻吉会:图表可视化;谢玲:实验指导、资源提供、论文修改;张艳:项目管理、审阅、论文修改。

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Physiological and Molecular Plant Pathology, 2026, 141: 102985., articleTitle=Identification and characterization of Streptomyces sp. strain NEAU-174 with biocontrol ability against tomato bacterial wilt, refAbstract=null)], funds=[Fund(id=1280925036726956824, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, awardId=AB241484049, language=EN, fundingSource=the Key Research and Development Program of Guangxi Zhuang Autonomous Region (Guinongke(AB241484049), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1280925028640338635, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, xref=1., ext=[AuthorCompanyExt(id=1280925028652921548, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, companyId=1280925028640338635, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.Key Laboratory of Green Prevention and Control on Fruits and Vegetables in South China, Ministry of Agriculture and Rural Affairs, Guangxi Key Laboratory of Biology for Crop Diseases and Insect Pests, Plant Protection Research Institute, Guangxi Academy of Agricultural Sciences, Nanning, Guangxi, China), AuthorCompanyExt(id=1280925028665504461, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, companyId=1280925028640338635, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.广西农业科学院植物保护研究所,农业农村部华南果蔬绿色防控重点实验室,广西作物病虫害生物学重点实验室,广西 南宁)]), AuthorCompany(id=1280925028749390542, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, xref=2., ext=[AuthorCompanyExt(id=1280925028761973455, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, companyId=1280925028749390542, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.School of Marine Sciences and Biotechnology, Guangxi Minzu University, Nanning, Guangxi, China), AuthorCompanyExt(id=1280925028774556368, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, companyId=1280925028749390542, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.广西民族大学 海洋与生物技术学院,广西 南宁)])], figs=[ArticleFig(id=1280925033518314243, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, language=EN, label=Figure 1, caption=Optimization of ratio and dosage of dispersants and wetting agents for HX2 chlamydospore wettable powder. A, B: Determination of wetting time, suspension rate and chlamydospore germination rate for ratio optimization of dispersants and wetting agents; C, D: Determination of wetting time, suspension rate and chlamydospore germination rate for dosage optimization of dispersants and wetting agents., figureFileSmall=/TX9sjdM3GP90crHfab8+Q==, figureFileBig=XlZGay9GqMnf1ExEImPOlg==, tableContent=null), ArticleFig(id=1280925033593811716, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, language=CN, label=图1, caption=HX2厚垣孢子可湿性粉剂分散剂和润湿剂的配比及用量优化, figureFileSmall=/TX9sjdM3GP90crHfab8+Q==, figureFileBig=XlZGay9GqMnf1ExEImPOlg==, tableContent=null), ArticleFig(id=1280925033702863621, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, language=EN, label=Figure 2, caption=Growth status of tomato plants under different treatments after 15 days., figureFileSmall=/33NcpTFLCyeJrxmysl1tA==, figureFileBig=XBSRxeBdAVsqJvmyZ3HwuQ==, tableContent=null), ArticleFig(id=1280925033765778182, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, language=CN, label=图2, caption=不同处理番茄15 d后生长状态, figureFileSmall=/33NcpTFLCyeJrxmysl1tA==, figureFileBig=XBSRxeBdAVsqJvmyZ3HwuQ==, tableContent=null), ArticleFig(id=1280925033849664263, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, language=EN, label=Figure 3, caption=Control effects of different treatments on tomato bacterial wilt. 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Effects of different carriers on the wettable powder of HX2 chlamydospores

, figureFileSmall=null, figureFileBig=null, tableContent=
Type of carriersWetting time/sSuspension rate/%Chlamydospore germination rate/%Adsorption capacity/(g/g)
Diatomite6.31±0.44e1.52±0.29b70.33±1.72bc1.33±0.09b
Silica24.97±1.16c18.13±0.72a78.71±2.47a2.52±0.10a
Talc powder67.13±2.58a2.45±0.25b73.27±2.48ab0.73±0.01d
Calcium carbonate44.86±2.32b4.69±0.42b65.93±2.05cd0.60±0.09d
Kaolin3.37±0.10e4.74±0.96b59.62±2.73d1.04±0.16c
Plant ash12.06±1.28d3.49±0.29b61.48±2.79d1.02±0.07c
CK--78.56±2.48a-
), ArticleFig(id=1280925035984565007, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, language=CN, label=表1, caption=

不同载体对HX2厚垣孢子可湿性粉剂的影响

, figureFileSmall=null, figureFileBig=null, tableContent=
Type of carriersWetting time/sSuspension rate/%Chlamydospore germination rate/%Adsorption capacity/(g/g)
Diatomite6.31±0.44e1.52±0.29b70.33±1.72bc1.33±0.09b
Silica24.97±1.16c18.13±0.72a78.71±2.47a2.52±0.10a
Talc powder67.13±2.58a2.45±0.25b73.27±2.48ab0.73±0.01d
Calcium carbonate44.86±2.32b4.69±0.42b65.93±2.05cd0.60±0.09d
Kaolin3.37±0.10e4.74±0.96b59.62±2.73d1.04±0.16c
Plant ash12.06±1.28d3.49±0.29b61.48±2.79d1.02±0.07c
CK--78.56±2.48a-
), ArticleFig(id=1280925036043285264, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, language=EN, label=Table 2, caption=

Screening of wetting agents and dispersants for HX2 chlamydospore wettable powder

, figureFileSmall=null, figureFileBig=null, tableContent=
TypeWetting agents and dispersants forWetting time/sSuspension rate/%Chlamydospore germination rate/%
Wetting agentTween-6030.43±0.33f21.05±1.22c71.52±2.54a
DispersantPEG800035.41±0.89e37.50±1.56ab71.60±1.07a
DispersantSLS47.72±0.82a37.77±0.77ab22.93±4.61e
DispersantSHP44.54±0.25b38.18±0.29a60.66±2.48b
Wetting agentSP48.86±0.06a35.92±0.41ab35.63±0.44d
DispersantSPP43.53±0.28b36.46±1.08ab47.61±2.64d
Wetting agentSDS37.89±0.14d35.01±1.09b49.88±1.59d
Wetting agentSTPP42.52±0.86bc37.12±0.89ab18.16±2.07e
DispersantCMC40.22±0.44c36.41±0.86ab59.68±2.77bc
CK28.91±1.40f23.85±0.57c78.56±2.90a
), ArticleFig(id=1280925036122977041, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, language=CN, label=表2, caption=

HX2厚垣孢子可湿性粉剂润湿剂和分散剂的筛选

, figureFileSmall=null, figureFileBig=null, tableContent=
TypeWetting agents and dispersants forWetting time/sSuspension rate/%Chlamydospore germination rate/%
Wetting agentTween-6030.43±0.33f21.05±1.22c71.52±2.54a
DispersantPEG800035.41±0.89e37.50±1.56ab71.60±1.07a
DispersantSLS47.72±0.82a37.77±0.77ab22.93±4.61e
DispersantSHP44.54±0.25b38.18±0.29a60.66±2.48b
Wetting agentSP48.86±0.06a35.92±0.41ab35.63±0.44d
DispersantSPP43.53±0.28b36.46±1.08ab47.61±2.64d
Wetting agentSDS37.89±0.14d35.01±1.09b49.88±1.59d
Wetting agentSTPP42.52±0.86bc37.12±0.89ab18.16±2.07e
DispersantCMC40.22±0.44c36.41±0.86ab59.68±2.77bc
CK28.91±1.40f23.85±0.57c78.56±2.90a
), ArticleFig(id=1280925036202668818, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, language=EN, label=Table 3, caption=

Screening of UV protectants for HX2 chlamydospore wettable powder

, figureFileSmall=null, figureFileBig=null, tableContent=
UV protectantConcentration/%Chlamydospore germination rate/%
Dextrin0.572.23±2.64abc
1.067.43±1.17def
1.572.90±1.93ab
2.064.93±2.26ef
Sodium alginate0.566.45±0.56ef
1.072.32±1.00abc
1.571.41±0.75abcd
2.073.28±1.63ab
Xanthan gum0.567.70±1.78def
1.068.93±2.59bcde
1.564.39±1.76f
2.065.94±1.91ef
Mineral-sourced sodium fulvate0.564.59±0.92ef
1.072.74±0.49ab
1.572.79±0.91ab
2.068.20±1.23cdef
Ascorbic acid0.574.43±2.55a
1.075.74±1.95a
1.575.10±0.82a
2.072.99±0.12ab
CK-56.91±0.43g
), ArticleFig(id=1280925036278166291, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, language=CN, label=表3, caption=

HX2厚垣孢子可湿性粉剂紫外保护剂筛选

, figureFileSmall=null, figureFileBig=null, tableContent=
UV protectantConcentration/%Chlamydospore germination rate/%
Dextrin0.572.23±2.64abc
1.067.43±1.17def
1.572.90±1.93ab
2.064.93±2.26ef
Sodium alginate0.566.45±0.56ef
1.072.32±1.00abc
1.571.41±0.75abcd
2.073.28±1.63ab
Xanthan gum0.567.70±1.78def
1.068.93±2.59bcde
1.564.39±1.76f
2.065.94±1.91ef
Mineral-sourced sodium fulvate0.564.59±0.92ef
1.072.74±0.49ab
1.572.79±0.91ab
2.068.20±1.23cdef
Ascorbic acid0.574.43±2.55a
1.075.74±1.95a
1.575.10±0.82a
2.072.99±0.12ab
CK-56.91±0.43g
), ArticleFig(id=1280925036349469460, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, language=EN, label=Table 4, caption=

Determination of quality indicators for HX2 chlamydospore wettable powder

, figureFileSmall=null, figureFileBig=null, tableContent=
IndexNational standardMeasuring value
Viable spore count/(CFU/g)≥2×1082.35×108
Contaminating microorganism rate/%≤20%0
Suspension rate/%≥7073.80±1.24
Wetting time/s≤18024.25±1.23
pH range5.5-8.55.71±0.07
Fineness/%≥8098.81±0.07
Water content/%≤3516.67±0.44
), ArticleFig(id=1280925036420772629, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, language=CN, label=表4, caption=

HX2厚垣孢子可湿性粉剂质量指标检测

, figureFileSmall=null, figureFileBig=null, tableContent=
IndexNational standardMeasuring value
Viable spore count/(CFU/g)≥2×1082.35×108
Contaminating microorganism rate/%≤20%0
Suspension rate/%≥7073.80±1.24
Wetting time/s≤18024.25±1.23
pH range5.5-8.55.71±0.07
Fineness/%≥8098.81±0.07
Water content/%≤3516.67±0.44
), ArticleFig(id=1280925036496270102, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, language=EN, label=Table 5, caption=

Growth-promoting effects of different treatments on tomato plants

, figureFileSmall=null, figureFileBig=null, tableContent=
TreatmentRoot length/cmPlant height/cmStem diameter/cmFresh weight/gDry weight/g
CK8.82±1.39d11.88±0.39c2.11±0.11b11.28±0.25c1.44±0.10c

T1

1×108 CFU/mL

12.01±0.86b16.57±0.51a2.37±0.15ab18.34±1.45b1.97±0.38b

T2

1×107 CFU/mL

13.20±1.07a15.66±0.55b2.63±0.03a21.37±1.24a2.75±0.20a

T3

1×106 CFU/mL

13.30±1.36a15.68±0.98b2.58±0.05a18.83±0.64b2.71±0.32a

T4

1×105 CFU/mL

10.91±0.37c11.17±0.75c2.16±0.10b9.73±0.99d1.49±0.25c
), ArticleFig(id=1280925036571767575, tenantId=1146029695717560320, journalId=1192105938417971205, articleId=1280817655670084423, language=CN, label=表5, caption=

不同处理对番茄的促生作用

, figureFileSmall=null, figureFileBig=null, tableContent=
TreatmentRoot length/cmPlant height/cmStem diameter/cmFresh weight/gDry weight/g
CK8.82±1.39d11.88±0.39c2.11±0.11b11.28±0.25c1.44±0.10c

T1

1×108 CFU/mL

12.01±0.86b16.57±0.51a2.37±0.15ab18.34±1.45b1.97±0.38b

T2

1×107 CFU/mL

13.20±1.07a15.66±0.55b2.63±0.03a21.37±1.24a2.75±0.20a

T3

1×106 CFU/mL

13.30±1.36a15.68±0.98b2.58±0.05a18.83±0.64b2.71±0.32a

T4

1×105 CFU/mL

10.91±0.37c11.17±0.75c2.16±0.10b9.73±0.99d1.49±0.25c
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广西枝孢瓶霉HX2厚垣孢子可湿性粉剂制备及盆栽效果
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王兴浩 1, 2 , 曾凤花 1 , 龙艳艳 1 , 刘红全 2 , 于延鑫 1 , 喻吉会 1 , 谢玲 1 , 张艳 1
微生物学报 | 研究报告 2026,66(7): 3544-3557
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微生物学报 |研究报告 2026 , 66 (7) : 3544 -3557
广西枝孢瓶霉HX2厚垣孢子可湿性粉剂制备及盆栽效果
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王兴浩1, 2, 曾凤花1, 龙艳艳1, 刘红全2, 于延鑫1, 喻吉会1, 谢玲1 , 张艳1
作者信息
  • 1.广西农业科学院植物保护研究所,农业农村部华南果蔬绿色防控重点实验室,广西作物病虫害生物学重点实验室,广西 南宁
  • 2.广西民族大学 海洋与生物技术学院,广西 南宁
作者简介:

作者贡献声明

王兴浩:实验设计、数据分析、初稿撰写、论文修改;曾凤花:数据收集;龙艳艳:监督管理;刘红全:实验指导;于延鑫:数据整理;喻吉会:图表可视化;谢玲:实验指导、资源提供、论文修改;张艳:项目管理、审阅、论文修改。

Preparation of chlamydospore wettable powder of Cladophialophoraguangxiense HX2 and pot experiment evaluation of its efficacy
Xinghao WANG1, 2, Fenghua ZENG1, Yanyan LONG1, Hongquan LIU2, Yanxin YU1, Jihui YU1, Ling XIE1 , Yan ZHANG1
Affiliations
  • 1.Key Laboratory of Green Prevention and Control on Fruits and Vegetables in South China, Ministry of Agriculture and Rural Affairs, Guangxi Key Laboratory of Biology for Crop Diseases and Insect Pests, Plant Protection Research Institute, Guangxi Academy of Agricultural Sciences, Nanning, Guangxi, China
  • 2.School of Marine Sciences and Biotechnology, Guangxi Minzu University, Nanning, Guangxi, China
出版时间: 2026-07-04 doi: 10.13343/j.cnki.wsxb.20251011
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目的 以深色有隔内生真菌(dark septate endophyte, DSE)广西枝孢瓶霉(Cladophialophora guangxiense) HX2为研究对象,制备对番茄青枯病具有显著防治效果的HX2厚垣孢子可湿性粉剂。 方法 通过单因素试验筛选可湿性粉剂的载体、润湿剂、分散剂及紫外保护剂种类与用量,采用盆栽试验评价了4种浓度(T1:1×108 CFU/mL;T2:1×107 CFU/mL;T3:1×106 CFU/mL;T4:1×105 CFU/mL)浸种番茄种子30 min后,对番茄植株生长、番茄青枯病防治效果以及过氧化物酶(peroxidase, POD)、多酚氧化酶(polyphenol oxidase, PPO)、苯丙氨酸解氨酶(phenylalanine ammonialyase, PAL)、过氧化氢酶(catalase, CAT)和超氧化物歧化酶(superoxide dismutase, SOD) 5种防御酶活性的影响。 结果 单因素试验结果表明,最佳配方为厚垣孢子浆25%,分散剂聚乙二醇(polyethylene glycol, PEG8000)和润湿剂吐温-60 (质量比为1:3)总用量8%,紫外保护剂抗坏血酸(ascorbic acid, VC) 0.5%-1.5%,并以白炭黑为载体配齐100%。按此配方制备的可湿性粉剂中厚垣孢子含量为2.35×108 CFU/g,润湿时间为24.25 s,悬浮率为73.8%,pH为5.71,含水量为16.67%,细度为98.81%,各项指标均符合《农用微生物菌剂》国家标准(GB 20287—2006)。盆栽试验结果显示,T2浓度处理促生效果显著,根长、株高、茎粗、鲜重、干重分别较对照增加47.39%、31.82%、24.64%、89.45%、90.97%;接种病原30 d后对番茄青枯病防效达50.9%,显著高于哈茨木霉处理,并能显著提升5种防御酶活性。 结论 本研究成功制备的HX2厚垣孢子可湿性粉剂对番茄青枯病具有显著防效,可为该制剂的规模化推广应用提供技术支撑。

厚垣孢子  /  可湿性粉剂  /  番茄青枯病  /  防御酶活性

Objective To prepare the chlamydospore wettable powder with significant control efficacy against tomato bacterial wilt from Cladophialophora guangxiense HX2, a dark septate endophyte (DSE). Methods Single-factor experiments were carried out to screen the types and dosages of carriers, wetting agents, dispersants, and ultraviolet protectants for the wettable powder. Pot experiments were conducted to evaluate the effects of soaking tomato seeds with four concentrations (T1: 1×108 CFU/mL; T2: 1×107 CFU/mL; T3: 1×106 CFU/mL; T4: 1×105 CFU/mL) for 30 min on tomato plant growth, tomato bacterial wilt, and activities of five defense enzymes—peroxidase (POD), polyphenol oxidase (PPO), phenylalanine ammonialyase (PAL), catalase (CAT), and superoxide dismutase (SOD). Results The formulation was optimized as follows: 25% chlamydospore suspension, 8% dispersant polyethylene glycol (PEG8000) and wetting agent Tween-60 at a mass ratio of 1:3, 0.5%-1.5% UV protectant ascorbic acid (VC), and white carbon black as the carrier to make up the remaining proportion to 100%. The wettable powder prepared according to this formulation had the chlamydospore content of 2.35×108 CFU/g, the wetting time of 24.25 s, a suspension rate of 73.8%, pH 5.71, the moisture content of 16.67%, and the fineness of 98.81%. All indicators met the requirements of the national standard GB 20287—2006 Microbial Inoculants in Agriculture. The results of pot experiments indicated that the T2 treatment exhibited a significant plant growth-promoting effect, increasing the root length, plant height, stem diameter, fresh weight, and dry weight by 47.39%, 31.82%, 24.64%, 89.45%, and 90.97%, respectively, compared with the control group. On day 30 after pathogen inoculation, the control efficacy of this treatment against tomato bacterial wilt reached 50.9%, which was significantly higher than that of the Trichoderma harzianum treatment. Moreover, the T2 treatment significantly enhanced the activities of the five defense enzymes. Conclusion The HX2 chlamydospore wettable powder prepared in this study has good control efficacy against tomato bacterial wilt. This study provides a technical basis for the large-scale popularization and application of this agent.

chlamydospores  /  wettable powder  /  tomato bacterial wilt  /  defense enzyme activity
王兴浩, 曾凤花, 龙艳艳, 刘红全, 于延鑫, 喻吉会, 谢玲, 张艳. 广西枝孢瓶霉HX2厚垣孢子可湿性粉剂制备及盆栽效果. 微生物学报, 2026 , 66 (7) : 3544 -3557 . DOI: 10.13343/j.cnki.wsxb.20251011
Xinghao WANG, Fenghua ZENG, Yanyan LONG, Hongquan LIU, Yanxin YU, Jihui YU, Ling XIE, Yan ZHANG. Preparation of chlamydospore wettable powder of Cladophialophoraguangxiense HX2 and pot experiment evaluation of its efficacy[J]. Acta Microbiologica Sinica, 2026 , 66 (7) : 3544 -3557 . DOI: 10.13343/j.cnki.wsxb.20251011
番茄作为我国乃至全球重要的经济作物,其产量与品质直接关系到农业生产安全和农民经济收益[1]。然而,在集约化、规模化种植模式下,土传病害频发与蔓延已成为制约其优质高产的关键问题[2]。其中,番茄青枯病在生产上危害严重,该病害由茄科罗尔斯通氏菌(Ralstonia solanacearum)引起,是一种典型的土传维管束病害,具有传播速度快、寄主范围广、防治难度大等特点,发病后番茄植株常快速萎蔫枯死,一般田块发病率可达20%-30%,严重地块甚至绝收,给番茄产业造成惨重经济损失[3-4]。由于化学防治效果不理想,加之抗病资源有限,环境友好的生物防治一直是研究热点。
目前,针对番茄青枯病的生防微生物多以芽孢杆菌(Bacillus spp.)等细菌为主[5-8]。刘玉敏等[6]分离得到的贝莱斯芽孢杆菌(B. velezensis) SB10菌体重悬液和发酵液均对茄科罗尔斯通氏菌具有预防和治疗作用。Mekonnen等[7]发现芽孢杆菌(Bacillus sp.)分离株BDUA1在温室条件下可显著降低番茄细菌性萎蔫病发病率。慕雪男等[8]从番茄根系中分离的暹罗芽孢杆菌(B. siamensis) A72对番茄青枯病具有较好的防治效果,并对番茄植株具有较好的促生功能。然而,由于田间土壤环境复杂且青枯病菌生理小种高度分化,生防菌普遍存在定殖能力弱、防效不稳定的问题,亟待开发定殖能力强、防效稳定的新型生防微生物[9]
深色有隔内生真菌(dark septate endophytes, DSE)是一类广泛定殖于植物根系内部或表面、形成深色有隔菌丝和微菌核的内生真菌类群,可与多种植物形成共生关系[10]。研究表明,DSE可通过促进养分吸收、分泌生长调节物质、诱导植物抗性及抑制病原菌生长等途径,增强植物对生物与非生物胁迫的耐受性,在作物病害生物防治中极具应用潜力[11-12]。张晓容等[13]发现,DSE甘瓶霉(Phialophora mustea)接种可增强植物抗氧化酶活性,缓解由尖镰孢菌(Fusarium oxysporum)导致的脂膜过氧化胁迫,提高番茄抗性。胡佳铭等[14]研究表明,DSE瓜朱姆泛孢霉(Pantospora guazumae)菌株能抑制番茄灰霉病菌灰葡萄孢(Botrytis cinerea)菌丝生长,抑制率达73.5%,且兼具促生作用。DSE因可定殖在植物体内与宿主形成共生体,受外界环境影响相对较小,其对多种植物病害的生防效果已得到证实。课题组在前期DSE资源调查中发现,枝孢瓶霉属(Cladophialophora)是DSE类群中常见的优势属,该属在全球范围内各类生境的DSE群落中均占据优势地位,且相关菌株通过种子或种苗处理对青枯病表现出良好的防效[15-16]。DSE菌株枝孢瓶霉(C. immunda)LC3菌液浸泡番茄种子和浸泡生姜种苗根部30 min对青枯病的盆栽防效分别达63.2%和52.4%[9];枝孢瓶霉属(Cladophialophora)合成菌群在生姜苗期施用1次,青枯病田间发病率和病情指数均显著低于市售木霉(Trichoderma spp.)、芽孢杆菌(Bacillus spp.)复合菌剂施用5次的处理,且显著提高了生姜产量[17]。可见,DSE的内生定殖特性和防效稳定性使其在作物青枯病防治上具有良好的应用前景。
研究发现,广西枝孢瓶霉(C. guangxiense) HX2对番茄的侵染可调控水杨酸代谢、乙烯激活信号通路、光合作用及苯丙烷生物合成等途径,显著上调IAA4ERF1Hqt等抗病基因的表达,并显著抑制致病基因Hsc70的表达,对番茄青枯病具有优良的防治效果[18]。同时,该菌株能够形成抗逆性极强的厚垣孢子,在逆境中可长期存活,为其在田间环境中的定殖和病害防控提供了有利条件,具备开发为生防菌剂的优良特性。目前,该菌株已完成液体厚垣孢子培养条件的优化。为满足生产应用需求,本研究拟研制可湿性粉剂,相较于水剂,其具有体积小、质量轻、便于长途运输的优势,可满足不同生产场景的需求。本研究同时探究了其对番茄青枯病防控的适宜施用条件,以期为番茄青枯病的绿色防控提供新的技术手段和菌剂产品,减少化学农药的使用,促进农业生态环境的可持续发展。
广西枝孢瓶霉(C. guangxiense) HX2菌株(专利保藏编号为CGMCC41498)由广西农业科学院植物保护研究所植物病害生物防治团队分离保存。
番茄茄科罗尔斯通氏菌(R. solanacearum) Gg24由广西大学农学院袁高庆教授团队惠赠。
供试番茄品种为‘艺丰一代’,购自南宁市艺丰农业科技有限公司。
PDA培养基(g/L):马铃薯200.0,葡萄糖20.0,琼脂15.0。
糖蜜液体培养基(g/L):糖蜜84.0,大豆粉25.0,玉米粉26.0。
LA培养基(g/L):胰蛋白胨10.0,酵母提取物5.0,NaCl 10.0,琼脂15.0。
LB培养基(g/L):胰蛋白胨10.0,酵母提取物5.0,NaCl 10.0。
硅藻土(diatomite)、白炭黑(silica)、滑石粉(talc powder)、碳酸钙(calcium carbonate)、高岭土(kaolin)、草木灰(plant ash)、木质素磺酸钠(sodium lignosulfonate, SLS)、六偏磷酸钠(sodium hexametaphosphate, SHP)、焦磷酸钠(sodium pyrophosphate, SPP)、聚乙二醇(polyethylene glycol, PEG8000)、吐温-60 (Tween-60)、皂苷(saponin, SP)、十二烷基硫酸钠(sodium dodecyl sulfate, SDS)、三聚磷酸钠(sodium tripolyphosphate, STPP)、羧甲基纤维素钠(sodium carboxymethyl cellulose, CMC)、糊精(dextrin)、海藻酸钠(sodium alginate)、黄原胶(xanthan gum)、矿源黄腐植酸钠(mineral-sourced sodium fulvate)、抗坏血酸(ascorbic acid, VC),广西南宁壹棵松生物科技有限公司;哈茨木霉(2×108 CFU/g,粉剂),垄沃园艺洛阳有限责任公司;过氧化物酶(peroxidase, POD)试剂盒、多酚氧化酶(polyphenol oxidase, PPO)试剂盒、苯丙氨酸解氨酶(phenylalnine ammonialyase, PAL)试剂盒、过氧化氢酶(catalase, CAT)试剂盒和超氧化物歧化酶(superoxide dismutase, SOD)试剂盒,苏州格锐思生物科技有限公司。
电热鼓风干燥箱,上海博迅医疗生物仪器股份有限公司医疗设备厂;洁净工作台,苏州安泰空气技术有限公司;霉菌培养箱,天津市泰斯特仪器有限公司;高温灭菌锅,致微(厦门)仪器有限公司;多功能微孔板检测仪,广西南宁市博美生物科技有限公司;制冰机,南京贝登医疗股份有限公司;台式微量高速冷冻离心机,Hettich公司;紫外可见分光光度计,赛默飞世尔科技(中国)有限公司。
将保存的HX2菌株接种于PDA培养基平板上,28 ℃培养活化10 d后,挑取菌落破碎稀释,孢子浓度调整为1×105个/mL,转接3 mL于115 mL糖蜜培养基中,置于恒温摇床中28 ℃、170 r/min条件下培养14 d,备用。将厚垣孢子发酵液通过3层无菌纱布过滤得到厚垣孢子浆,利用血球计数法计算厚垣孢子含量,备用。
将质量分数为25%的厚垣孢子浆分别与下述质量分数为75%的载体材料混合均匀:硅藻土、白炭黑、滑石粉、碳酸钙、高岭土及草木灰。所有载体在使用前均经121 ℃灭菌30 min。将混合后的样品置于(40±1) ℃的烘箱中6 h,以不添加载体的厚垣孢子浆作为对照。干燥后的混合物研磨成粉末(过200目筛)得到厚垣孢子母粉,随后进行厚垣孢子萌发率、悬浮率、润湿时间和吸附率的测定。
厚垣孢子萌发率测定方法:称取1 g母粉稀释10倍后,吸取0.2 mL涂布于PDA平板上,28 ℃下培养30 h后置于显微镜下,记录厚垣孢子萌发个数(萌发标准为芽管长度超过本身直径的一半),每个处理设置3个重复,取其平均值作为结果分析,以每平板100个涂布个数为基数,萌发率以百分数表示。
悬浮率测定方法:参考GB/T 14825—2006进行测定[19]
润湿时间测定方法:参考GB/T 5451—2001进行测定[19]
吸附率测定方法:准确称取各载体2 g,放入洁净的杯中,使用胶头滴管,将浓度为1.0×108 CFU/mL的HX2厚垣孢子液逐滴加入不同载体中,用玻璃棒搅拌至载体聚成一团,再称量吸附厚垣孢子液后的总质量,计算载体对HX2厚垣孢子液的吸附率,如公式(1)所示。
载体吸附率(g/g)=(吸附厚垣孢子液后载体的质量-吸附前载体的质量)/吸附前载体的质量
将分散剂(SLS、SHP、SPP、PEG8000、CMC)与润湿剂(吐温-60、SP、SDS、STPP)按1%的质量分数与母粉混合,以不添加任何助剂的母粉作为对照(CK),根据1.3节方法进行厚垣孢子萌发率、润湿时间和悬浮率等各项性能指标的测定(下同),初步筛选出最佳分散剂和润湿剂。
将初步筛选确定的最佳分散剂与最佳润湿剂,分别按质量比5:1、4:1、3:1、2:1、1:1、1:2、1:3、1:4、1:5的比例与母粉进行复配,每组设置3个平行重复。复配后烘干制成粉剂,通过测定上述已明确的各项性能指标,结合综合评价结果确定分散剂与润湿剂二者之间的最佳质量配比。基于上述试验确定的分散剂与润湿剂最佳质量比,固定二者配比不变,分别按母粉质量的1%、2%、3%、4%、5%、6%、7%、8%、9%和10%依次与母粉充分混合均匀,每组设置3个平行重复,烘干制成粉剂后测定其各项性能指标,最终确定复配助剂体系的最佳总用量。
分别选取糊精、海藻酸钠、黄原胶、矿源黄腐植酸钠和抗坏血酸(VC)作为菌剂的紫外保护剂,按0.5%、1.0%、1.5%和2.0%的比例与加入复配助剂后的母粉混合,称取1 g稀释10倍后,吸取0.2 mL涂布于PDA平板上,打开培养皿盖,与CK (不添加紫外保护剂) 一同置于无菌环境中的20 W紫外灯(光强120 lx)下30 cm处,照射3 min,记录厚垣孢子萌发率(方法同1.3节)。
根据单因素试验结果,可湿性粉剂的最终配方为厚垣孢子浆25%,分散剂PEG8000和润湿剂吐温-60 (质量比为1:3)总用量8%,紫外保护剂VC 0.5%-1.5%,以白炭黑为载体配齐100%。将按上述配方配制完成的可湿性粉剂置于(40±1) ℃的烘箱中恒温放置6 h,随后研磨粉碎,过200目筛,即得到厚垣孢子可湿性粉剂最终产品。
含孢量、杂菌率、水分含量、细度及pH测定参照GB 20287—2006标准[19]。润湿时间和悬浮率测定方法同1.3节。
将番茄种子经0.5%次氯酸钠溶液表面消毒15-20 min,随后用无菌水冲洗3-5次,以去除外源微生物。将消毒完成的种子置于无菌水中,于(28±1) ℃下催芽72 h,厚垣孢子可湿性粉剂设置4个处理组:T1、T2、T3和T4,依次分别将上述制备的HX2厚垣孢子可湿性粉剂稀释成浓度为1×108、1×107、1×106、1×105 CFU/mL的悬浊液浸种30 min;对照组(CK)用清水浸种30 min,试验共设5个处理。经上述处理后的番茄种子播种于规格为55 cm×25 cm、含50个育苗孔的育苗盘中,各孔内均装填等量灭菌育苗基质,每孔播1粒种子,每个重复15株苗,每处理3个重复,15 d后测量其根长、株高、茎粗、鲜重、干重。
分别设置6个处理组,T5:用2 g/L的哈茨木霉稀释液在幼苗期(3-5片真叶)时喷施第一次,在接种病原后喷施第二次,其余处理组设置同1.8节。
番茄青枯菌菌液制备:将病原菌Gg24接种于LA培养基上活化,2 d后挑取单菌落接种至LB培养基,于30 ℃、180 r/min培养2 d后,采用紫外可见分光光度计测量菌液浓度,配成OD600=1.0的菌液备用,利用直径为0.4 mm的大头针刺伤番茄根部后,每株灌根接种10 mL Gg24菌液,分别于15 d和30 d后调查发病情况。参照《农药田间药效试验准则第32-部分:杀菌剂防治番茄青枯病》NYT1464.32—2010[9],计算病情指数和防效,如公式(2)、(3)所示。
病情指数=∑(各病级株数×各级代表值)/(调查总株数×最高级代表值)×100
防效=(对照病情指数-处理病情指数)/对照病情指数×100%
分级标准为0级:无病,植株生长正常,无任何萎蔫症状,无可见病害表现;1级:1-2片叶萎蔫,植株其余部分生长正常,仅少数叶片出现萎蔫,不影响整体生长;2级:3-4片叶萎蔫,植株生长受到一定影响,部分叶片萎蔫,植株生长势减弱;3级:全株大部分叶片萎蔫,植株生长严重受阻,无恢复能力重度发病,多数叶片萎蔫,植株基本失去正常生长能力;4级:全株枯死,植株完全死亡。
试验处理后,分别于第1、7、14天采集番茄植株相同部位叶片样本,选择最优HX2厚垣孢子可湿性粉剂处理组、CK和T5测定其防御酶(POD、PPO、PAL、CAT、SOD)活性,试验方法及原理详见试剂盒使用说明书。
数据采用Microsoft Excel 2021和DPS 7.05软件进行统计分析。采用单因素方差分析中的Duncan’s新复极差法进行差异显著性检验,作图软件使用Origin 2024。
表1所示,综合比较HX2厚垣孢子浆加入不同载体后的悬浮率、吸附率、润湿时间及对厚垣孢子萌发的影响,最终选择白炭黑作为粉剂的最佳载体。以白炭黑为载体时,HX2厚垣孢子可湿性粉剂的悬浮率(18.13%)、厚垣孢子萌发率(78.71%)和吸附率(2.52 g/g)均为最高,润湿时间(24.97 s)在6种载体中处于中等水平。
表2可知,最佳分散剂和润湿剂分别为PEG8000和吐温-60,其悬浮率分别为37.50%和21.05%,润湿时间分别为35.41 s和30.43 s,HX2厚垣孢子可湿性粉剂的厚垣孢子萌发率分别为71.60%和71.52%。进一步将PEG8000、吐温-60与母粉按一定比例混合,结果表明,当PEG8000与吐温-60的质量比在2:1至1:3区间内时润湿时间较短,润湿性能较好;当比例过高(1:4、1:5)或过低(5:1)时,润湿时间显著增加,润湿性能下降。悬浮率随吐温-60比例升高逐渐提升,厚垣孢子萌发率则在1:3时达到峰值(74.67%),过高或过低的比例均导致萌发率下降,说明该比例更有利于厚垣孢子维持活性。因此,综合上述性能评价选用1:3的质量比进行用量优化。当助剂总用量为8%时,润湿时间最短(11.07 s)、悬浮率最高(63.33%),且厚垣孢子萌发率(81.49%)效果最优,因此选用此条件进行后续筛选(图1)。
表3可知,紫外照射条件下,不同紫外保护剂对HX2厚垣孢子可湿性粉剂中厚垣孢子萌发的影响存在差异。以抗坏血酸(VC)作为保护剂时,添加浓度为0.5%-1.5%条件下,厚垣孢子萌发率高于其他保护剂处理,且萌发效果更稳定、更具经济价值,与对照CK (56.91%)差异显著,表明VC对厚垣孢子萌发具有良好的紫外保护效果。
表4所示,HX2厚垣孢子可湿性粉剂厚垣孢子含量为2.35×108 CFU/g,不含杂菌,悬浮率为73.8%,润湿时间为24.25 s,pH为5.71,细度为98.81%,含水量16.67%,各项指标均符合《农用微生物菌剂》国家标准(GB 20287—2006)。
种植15 d后对番茄植株各项生长指标进行统计分析,结果表明不同浓度HX2厚垣孢子可湿性粉剂处理对番茄植株皆有不同程度的促进作用。由表5可知,T2 (1×107 CFU/mL)的促生效果最为显著,番茄根长、株高、茎粗、鲜重、干重分别较CK增加了47.39%、31.82%、24.64%、89.45%、90.97%,且与对照差异显著(图2)。
图3可知,接种病原15 d和30 d时,不同浓度HX2厚垣孢子可湿性粉剂处理的病情指数均低于CK。其中T2处理的病情指数分别为0.49和0.16,防效分别为28.67%和50.90%,在接种病原15 d时,T2防效略低于T4 (43.31%)和T5 (38.85%);但接种病原30 d后防治效果明显提升,呈随时间递增的趋势(图4)。
为明确HX2厚垣孢子是否能激活番茄植株的防御系统,测定了5种防御酶(PAL、PPO、POD、CAT和SOD)在接种病原后第1、7、14天的活性。由图5可知,与CK相比,T2处理防御酶活性显著升高,且第7天时的5种酶活性均高于T5 (哈茨木霉)。其中PPO、POD和CAT活性在第7天达到峰值,较CK分别提高了1.69倍、3.39倍和0.25倍;PAL和SOD活性在第14天达到最高,分别提高了0.44倍和1.17倍。上述结果表明,HX2厚垣孢子可显著提高番茄植株防御酶活性,从而增强其对R. solanacearum的抗性。
微生物可湿性粉剂的理化性质直接决定其田间应用效果,载体、分散剂、润湿剂及紫外保护剂的筛选是配方优化的核心环节[20]。本研究通过综合评价吸附率、悬浮率、润湿时间及厚垣孢子萌发率等指标,筛选出白炭黑作为最优载体,PEG8000与吐温-60作为最佳分散剂和润湿剂,按1:3比例混合、总添加量8%时各项指标最优,此时悬浮率提升至63.33%,润湿时间缩短至11.07 s,厚垣孢子萌发率达81.49%。这与白炭黑的多孔结构特性密切相关,其高比表面积不仅能为厚垣孢子提供稳定的附着位点、减少孢子损伤,还能提升制剂的分散性能[21]。分散剂PEG8000与润湿剂吐温-60的协同作用可有效防止孢子团聚,快速降低制剂表面张力,提升其润湿铺展能力,显著优化了制剂的理化特性[22-23]。此外,紫外保护剂筛选结果表明,0.5%-1.5%抗坏血酸(VC)可使厚垣孢子萌发率维持在70%以上,显著高于对照(56.91%)。这是因为VC具有较强的抗氧化性,能有效清除紫外照射产生的活性氧,减少孢子DNA损伤,为制剂在田间紫外环境下的稳定性提供保障[24]。王喜刚等[25]报道,以20%厚垣孢子粉、68%凹凸棒土为载体、4%十二烷基硫酸钠为润湿剂、7%羧甲基纤维素钠为分散剂、1%抗坏血酸为紫外保护剂时,哈茨木霉(T. harzianum) M-17的各项指标最优,厚垣孢子含量为3.1×108 CFU/g,润湿时间为56.5 s,悬浮率为82.27%。张晶晶等[26]以硅藻土为载体,加入25%厚垣孢子粉、4%十二烷基硫酸钠为润湿剂、5% CMC为分散剂、1%糊精为紫外保护剂,配齐100%时,木霉(Trichoderma)厚垣孢子可湿性粉剂的活孢子数为2.5×109 CFU/g,润湿时间为58 s,悬浮率为78%。本研究以厚垣孢子浆25%、分散剂PEG8000和润湿剂吐温-60 (质量比为1:3)总用量8%、紫外保护剂VC 0.5%-1.5%,并以白炭黑为载体配齐100%所制备的HX2厚垣孢子可湿性粉剂,活孢子含量达2.35×108 CFU/g,悬浮率为73.8%,润湿时间为24.25 s。与上述两者相比,本研究制备的可湿性粉剂悬浮率和活孢子含量略低,但润湿时间明显缩短,可显著提升其润湿铺展能力。这可能与真菌种类不同、所用助剂差异有关。
本研究将DSE广西枝孢瓶霉HX2厚垣孢子制备为可湿性粉剂。研究表明,该可湿性粉剂在T2 (1×107 CFU/mL)处理下能显著促进番茄生长,其根长、株高、茎粗、鲜重、干重分别较CK增加了47.39%、31.82%、24.64%、89.45%、90.97%。在番茄青枯病防治方面,T2处理接种病原15 d和30 d后防效分别为28.67%和50.90%,显著优于低浓度处理T4 (1×105 CFU/mL),且后期防效接近甚至优于T5 (哈茨木霉处理)。T2处理防效随时间递增,可能与厚垣孢子在番茄根际的定殖能力相关。HX2厚垣孢子抗逆性强,可在根系中长期存活并逐渐定殖,随着定殖量增加,其生防作用持续增强。前期研究显示将HX2孢子悬浮液(1×106 CFU/mL)与R. solanacearum同时接种可降低病情指数,防效达76.7%,且该处理能促进番茄生长[18]。这一结果与本研究一致,但防效存在差异,可能是由于病原菌与HX2菌株的接种方式和浓度不同所致,总体而言,二者均表现出显著防效,而本研究构建的接种技术更贴合农业生产实际,在操作可行性、田间适配性及规模化应用潜力方面具有明显优势。番茄青枯病由R. solanacearum侵入维管束引起,需要较高浓度的生防菌形成优势菌群,抑制病原菌侵染[6]。赵琳琳等[27]发现,链霉菌(Streptomyces sp.) TOR3209以1×107 CFU/mL浓度接种对番茄枯萎病防治效果最佳。Bing等[28]研究表明,贝莱斯芽孢杆菌(B. velezensis) NEAU-CP5以108 CFU/mL浓度处理番茄植株可显著降低番茄青枯病发病率。以上研究均表明HX2菌株具有同等优异的生防潜力。
内生真菌的生防机制主要包括直接抑制病原菌生长和诱导植物系统抗性(induced systemic resistance, ISR)两大途径[29]。本研究通过测定殖株防御酶活性发现,HX2可湿性粉剂处理能显著激活番茄的抗氧化防御系统,这是其发挥生防作用的关键机制之一。防御酶系统中,POD和PPO参与酚类物质氧化和细胞壁木质化,增强植株机械屏障[30];PAL是植保素合成的关键酶,可提升植株抗菌和抗病毒能力[31];SOD和CAT则通过清除活性氧,减轻病原菌侵染导致的氧化损伤[32]。番茄植株经T2处理后,PPO、POD和CAT活性在接种病原菌第7天达峰值,PAL和SOD活性在第14天达最高,与CK相比分别提高了1.69倍、3.39倍、0.25倍、0.44倍和1.17倍。Kaari等[33]发现链霉菌(Streptomyces sp.) UP1A-1处理的番茄植株在接种病原菌1-9 d时POD、PPO和PAL积累量最高,前2种酶的变化趋势与本研究一致,而PAL的峰值有所提前。Luo等[34]研究表明,链霉菌(Streptomyces sp.) NEAU-174菌株能显著降低番茄青枯病发病率,增强番茄植株中CAT、POD和SOD等防御酶活性,并降低丙二醛含量。以上研究均说明番茄植株防御酶活性变化与其对R. solanacearum的抗性密切相关。
综上所述,本研究将广西枝孢瓶霉HX2菌株厚垣孢子制备为可湿性粉剂,可促进番茄植株生长,并对番茄青枯病具有显著的防治效果,为该菌株的开发利用与规模化应用提供了科学依据。
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2026年第66卷第7期
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doi: 10.13343/j.cnki.wsxb.20251011
  • 接收时间:2025-12-31
  • 首发时间:2026-07-06
  • 出版时间:2026-07-04
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  • 收稿日期:2025-12-31
  • 录用日期:2026-03-19
基金
the Key Research and Development Program of Guangxi Zhuang Autonomous Region (Guinongke(AB241484049)
作者信息
    1.广西农业科学院植物保护研究所,农业农村部华南果蔬绿色防控重点实验室,广西作物病虫害生物学重点实验室,广西 南宁
    2.广西民族大学 海洋与生物技术学院,广西 南宁

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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

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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