Article(id=1266470568367907817, tenantId=1146029695717560320, journalId=1266358857061122103, issueId=1266470523241382909, articleNumber=null, orderNo=null, doi=10.13802/j.cnki.zwbhxb.2026.2025154, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1762185600000, receivedDateStr=2025-11-04, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1779879713383, onlineDateStr=2026-05-27, pubDate=1777478400000, pubDateStr=2026-04-30, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1779879713383, onlineIssueDateStr=2026-05-27, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1779879713383, creator=13701087609, updateTime=1779879713383, updator=13701087609, issue=Issue{id=1266470523241382909, tenantId=1146029695717560320, journalId=1266358857061122103, year='2026', volume='53', issue='2', pageStart='301', pageEnd='586', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1779879702622, creator=13701087609, updateTime=1779879723857, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1266470612705890690, tenantId=1146029695717560320, journalId=1266358857061122103, issueId=1266470523241382909, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1266470612705890691, tenantId=1146029695717560320, journalId=1266358857061122103, issueId=1266470523241382909, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=356, endPage=366, ext={EN=ArticleExt(id=1266470569726862315, articleId=1266470568367907817, tenantId=1146029695717560320, journalId=1266358857061122103, language=EN, title=Isolation, identification of antagonistic bacterial strains, and the antagonistic mechanisms against tomato late blight pathogen, columnId=1266470561661206635, journalTitle=Journal of Plant Protection, columnName=Research reports, runingTitle=null, highlight=null, articleAbstract=

To screen microbial resources for the control of tomato late blight caused by pathogen Phytophthora infestans, strains with strong antagonistic activity were isolated from soil using the plate confrontation method. Their taxonomic status was determined using integrating morphological, physiological, and biochemical characteristics, combined with the 16S rDNA gene sequence analysis. The antifungal activity, action mechanisms, and control effect of the antagonistic strains were investigated via the mycelial growth inhibition method, spore germination assay, and pot experiment. The results showed that a total of 11 strains exhibited antagonistic activities against P. infestans. Among these antagonistic strains, strain C-21 exhibited the highest mycelial growth inhibition rate of 84.41%, and it was identified as Streptomyces flavogriseus. This strain could produce extracellular enzymes including cellulase, protease and amylase. Its fermentation broth could disrupt the mycelial morphology and cell membrane integrity of P. infestans, and significantly inhibited spore germination and zoospore release, with inhibition rates of 69.31% and 57.94%, respectively. The preventive and curative control effects of strain C-21 against tomato late blight were 43.57% and 59.28%, respectively. These findings indicated that strain C-21 had an excellent control effect on tomato late blight, and exhibited promising biocontrol potential for further development and application.

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为筛选防治由致病疫霉Phytophthora infestans引起的番茄晚疫病的菌种资源,采用平板对峙法从土壤中分离筛选出对致病疫霉拮抗作用较强的菌株,结合形态学、生理生化特征及16S rDNA基因序列分析其分类地位,通过菌丝生长、孢子萌发、盆栽试验等方法对其抑菌作用、抑菌机制和防治效果进行探究。结果表明:共筛选出11株菌株对致病疫霉具有拮抗作用,其中菌株C-21对致病疫霉的菌丝生长抑制率最高,为84.41%;菌株C-21最终被鉴定为链霉菌属Streptomyces的黄灰链霉菌S. flavogriseus;菌株C-21能产生纤维素酶、蛋白酶和淀粉酶等胞外酶;其发酵液能破坏致病疫霉的菌丝形态及细胞膜的完整性,显著抑制致病疫霉孢子囊萌发及释放游动孢子,抑制率分别为69.31%和57.94%;菌株C-21对番茄晚疫病的预防效果和治疗效果分别为43.57%和59.28%。表明菌株C-21对番茄晚疫病的防治效果较好,具有良好的开发应用潜力。

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Acta Agriculturae Zhejiangensis, 37(12): 2554-2562 (in Chinese), articleTitle=null, refAbstract=null), Reference(id=1266746618515706261, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, doi=null, pmid=null, pmcid=null, year=2025, volume=37, issue=12, pageStart=2554, pageEnd=2562, url=null, language=null, rfNumber=null, rfOrder=44, authorNames=张玉, 丁一秀, 李俊达, 尚少杰, 赵晓静, 候可心, 高艳东, 刘惠荣, journalName=浙江农业学报, refType=null, unstructuredReference=张玉, 丁一秀, 李俊达, 尚少杰, 赵晓静, 候可心, 高艳东, 刘惠荣. 2025. 一株拮抗致病疫霉的黏细菌菌株的分离、鉴定与发酵条件优化. 浙江农业学报, 37(12): 2554-2562, articleTitle=一株拮抗致病疫霉的黏细菌菌株的分离、鉴定与发酵条件优化, refAbstract=null), Reference(id=1266746618574426518, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, doi=null, pmid=null, pmcid=null, year=2019, volume=136, issue=null, pageStart=104004, pageEnd=null, url=null, language=null, rfNumber=null, rfOrder=45, authorNames=Zheng XF, Wang JP, Chen Z, Zhang HF, Wang ZR, Zhu YJ, Liu B, journalName=Biological Control, refType=null, unstructuredReference=Zheng XF, Wang JP, Chen Z, Zhang HF, Wang ZR, Zhu YJ, Liu B. 2019. A Streptomyces sp. strain: isolation, identification, and potential as a biocontrol agent against soilborne diseases of tomato plants. Biological Control, 136: 104004, articleTitle=A Streptomyces sp. strain: isolation, identification, and potential as a biocontrol agent against soilborne diseases of tomato plants, refAbstract=null)], funds=[Fund(id=1266746614929576295, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, awardId=2025CYJSTX08, language=CN, fundingSource=山西省现代农业产业技术体系建设项目(2025CYJSTX08), fundOrder=null, country=null), Fund(id=1266746615026045288, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, awardId=20210302123404, language=CN, fundingSource=山西省基础研究计划(自由探索类)面上项目(20210302123404), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1266746603110027541, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, xref=null, ext=[AuthorCompanyExt(id=1266746603118416150, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, companyId=1266746603110027541, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Shanxi Key Laboratory of Bioagent Utilization and Eco-Pesticide Innovation in Agriculture, College of Plant Protection, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China), AuthorCompanyExt(id=1266746603126804759, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, companyId=1266746603110027541, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=山西农业大学植物保护学院,农业有益生物利用与生态农药创制山西省重点实验室,太谷 030801)])], figs=[ArticleFig(id=1266746612559794507, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=EN, label=Fig. 1, caption=Inhibitory effect of strain C-21 on the growth of Phytophthora infestans, figureFileSmall=CL9bR+jTnHxpdByToC9aCQ==, figureFileBig=kCrH9T0oZR/I9QPQ93g7xQ==, tableContent=null), ArticleFig(id=1266746612622709068, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=CN, label=图1, caption=菌株C-21对致病疫霉生长的抑制效果, figureFileSmall=CL9bR+jTnHxpdByToC9aCQ==, figureFileBig=kCrH9T0oZR/I9QPQ93g7xQ==, tableContent=null), ArticleFig(id=1266746612723372365, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=EN, label=Fig. 2, caption=Spore chain and spore morphology of strain C-21, figureFileSmall=wKxe6ca3mzelQkyrSoDBTA==, figureFileBig=e6hNO3uwitvknvZb4Zrpvw==, tableContent=null), ArticleFig(id=1266746612773704014, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=CN, label=图2, caption=菌株C-21孢子链及孢子形态特征

A:菌株C-21的孢子链;B:孢子;C:孢子分裂后的断面,箭头所示为鞘套。A: Spore chain morphology of strain C-21; B: spore morphology; C: cross-section of spores after division, with arrows indicating the sheath.

, figureFileSmall=wKxe6ca3mzelQkyrSoDBTA==, figureFileBig=e6hNO3uwitvknvZb4Zrpvw==, tableContent=null), ArticleFig(id=1266746612832424271, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=EN, label=Fig. 3, caption=Phylogenetic tree of strain C-21 and related strains based on 16S rDNA sequences using the neighbor-joining method, figureFileSmall=CmiM1WwH66kfeXviF21wag==, figureFileBig=esGmrH310Zv8iCAUJpZIvg==, tableContent=null), ArticleFig(id=1266746612895338832, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=CN, label=图3, caption=基于 16S rDNA 序列采用邻接法构建菌株C-21与相关菌株的系统发育树

T:模式菌株。T: Type strain.

, figureFileSmall=CmiM1WwH66kfeXviF21wag==, figureFileBig=esGmrH310Zv8iCAUJpZIvg==, tableContent=null), ArticleFig(id=1266746612962447697, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=EN, label=Fig. 4, caption=Inhibition rate of strain C-21 against Phytophthora infestans after being cultured in different fermentation media, figureFileSmall=G0Wh3Y8te4hn5TN97/8iuw==, figureFileBig=qAuilTRNWtBdIx0QILBdSQ==, tableContent=null), ArticleFig(id=1266746613029556562, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=CN, label=图4, caption=菌株C-21经不同发酵培养基培养后对致病疫霉的抑制率

图中数据为平均数±标准差。不同小写字母表示经Duncan氏新复极差法检验差异显著(P<0.05)。Data are mean±SD. Different lowercase letters indicate significant difference by Duncan’s new multiple range test (P<0.05).

, figureFileSmall=G0Wh3Y8te4hn5TN97/8iuw==, figureFileBig=qAuilTRNWtBdIx0QILBdSQ==, tableContent=null), ArticleFig(id=1266746613096665427, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=EN, label=Fig. 5, caption=Detection of cellulase (A), amylase (B), and protease (C) produced by strain C-21, figureFileSmall=5FtlLmiGXViM9OwB3upO1g==, figureFileBig=xJ30gTZZAhku1pqEjH6Ouw==, tableContent=null), ArticleFig(id=1266746613163774292, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=CN, label=图5, caption=菌株C-21产纤维素酶(A)、淀粉酶(B)和蛋白酶(C)的检测, figureFileSmall=5FtlLmiGXViM9OwB3upO1g==, figureFileBig=xJ30gTZZAhku1pqEjH6Ouw==, tableContent=null), ArticleFig(id=1266746613226688853, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=EN, label=Fig. 6, caption=Effect of strain C-21 on the mycelial morphology of Phytophthora infestans, figureFileSmall=Msd/tlS7GM95+VJxFHI0IA==, figureFileBig=it6dz6gq0z0+Mblw5c2gfw==, tableContent=null), ArticleFig(id=1266746613293797718, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=CN, label=图6, caption=菌株C-21对致病疫霉菌丝形态的影响

A:对照组;B、C、D:处理组;1~4:分别示扭曲变形、细胞壁破裂及内容物外溢、内容物分布不均、内容物分布不均。A: Control group; B, C, D: treatment groups; 1-4: indicate distortion and deformation, cell wall rupture with cytoplasmic leakage, uneven distribution of cytoplasmic contents, and uneven distribution of cytoplasmic contents, respectively.

, figureFileSmall=Msd/tlS7GM95+VJxFHI0IA==, figureFileBig=it6dz6gq0z0+Mblw5c2gfw==, tableContent=null), ArticleFig(id=1266746613369295191, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=EN, label=Fig. 7, caption=Detection of effect of strain C-21 on cell membrane integrity of Phytophthora infestans, figureFileSmall=YYxwpQvLs+BEyXX6vvi90w==, figureFileBig=7R4SQZulxBT6VYcxQBKqEQ==, tableContent=null), ArticleFig(id=1266746613432209752, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=CN, label=图7, caption=菌株C-21对致病疫霉细胞膜完整性的影响, figureFileSmall=YYxwpQvLs+BEyXX6vvi90w==, figureFileBig=7R4SQZulxBT6VYcxQBKqEQ==, tableContent=null), ArticleFig(id=1266746613516095833, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=EN, label=Table 1, caption=

Screening of antagonistic strains against Phytophthora infestans

, figureFileSmall=null, figureFileBig=null, tableContent=

菌株编号

Strain no.

菌落半径

Colony radius/mm

抑制率

Inhibition rate/%

C-218.72±0.94 a84.41±3.10 a
L-19.27±0.02 a82.61±0.05 a
C-C15.26±2.71 bcd62.84±8.93 bc
C-E15.81±0.48 bcd57.40±3.54 cd
C-2014.44±0.16 bc65.54±0.54 bc
C-1017.04±0.09 de56.97±0.29 cd
C-813.94±0.58 b67.20±1.92 b
C-1716.28±0.11 bcde59.47±0.35 bcd
C-H16.54±0.25 cde58.60±0.82 bcd
C-3416.86±1.87 cde57.54±6.17 cd
C-3518.35±0.75 e52.62±2.47 d
对照CK34.29±0.11 f-
), ArticleFig(id=1266746613595787610, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=CN, label=表1, caption=

致病疫霉拮抗菌株的筛选

, figureFileSmall=null, figureFileBig=null, tableContent=

菌株编号

Strain no.

菌落半径

Colony radius/mm

抑制率

Inhibition rate/%

C-218.72±0.94 a84.41±3.10 a
L-19.27±0.02 a82.61±0.05 a
C-C15.26±2.71 bcd62.84±8.93 bc
C-E15.81±0.48 bcd57.40±3.54 cd
C-2014.44±0.16 bc65.54±0.54 bc
C-1017.04±0.09 de56.97±0.29 cd
C-813.94±0.58 b67.20±1.92 b
C-1716.28±0.11 bcde59.47±0.35 bcd
C-H16.54±0.25 cde58.60±0.82 bcd
C-3416.86±1.87 cde57.54±6.17 cd
C-3518.35±0.75 e52.62±2.47 d
对照CK34.29±0.11 f-
), ArticleFig(id=1266746613683867995, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=EN, label=Table 2, caption=

Cultural characteristics of strain C-21 on different media

, figureFileSmall=null, figureFileBig=null, tableContent=

培养基种类

Medium type

生长状况

Growth status

气生菌丝

Aerial mycelium

基内菌丝

Substrate mycelium

可溶色素

Soluble pigment

ISP 1++浅灰色 Light gray灰黄色 Gray-yellow无 None
ISP 2+++浅灰色 Light gray黄色、橙黄色 Yellow, orange-yellow黄色 Yellow
ISP 3+++浅灰色 Light gray黄色、浅黄色 Yellow, light yellow黄色 Yellow
ISP 4+++浅灰色 Light gray棕黄色 Brown-yellow浅棕色 Light brown
ISP 5+++浅灰色 Light gray黄色 Yellow浅黄色 Light yellow
ISP 6+++浅灰色 Light gray棕黄色 Brown-yellow无 None
ISP 7+++浅灰色 Light gray橙黄色 Orange-yellow黄色 Yellow

高氏1号培养基

Gauze’s medium #1

+浅灰色 Light gray米白色 Cream white浅黄色 Light yellow
), ArticleFig(id=1266746613767754076, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=CN, label=表2, caption=

菌株 C-21 在不同培养基上的培养性状

, figureFileSmall=null, figureFileBig=null, tableContent=

培养基种类

Medium type

生长状况

Growth status

气生菌丝

Aerial mycelium

基内菌丝

Substrate mycelium

可溶色素

Soluble pigment

ISP 1++浅灰色 Light gray灰黄色 Gray-yellow无 None
ISP 2+++浅灰色 Light gray黄色、橙黄色 Yellow, orange-yellow黄色 Yellow
ISP 3+++浅灰色 Light gray黄色、浅黄色 Yellow, light yellow黄色 Yellow
ISP 4+++浅灰色 Light gray棕黄色 Brown-yellow浅棕色 Light brown
ISP 5+++浅灰色 Light gray黄色 Yellow浅黄色 Light yellow
ISP 6+++浅灰色 Light gray棕黄色 Brown-yellow无 None
ISP 7+++浅灰色 Light gray橙黄色 Orange-yellow黄色 Yellow

高氏1号培养基

Gauze’s medium #1

+浅灰色 Light gray米白色 Cream white浅黄色 Light yellow
), ArticleFig(id=1266746613847445853, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=EN, label=Table 3, caption=

Physiological and biochemical characteristics of strain C-21

, figureFileSmall=null, figureFileBig=null, tableContent=
检测项目 Test item结果 Result检测项目 Test item结果 Result
革兰氏染色 Gram staining+D-果糖 D-fructose±
接触酶反应 Catalase reaction+葡萄糖 Glucose+
纤维素水解 Cellulose hydrolysis+乳糖 Lactose+
淀粉水解 Starch hydrolysis+L-半乳糖 L-galactose+
产H2S H2S production+肌醇 Inositol+
牛奶胨化 Milk peptonization+棉子糖 Raffinose+
明胶液化 Gelatin liquefaction+L-阿拉伯糖 L-arabinose+
产类黑色素Melanin-like substance production-鼠李糖 Rhamnose+
蔗糖 Sucrose+麦芽糖 Maltose+
), ArticleFig(id=1266746613922943326, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=CN, label=表3, caption=

菌株C-21的生理生化特性

, figureFileSmall=null, figureFileBig=null, tableContent=
检测项目 Test item结果 Result检测项目 Test item结果 Result
革兰氏染色 Gram staining+D-果糖 D-fructose±
接触酶反应 Catalase reaction+葡萄糖 Glucose+
纤维素水解 Cellulose hydrolysis+乳糖 Lactose+
淀粉水解 Starch hydrolysis+L-半乳糖 L-galactose+
产H2S H2S production+肌醇 Inositol+
牛奶胨化 Milk peptonization+棉子糖 Raffinose+
明胶液化 Gelatin liquefaction+L-阿拉伯糖 L-arabinose+
产类黑色素Melanin-like substance production-鼠李糖 Rhamnose+
蔗糖 Sucrose+麦芽糖 Maltose+
), ArticleFig(id=1266746614006829407, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=EN, label=Table 4, caption=

Inhibitory effects of various concentrations of strain C-21 fermentation broth on the mycelial growth of Phytophthora infestans

, figureFileSmall=null, figureFileBig=null, tableContent=

发酵液浓度

Concentration of strain

C-21 fermentation/%

菌落直径

Colony

diameter/mm

抑制率

Inhibition rate/%

0.155.48±0.37 a9.75±0.71 a
0.547.27±0.53 b25.35±1.01 b
1.037.59±2.15 c43.76±4.09 c
2.028.06±1.24 d61.86±2.36 d
4.020.13±1.49 e76.95±2.84 e
对照CK60.61±3.21 f-
), ArticleFig(id=1266746614090715488, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=CN, label=表4, caption=

不同浓度菌株C-21发酵液对致病疫霉菌丝生长的抑制作用

, figureFileSmall=null, figureFileBig=null, tableContent=

发酵液浓度

Concentration of strain

C-21 fermentation/%

菌落直径

Colony

diameter/mm

抑制率

Inhibition rate/%

0.155.48±0.37 a9.75±0.71 a
0.547.27±0.53 b25.35±1.01 b
1.037.59±2.15 c43.76±4.09 c
2.028.06±1.24 d61.86±2.36 d
4.020.13±1.49 e76.95±2.84 e
对照CK60.61±3.21 f-
), ArticleFig(id=1266746614438842721, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=EN, label=Table 5, caption=

Inhibitory effect of fermentation broth of strain C-21 on sporangium germination of Phytophthora infestans

, figureFileSmall=null, figureFileBig=null, tableContent=

浓度

Concentration/

%

孢子囊萌发率

Sporangium germination

rate/%

抑制率

Inhibition rate/%

对照组

Control group

处理组

Test group

1036.96±6.51 a24.85±7.48 a33.77±8.74 a
2039.25±5.34 a19.75±6.67 ab50.33±13.02 ab
5040.56±4.19 a12.19±2.49 b69.31±9.09 b
), ArticleFig(id=1266746614510145890, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=CN, label=表5, caption=

菌株C-21发酵液对致病疫霉孢子囊萌发的抑制作用

, figureFileSmall=null, figureFileBig=null, tableContent=

浓度

Concentration/

%

孢子囊萌发率

Sporangium germination

rate/%

抑制率

Inhibition rate/%

对照组

Control group

处理组

Test group

1036.96±6.51 a24.85±7.48 a33.77±8.74 a
2039.25±5.34 a19.75±6.67 ab50.33±13.02 ab
5040.56±4.19 a12.19±2.49 b69.31±9.09 b
), ArticleFig(id=1266746614573060451, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=EN, label=Table 6, caption=

Inhibitory effect of fermentation broth of strain C-21 on zoospore release from sporangia of Phytophthora infestans

, figureFileSmall=null, figureFileBig=null, tableContent=

浓度

Concentration/%

游动孢子释放率

Zoospore release rate/%

抑制率

Inhibition rate/%

对照组

Control group

处理组

Test group

1039.66±2.58 a33.31±1.85 a15.85±5.83 a
2037.36±2.06 a18.06±1.52 b51.71±1.68 b
5038.46±1.44 a16.16±0.63 b57.94±2.45 b
), ArticleFig(id=1266746614648557924, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=CN, label=表6, caption=

菌株C-21发酵液对致病疫霉孢子囊释放游动孢子的抑制作用

, figureFileSmall=null, figureFileBig=null, tableContent=

浓度

Concentration/%

游动孢子释放率

Zoospore release rate/%

抑制率

Inhibition rate/%

对照组

Control group

处理组

Test group

1039.66±2.58 a33.31±1.85 a15.85±5.83 a
2037.36±2.06 a18.06±1.52 b51.71±1.68 b
5038.46±1.44 a16.16±0.63 b57.94±2.45 b
), ArticleFig(id=1266746614724055397, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=EN, label=Table 7, caption=

Control effect of strain C-21 fermentation broth on tomato late blight

, figureFileSmall=null, figureFileBig=null, tableContent=

处理

Treatment

预防试验 Preventive assay治疗试验 Curative assay

病情指数

Disease index

预防效果

Preventive effect/%

病情指数

Disease index

治疗效果

Therapeutic effect/%

菌株C-21发酵液

Strain C-21 fermentation broth

24.65±2.78 a43.57±6.35 a19.04±2.6 ab59.28±5.56 ab
29.40±1.22 b32.71±2.79 b21.82±2.35 b53.34±5.02 b
10×36.86±2.75 c15.63±6.30 c31.02±3.49 c33.66±7.47 c

25%甲霜·霜霉威WP

25% metalaxyl·propamocarb WP

800×24.36±1.57 a44.24±3.60 a14.87±2.33 a68.20±4.98 a
清水对照 Water control43.69±1.02 d46.76±2.63 d
), ArticleFig(id=1266746614795358566, tenantId=1146029695717560320, journalId=1266358857061122103, articleId=1266470568367907817, language=CN, label=表7, caption=

菌株C-21发酵液对番茄晚疫病的防治效果

, figureFileSmall=null, figureFileBig=null, tableContent=

处理

Treatment

预防试验 Preventive assay治疗试验 Curative assay

病情指数

Disease index

预防效果

Preventive effect/%

病情指数

Disease index

治疗效果

Therapeutic effect/%

菌株C-21发酵液

Strain C-21 fermentation broth

24.65±2.78 a43.57±6.35 a19.04±2.6 ab59.28±5.56 ab
29.40±1.22 b32.71±2.79 b21.82±2.35 b53.34±5.02 b
10×36.86±2.75 c15.63±6.30 c31.02±3.49 c33.66±7.47 c

25%甲霜·霜霉威WP

25% metalaxyl·propamocarb WP

800×24.36±1.57 a44.24±3.60 a14.87±2.33 a68.20±4.98 a
清水对照 Water control43.69±1.02 d46.76±2.63 d
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番茄晚疫病菌拮抗菌株筛选鉴定及其作用机制
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彭佳乐 , 杨平鸽 , 董欣杰 , 刘书剑 , 王磊 , 姚艳平 , 郝晓娟 , 王美琴
植物保护学报 | 研究论文 2026,53(2): 356-366
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植物保护学报 | 研究论文 2026, 53(2): 356-366
番茄晚疫病菌拮抗菌株筛选鉴定及其作用机制
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彭佳乐, 杨平鸽, 董欣杰, 刘书剑, 王磊, 姚艳平, 郝晓娟, 王美琴
作者信息
  • 山西农业大学植物保护学院,农业有益生物利用与生态农药创制山西省重点实验室,太谷 030801

通讯作者:

Isolation, identification of antagonistic bacterial strains, and the antagonistic mechanisms against tomato late blight pathogen
Jiale Peng, Pingge Yang, Xinjie Dong, Shujian Liu, Lei Wang, Yanping Yao, Xiaojuan Hao, Meiqin Wang
Affiliations
  • Shanxi Key Laboratory of Bioagent Utilization and Eco-Pesticide Innovation in Agriculture, College of Plant Protection, Shanxi Agricultural University, Taigu 030801, Shanxi Province, China
出版时间: 2026-04-30 doi: 10.13802/j.cnki.zwbhxb.2026.2025154
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为筛选防治由致病疫霉Phytophthora infestans引起的番茄晚疫病的菌种资源,采用平板对峙法从土壤中分离筛选出对致病疫霉拮抗作用较强的菌株,结合形态学、生理生化特征及16S rDNA基因序列分析其分类地位,通过菌丝生长、孢子萌发、盆栽试验等方法对其抑菌作用、抑菌机制和防治效果进行探究。结果表明:共筛选出11株菌株对致病疫霉具有拮抗作用,其中菌株C-21对致病疫霉的菌丝生长抑制率最高,为84.41%;菌株C-21最终被鉴定为链霉菌属Streptomyces的黄灰链霉菌S. flavogriseus;菌株C-21能产生纤维素酶、蛋白酶和淀粉酶等胞外酶;其发酵液能破坏致病疫霉的菌丝形态及细胞膜的完整性,显著抑制致病疫霉孢子囊萌发及释放游动孢子,抑制率分别为69.31%和57.94%;菌株C-21对番茄晚疫病的预防效果和治疗效果分别为43.57%和59.28%。表明菌株C-21对番茄晚疫病的防治效果较好,具有良好的开发应用潜力。

番茄晚疫病  /  致病疫霉  /  黄灰链霉菌  /  生物防治  /  作用机制

To screen microbial resources for the control of tomato late blight caused by pathogen Phytophthora infestans, strains with strong antagonistic activity were isolated from soil using the plate confrontation method. Their taxonomic status was determined using integrating morphological, physiological, and biochemical characteristics, combined with the 16S rDNA gene sequence analysis. The antifungal activity, action mechanisms, and control effect of the antagonistic strains were investigated via the mycelial growth inhibition method, spore germination assay, and pot experiment. The results showed that a total of 11 strains exhibited antagonistic activities against P. infestans. Among these antagonistic strains, strain C-21 exhibited the highest mycelial growth inhibition rate of 84.41%, and it was identified as Streptomyces flavogriseus. This strain could produce extracellular enzymes including cellulase, protease and amylase. Its fermentation broth could disrupt the mycelial morphology and cell membrane integrity of P. infestans, and significantly inhibited spore germination and zoospore release, with inhibition rates of 69.31% and 57.94%, respectively. The preventive and curative control effects of strain C-21 against tomato late blight were 43.57% and 59.28%, respectively. These findings indicated that strain C-21 had an excellent control effect on tomato late blight, and exhibited promising biocontrol potential for further development and application.

tomato late blight  /  Phytophthora infestans  /  Streptomyces flavogriseus  /  biological control  /  action mechanism
彭佳乐, 杨平鸽, 董欣杰, 刘书剑, 王磊, 姚艳平, 郝晓娟, 王美琴. 番茄晚疫病菌拮抗菌株筛选鉴定及其作用机制. 植物保护学报, 2026 , 53 (2) : 356 -366 . DOI: 10.13802/j.cnki.zwbhxb.2026.2025154
Jiale Peng, Pingge Yang, Xinjie Dong, Shujian Liu, Lei Wang, Yanping Yao, Xiaojuan Hao, Meiqin Wang. Isolation, identification of antagonistic bacterial strains, and the antagonistic mechanisms against tomato late blight pathogen[J]. Journal of Plant Protection, 2026 , 53 (2) : 356 -366 . DOI: 10.13802/j.cnki.zwbhxb.2026.2025154
近年来,随着设施蔬菜产业不断发展,番茄种植面积逐年增加,其产量与品质直接关系到农业经济发展和民生需求。由致病疫霉Phytophthora infestans引起的晚疫病是全球范围内制约番茄生产的重要因素之一(Yerasu et al.,2025)。该病具有传播速度快、流行范围广、危害严重的特点,尤其是北方设施蔬菜低温高湿的环境加重了该病害的发生,发病后果实腐败给种植户造成较大的经济损失。传统的病害防控策略频繁使用化学农药,也促进了耐药病原群体的出现,同时随着人们食品安全意识不断提高,农药残留问题显现,因此,筛选和开发环境友好型的生物菌株和生物农药尤为重要(Wei et al.,2025)。
链霉菌Streptomyces种类繁多,代谢产物丰富,防治病害的作用方式多样,目前已被广泛用作生物防治资源。张梅琳等(2025)研究发现孔雀石刺链霉菌Streptomyces malachitospinus可有效防治草莓炭疽病,并显著促进草莓根系生长。石妞妞等(2024)研究发现野尻链霉菌Streptomyces nojiriensis 9-13发酵液高效拮抗平头炭疽菌Colletotrichum truncatum,能够破坏炭疽病菌菌丝结构,对炭疽病菌分生孢子萌发抑制率达100%。Van Minh et al.(2015)发现链霉菌A3265菌株产生抗菌活性物质,对灰葡萄孢菌Botrytis cinerea、尖镰孢菌Fusarium oxysporum等多种植物病原真菌具有抑制活性。Zheng et al.(2019)发现链霉菌FJAT-31547菌株发酵液粗提物对番茄枯萎病和青枯病有较高的防治效果,且对植株有促生作用。Khandan & Janardhana(2015)从土壤中分离出的黄灰链霉菌Streptomyces flavogriseus ACTK2菌株对层出镰孢菌Fusarium proliferatum具有拮抗作用。Kuzikova & Medvedeva(2022)从土壤中分离得到的黄灰链霉菌MK17菌株对灰葡萄孢菌、尖镰孢菌和链格孢菌Alternaria alternata以及其他多种病原菌具有较好的抑制作用,并且能调节植物生长。Oh et al.(2016)研究发现黄灰链霉菌KRA15-528菌株的发酵滤液对南方根结线虫Meloidogyne incognita二龄幼虫及卵均具有较强的抑制与致死作用。因此,深入开展黄灰链霉菌资源挖掘与生防机制研究具有重要意义。
近年来,针对番茄晚疫病的拮抗菌株筛选研究已取得一定进展,从土壤、植物体中等筛选出的木霉菌Trichoderma spp.、荧光假单胞菌Pseudomonas fluorescens和贝莱斯芽胞杆菌Bacillus velezensis等菌种在室内试验或田间小区试验中表现出较好的防治效果(Elhjouji et al.,2025Rana et al.,2025),而目前针对链霉菌防治番茄晚疫病的相关研究仍相对有限。本研究拟从健康植物的根际土壤中分离筛选获得能够有效抑制致病疫霉的菌株,并对该菌株的分类地位、防治效果及抑菌机制进行研究,以期为番茄晚疫病的生物防治提供优质的微生物菌种资源。
供试病原菌:致病疫霉菌种由山西农业大学植物病害生物防治实验室保存并提供。
供试番茄、育苗基质及土壤样品:番茄品种为白果强丰,由青县纯丰蔬菜育种繁殖场提供。育苗基质购自山东昊喆农业科技有限公司。供试土样采集于山西省晋中市太谷区健康西葫芦植株的根际土壤。
供试培养基:黑麦(rye sucrose agar,RSA)培养基成分为黑麦60 g(研磨成粉,加水煮沸,8层纱布过滤取滤液)、蔗糖20 g、琼脂18 g、蒸馏水1 L;燕麦(oatmeal agar,OA)培养基成分为燕麦20 g、微量盐溶液(1 mg/L FeSO4·7H2O、1 mg/L MnCl2·4H2O、1 mg/L ZnSO4·7H2O)、琼脂18 g、蒸馏水1 L;不添加琼脂为燕麦液体(oatmeal broth,OMB)培养基;胰酪大豆胨(trypticase soy agar,TSA)培养基成分为胰蛋白胨15 g、NaCl 5 g、大豆蛋白胨5 g、琼脂18 g、蒸馏水1 L;不添加琼脂为胰酪大豆胨液体(trypticase soy broth,TSB)培养基;国际链霉菌计划(international streptomyces project 1,ISP1)~ISP7培养基成分参考葛米红(2022)方法配制;高氏1号(Gauze’s medium #1,GM)培养基成分为可溶性淀粉20 g、KNO3 1 g、NaCl 0.5 g、K2HPO4 0.5 g、MgSO4·7H2O 0.5 g、FeSO4·7H2O 0.01 g、琼脂15 g、蒸馏水1 L;不添加琼脂为GM液体培养基;小米浸汁培养基成分为小米10 g、葡萄糖10 g、蛋白胨3 g、NaCl 2.5 g、CaCO3 2 g、蒸馏水1 L;葡萄糖酵母提取物(glucose yeast extract,GY)培养基成分为麦芽提取物10 g、酵母提取物10 g、葡萄糖4 g、蒸馏水1 L;酵母麦芽提取物(yeast malt extract,YM)培养基成分为麦芽提取物10 g、酵母提取物10 g、葡萄糖4 g、蒸馏水1 L;酵母浸出粉胨葡萄糖(yeast extract peptone dextrose,YPD)培养基成分为葡萄糖20 g、蛋白胨20 g、酵母粉10 g、蒸馏水1 L;羧甲基纤维素钠(carboxy methyl cellulose-Na,CMC-Na)培养基成分为CMC-Na 10 g、酵母提取物2 g、MgSO4·7H2O 0.2 g、NaCl 0.2 g、琼脂20 g、蒸馏水1 L;淀粉酶检测培养基成分为可溶性淀粉10 g、KNO3 1 g、MgCO3 1 g、NaCl 0.5 g、K2HPO4 0.3 g、琼脂粉15 g、蒸馏水1 L;蛋白酶检测培养基成分为脱脂奶粉15 g、琼脂16 g、蒸馏水1 L。
药剂、试剂及仪器:25%甲霜·霜霉威(metalaxyl·propamocarb)可湿性粉剂,江苏宝灵化工股份有限公司。细菌基因组提取试剂盒、2×EasyTaq SuperMix,北京全式金生物技术有限公司;其他试剂均为国产分析纯。T100 Thermal Cycler型PCR仪,上海伯乐生命医学产品有限公司;DYY-7C型电泳仪、WD-9413B型凝胶成像仪,北京六一生物科技有限公司;SK200型生物显微镜,麦克奥迪实业集团有限公司;GMF-520型荧光显微镜,上海光密仪器有限公司。
参考Zhang et al.(2021)方法从供试土壤中分离菌株。称取0.02 g土壤样品,倒入无菌10 mmol/L MgCl2溶液25 mL,混匀后室温静置15 min制备成匀浆液。分别吸取4 500、1 500、500、167、56、19 μL匀浆至1 L 10% TSB培养基中,制备222、666、2 000、6 000、18 000、54 000倍系列稀释液。各稀释液以每孔160 μL分装至96孔细胞培养板,封口膜密封后30 ℃避光培养7 d。观察细菌生长,选取约30%孔出现可见生长的稀释度作为最佳稀释浓度(optimal dilution concentration,ODC),此条件下阳性孔多为单克隆(源自单个细菌细胞)。试验设ODC、ODC/3及3×ODC梯度,每稀释度分装30~45块96孔细胞培养板,设3块无菌培养基作阴性对照。将致病疫霉培养7 d后用直径8 mm的打孔器打取菌饼,接种于RSA培养基平板中央,在距离菌饼2.5 cm处点接上述分离到的菌株,每株菌株重复6个皿,以仅接种致病疫霉的平板作为对照。于20 ℃恒温暗培养7 d,测量致病疫霉菌落半径,计算抑菌率。抑菌率=(对照组菌落半径-处理组菌落半径)/(对照组菌落半径-菌饼半径)×100%。筛选出对致病疫霉的抑制效果最好的菌株进行后续试验。
参考阎逊初(1992)方法将菌株C-21划线接种于ISP 1~ISP 7七种培养基及GM培养基上,于28 ℃恒温暗培养7 d,每种培养基重复3次,观察并记录菌株C-21在8种培养基上的菌丝生长情况、气生菌丝和基内菌丝颜色、可溶性色素颜色等。将菌株C-21划线培养在OA培养基平板上,用扫描电镜观察菌株C-21孢子形态,试验设置3个平板,共观察12个视野。参考《伯杰细菌鉴定手册》测定菌株C-21的生理生化特性,包括碳源利用、产类黑色素、牛奶分解、H2S产生及明胶液化等。
采用细菌基因组提取试剂盒提取菌株C-21的基因组DNA,利用27F(5′-AGAGTTTGATCTGGC-TCAG-3′)/1492R(5′-TACGGCTACCTTGTTACG-ACTT-3′)引物,以DNA为模板扩增16S rDNA基因。30 μL PCR扩增体系:2×EasyTaq SuperMix 15 μL、DNA模板2 μL、正反向引物各1 μL、ddH2O 11 μL。PCR反应程序:94 ℃预变性5 min;94 ℃变性30 s,57 ℃退火30 s,72 ℃延伸90 s,25个循环;72 ℃再延伸7 min。扩增产物经1%琼脂糖凝胶电泳检测后,将扩增产物送至北京诺赛基因组研究中心有限公司测序。将测序得到的序列在NCBI网站上进行BLAST比对,并下载具有高同源性的菌株序列;采用MEGA 12.0软件以邻接法构建系统发育树,bootstrap检验值为1 000。
参考李桂花等(2024)的方法,选用小米浸汁培养基、TSB培养基、GY培养基、YM培养基、OMB培养基、YPD培养基和GM液体培养基7种链霉菌常用发酵培养基制备相应的发酵液。将菌株C-21划线接种于OA培养基平板,28 ℃下暗培养7 d后,刮取培养基表面孢子,加入无菌水制成孢子悬浮液,稀释到浓度为108 CFU/mL,然后以2%接种量接入上述不同的发酵培养基中,28 ℃、180 r/min条件下培养7 d后以12 000 r/min离心30 min,上清液经0.22 μm微孔滤膜过滤得到菌株C-21发酵液。
采用菌丝生长速率法测定上述不同发酵液对致病疫霉菌丝生长的影响(刘仕达等,2024)。待已灭菌的RSA培养基冷却至55 ℃左右,按体积比1∶19将不同发酵液与RSA培养基混合并摇匀,制成含药平板。在每种含药平板中央接种1个直径为8 mm的致病疫霉菌饼,以接种至不含发酵液的RSA培养基平板作为对照,于20 ℃恒温避光培养,每种发酵液重复3个皿。培养7 d后采用十字交叉法测量菌落直径,计算菌丝生长抑制率。抑制率=(对照组菌落直径-处理组菌落直径)/(对照组菌落直径-菌饼直径)×100%。
对致病疫霉菌丝生长速率的影响:使用1.2.4筛选出的小米浸汁培养基制备C-21发酵液,按照不同体积比加到RSA培养基中配制成浓度为0.1%、0.5%、1%、2%和4%的含药平板,平板中央接种1个直径为8 mm的致病疫霉菌饼,于20 ℃恒温避光培养。以接种至不含发酵液的RSA培养基平板作空白对照,每个处理重复3个皿,培养7 d后采用十字交叉法测量菌落直径,计算抑制率。方法同1.2.4。
对致病疫霉孢子囊萌发的影响:刮下培养7 d的致病疫霉菌丝,加入3 mL无菌水振荡,经孔径约为29.4 μm滤网过滤后得到孢子囊悬浮液。用1.2.4筛选出的小米浸汁培养基制备菌株C-21发酵液,将其与致病疫霉孢子囊悬浮液按照1∶1体积比混合于96孔细胞培养板中,每孔总体积为300 μL,混合后体系中菌株C-21发酵液的终浓度分别为10%、20%和50%,致病疫霉孢子囊悬浮液的终浓度为106个/mL,以不同浓度发酵液为处理组,对应浓度发酵培养基的上清液作空白对照,于25 ℃恒温避光培养24 h,每处理随机选取10个视野,共100~150个孢子囊,在光学显微镜下观察孢子囊的萌发情况,其中芽管长度不小于孢子囊长度的1/2记为萌发,并计算抑制率。试验重复3次。萌发率=萌发孢子囊数/总孢子囊数×100%,抑制率=(对照组孢子囊萌发率-处理组孢子囊萌发率)/对照组孢子囊萌发率×100%。
对致病疫霉孢子囊释放游动孢子的影响:参考上述方法将菌株C-21发酵液与致病疫霉孢子囊悬浮液于96孔细胞培养板混合后置于4 ℃下避光静置2 h,于20 ℃避光静置1 h。在光学显微镜下观察孢子囊释放游动孢子情况,每处理随机选取10个视野,共100~150个孢子囊,其中空孢子囊记为孢子囊已释放游动孢子。试验重复3次。游动孢子释放率=空孢子囊数/总孢子囊数×100%,抑制率=(对照组释放率-处理组释放率)/对照组释放率×100%。
菌株C-21产生胞外酶检测:纤维素酶、蛋白酶检测参考张强等(2024)方法,用接种环将菌株C-21点接在CMC-Na平板中央,于28 ℃恒温培养14 d,滴加0.5%刚果红染色剂染色15 min,然后用1 mol/L NaCl溶液脱色,观察菌株C-21菌落周围是否产生透明圈,若有透明圈则表明菌株C-21产生纤维素酶。将菌株C-21点接在蛋白酶检测平板中央,于28 ℃恒温培养14 d,观察菌株C-21菌落周围是否产生透明圈,若有透明圈则表明菌株C-21产生蛋白酶。淀粉酶检测参考谢晓莹等(2015)的方法,将菌株C-21点接在淀粉酶检测平板中央,于28 ℃恒温培养14 d,将碘液滴加在平板中,若菌株C-21菌落产生透明圈则表明菌株C-21产生淀粉酶。每种检测均设置3次重复。
对致病疫霉菌丝形态的影响:参考1.2.5方法,轻轻挑取适量经菌株C-21发酵液浓度为4%处理后的致病疫霉菌丝,于光学显微镜下观察形态,以小米浸汁培养基的上清液作空白对照,观察5个视野,试验3次重复。
对致病疫霉细胞膜完整性的影响:参考熊诗洁等(2024)方法,利用碘化丙啶染色法观察菌株C-21发酵液对致病疫霉菌丝细胞膜完整性的影响。轻轻挑取适量经C-21发酵液浓度为4%处理后的致病疫霉菌丝,接入装有1 mL 10 μg/mL碘化丙啶的2 mL离心管中,25 ℃避光染色20 min后,用0.01 mol/L、pH 7.2~7.4的磷酸缓冲盐溶液漂洗3次以除去残留的染液。挑取菌丝于荧光显微镜下随机观察菌丝荧光情况,观察5个视野,试验3次重复。
选取饱满的番茄种子,经消毒、浸种、催芽后,播种于装有基质的长18.5 cm、宽15.5 cm、高12.5 cm的营养钵中,每钵播种2粒,置于温室中培养,待幼苗长出6片复叶时,选取长势一致的番茄苗进行试验处理。用1.2.4筛选出的小米浸汁培养基制备菌株C-21发酵液,用无菌水将其稀释为2倍、5倍和10倍稀释液,分别于番茄叶面喷洒,待叶片表面自然干燥后,采用注射法接种病原菌(董汉松,2012),用无菌注射器针头刺破叶片表皮,注射浓度为106个/mL的致病疫霉孢子囊悬浮液,每片叶子20 μL,套袋保持湿度,3 d后取下袋子正常管理。以喷洒800倍25%甲霜·霜霉威可湿性粉剂和无菌水处理分别作阳性对照和阴性对照。每处理8株,3次重复。每隔7 d施药1次,共施药2次。第2次施药后1周进行调查,每株从下到上调查8片叶子。参考张蕊等(2024)方法进行病情分级,具体标准为:0级,无病斑;1级,病斑面积占叶片面积的5%及以下;3级,病斑面积占叶片面积的6%~10%;5级,病斑面积占叶片面积的11%~20%;7级,病斑面积占叶片面积的21%~50%;9级,病斑面积占叶片面积的50%以上。记录各级病叶数,计算病情指数以及预防效果。病情指数=[∑(各级病叶数×相对级代表值)/(调查总叶数×9)]×100;预防效果=(对照组病情指数-处理组病情指数)/对照病情指数×100%。
参考上述方法进行病原菌接种和制备不同稀释倍数菌株C-21发酵液。先注射接种致病疫霉病原菌,后施用不同稀释倍数的菌株C-21发酵液,套袋保湿后,取下袋子正常管理。方法同1.2.7。以喷洒800倍25%甲霜·霜霉威可湿性粉剂和无菌水处理分别作阳性对照和阴性对照。每处理8株,3次重复。每隔7 d施药1次,共施药2次。第2次施药后1周进行调查,计算病情指数以及治疗效果。
采用Microsoft Excel 2024软件对试验数据进行统计分析,使用SPSS 26.0软件进行单因素方差分析检验,应用Duncan氏新复极差法进行差异显著性检验。
将分离纯化的菌株和致病疫霉对峙培养,初步筛选得到11株对致病疫霉有拮抗作用的菌株,经过进一步筛选发现菌株C-21抑菌效果最好(图1),对菌丝的生长抑制率达84.41%(表1)。
在光学显微镜下观察到菌株C-21的孢子链直形或柔曲形(图2-A)。在扫描电镜下可以观察到菌株C-21的孢子链表面粗糙,类似黄瓜或苦瓜表皮的瘤状凸起,凸起顶部较圆,排列无明显规律,相邻凸起间有浅凹陷;孢子呈椭圆形,表面较光滑(图2-B~C)。孢子之间鞘套连接形成孢子链(图2-C)。根据C-21形态学特征可初步确定其属于链霉菌属Streptomyces
拮抗菌株C-21在不同培养基上的培养特征不同,在ISP 1~ISP 7培养基上生长旺盛,在高氏1号培养基上生长较差。菌株C-21在8种不同培养基上的菌落底色多呈现黄色、浅黄色,后期叠加浅灰色的孢子层,整体呈现黄底灰表的特征(表2)。确定菌株C-21属于链霉菌属的灰色系列。
生理生化试验结果表明,菌株C-21的革兰氏染色为阳性,明胶液化、牛奶胨化、产H2S、接触酶反应、淀粉水解、分解纤维素试验均为阳性,产类黑色素试验为阴性。能以蔗糖、葡萄糖、麦芽糖等为碳源,但利用D-果糖为弱阳性(表3)。通过形态学特征、培养性状以及生理生化特征,可将菌株C-21鉴定为黄灰链霉菌Streptomyces flavogriseus
系统进化树结果显示,菌株C-21与黄灰链霉菌、黄微绿链霉菌Streptomyces flavovirens聚为一簇(图3)。但是黄微绿链霉菌不能利用蔗糖和肌醇等碳源。结合形态学特征、培养性状、生理生化特性和分子生物学分析结果,最终将菌株C-21鉴定为黄灰链霉菌Streptomyces flavogriseus
小米浸汁培养基、TSB培养基、GY培养基对应的菌株C-21发酵液对致病疫霉的抑制率最高,均达到100.00%。GM液体培养基对应的C-21发酵液的抑制率最低,为12.15%(图4)。综合经济成本和抑菌效果,选取小米浸汁培养基作为菌株C-21的最佳发酵培养基进行后续试验。
菌株C-21对致病疫霉的抑制效果随着发酵液浓度增加而增强,发酵液浓度为0.5%时,对致病疫霉的菌丝生长抑制率为25.35%,浓度为4%时,对致病疫霉的菌丝生长抑制率可达76.95%(表4)。
3种不同浓度的菌株C-21发酵液对致病疫霉孢子囊萌发均表现出抑制作用(表5)。随着菌株C-21发酵液浓度的升高,致病疫霉孢子囊萌发率逐渐降低。浓度为10%的发酵液对致病疫霉孢子囊萌发的抑制率为33.77%;浓度为50%的发酵液对致病疫霉孢子囊萌发的抑制率为69.31%。表明菌株C-21发酵液能抑制致病疫霉孢子囊萌发。
3种不同浓度的菌株C-21发酵液对致病疫霉孢子囊释放游动孢子均表现出抑制作用(表6)。随着发酵液浓度的升高,致病疫霉游动孢子的释放率呈逐渐下降趋势。浓度为10%的发酵液对游动孢子释放的抑制率为15.85%,浓度为50%的发酵液对游动孢子释放的抑制率为57.94%。表明菌株C-21发酵液能抑制致病疫霉孢子囊释放游动孢子。
菌株C-21在纤维素酶、淀粉酶和蛋白酶检测培养基上均能产生明显且直径较大的透明圈(图5),说明菌株C-21有产蛋白酶、纤维素酶和淀粉酶的能力。
显微镜观察结果显示,对照组和处理组致病疫霉菌丝形态出现了明显的差异,对照组菌丝光滑,无变形、内容物分布不均及外泄等现象(图6-A);处理组的致病疫霉菌丝出现扭曲、弯曲变形、细胞壁破裂、内容物外泄及分布不均等现象(图6-B~D)。表明菌株C-21可显著破坏致病疫霉的菌丝形态。
菌株C-21处理后的致病疫霉菌丝经碘化丙啶染色后,可观察到明显的红色荧光信号;而在相同染色条件下,对照组则未检测到明显红色荧光,仅呈现微弱背景信号或无特异性荧光(图7)。表明菌株C-21能破坏致病疫霉菌丝细胞膜的完整性。
不同稀释倍数下菌株C-21发酵液对番茄晚疫病均有一定防治效果。在预防试验中,2倍稀释浓度下的发酵液对番茄晚疫病的预防效果为43.57%,25%甲霜·霜霉威处理的预防效果为44.24%,两者间无显著差异(表7)。在治疗试验中,2倍稀释浓度下的发酵液处理的治疗效果为59.28%,25%甲霜·霜霉威处理的治疗效果为68.20%,两者间无显著差异(表7)。随着发酵液浓度的升高,其病情指数逐步下降,防治效果逐渐提高,表明菌株C-21发酵液对番茄晚疫病具有明显的防治效果。
番茄作为全球广泛种植的重要经济作物,晚疫病是影响其产业发展的主要病害之一。相较于化学杀菌剂,利用微生物资源防治病害更具备环境友好、残留低、安全性高等优势,适合用于绿色和可持续农业(何烈干等,2021)。本研究从植物根际土壤中筛选到1株对致病疫霉具有较强抑制作用的黄灰链霉菌菌株C-21。已有研究报道黄灰链霉菌具有良好的抑菌活性,能够抑制层出镰孢菌、灰葡萄孢菌、尖镰孢菌和链格孢菌等多种病原菌生长(Khandan & Janardhana,2015Kuzikova & Medvedeva,2022)。
张玉等(2025)刘雅萍等(2024)分别报道了珊瑚状珊瑚球菌Corallococcus coralloides和弱小珊瑚球菌Corallococcus exiguus的发酵浓缩液能显著抑制致病疫霉的生长;程海洋等(2021)报道荧光假单胞菌Pseudomonas fluorescens菌株KBL17的发酵滤液对致病疫霉生长及孢子囊萌发的抑制率均达90%以上。本研究结果表明菌株C-21发酵液浓度为4%时,对致病疫霉的菌丝生长抑制率为76.95%,浓度为50%时对孢子囊萌发和游动孢子释放的抑制率分别为69.31%和57.94%,同时可破坏菌丝的形态结构及细胞膜的完整性,表明菌株C-21发酵液中的活性物质对菌丝生长的抑制作用较强。
拮抗菌株分泌的水解酶在抑制病原菌生长中具有关键作用,黄馨玉等(2025)研究发现贝莱斯芽胞杆菌Sneb2570能分泌淀粉酶和蛋白酶。肖珂雨等(2024)筛选出的小麦条锈菌Puccinia striiformis f. sp. tritici高效拮抗菌株直丝紫链霉菌Streptomyces rectiviolaceus能分泌淀粉酶、蛋白酶、纤维素酶。本研究发现菌株C-21可以产纤维素酶、蛋白酶、淀粉酶等胞外酶。而致病疫霉的细胞壁主要成分是纤维素。因此,菌株C-21通过产生的纤维素酶溶解菌丝细胞壁,通过代谢产物破坏致病疫霉菌丝形态及细胞膜完整性,从而抑制菌丝的生长发育,使其丧失侵染力。还有研究报道,黄灰链霉菌可产生几丁质酶(Ekundayo et al.,2022),许多植物病原菌细胞壁的主要成分为几丁质。菌株C-21是否能产生几丁质酶需要进一步研究。
近年来,已有研究针对番茄晚疫病展开生防试验,Elhjouji et al.(2025)研究表明,经贝莱斯芽胞杆菌菌株RS65菌悬液处理后能显著降低番茄晚疫病的发病程度,相比对照降低了14.72%。Di Francesco et al.(2017)研究表明,出芽短梗霉菌Aureobasidium pullulans菌株L8对番茄晚疫病具有显著的防治效果,且预防效果优于治疗效果,预防处理使病害降低了60%。本研究通过温室防治试验发现,菌株C-21发酵液对番茄晚疫病的预防效果低于治疗效果,分别为43.57%和59.28%,与上述研究不同,推测这可能是因为菌丝相较于孢子囊对发酵液更敏感,使得发酵液在病原菌侵染后期菌丝生长阶段能更有效地抑制病害扩展。菌株C-21发酵液中活性物质后续还需进一步检测和深入分析。
  • 山西省现代农业产业技术体系建设项目(2025CYJSTX08)
  • 山西省基础研究计划(自由探索类)面上项目(20210302123404)
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2026年第53卷第2期
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doi: 10.13802/j.cnki.zwbhxb.2026.2025154
  • 接收时间:2025-11-04
  • 首发时间:2026-05-27
  • 出版时间:2026-04-30
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山西省现代农业产业技术体系建设项目(2025CYJSTX08)
山西省基础研究计划(自由探索类)面上项目(20210302123404)
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    山西农业大学植物保护学院,农业有益生物利用与生态农药创制山西省重点实验室,太谷 030801

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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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