Article(id=1256263564730446058, tenantId=1146029695717560320, journalId=1255847803461844995, issueId=1256263559323967535, articleNumber=null, orderNo=null, doi=10.13346/j.mycosystema.250162, pmid=null, cstr=32115.14.j.mycosystema.250162, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=research-article, receivedDate=1748880000000, receivedDateStr=2025-06-03, revisedDate=null, revisedDateStr=null, acceptedDate=1759766400000, acceptedDateStr=2025-10-07, onlineDate=1777446174080, onlineDateStr=2026-04-29, pubDate=1771689600000, pubDateStr=2026-02-22, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1777446174080, onlineIssueDateStr=2026-04-29, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1777446174080, creator=13701087609, updateTime=1777446174080, updator=13701087609, issue=Issue{id=1256263559323967535, tenantId=1146029695717560320, journalId=1255847803461844995, year='2026', volume='45', issue='2', pageStart='250058', pageEnd='250280', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1777446172791, creator=13701087609, updateTime=1777447435276, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1256268854674710546, tenantId=1146029695717560320, journalId=1255847803461844995, issueId=1256263559323967535, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1256268854678904851, tenantId=1146029695717560320, journalId=1255847803461844995, issueId=1256263559323967535, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=250162, endPage=, ext={EN=ArticleExt(id=1256263569180602619, articleId=1256263564730446058, tenantId=1146029695717560320, journalId=1255847803461844995, language=EN, title=Fusarium foetens, the pathogen of Fusarium root rot of Vernicia fordii, columnId=1256263562373226548, journalTitle=Mycosystema, columnName=Research paper, runingTitle=null, highlight=null, articleAbstract=

A new disease was found on Vernicia fordii with symptom of root rot in Dushan, Guizhou Province. The disease occurred most seriously at growth stage (May to July), primarily affecting the roots. The diseased plants initially showed wilting and leaves fade green to withered until plant death. The fungal community composition of the rhizosphere soil was analyzed from healthy and diseased plants. The result showed that mainly dominant phyla and genera in the rhizosphere soil of diseased and healthy plants were similar, but the relative abundance was absolutely different. Moreover, the genus Fusarium causing a wide range of plant diseases only appeared in the fungal community of rhizosphere soil of diseased V. fordii, with relative abundance accounting for 2.44%. The pathogens were isolated and purified through tissue isolation. Ten strains were isolated from the healthy junction of V. fordii rhizome suffering from typical root rot. Among them, strain AX-2 induced typical symptoms of root rot after being back-inoculated to the seedlings of V. fordii. The pathogen was subsequently isolated from the roots and stems of diseased plants. Its morphological characteristics were found to be consistent with those of the pathogen isolated from the field. Molecular identification further verified that this strain was identical to AX-2. Morphological characteristics and multi-loci (ITS+tef1-α+rpb2+CaM+tub2) phylogeny classified AX-2 as Fusarium foetens. The species is firstly reported as the pathogen of root rot in V. fordii. This study provides a theoretical foundation for investigating the infection characteristics and control strategies of root rot disease in V. fordii.

, correspAuthors=Yicun CHEN, Yangdong WANG, authorNote=null, correspAuthorsNote=
*E-mail: WANG Yangdong, ;
CHEN Yicun,
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CHEN Yicun (0000-0001-6625-8002)

ORCID: WANG Yangdong (0000-0003-0160-5813),

, authorsList=Jia WANG, Anren YANG, Beiping LIU, Ming GAO, Yunxiao ZHAO, Yicun CHEN, Yangdong WANG), CN=ArticleExt(id=1256263581524439425, articleId=1256263564730446058, tenantId=1146029695717560320, journalId=1255847803461844995, language=CN, title=由臭镰孢Fusarium foetens侵染引起的油桐根腐病, columnId=1256263563312771301, journalTitle=菌物学报, columnName=研究论文, runingTitle=null, highlight=null, articleAbstract=

在贵州独山发现一种油桐根腐病,以生长旺盛期5月至7月发生最严重,发病部位在油桐根部,罹病植株叶片萎蔫、失水褪绿,最后干枯死亡。采用微生物群落真菌ITS高通量测序对健康油桐和发病油桐根际土壤的真菌群落组成进行了分析,它们的主要优势菌门和菌属组成相近,但相对丰度却大不相同,并且能广泛引起植物病害的镰孢属Fusarium只在发病油桐根际土壤菌群中出现,相对丰度占比为2.44%;采用组织分离法对引起油桐根腐病的病原菌进行分离纯化,从发病油桐根茎的病健交界处获得10株分离菌株,其中菌株AX-2回接到油桐幼苗后,呈典型的根腐病发病症状,进一步从发病植株的根茎部重新分离获得病原菌,与田间采集分离的病原菌形态特征一致,通过分子鉴定确定了该菌株与AX-2是同一种菌株。经形态学观察和多基因联合建树分析(ITS+tef1-a+rpb2+CaM+tub2),菌株AX-2被鉴定为臭镰孢Fusarium foetens。这是该菌所致油桐根腐病的首次发现,为油桐根腐病的侵染特性及防控技术研究提供了理论依据。

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tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, doi=null, pmid=null, pmcid=null, year=2020, volume=49, issue=1, pageStart=27, pageEnd=30, url=null, language=null, rfNumber=[52], rfOrder=51, authorNames=徐红梅, 邓先珍, 杜洋文, journalName=湖北林业科技, refType=null, unstructuredReference=徐红梅, 邓先珍, 杜洋文, 2020. 一种新的油桐黑斑病病原及其生物学特性研究. 湖北林业科技, 49(1): 27-30, articleTitle=一种新的油桐黑斑病病原及其生物学特性研究, refAbstract=null), Reference(id=1256263618627253144, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, doi=null, pmid=null, pmcid=null, year=2014, volume=27, issue=6, pageStart=752, pageEnd=757, url=null, language=null, rfNumber=[53], rfOrder=52, authorNames=杨素素, 高暝, 朱慧萍, 刘英冠, 汪阳东, 陈益存, journalName=林业科学研究, refType=null, unstructuredReference=杨素素, 高暝, 朱慧萍, 刘英冠, 汪阳东, 陈益存, 2014. 三年桐、千年桐感染枯萎病病原菌后的生理反应. 林业科学研究, 27(6): 752-757, articleTitle=三年桐、千年桐感染枯萎病病原菌后的生理反应, refAbstract=null), Reference(id=1256263618727916443, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, doi=null, pmid=null, pmcid=null, year=2024, volume=26, issue=7, pageStart=121, pageEnd=135, url=null, language=null, rfNumber=[54], rfOrder=53, authorNames=杨娅琳, 吴峰婧琳, 陈健鑫, 武自强, 刘丽, 张东华, 马焕成, 伍建榕, journalName=中国农业科技导报, refType=null, unstructuredReference=杨娅琳, 吴峰婧琳, 陈健鑫, 武自强, 刘丽, 张东华, 马焕成, 伍建榕, 2024. 油茶根腐病根际土壤、根系内真菌群落结构和多样性分析. 中国农业科技导报, 26(7): 121-135, articleTitle=油茶根腐病根际土壤、根系内真菌群落结构和多样性分析, refAbstract=null), Reference(id=1256263618795025310, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, doi=null, pmid=null, pmcid=null, year=2024, volume=null, issue=null, pageStart=null, pageEnd=null, url=https://link.cnki.net/urlid/46.1068.S.20231017.1039.004, language=null, rfNumber=[55], rfOrder=54, authorNames=杨阳, 孔维宝, 牛世全, journalName=分子植物育种, refType=null, unstructuredReference=杨阳, 孔维宝, 牛世全, 2024. 党参根腐病病原菌的分离鉴定及其特性研究. 分子植物育种, https://link.cnki.net/urlid/46.1068.S.20231017.1039.004, articleTitle=党参根腐病病原菌的分离鉴定及其特性研究, refAbstract=null), Reference(id=1256263618857939873, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, doi=null, pmid=null, pmcid=null, year=2011, volume=30, issue=4, pageStart=658, pageEnd=662, url=null, language=null, rfNumber=[56], rfOrder=55, authorNames=袁志林, 陈益存, 汪阳东, journalName=菌物学报, refType=null, unstructuredReference=袁志林, 陈益存, 汪阳东, 2011. 一种新发生的油桐叶枯病病原真菌. 菌物学报, 30(4): 658-662, articleTitle=一种新发生的油桐叶枯病病原真菌, refAbstract=null)], funds=[Fund(id=1256263610574189247, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, awardId=32572095, language=EN, fundingSource=National Natural Science Foundation of China(32572095), fundOrder=null, country=null), Fund(id=1256263610674852549, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, awardId=32572095, language=CN, 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Ltd., Qiannan 558200, Guizhou, China), AuthorCompanyExt(id=1256263586473718201, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, companyId=1256263586096230836, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=3 贵州鸿发生态农业科技有限责任公司,贵州 黔南 558200)])], figs=[ArticleFig(id=1256263608141492856, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, language=EN, label=Fig. 1, caption=The field symptom of Fusarium root rot in Vernicia fordii.

A: Infected plants; B: Comparison of longitudinal sections of the main roots of diseased plants and healthy plants; C: Comparison of cross sections of the lateral roots of diseased plants and healthy plants; D: Leaf symptoms of infected plants.

, figureFileSmall=MUxJiJQH53fIM6ENqw1H5g==, figureFileBig=iGinnrFigkiiLP+t2GqaVg==, tableContent=null), ArticleFig(id=1256263608300876412, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, language=CN, label=图1, caption=油桐根腐病的田间症状

A:发病植株;B:发病植株与健康植株主根纵截面的对比图;C:发病植株与健康植株侧根横截面的对比图;D:发病植株叶片症状

, figureFileSmall=MUxJiJQH53fIM6ENqw1H5g==, figureFileBig=iGinnrFigkiiLP+t2GqaVg==, tableContent=null), ArticleFig(id=1256263608737084038, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, language=EN, label=Fig. 2, caption=Fungal diversity in rhizosphere soil of the healthy and diseased Vernicia fordii.

A: The ASV diversity in rhizosphere soil of the healthy and diseased V. fordii. B: The relative abundance in rhizosphere soil of the healthy and diseased V. fordii at the phylum level. C: The TOP30 relative abundance in rhizosphere soil of the healthy and diseased V. fordii at the genus level. D: The differential abundance in rhizosphere soil of the healthy and diseased V. fordii at the TOP30 genus level. Single asterisk and double asterisk respectively indicated significant (P<0.05) and extremely significant (P<0.01) level difference. The “ns” represents no significant difference.

, figureFileSmall=MTMH1zPlCe0vsOfY+HPfHg==, figureFileBig=aD++qLFGx4hez1owSKhiPA==, tableContent=null), ArticleFig(id=1256263608984547978, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, language=CN, label=图2, caption=健康油桐和发病油桐的根际土壤菌群多样性分析

A:健康油桐和发病油桐的根际土壤ASV多样性比较;B:健康油桐和发病油桐的根际土壤样本在门水平上的相对丰度;C:健康油桐和发病油桐的根际土壤样本在TOP30属水平上的相对丰度;D:健康油桐和发病油桐的根际土壤样本在TOP30属水平上相对丰度的差异分析;*代表差异显著(P<0.05),**代表差异极显著(P<0.01),ns代表无显著差异

, figureFileSmall=MTMH1zPlCe0vsOfY+HPfHg==, figureFileBig=aD++qLFGx4hez1owSKhiPA==, tableContent=null), ArticleFig(id=1256263609114571407, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, language=EN, label=Fig. 3, caption=Ten pure cultures isolated from the interface area of the diseased and healthy parts in diseased Vernicia fordii., figureFileSmall=MQgXWl13HJFlQbtsiFF3sw==, figureFileBig=6VWdibe1gV1EPO/PP2Gvzg==, tableContent=null), ArticleFig(id=1256263609357841045, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, language=CN, label=图3, caption=发病油桐病健交界处分离到的10个纯培养物, figureFileSmall=MQgXWl13HJFlQbtsiFF3sw==, figureFileBig=6VWdibe1gV1EPO/PP2Gvzg==, tableContent=null), ArticleFig(id=1256263609529807513, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, language=EN, label=Fig. 4, caption=The changing phenotype of Vernicia fordii after pathogen AX-2 infection., figureFileSmall=Pv8b3tL3MYvKQ1KYh+YHyQ==, figureFileBig=MZGbKN/b1ZydpjNwcBEqsA==, tableContent=null), ArticleFig(id=1256263609680802460, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, language=CN, label=图4, caption=油桐幼苗接种病原菌AX-2后的表型变化, figureFileSmall=Pv8b3tL3MYvKQ1KYh+YHyQ==, figureFileBig=MZGbKN/b1ZydpjNwcBEqsA==, tableContent=null), ArticleFig(id=1256263609781465760, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, language=EN, label=Fig. 5, caption=Morphological characteristics of pathogen AX-2.

A: Colony morphology; B: Morphology of macroconidia and microconidia; C: Conidiophore morphology; D: Spore morphology.

, figureFileSmall=VI41UzZWvQpEKlf2jNfIWQ==, figureFileBig=iQKSRccSTDq5D+6KCZ7B3A==, tableContent=null), ArticleFig(id=1256263609982792361, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, language=CN, label=图5, caption=病原菌AX-2的形态学特征

A:菌落形态;B:大、小型分生孢子;C:分生孢子梗;D:分生孢子

, figureFileSmall=VI41UzZWvQpEKlf2jNfIWQ==, figureFileBig=iQKSRccSTDq5D+6KCZ7B3A==, tableContent=null), ArticleFig(id=1256263610070872747, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, language=EN, label=Fig. 6, caption=The phylogenetic tree of the multi-gene sequences of pathogen AX-2 based on the maximum likelihood.

Strain of this study is in bold.

, figureFileSmall=WCvU9tTJiBiOcmoOi5FD9w==, figureFileBig=OfjGiB9UxFh74Pa4Xi9Wug==, tableContent=null), ArticleFig(id=1256263610196701871, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, language=CN, label=图6, caption=基于邻接法构建的病原菌AX-2的多基因序列系统发育树

本研究菌株用粗体表示

, figureFileSmall=WCvU9tTJiBiOcmoOi5FD9w==, figureFileBig=OfjGiB9UxFh74Pa4Xi9Wug==, tableContent=null), ArticleFig(id=1256263610330919604, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, language=EN, label=Table 1, caption=

The information of high-throughput sequencing data of rhizosphere soil in the healthy and diseased Vernicia fordii

, figureFileSmall=null, figureFileBig=null, tableContent=
样本
Sample
原始读序
Raw data
有效读序
Valid data
有效读序占原始读序的比例
Valid ratio/%

Phylum

Genus
Healthy1 84 882 81 399 95.90 6 138
Healthy2 83 925 81 703 97.35
Healthy3 84 270 77 241 91.66
Diseased1 81 662 77 150 94.47 8 209
Diseased2 82 696 77 430 93.63
Diseased3 83 935 78 049 92.99
), ArticleFig(id=1256263610452554426, tenantId=1146029695717560320, journalId=1255847803461844995, articleId=1256263564730446058, language=CN, label=表1, caption=

健康油桐和发病油桐的根际土壤高通量测序数据信息

, figureFileSmall=null, figureFileBig=null, tableContent=
样本
Sample
原始读序
Raw data
有效读序
Valid data
有效读序占原始读序的比例
Valid ratio/%

Phylum

Genus
Healthy1 84 882 81 399 95.90 6 138
Healthy2 83 925 81 703 97.35
Healthy3 84 270 77 241 91.66
Diseased1 81 662 77 150 94.47 8 209
Diseased2 82 696 77 430 93.63
Diseased3 83 935 78 049 92.99
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由臭镰孢Fusarium foetens侵染引起的油桐根腐病
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王嘉 1, 2 , 杨安仁 3 , 刘北平 1 , 高暝 1 , 赵耘霄 1 , 陈益存 1, * , 汪阳东 1, *
菌物学报 | 研究论文 2026,45(2): 250162
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菌物学报 | 研究论文 2026, 45(2): 250162
由臭镰孢Fusarium foetens侵染引起的油桐根腐病
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王嘉1, 2, 杨安仁3, 刘北平1, 高暝1, 赵耘霄1, 陈益存1, * , 汪阳东1, *
作者信息
  • 1 中国林业科学研究院亚热带林业研究所 全省林木育种重点实验室,浙江 杭州 311400
  • 2 南京林业大学,江苏 南京 210037
  • 3 贵州鸿发生态农业科技有限责任公司,贵州 黔南 558200
Fusarium foetens, the pathogen of Fusarium root rot of Vernicia fordii
Jia WANG1, 2, Anren YANG3, Beiping LIU1, Ming GAO1, Yunxiao ZHAO1, Yicun CHEN1, * , Yangdong WANG1, *
Affiliations
  • 1 Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Zhejiang Key Laboratory of Forest Genetics and Breeding, Hangzhou 311400, Zhejiang, China
  • 2 Nanjing Forestry University, Nanjing 210037, Jiangsu, China
  • 3 Hongfa Ecological Agriculture Co. Ltd., Qiannan 558200, Guizhou, China
  • CHEN Yicun (0000-0001-6625-8002)

    ORCID: WANG Yangdong (0000-0003-0160-5813),

出版时间: 2026-02-22 doi: 10.13346/j.mycosystema.250162
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在贵州独山发现一种油桐根腐病,以生长旺盛期5月至7月发生最严重,发病部位在油桐根部,罹病植株叶片萎蔫、失水褪绿,最后干枯死亡。采用微生物群落真菌ITS高通量测序对健康油桐和发病油桐根际土壤的真菌群落组成进行了分析,它们的主要优势菌门和菌属组成相近,但相对丰度却大不相同,并且能广泛引起植物病害的镰孢属Fusarium只在发病油桐根际土壤菌群中出现,相对丰度占比为2.44%;采用组织分离法对引起油桐根腐病的病原菌进行分离纯化,从发病油桐根茎的病健交界处获得10株分离菌株,其中菌株AX-2回接到油桐幼苗后,呈典型的根腐病发病症状,进一步从发病植株的根茎部重新分离获得病原菌,与田间采集分离的病原菌形态特征一致,通过分子鉴定确定了该菌株与AX-2是同一种菌株。经形态学观察和多基因联合建树分析(ITS+tef1-a+rpb2+CaM+tub2),菌株AX-2被鉴定为臭镰孢Fusarium foetens。这是该菌所致油桐根腐病的首次发现,为油桐根腐病的侵染特性及防控技术研究提供了理论依据。

油桐  /  根腐病  /  臭镰孢  /  形态特征  /  分子鉴定

A new disease was found on Vernicia fordii with symptom of root rot in Dushan, Guizhou Province. The disease occurred most seriously at growth stage (May to July), primarily affecting the roots. The diseased plants initially showed wilting and leaves fade green to withered until plant death. The fungal community composition of the rhizosphere soil was analyzed from healthy and diseased plants. The result showed that mainly dominant phyla and genera in the rhizosphere soil of diseased and healthy plants were similar, but the relative abundance was absolutely different. Moreover, the genus Fusarium causing a wide range of plant diseases only appeared in the fungal community of rhizosphere soil of diseased V. fordii, with relative abundance accounting for 2.44%. The pathogens were isolated and purified through tissue isolation. Ten strains were isolated from the healthy junction of V. fordii rhizome suffering from typical root rot. Among them, strain AX-2 induced typical symptoms of root rot after being back-inoculated to the seedlings of V. fordii. The pathogen was subsequently isolated from the roots and stems of diseased plants. Its morphological characteristics were found to be consistent with those of the pathogen isolated from the field. Molecular identification further verified that this strain was identical to AX-2. Morphological characteristics and multi-loci (ITS+tef1-α+rpb2+CaM+tub2) phylogeny classified AX-2 as Fusarium foetens. The species is firstly reported as the pathogen of root rot in V. fordii. This study provides a theoretical foundation for investigating the infection characteristics and control strategies of root rot disease in V. fordii.

Vernicia fordii  /  Fusarium root rot  /  Fusarium foetens  /  morphological characteristics  /  molecular identification
王嘉, 杨安仁, 刘北平, 高暝, 赵耘霄, 陈益存, 汪阳东. 由臭镰孢Fusarium foetens侵染引起的油桐根腐病. 菌物学报, 2026 , 45 (2) : 250162 - . DOI: 10.13346/j.mycosystema.250162
Jia WANG, Anren YANG, Beiping LIU, Ming GAO, Yunxiao ZHAO, Yicun CHEN, Yangdong WANG. Fusarium foetens, the pathogen of Fusarium root rot of Vernicia fordii[J]. Mycosystema, 2026 , 45 (2) : 250162 - . DOI: 10.13346/j.mycosystema.250162
油桐Vernicia fordii是大戟科Euphorbiaceae落叶乔木,是世界著名的木本非食用油料植物,在工业、生物、能源、医药和生态环境等方面具有重要的作用(方嘉兴等 2017)。从油桐果实中提取的桐油含有大量的不饱和脂肪酸,是一种非常好的生物质燃料油和优良的干燥植物油,广泛用于油漆、塑料、人造橡胶、油墨、润滑油和生物柴油等,具有可再生、清洁、安全的特点(Chen et al. 2016),在能源转型背景下,油桐作为生物质燃料油的潜力日益凸显,全球需求持续增长。不仅如此,油桐树皮可制胶(Shockey et al. 2006),其根、叶、果均可以药用,特别是油桐叶(Dyer et al. 2002)。此外,油桐因其花期长、颜色绚丽,也被视为重要的园艺作物,又有“五月雪”之称,以油桐为支撑的旅游相关产业已经在我国南方多地兴盛(王承南等 2014)。中国作为油桐主要生产国,现有种植面积近百万公顷,主要集中在湖南、贵州、广西等南方省份和地区,形成了完整的产业链,对山区经济发展和乡村振兴具有支撑作用。然而,油桐的产业化发展面临严峻挑战。该树种虽生长迅速,但遗传抗性较弱,在生长过程中极易遭受多种病原菌的侵害,严重影响油桐的生长和产量,给我国的油桐种植及产业发展带来了毁灭性损失。
油桐病害种类繁多,主要以真菌性病害为主,其中被报道最典型、发病较严重的包括由尖镰孢Fusarium oxysporum Schltdl.引起的油桐枯萎病(又称“桐瘟”) (Zhang et al. 2021);由小新壳梭孢Neofusicoccum parvum (Pennycook & Samuels) Crous et al. 引起的叶枯病(袁志林等 2011);由茄病新孢子菌Neocosmospora solani (Mart.) L. Lombard & Crous引起的油桐根腐病(陈守常和肖育贵 1990);由油桐尾孢Cercospora aleuritis I. Miyake和小新壳梭孢Neofusicoccum parvum (Pennycook & Samuels) Crous侵染所致的油桐黑斑病等(陈鹏等 2006;徐红梅等 2020),由菲律宾灵芝Ganoderma philippii (Bres. & Henn. ex Sacc.) Bres.引起的根腐病(Yuan et al. 2023),其中,油桐根腐病依赖于土壤传播,会造成根部腐烂,使根部吸收养分和水分的功能逐渐丧失,最后导致整株死亡,严重发生时会导致大面积油桐林枯萎,对油桐产业的可持续发展造成巨大影响。
笔者在2023年至2024年对贵州省独山县百泉镇油桐主产区实地调研中发现油桐受到某种土传病原菌的侵害,在根部组织可明显看到红色物质沉积,发病后期表现为叶片枯黄、根部腐烂,进而影响坐果率和产量。这些发病症状与已报道的油桐真菌病害存在一定区别,并且多年的栽培可能导致在局部区域传播一些新的病原物。为此,本研究对健康油桐和发病油桐的根际土壤样品进行真菌群落组成分析,通过病原菌分离纯化,采用病原菌形态学特征和分子生物学方法鉴定病原菌的种类,旨在为油桐病害的防控提供依据。
于2024年7月在贵州省独山县百泉镇油桐主产区采集具有典型根腐病症状的油桐根茎部样品,同时采集健康油桐和发病油桐的根际土壤。根茎部样品存放在封口袋中,置于低温保鲜盒,带回实验室通过组织分离法进行病原菌的分离纯化工作;根际土壤从油桐根部获得,首先去除根表面的松散土壤,将含有一层薄的剩余土壤聚集体的根转移到100 mmol/L磷酸盐缓冲溶液(PBS, pH 7.2)中,置于50 mL无菌离心管中以最大速度涡旋20 s,使其释放大部分根际土壤,将得到的混浊溶液在4 ℃、6 000×g离心15 min,形成的沉淀即为根际土壤样品。
利用E.Z.N.A.®土壤DNA提取试剂盒提取健康油桐和发病油桐的根际土DNA样品,分别设置3个重复。按照如下反应体系进行PCR扩增:98 ℃预变性30 s;98 ℃变性10 s,54 ℃退火30 s,72 ℃延伸45 s,共32个循环;最后在72 ℃下延伸10 min,再由AMPure XT beads和Qubit进行PCR产物的纯化和定量。对纯化后的PCR产物使用Agilent 2100生物分析仪和Illumina的文库定量试剂盒进行评估,合格的文库浓度应在2 nmol/L 以上,使用NovaSeq 6000测序仪进行2×250 bp的双端测序,相应试剂为NovaSeq 6000 SP Reagent Kit (500 cycles)。对测序获得的双端数据,根据barcode信息对样品进行数据拆分,并去除接头和barcode序列,进行数据拼接、过滤和DADA2去噪等处理,质控后产生的每个去重序列称为ASVs (amplicon sequence variants)。根据ASV序列文件采用SILVA和NT-16S数据库进行物种注释,并根据ASV丰度表对各层级物种在各样本中的相对丰度进行统计,基于得到的物种相对丰度统计信息,进行各比较组之间的差异分析。
采集的发病油桐根茎部的病健交界处带回实验室后,用流动的自来水将表面的颗粒污染物清洗干净,分离其主根和茎部的韧皮部、木质部及侧根,切成0.5 cm × 0.5 cm的组织块,在75%乙醇中浸泡约30 s,随后在1%次氯酸钠溶液中表面消毒2 min,最后用无菌蒸馏水清洗多次(3次以上),并用无菌滤纸吸干水分,移植于麦芽粉琼脂培养基(malt extract agar, MEA)平板上,28 ℃黑暗培养。培养基中预先加入硫酸链霉素(50 mg/L)和盐酸四环素(20 mg/L)以防止细菌的污染。培养4 d后,将从组织块边缘长出的菌丝转接至新鲜马铃薯葡萄糖琼脂培养基(potato dextrose agar, PDA)中进行纯化,培养7 d后,4 ℃冰箱保存备用。
参考油桐枯萎病的致病模型(杨素素等 2014;Zhang et al. 2018)进行致病性检测。取在PDA培养基上生长良好的用于致病性测定的菌块,将其接种于合成低营养琼脂培养基(synthetic low nutrient medium, SNA),28 ℃培养7-10 d,在平板上加少量无菌水,用灭菌的软毛刷将孢子刷下,并用3层纱布过滤掉多余的培养基,配制浓度为2×106个/mL的孢子悬浮液。挑取生长良好的含有4-5片叶子的油桐一年生幼苗,将根周围的土小心拂去,用大头针将主根和侧根轻轻划几个伤口后,浸泡于孢子悬浮液中,以等量无菌水为对照,每个处理5株苗。浸泡30 min后将植株重新种植于无菌土中,置于28 ℃的人工气候箱中培养,光照周期设置为16 h光照,8 h黑暗,光照强度为5 000 lx。定期观察油桐幼苗的发病情况,分别从接种后发病的根茎部重新分离病原菌并进行柯赫法则验证。
将分离纯化获得的菌株接种到PDA培养基上,在28 ℃恒温培养箱内黑暗培养5-7 d,观察和记录菌落的形态特征;接种在SNA产孢培养基培养8 d后,使用0.85%生理盐水清洗菌落平板并在光学显微镜下(ZEISS LSM 900 with Airyscan 2, Carl Zeiss)观察菌丝和分生孢子形态。
采用CTAB法提取病原菌的总DNA,参考White et al. (1990)的方法扩增核糖体转录间隔区(ITS),引物为ITS1-F (5′-TCCGTAGGTGAAC CTGCGG-3′)和ITS4 (5′-TCCTCCGCTTATTGAT ATGC-3′);采用RPB2-5f (5′-GAYGAYMGWGA TCAYTTYGG-3′)、RPB2-7cr (5′-CCCATRGCTT GYTTRCCCAT-3′)引物对扩增RNA聚合酶次大亚基(rpb2) (Liu et al. 1999);采用引物EF1-983F (5′-GCYCCYGGHCAYCGTGAYTTYAT-3′)和EF1- 2218R (5′-ATGACACCRACRGCRACRGTYTG-3′)扩增翻译延长因子1-a (tef1-a) (Rehner & Buckley 2005)的基因片段;参考Quaedvlieg et al. (2011)的方法扩增钙调蛋白(CaM),引物为CAL-228f (5′-GAGTTCAAGGAGGCCTTCTCCC-3′)和CAL- 2Rd (5′-TGRTCNGCCTCDCGGATCATCTC-3′);参考Woudenberg et al. (2009)的方法扩增β-微管蛋白(tub2),引物为T1 (5′-AACATGCGTGAGAT TGTAAGT-3′)和TUB4RD (5′-CCRGAYTGRCC RAARACRAAGTTGTC-3′)。
扩增上述基因的PCR体系反应条件为95 ℃预变性5 min;95 ℃变性30 s,58 ℃退火30 s,72 ℃延伸2.5 min,共35个循环。PCR产物经过1 %的琼脂糖凝胶电泳检测后,切胶送交杭州有康生物科技有限公司进行测序。
首先根据上述5个基因进行BLAST比对分析,确定大致的分类地位;根据Waalwijk et al. (2018)发表的有关镰孢属详细的系统发育关系,从NCBI数据库下载了28个与镰孢属菌株AX-2亲缘关系相近的基因序列。基于邻接法(neighbor- joining, NJ)构建多基因系统发育树,以确定病原菌种类。
病害发生时间一般在3月至4月开始,5月至7月出现发病高峰期,之后发病趋缓。病原菌沿土壤传播,菌丝定殖于油桐根部表皮,进一步深入木质部导致根部腐烂,并沿着根部维管束组织向上传播至根茎连接处,病斑沿茎进一步扩展,严重时树体叶片萎蔫、失水褪绿、叶片枯黄,最终导致整个植株的坏死(图 1),与健康油桐相比,在发病油桐根部组织可明显看到有红色物质沉积(图1B, 1C)。
对健康油桐(Healthy)和发病油桐(Diseased)的根际土壤进行高通量测序,得到的原始读序、有效读序、门和属水平的聚类数量见表1,其中,有效读序占原始读序的比例均超过了 90%,进一步地,利用SPSS 22.0对健康油桐和发病油桐根际土壤的ASV相对丰度进行统计分析,结果表明,两者在真菌ASV相对丰度上具有显著差异(图2A)。
在门水平上,去除未分类的真菌门类后,Healthy和Diseased样本分别聚类到6门和8门,门水平上的物种组成见图2B,两者的主要优势菌门均为子囊菌门Ascomycota、担子菌门Basidiomycota、接合菌门Zygomycota,但这3个门在Healthy和Diseased样本中的相对丰度却不同。子囊菌门、担子菌门、接合菌门在Healthy、Diseased样本中的相对丰度占比分别为56.59%和73.26%;38.37%和13.31%;4.81%和11.46%。与Healthy样本相比,在Diseased样本中子囊菌门和接合菌门相对丰度占比上升了 16.67%和6.65%,担子菌门相对丰度占比下降了25.06%。
在属水平上,Healthy和Diseased样本共检测到138和209属,Diseased样本中属的数量有所增加。根据Healthy和Diseased样本的ASV相对丰度值,选取相对丰度TOP30的真菌属,比较在2个样本之间的相对丰度差异(图2C),其中,Healthy样本中生赤壳属Bionectria相对丰度占比21.66%、隐球酵母属Cryptococcus相对丰度占比17.43%、肉座菌属Hypocrea相对丰度占比8.42%、被孢霉属Mortierella相对丰度占比5.22%,而在Diseased样本中肉座菌属Hypocrea相对丰度占比13.11%、丛赤壳属Nectria相对丰度占比12.00%、被孢霉属Mortierella相对丰度占比7.86%、隐球酵母属Cryptococcus相对丰度占比5.35%,与Healthy样本相比,丛赤壳属Nectria、肉座菌属Hypocrea、被孢霉属Mortierella在Diseased样本中的相对丰度占比分别上升了11.98%、9.65%、4.77%,而生赤壳属Bionectria和隐球酵母属Cryptococcus相对丰度占比下降了24.01%、18.92%。需要特别关注的是,能广泛引起植物病害的镰孢属Fusarium只在发病油桐根际土壤菌群中出现,相对丰度占比为2.44%。
进一步分析Healthy和Diseased样本在TOP30属水平上相对丰度的差异,结果表明,Diseased样本中肉座菌属Hypocrea、镰孢属Fusarium、角担菌属Ceratobasidium、红酵母属RhodotorulaConlarium属的相对丰度极显著高于Healthy样本,而生赤壳属Bionectria的相对丰度显著低于Healthy样本(图2D)。
将病害发生地采集到的病株,分离其病健交界处的主根和茎部的木质部、韧皮部及侧根,在MEA培养基上培养3-5 d后即可见菌丝在边缘切口处长出,经PDA培养基纯化后共获得10个纯培养物(图3),通过形态学和ITS初步鉴定,有6个纯培养物属于木霉属Trichoderma真菌,它们分别是Trichoderma pubescens (AX-1、AP-2)、螺旋木霉T. spirale (AP-1)、云南木霉T. yunnanense (FR-3)、拟康宁木霉T. koningiopsis (AP-3)、深绿木霉T. atroviride (AX-3),有4个纯培养物属于镰孢属Fusarium真菌,分别是F. pseudoanthophilum (FR-1、FR-2、AR-1)和F. foetens (AX-2)。根据笔者所在课题组的相关研究,油桐根际土壤中的木霉菌可增强植物的抗病能力,包括T. koningiopsisT. spiraleT. atrovirideT. harzianum,对油桐尖镰孢均具有拮抗作用,其中,T. koningiopsis对尖镰孢的抑菌效果最好(梁晓洁等 2020)。结合田间观察,发病植株根部可明显看到红色物质沉积,因此,我们重点研究在培养基上表现为红色色素沉积的4个镰孢属菌株(图3),包括菌株FR-1、FR-2、AX-2、AR-1。
利用这4种菌株进行致病性测定(回接试验),结果显示,只有菌株AX-2能引起根部腐烂、叶片变黄枯萎(图4)。与未接种真菌的对照油桐相比,接种第1天,植株的表型没有明显变化;接种第4天,叶片开始有萎蔫的症状,茎干没有变化;接种第8天,叶片萎蔫更加严重,茎干开始变软,侧根根系已经腐烂;接种第11天,根部腐烂,茎干变软,根茎连接处的颜色已变为深褐色,叶片变为暗褐色并且叶片开始脱落,而对照植株始终保持健康状态。对发病植株的根茎部以及叶片重新分离获得病原菌,与田间采集分离的病原菌形态特征一致,进一步通过分子鉴定确定了该菌株与AX-2是同一种菌株。
菌株AX-2在PDA培养基上生长较快,培养7 d后菌落直径为8.6 cm,呈毡状,颜色为粉色(图 5A),气生菌丝致密;光学显微镜下可观察到大、小型分生孢子(图 5B)及分生孢子梗(图 5C),其中,大型分生孢子呈镰刀状,具隔膜,有产孢梗,大小为1.6-4.2 × 21.0-42.0 μm;小型分生孢子呈卵形或肾形,单胞,大小为1.3-3.4 × 5.8-10.9 μm,这些形态特征与镰孢属Fusarium的描述相符。
利用ITS、tef1-arpb2CaMtub2的引物对菌株AX-2的ITS、tef1-arpb2CaMtub2基因进行扩增,经测序分析,确定片段长度分别为530、993、1 175、594、532 bp。将ITS、tef1-arpb2CaMtub2基因序列提交到NMDC (http://nmdc.cn),核酸序列编号分别为NMDCN- 0008I06、NMDCN0008I07、NMDCN0008I08、NMDCN0008QF3、NMDCN0008QF4。基于ITS、tef1-arpb2CaMtub2联合基因序列,采用邻接法构建菌株AX-2以及GenBank中同源性较高的28种Fusarium属近缘菌株的系统发育树,结果显示,菌株AX-2与臭镰孢F. foetens聚为一支,支持率达到100% (图6)。
在分类学上,镰孢属隶属于真菌界、子囊菌门、粪壳菌纲、肉座菌目、丛赤壳科,是农作物及经济作物最重要的寄生病原真菌之一,其寄主范围广泛,可引起瓜类、茄科、香蕉、棉、豆科及花卉等100多种植物枯萎病(林文凤等 2024)。油桐是我国重要的木本生物质能源树种,它的栽培生产也易受到镰孢属病原菌的影响。目前,报道最多的是由尖镰孢油桐专化型Fusarium oxysporum f. sp. fordii (Fof-1)引起的油桐枯萎病害(又称“桐瘟”) (Zhang et al. 2018)和茄病新孢子菌Neocosmospora solani引起的油桐根腐病(陈守常和肖育贵 1990),其中,关于引起油桐根腐病的病原菌的报道只限于腐皮新赤壳菌,而且相关的研究多集中于油桐根腐病的发生与防治(宋强和赵伟 2017),国内外未见腐皮新赤壳菌引起的油桐根腐病致病机理的研究报道。
我们的研究首次在油桐根腐病的组织中分离到臭镰孢F. foetens,油桐幼苗接种病原菌F. foetens后,可导致根部褐变腐烂、叶片变黄枯萎直至整个植株死亡,该菌最初是在秋海棠属Begonia植物中发现的,可导致植物茎基部腐烂,维管束变色、叶脉褪绿、叶片发黄萎蔫(Elmer et al. 2004;Schroers et al. 2004),这与病原菌F. foetens导致的油桐根腐病的发病症状具有一定的相似性。近年来,已有文献报道该菌也可侵染其他植物,梁忠厚和李静纳(2021)发现,F. foetens是引起湖南多花黄精Polygonatum cyrtonema根腐病的主要致病菌之一;杨阳等(2024)的研究明确了F. foetens是党参Codonopsis pilosula根腐病的主要病原菌之一;在薰衣草Lavandula angustifolia中也首次报道了由F. foetens引起的根腐病(Wei et al. 2023)。
本研究对健康油桐(Healthy)和发病油桐(Diseased)的根际土壤进行测序,分析了在门水平和属水平上的土壤菌群组成,我们发现,健康油桐和发病油桐的主要优势菌门和菌属组成相近,但相对丰度却大不相同,这可能是导致油桐根腐病发生的原因之一。子囊菌门Ascomycota在Diseased样本相对丰度占比为73.26%,且比Healthy样本中的占比增加了16.67%,这可能与该类群包含大量腐生真菌有关(邓娇娇等 2019),油桐患病后,根系内和根际土壤中的腐生真菌增加,导致子囊菌门的相对丰度增加,研究表明,子囊菌门在烟草Nicotiana tabacum镰孢根腐病根际土壤的菌群占比较高,均值68.29%-73.44% (何晓冰等 2025);子囊菌门也是油茶Camellia oleifera根腐病中的优势菌门(杨娅琳等 2024)。同时,担子菌门Basidiomycota在Healthy样本中的相对丰度增加了25.06%,猜测可能是该类群可与植物共生形成菌根,有利于植物生长发育(Miguel et al. 2020)。与Healthy相比,丛赤壳属Nectria、被孢霉属Mortierella在Diseased样本中的相对丰度占比上升了11.98%、4.77%,被孢霉属由于其超强的纤维素分解能力,可能在抵抗根腐病中发挥作用(Papanikolaou & Aggelis 2019),已报道被孢霉属是三七Panax notoginseng植物黄腐植株中的优势菌群(丁子元等 2024),在猕猴桃Actinidia chinensis根腐病植株根际土壤中,被孢霉属的相对丰度明显低于未感病植株(陈海生等 2024),而谢珺等(2022)的研究表明,灯盏花Erigeron breviscapus根腐病病原主要集中在丛赤壳属。在我们的研究中,作为能导致多种植物根腐病发生的镰孢属(Chang et al. 2018)只在发病油桐根际土壤菌群中出现,相对丰度占比为2.44%。
根据我们在2023-2024年对贵州省独山县百泉镇油桐主产区发病油桐的连续观察,该病一般3月至4月开始发病,5月至7月出现发病高峰期,之后发病趋缓,因此,在发病潜伏期和高峰期要特别注意病原菌F. foetens的预防与防治。在农业防治方面,首先选用品质好、抗病抗逆能力强的油桐品种;其次,在种植过程中要避免积水,深翻土壤,保持土壤透气良好,并做好水肥管理;最后,枯死病株应挖除病根烧毁,病土用石灰消毒(宋强和赵伟 2017)。在化学防治方面,定植前要做好种子消毒、土壤消毒以及药剂防治(刘燕 2024)。
本研究通过健康油桐和发病油桐的根际土壤菌群组成分析、形态学观察、分子鉴定以及致病性测定等研究,明确了引起贵州省独山县百泉镇油桐主产区油桐根腐病的病原菌为臭镰孢F. foetens,这是首次在油桐根部发现的由该病菌引起的根腐病,研究结果可为该病害的发生流行和综合防治提供参考。
王嘉:实验实施、数据分析、论文撰写及修改;杨安仁:调查采样、数据管理;刘北平:数据分析、协助实验;高暝:调查采样、仪器使用指导及数据分析;赵耘霄:调查采样、软件应用及多样性分析;陈益存:论文修改、实验指导;汪阳东:选题、试验方案设计、论文修改。
该研究不存在任何潜在利益冲突的商业或财务关系。
  • 国家自然科学基金(32572095)
  • 浙江省万人计划科技创新领军人才项目(2022R52028)
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2026年第45卷第2期
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doi: 10.13346/j.mycosystema.250162
  • 接收时间:2025-06-03
  • 首发时间:2026-04-29
  • 出版时间:2026-02-22
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  • 收稿日期:2025-06-03
  • 录用日期:2025-10-07
基金
National Natural Science Foundation of China(32572095)
国家自然科学基金(32572095)
Ten Thousand People Plan of Science and Technology Innovation Leading Talent of Zhejiang, China(2022R52028)
浙江省万人计划科技创新领军人才项目(2022R52028)
作者信息
    1 中国林业科学研究院亚热带林业研究所 全省林木育种重点实验室,浙江 杭州 311400
    2 南京林业大学,江苏 南京 210037
    3 贵州鸿发生态农业科技有限责任公司,贵州 黔南 558200

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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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