Article(id=1266801747608621235, tenantId=1146029695717560320, journalId=1266358746360856629, issueId=1266801713492153305, articleNumber=null, orderNo=null, doi=10.13324/j.cnki.jfcf.202506014, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1750089600000, receivedDateStr=2025-06-17, revisedDate=1756828800000, revisedDateStr=2025-09-03, acceptedDate=null, acceptedDateStr=null, onlineDate=1779958672668, onlineDateStr=2026-05-28, pubDate=1763136000000, pubDateStr=2025-11-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1779958672668, onlineIssueDateStr=2026-05-28, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1779958672668, creator=13701087609, updateTime=1779958672668, updator=13701087609, issue=Issue{id=1266801713492153305, tenantId=1146029695717560320, journalId=1266358746360856629, year='2025', volume='45', issue='6', pageStart='570', pageEnd='672', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=1, specialIssue=null, createTime=1779958664534, creator=13701087609, updateTime=1779959653473, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1266805862552199994, tenantId=1146029695717560320, journalId=1266358746360856629, issueId=1266801713492153305, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1266805862552199995, tenantId=1146029695717560320, journalId=1266358746360856629, issueId=1266801713492153305, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=649, endPage=656, ext={EN=ArticleExt(id=1266801749248594101, articleId=1266801747608621235, tenantId=1146029695717560320, journalId=1266358746360856629, language=EN, title=Identification of pathogens causing branch canker on Cinnamomum chekiangense, columnId=1266801745758937344, journalTitle=Journal of Forest and Environment, columnName=Forest protection, runingTitle=null, highlight=null, articleAbstract=

Canker was a newly found disease causing branch canker, branch dieback, and even death of Cinnamomum chekiangense. The study aimed to identify the causal agents of branch canker on C. chekiangense, thus providing theoretical support for forecasting and controlling this disease scientifically and effectively in practice. A total of 15 fungal isolates were obtained from diseased samples via the tissue isolation method. Pathogenicity assays demonstrated that strains JYZJZ2 and JYZJZ6 could infect the branches of C. chekiangense and reproduce typical canker symptoms, while they showed a difference in pathogenicity, with JYZJZ2 showing stronger pathogenicity. JYZJZ2 and JYZJZ6 were identified as Lasiodiplodia pseudotheobromae and L.iranensis, respectively, by comparing the DNA sequence data of internal transcribed spacer (ITS), translation elongation factor 1-alpha (tef1-α), β-tubulin (tub2), and largest subunit of RNA polymerase Ⅱ (rpb2) gene regions using the PhyloSuite platform, and by Ⅱ the morphological features. This is the first report of L. pseudotheobromae and L. iranensis causing canker on C. chekiangense worldwide.

, correspAuthors=Xiaozhen YE, authorNote=null, correspAuthorsNote=null, copyrightStatement=Copyright ©2025 Journal of Forest and Environment. All rights reserved., copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Quanzhu CHEN, Senhui HAN, Zhongpeng JIANG, Weican LI, Youwei WU, Xiaozhen YE), CN=ArticleExt(id=1266801750322335937, articleId=1266801747608621235, tenantId=1146029695717560320, journalId=1266358746360856629, language=CN, title=浙江樟枝干溃疡病病原鉴定, columnId=1266801745935098114, journalTitle=森林与环境学报, columnName=森林保护学, runingTitle=null, highlight=null, articleAbstract=

为科学有效地指导浙江樟(Cinnamomum chekiangense)枝干溃疡病的监测预报与防治, 通过组织分离法辅以致病性测定验证, 从病样组织中分离获得2株能够侵染浙江樟枝干引起典型溃疡病症状的致病菌株JYZJZ2和JYZJZ6, 基于PhyloSuite平台进行菌株核糖体转录间隔区(ITS)、翻译延伸因子1-α (tef1-α)、β-微管蛋白(tub2)和RNA聚合酶Ⅱ大亚基(rpb2)多个基因联合系统发育分析, 结合形态学特征描述, 对病原种类进行分类鉴定, 明确其分类地位。结果表明, 供试菌株JYZJZ2和JYZJZ6分别与假可可毛色二孢(Lasiodiplodia pseudotheobromae)和伊朗毛色二孢(L. iranensis)的形态特征基本一致, 亲缘关系相近, 分别将其鉴定为假可可毛色二孢和伊朗毛色二孢, 该致病菌能够侵染浙江樟引发溃疡病属首次发现。致病性测定结果还表明, 假可可毛色二孢JYZJZ2菌株与伊朗毛色二孢JYZJZ6菌株接种后致病力存在明显差异, JYZJZ2菌株致病力较强, 而JYZJZ6菌株致病力较弱。

, correspAuthors=叶小真, authorNote=null, correspAuthorsNote=
叶小真(1983-), 女, 副教授, 从事林木病理学研究。Email:
, copyrightStatement=版权所有©《森林与环境学报》编辑部2025, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=xZi5/hYbWMtlbdeAI7yXQQ==, magXml=oO11EP8o5Me+q8LpDtPWMw==, pdfUrl=null, pdf=CF+2uDi8SYVdZYqlwvIazg==, pdfFileSize=6572997, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=N8e85gb0W3+GeEZDPxaA4Q==, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=dYaRvnL+K8vwBUpiJnfw1Q==, mapNumber=null, authorCompany=null, fund=null, authors=

陈全助(1981-), 男, 副教授, 从事林木病理学研究。Email:

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陈全助(1981-), 男, 副教授, 从事林木病理学研究。Email:

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注:A. 受害部位;B. 受害部位树皮开裂;C. 产生子实体;D~E. 受害部位肿胀呈瘤状;F. 植株枯萎,箭头指示病斑环绕枝干部位。

, figureFileSmall=yXY3nRqwOrwn+MUsehItZw==, figureFileBig=N8e85gb0W3+GeEZDPxaA4Q==, tableContent=null), ArticleFig(id=1266801759415587070, tenantId=1146029695717560320, journalId=1266358746360856629, articleId=1266801747608621235, language=EN, label=Fig.2, caption=Symptom development of Cinnamomum chekiangense inoculated with JYZJZ2 and JYZJZ6, figureFileSmall=/o6GPWBwUpLx4FB7ddVDVA==, figureFileBig=1Qa4sw5sTpdZtGyAZtpxLQ==, tableContent=null), ArticleFig(id=1266801759503667455, tenantId=1146029695717560320, journalId=1266358746360856629, articleId=1266801747608621235, language=CN, label=图2, caption=接种溃疡病菌浙江樟症状发展

注:A. 对照;B. 接种JYZJZ2菌株;C. 接种JYZJZ6菌株。

, figureFileSmall=/o6GPWBwUpLx4FB7ddVDVA==, figureFileBig=1Qa4sw5sTpdZtGyAZtpxLQ==, tableContent=null), ArticleFig(id=1266801759574970624, tenantId=1146029695717560320, journalId=1266358746360856629, articleId=1266801747608621235, language=EN, label=Fig.3, caption=Morphological characteristics of L. pseudotheobromae, figureFileSmall=dMeSP7cig1fL61AvXNbWuw==, figureFileBig=KgjAHmq7fLfPYC8IQYEZHA==, tableContent=null), ArticleFig(id=1266801759780491521, tenantId=1146029695717560320, journalId=1266358746360856629, articleId=1266801747608621235, language=CN, label=图3, caption=假可可毛色二孢的形态特征

注:A~B. PDA培养基前后期正背面菌落形态;C. 分生孢子器;D~E. 分生孢子、产孢细胞及侧丝;F. 分生孢子着生产孢细胞;G~H. 无隔分生孢子;I. 具1隔膜分生孢子;J. 具1隔膜和纵条纹分生孢子。

, figureFileSmall=dMeSP7cig1fL61AvXNbWuw==, figureFileBig=KgjAHmq7fLfPYC8IQYEZHA==, tableContent=null), ArticleFig(id=1266801759839211778, tenantId=1146029695717560320, journalId=1266358746360856629, articleId=1266801747608621235, language=EN, label=Fig.4, caption=Morphological characteristics of L. iranensis, figureFileSmall=9ajo4grdW1/OCf1EvDS0Lw==, figureFileBig=zcj9o2XeipzSx8UOC2j1Ug==, tableContent=null), ArticleFig(id=1266801759914709251, tenantId=1146029695717560320, journalId=1266358746360856629, articleId=1266801747608621235, language=CN, label=图4, caption=伊朗毛色二孢的形态学特征

注:A~B. PDA培养基前后期正反面菌落形态;C. 分生孢子器;D~E. 分生孢子、产孢细胞及侧丝;F~G. 无隔分生孢子;H. 具1隔膜分生孢子;I. 具1隔膜和纵条纹分生孢子。

, figureFileSmall=9ajo4grdW1/OCf1EvDS0Lw==, figureFileBig=zcj9o2XeipzSx8UOC2j1Ug==, tableContent=null), ArticleFig(id=1266801759986012420, tenantId=1146029695717560320, journalId=1266358746360856629, articleId=1266801747608621235, language=EN, label=Fig.5, caption=Maximum likelihood tree based on multi-gene combination of ITS-tef1-α-tub2-rpb2, figureFileSmall=OytFNorM2N71h3XYdLnw7w==, figureFileBig=oT0FdkF6xaPGfsmBLTRmcw==, tableContent=null), ArticleFig(id=1266801760065704197, tenantId=1146029695717560320, journalId=1266358746360856629, articleId=1266801747608621235, language=CN, label=图5, caption=基于ITS-tef1-α-tub2-rpb2多基因系统发育分析最大似然树

注:该发育树以平截色二孢为外群;图中分支处数值分别代表SH近似似然比检验(左侧) 和超快自展支持率(右侧)。

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浙江樟枝干溃疡病病原鉴定
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陈全助 1 , 韩森汇 1, 2 , 江仲鹏 1, 3 , 李伟灿 1 , 吴有为 1 , 叶小真 4, *
森林与环境学报 | 森林保护学 2025,45(6): 649-656
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森林与环境学报 | 森林保护学 2025, 45(6): 649-656
浙江樟枝干溃疡病病原鉴定
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陈全助1 , 韩森汇1, 2, 江仲鹏1, 3, 李伟灿1, 吴有为1, 叶小真4, *
作者信息
  • 1.福建农林大学林学院, 福建 福州 350002
  • 2.凉山彝族自治州林业草原科学研究院, 四川 西昌 615000
  • 3.绵阳市安州区自然资源局, 四川 绵阳 622600
  • 4.福建农林大学金山学院, 福建 福州 350002
  • 陈全助(1981-), 男, 副教授, 从事林木病理学研究。Email:

通讯作者:

叶小真(1983-), 女, 副教授, 从事林木病理学研究。Email:
Identification of pathogens causing branch canker on Cinnamomum chekiangense
Quanzhu CHEN1 , Senhui HAN1, 2, Zhongpeng JIANG1, 3, Weican LI1, Youwei WU1, Xiaozhen YE4, *
Affiliations
  • 1.College of Forestry, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China
  • 2.Liangshan Yi Autonomous Prefecture Academy of Forestry and Grassland Sciences, Xichang, Sichuan 615000, China
  • 3.Natural Resources Bureau of Anzhou District, Mianyang, Sichuan 622600, China
  • 4.Jinshan College, Fujian Agriculture and Forestry University, Fuzhou, Fujian 350002, China
出版时间: 2025-11-15 doi: 10.13324/j.cnki.jfcf.202506014
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为科学有效地指导浙江樟(Cinnamomum chekiangense)枝干溃疡病的监测预报与防治, 通过组织分离法辅以致病性测定验证, 从病样组织中分离获得2株能够侵染浙江樟枝干引起典型溃疡病症状的致病菌株JYZJZ2和JYZJZ6, 基于PhyloSuite平台进行菌株核糖体转录间隔区(ITS)、翻译延伸因子1-α (tef1-α)、β-微管蛋白(tub2)和RNA聚合酶Ⅱ大亚基(rpb2)多个基因联合系统发育分析, 结合形态学特征描述, 对病原种类进行分类鉴定, 明确其分类地位。结果表明, 供试菌株JYZJZ2和JYZJZ6分别与假可可毛色二孢(Lasiodiplodia pseudotheobromae)和伊朗毛色二孢(L. iranensis)的形态特征基本一致, 亲缘关系相近, 分别将其鉴定为假可可毛色二孢和伊朗毛色二孢, 该致病菌能够侵染浙江樟引发溃疡病属首次发现。致病性测定结果还表明, 假可可毛色二孢JYZJZ2菌株与伊朗毛色二孢JYZJZ6菌株接种后致病力存在明显差异, JYZJZ2菌株致病力较强, 而JYZJZ6菌株致病力较弱。

浙江樟  /  溃疡病  /  病原鉴定  /  毛色二孢属

Canker was a newly found disease causing branch canker, branch dieback, and even death of Cinnamomum chekiangense. The study aimed to identify the causal agents of branch canker on C. chekiangense, thus providing theoretical support for forecasting and controlling this disease scientifically and effectively in practice. A total of 15 fungal isolates were obtained from diseased samples via the tissue isolation method. Pathogenicity assays demonstrated that strains JYZJZ2 and JYZJZ6 could infect the branches of C. chekiangense and reproduce typical canker symptoms, while they showed a difference in pathogenicity, with JYZJZ2 showing stronger pathogenicity. JYZJZ2 and JYZJZ6 were identified as Lasiodiplodia pseudotheobromae and L.iranensis, respectively, by comparing the DNA sequence data of internal transcribed spacer (ITS), translation elongation factor 1-alpha (tef1-α), β-tubulin (tub2), and largest subunit of RNA polymerase Ⅱ (rpb2) gene regions using the PhyloSuite platform, and by Ⅱ the morphological features. This is the first report of L. pseudotheobromae and L. iranensis causing canker on C. chekiangense worldwide.

Cinnamomum chekiangense  /  canker  /  pathogen identification  /  Lasiodiplodia
陈全助, 韩森汇, 江仲鹏, 李伟灿, 吴有为, 叶小真. 浙江樟枝干溃疡病病原鉴定. 森林与环境学报, 2025 , 45 (6) : 649 -656 . DOI: 10.13324/j.cnki.jfcf.202506014
Quanzhu CHEN, Senhui HAN, Zhongpeng JIANG, Weican LI, Youwei WU, Xiaozhen YE. Identification of pathogens causing branch canker on Cinnamomum chekiangense[J]. Journal of Forest and Environment, 2025 , 45 (6) : 649 -656 . DOI: 10.13324/j.cnki.jfcf.202506014
浙江樟(Cinnamomum chekiangense) 属樟科(Lauraceae) 樟属(Cinnamomum) 常绿乔木,系地带性常绿阔叶林建群或伴生树种,亦为我国特有珍稀树种[1-2]。浙江樟木材材质优良,树形优美端庄,植株富含芳香油,对二氧化硫(SO2) 等有害物耐受性较强,可分泌释放空气净化物质,是优良的用材、绿化、观赏、经济和生态防护树种[1-3]。2022年,本课题组在福建省南平市建阳区浙江樟种植区首次发现溃疡病为害,该病害多发生在枝条分杈部位,严重时病变部位瘤状肿大开裂致枝条枯萎,植株枯死,发病株率达100%,病情指数47.96。经形态观察和核糖体转录间隔区(internal transcribed spacer,ITS) 比对分析,初步判断浙江樟溃疡病可能由毛色二孢属真菌(Lasiodiplodia spp.) 引起,目前国内外尚无该病害及其病原鉴定的研究报道。毛色二孢属真菌属葡萄座腔菌科(Botryosphaeriaceae),是热带和亚热带地区多种植物病害重要的病原菌,因寄主广泛、形态多变,以及生长因子各异,导致该属真菌种间关键形态特征显著重叠,仅仅依靠形态特征难以实现其种间有效鉴定识别[4-5]。鉴于此,学者们应用形态特征识别和系统发育分析相结合的方法成功实现了毛色二孢属真菌的种间识别,然而,系统发育分析结果是否准确、可靠与所选基因种类和数量有关[5-6]。本研究应用PhyloSuite平台[7]对ITS、翻译延伸因子1-α (translation elongation factor 1-alpha,tef1-α)、β-微管蛋白(β-tubulin,tub2) 和RNA聚合酶Ⅱ大亚基(largest subunit of RNA polymerase Ⅱ,rbp2) 等基因序列进行多基因联合系统发育分析[6, 8],结合形态特征,对浙江樟溃疡病病原菌株进行鉴定,以期明确该病原菌的分类地位,为浙江樟溃疡病防治提供理论支撑。
2023年5月,从福建省南平市建阳区国有林场坤中场(27°28′14″N,118°23′10″E) 采集具有溃疡病典型症状的浙江樟枝干组织。
取受害浙江樟病健交界处枝干树皮切成2 mm×5 mm大小的组织块,依次经体积分数为75%的乙醇溶液消毒30 s,质量分数为1%的次氯酸钠溶液浸泡2 min,无菌水漂洗3次,无菌吸水纸吸干表面水分,备用。将消毒后的组织块平铺接种至马铃薯葡萄糖琼脂培养基(potato dextrose agar,PDA) 平板表面,28 ℃恒温箱避光培养,待其产生菌丝,挑取边缘菌丝转移接种至新PDA平板,重复转接至获得纯化菌落,依菌落形态特征进行编号保存。
选取经温室[室温28 ℃,光周期设置为14 h (光照):10 h (黑暗)] 预先适应性培养7 d、健康、长势一致的2 a生浙江樟盆栽苗(苗高60 cm,地径8 mm),枝干接种处用体积分数为75%的乙醇溶液消毒清洗后,取直径为6 mm的菌饼进行针刺有伤回接处理,脱脂棉蘸无菌水保湿,封口膜呈漏斗状缠绕固定,置温室培养观察发病情况,对发病植株再次取样分离,确定病原。以接种无菌PDA作对照(CK),每处理各重复3次。
将病原菌接种至PDA平板,28 ℃暗培养,逐日观察记录菌落变化。取菌株菌饼接种浙江樟枝干进行诱导产孢,待枝干表面形成子实体,徒手切片后使用尼康Nikon Ni-U显微镜镜检观察分生孢子器、侧丝和分生孢子等各种结构的形态特征,随机选择50个分生孢子测定大小。
刮取病原菌新鲜菌丝,使用SP Fungal DNA Kit试剂盒(OMEGA) 并参照其操作程序提取DNA。利用引物ITS1/ITS4、EF1-688F/EF1-1251R、Bt2a/Bt2b和RPB2-LasF/RPB2-LasR分别扩增ITS、tef1-αtub2和rbp2基因序列[9],聚合酶链式反应(polymerase chain reaction,PCR) 扩增参照LIANG et al[8]的操作方案进行。使用FastPure Gel DNA Extraction Mini Kit胶回收试剂盒(南京诺唯赞) 回收PCR产物,将回收的PCR产物连接到pMC100-MB载体(TaKaRa) 后,转化到大肠杆菌感受态细胞DH5α,筛选阳性克隆送生工生物工程(上海) 股份有限公司测序。测序所获序列经BioEdit软件检查修正后,上传提交至美国国家生物信息中心(National Center for Biotechnology Information,NCBI) 数据库进行BLAST (https://blast.ncbi.nlm.nih.gov/Blast.cgi) 比对分析,检索下载相似度和可信度较高、已发表菌株序列作为参考基因,结合本研究所获浙江樟溃疡病致病菌株序列进行系统发育分析。用PhyloSuite v1.2. 3软件平台构建系统发育树[7],该平台整合了多基因序列比对修剪、数据串联、进化模型筛选和系统发育树构建等分析功能,实现一站式流程化操作,便于系统发育分析初学者使用。具体步骤如下:首先,使用MAFFT功能选项进行基因序列的多重序列比对,随后利用TrimAI功能选项对结果进行修剪;接着,使用Concatenate Sequence功能选项将修剪后的序列按ITS-tef1-α-tub2-rpb2的顺序进行串联拼接;然后,采用IQ-TREE[10]功能选项,以平截色二孢(Diplodia mutila,CMW 7060) 为外群[8],基于最大似然法构建系统发育树,选择GTR+F+R5模型,并分别设置超快自举(Ultrafast Bootstrap) 和SH近似似然比检验(Shimodaira-Hasegawa approximate likelihood ratio test,SH-aLRT), 重复次数为1 000;最后,用FigTree v1.4. 4软件对生成的系统发育树进行读取注释。
随着雨季来临和气温升高,浙江樟溃疡病开始发生流行。病原菌最初主要从枝干分叉处易积水部位开始侵染(图 1AB),受害严重植株分布密集区其侵染位点可扩展至分杈处以外枝干部位(图 1C)。病斑初为黑褐色近圆形,随后逐渐加深变黑,呈梭形扩展至环绕枝干(图 1BCDE),潮湿环境下呈腐烂状凹陷,伴有红褐色或黑色子实体(图 1C)。随着病斑扩展变大,受害部位周围伴有愈伤组织异常增生,皮层突起(图 1BCE),树皮开裂,甚至肿大呈瘤状(图 1DE),输导组织病变造成水分和养分通路被阻断,病斑以上枝条逐渐枯死,严重时病斑遍布植株分杈处,树势明显衰弱,最终导致植株枯死(图 1F)。该病害林间蔓延速度快,短时间内造成林分成片枝干枯死。
采集的浙江樟溃疡病组织经分离纯化,获得菌株15株,编号分别为JYZJZ1~JYZJZ15。致病性测定结果表明,分离菌株中仅JYZJZ2和JYZJZ6能够侵染浙江樟引起溃疡病。接种溃疡病菌浙江樟症状发展结果(图 2) 表明,接种点初呈水渍状,近圆形;随后病斑不断扩大,呈近椭圆形或梭形,黑褐色,略凹陷,出现大量黑色子实体;后期病斑环绕枝干,病斑上部枝干逐步枯死。这与野外自然发病症状表现一致,接种发病部位可再次分离获得相应的接种菌,可确定该菌株为浙江樟溃疡病致病菌。同时,JYZJZ2和JYZJZ6菌株的致病力存在差异,JYZJZ2明显强于JYZJZ6。
JYZJZ2和JYZJZ6菌株的致病性、菌落形态和显微结构特征存在差异,从属不同分类群。
JYZJZ2菌株的形态特征如图 3所示。28 ℃暗培养48 h,菌落铺满培养皿,菌落正反面初为白色,气生菌丝绒毛状,较浓密发达(图 3A);后期菌落正反两面均呈灰黑色至深棕黑色,菌丝较初期更为致密、发达(图 3B)。浙江樟枝条接种JYZJZ2菌株发病后期,分生孢子器单生,单腔室,球形或近球形,初埋藏在寄生主枝干表皮下方,后突破表皮外露,呈深棕色至黑色颗粒状,个别表面附有菌丝,分生孢子器壁呈暗褐色(图 3C)。菌株侧丝呈丝状圆柱形,无色,透明,不具隔膜,少有分枝(图 3D~E)。初期未成熟分生孢子无色,透明,单胞不具隔膜,椭球形或卵球形,两端钝圆(图 3F~H);后期成熟分生孢子呈棕褐色,具1中横膈膜呈双胞,以及纵向条纹(图 3I~J);分生孢子大小(长×宽) 为(23.78~29.14) μm× (13.24~16.20) μm,均值为(26.46±2.68) μm× (14.72±1.48) μm (n=50)。未发现该病原菌株的有性态阶段。根据菌株菌落形态及其显微结构特征,可初步确定JYZJZ2菌株为假可可毛色二孢(L. pseudotheobromae) [5, 8, 11]
JYZJN6菌株的形态特征如图 4所示。PDA培养基菌落初期正反面亦均为白色,气生菌丝较稀薄(图 4A),后期两面均呈灰黑色,气生菌丝较初期稠密(图 4B)。菌株JYZJZ6接种浙江樟枝条发病后期,分生孢子器单生,初埋生枝条表皮下后突破表皮外露,深棕色至黑色,球形,大部分单腔室,壁厚,上被致密菌丝(图 4C)。侧丝圆柱形,无色透明,初无隔,成熟后有隔膜形成,几乎不分枝,偶尔基部或顶端细胞肿大,顶端细胞圆钝(图 4D~E)。初期未成熟分生孢子透明,单胞无隔膜,椭球形、近球形或近圆柱形,两端钝圆(图 4F~G);后期成熟分生孢子不断着色,呈深褐色,形成1中横膈膜,以及纵向条纹,双胞(图 4H~I);分生孢子大小(长×宽) 为(20.18~23.52) μm× (12.36~14.84) μm,均值为(21.85±1.67) μm× (13.60±1.24) μm (n=50)。未发现该病原菌株的有性态阶段。根据菌株菌落形态及其显微结构特征,初步确定JYZJN6菌株为伊朗毛色二孢(L. iranensis) [12-13]
基于ITS-tef1-α-tub2-rpb2多基因联合构建系统发育树,结果(图 5) 表明,菌株JYZJZ2与假可可毛色二孢(L. pseudotheobromae CBS 116459) 聚在同一分支,SH近似似然比检验和超快自举支持率值分别为97%和100%;菌株JYZJZ6与伊朗毛色二孢(L. iranensis IRAN 1520C) 聚在同一分支,SH近似似然比检验和超快自举支持率值分别为91%和98%。结合形态学特征和系统发育分析结果,最终将菌株JYZJZ2和JYZJZ6分别鉴定为假可可毛色二孢和伊朗毛色二孢。
毛色二孢属(Lasiodiplodia) 真菌隶属于葡萄座腔菌科(Botryosphaeriaceae) 无性态菌物类群,地理分布区域广[5-6, 8-9],可侵染多种木本植物造成植株根腐、叶斑枯、枝枯、梢枯、顶枯、流胶、溃疡、蒂腐和果腐等各种症状[5, 11-17],其中能够引起溃疡病的有假可可毛色二孢[5-6, 8, 11, 14-18]、可可毛色二孢(L. theobromae) [6, 14, 16-18]、伊朗毛色二孢[14, 16]、柑橘毛色二孢(L. citricola) [6, 14, 17]L. subglobosa[17]、巴西毛色二孢(L. brasiliense) [5, 17]、信阳毛色二孢(L. xinyangensis) [6]、黄岩毛色二孢(L. huangyanensis) [6, 14]、临海毛色二孢(L. linhaiensis) [14]、微小毛色二孢(L. microconidia) [14]、椪柑毛色二孢(L. ponkanicola) [14]、枣毛色二孢(L. ziziphi) [6]L. chiangraiensis[6]L. mitidjana[19]L. mediterranea[19]、核桃毛色二孢(L. regiae) [20]L. crassispora[21]、L. cinnamomi[22]等,致病菌种类之多,受害寄主涉及多种用材树种、经济树种和观赏树种[5-6, 8, 15-22],给农林业发展造成了不可估量的经济损失。
溃疡病是福建省南平市建阳区浙江樟种植过程中新出现的枝干病害。由于毛色二孢属真菌寄主种类丰富多样,生长环境各异,以致该属真菌的形态出现变异,形态特征有所差异,且存在形态特征重叠现象,因而仅凭形态特征识别难以得到准确可靠的鉴定结果[4-6]。目前,学者们已形成共识,将多基因联合系统发育分析结合传统形态学识别作为毛色二孢属真菌鉴定的有效手段[6, 9]。鉴于此,本研究借鉴了ZHANG et al[7]研发的PhyloSuite平台,成功构建了浙江樟溃疡病病原菌的ITS-tef1-α-tub2-rpb2多基因串联系统发育分析最大似然树,同时,基于形态学特征观察和系统发育关系分析,明确了浙江樟溃疡病主要由假可可毛色二孢(L. pseudotheobromae) 和伊朗毛色二孢(L. iranensis) 侵染引起,这是国内外关于浙江樟溃疡病病原菌的首次报道。
假可可毛色二孢(L. pseudotheobromae) 是全球农林业重要的强致病性病原菌,寄主范围广泛,在亚洲、大洋洲、美洲和非洲等地肆意侵染为害桉树(Eucalyptus spp.)、柑橘树(Citrus spp.)、橡胶树(Hevea brasiliensis)、台湾相思(Acacia confusa)、樱桃(Cerasus pseudocerasus)、楠木(Phoebe spp.)、樟树(Cinnamomum camphora) 和腰果(Anacardium occidentale) 等多种经济植物,引发溃疡病、枝枯病、梢枯病和叶斑病等多种病害[8, 11-13]。假可可毛色二孢通过茎干侵入橡胶树后,引发维管束褐变,短时间内造成植株叶片萎蔫下垂,继而枝条脱水、树皮干缩,最后植株枯萎[23]。与假可可毛色二孢相比,伊朗毛色二孢(L. iranensis) 的寄主谱相对较窄,但对特定树种造成的危害不容忽视。在伊朗,伊朗毛色二孢可侵染胡桃属(Juglans sp.)、柑橘属(Citrus sp.) 和杧果(Mangifera indica) 引起枝枯病和溃疡病[24]。在印度,伊朗毛色二孢能够侵染火龙果(Selenicereus spp.) 引起茎腐病,表现出比可可毛色二孢更强的致病力[13]。假可可毛色二孢、巴西毛色二孢和伊朗毛色二孢是引起印度主产区鳄梨(Persea americana) 根腐、茎溃疡与枝枯的主要病原菌,各病原菌之间的致病毒力差异显著,以假可可毛色二孢的致病毒力最高;受害植株表现出韧皮部和木质部维管组织褐变,以及根系腐烂,导致枝条枯死、植株萎蔫,病害发病率为20%~57%[5]
毛色二孢属溃疡病致病菌在不同寄主间存在优势种和致病力强弱不一的现象[6, 13, 16, 19]。本研究结果表明,假可可毛色二孢JYZJZ2菌株对浙江樟的致病力明显强于伊朗毛色二孢JYZJZ6菌株,与鳄梨溃疡病菌致病力的测定结果[5]相吻合,而张晓阳等[16]对樟树溃疡病菌的测定结果认为,两者之间的致病力差异不显著。病原菌的致病力强弱可能受地域、环境条件和寄主不同的影响,例如,本研究中弱致病性菌株JYZJZ6可与致病性菌株JYZJZ2以非致病性内生菌形式共存,这种致病力差异可能在寄主受外界胁迫时发生转变[5-6, 13]
野外病害调查发现,福建省南平市建阳区每年6—8月雨季来临,林间形成高温高湿环境条件,导致浙江樟溃疡病快速暴发,受害最为严重。同样,印度西高止山脉高温多雨环境为毛色二孢属真菌侵入鳄梨并快速扩散创造了有利条件,加剧病害流行为害[5]。可见,夏季多雨形成的高温高湿环境是促进浙江樟溃疡病暴发流行的主要因素。福建省气候温暖湿润,是浙江樟溃疡病菌适生分布区,病害的暴发流行潜在风险高。因此,在浙江樟栽培过程中,应根据树种生长习性合理控制林分密度,保持林分通风透气,降低林间温湿度,从营林措施角度预防溃疡病。
在浙江樟溃疡病被发现之前,国内外已陆续报道了毛色二孢属真菌侵染为害樟科(Lauraceae) 类群树种的情况,主要包括刨花润楠(Machilus pauhoi) 溃疡病(L. margaritacea) [25]、楠木枝枯病(L. pseudotheobromae) [15]、樟树溃疡病(L. pseudotheobromaeL. theobromaeL. iranensisL. cinnamomi) [16, 22]。这些病害分布地域相邻,寄主树种亲缘关系较近,且均为林业生产中的主要造林绿化树种,因此,存在较高的潜在交叉感染风险,应加强种苗产地检疫,杜绝使用带病种苗。
毛色二孢属真菌是热带和亚热带地区多种木本植物病害重要的病原菌。本研究结合多基因(ITS-tef1-α-tub2-rpb2) 串联的系统发育树分析与菌株形态特征观察,将引起浙江樟溃疡病的病原菌鉴定为假可可毛色二孢和伊朗毛色二孢,这是国内外关于毛色二孢属真菌可可毛色二孢和伊朗毛色二孢侵染为害浙江樟的首次报道,初步明确了该病害病原分类地位,研究结果为今后该病害的监测预警、病原菌致病机理和科学防治奠定理论基础。毛色二孢属真菌种类繁多,常以内生菌的形式隐蔽有效地寄生在寄主植物体内[6, 19],其扩散传播可能与植物材料人为调运活动有关[9],今后应加强该类病原菌的收集鉴定,对其开展快速检测技术研究,强化植物检疫监测,降低人为扩散风险。在明确病原的基础上,还需进一步开展浙江樟溃疡病流行规律观测和防治药剂筛选,并从效应蛋白角度探索病原侵染机理,从而为预防和控制病害发生流行提供有效的治理策略。
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2025年第45卷第6期
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doi: 10.13324/j.cnki.jfcf.202506014
  • 接收时间:2025-06-17
  • 首发时间:2026-05-28
  • 出版时间:2025-11-15
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  • 收稿日期:2025-06-17
  • 修回日期:2025-09-03
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    1.福建农林大学林学院, 福建 福州 350002
    2.凉山彝族自治州林业草原科学研究院, 四川 西昌 615000
    3.绵阳市安州区自然资源局, 四川 绵阳 622600
    4.福建农林大学金山学院, 福建 福州 350002

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叶小真(1983-), 女, 副教授, 从事林木病理学研究。Email:
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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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