Article(id=1276238142671819249, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276238094940639467, articleNumber=null, orderNo=null, doi=10.3969/j.issn.1000-2561.2025.02.011, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1723996800000, receivedDateStr=2024-08-19, revisedDate=1729008000000, revisedDateStr=2024-10-16, acceptedDate=null, acceptedDateStr=null, onlineDate=1782208484570, onlineDateStr=2026-06-23, pubDate=1740412800000, pubDateStr=2025-02-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1782208484570, onlineIssueDateStr=2026-06-23, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1782208484570, creator=13701087609, updateTime=1782208484570, updator=13701087609, issue=Issue{id=1276238094940639467, tenantId=1146029695717560320, journalId=1235980609244409860, year='2025', volume='46', issue='2', pageStart='247', pageEnd='489', issueExtLink='null', onlineDate='null', pubDate='1740412800000', pubDateStr='2025-02-25', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1782208473191, creator='13701087609', updateTime=1782208527160, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1276238321386909890, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276238094940639467, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1276238321386909891, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276238094940639467, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=359, endPage=368, ext={EN=ArticleExt(id=1276238144425038323, articleId=1276238142671819249, tenantId=1146029695717560320, journalId=1235980609244409860, language=EN, title=Isolation and Identification of Pathogens Causing Foliar Diseases in Coffea arabica in the Nujiang Dry-hot Valley, Yunnan, China, columnId=1236292524264968282, journalTitle=Chinese Journal of Tropical Crops, columnName=Plant Protection & Bio-safety, runingTitle=null, highlight=null, articleAbstract=

The Nujiang Dry-hot Valley region in Yunnan is an ideal area for the cultivation of Arabica coffee (Coffea arabica L.). However, the continuous occurrence of coffee leaf diseases has a significant impact on the health, yield and quality of coffee trees, causing considerable losses to local agricultural production. To effectively manage the diseases, this study focused on the isolation and identification of the pathogens associated with coffee leaf diseases in the Nujiang Dry-hot Valley region. Typical diseased leaf samples were collected from various coffee plantations, and through pathogen isolation, purification, verification using Koch’s postulates, morphological observation and ITS gene sequence analysis. Seven pathogenic strains, Lu 11-2, Lu 9-1, Xin 5-2, Xin 7-5, Xin 8-2, Xin 15-2 and Lu 23-1, were identified. Lu 11-2, Lu 9-1, Xin 5-2 and Xin 7-5 were identified as Alternaria sp., Xin 15-2 and Lu 23-1 were identified as Colletotrichum sp., and Xin 8-2 was identified as Pestalotiopsis sp. The primary pathogens causing coffee leaf diseases in this region could be categorized into three types, Alternaria sp., Pestalotiopsis sp. and Colletotrichum sp.. Specifically, Alternaria sp. and Pestalotiopsis sp. were the primary pathogens responsible for coffee leaf spot disease, while Colletotrichum sp. was the causative agent of coffee anthracnose disease. The findings of this study would not only provide a scientific basis for a deeper understanding of the types of coffee leaf diseases in the Nujiang Dry-hot Valley region of Yunnan but also offer theoretical support for effective disease management in the area. This research plays a significant guiding role for local coffee cultivation and serves as a reference for disease control in coffee-growing regions elsewhere.

, authors=null, authorsList=Na WANG, Wei YAN, Dongxiao MA, Yang YANG, Faguang HU, Xingfei FU, Guiping LI, Haohao YU, Yuqiang LOU, Yanan LI, authorCompany=null, correspAuthors=Yuqiang LOU, Yanan LI, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, 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, fund=null), CN=ArticleExt(id=1276238145658163711, articleId=1276238142671819249, tenantId=1146029695717560320, journalId=1235980609244409860, language=CN, title=怒江干热河谷流域小粒咖啡叶部病害病原分离与鉴定, columnId=1236292524520820846, journalTitle=热带作物学报, columnName=植物保护与生物安全, runingTitle=null, highlight=null, articleAbstract=

云南怒江干热河谷流域是小粒咖啡(Coffea arabica L.)种植的理想区域,但是咖啡叶部病害的持续发生,对咖啡树的健康、产量和品质造成了严重影响,给当地农业生产带来一定损失。为了有效防控这些病害,本研究采集怒江干热河谷流域不同咖啡种植地的典型病叶样本,对咖啡叶部病害进行病原菌分离与鉴定。通过病原分离、纯化、柯赫氏法则验证、形态学观察、ITS基因序列比对分析,鉴定到7个致病菌株,分别为Lu 11-2、Lu 9-1、Xin 5-2、Xin 7-5、Xin 8-2、Xin 15-2和Lu 23-1,其中Lu 11-2、Lu 9-1、Xin 5-2和Xin 7-5鉴定为链格孢菌(Alternaria sp.),Xin 15-2和Lu 23-1菌株鉴定为炭疽菌(Colletotrichum sp.),Xin 8-2菌株鉴定为拟盘多毛孢菌(Pestalotiopsis sp.)。该地区的咖啡叶部病害主要致病菌有3类:链格孢菌、拟盘多毛孢菌和炭疽菌,其中链格孢菌、拟盘多毛孢菌为咖啡褐斑病的主要致病菌,炭疽菌为咖啡炭疽病的致病菌。研究结果不仅为深入了解云南怒江干热河谷流域的咖啡叶部病害类型提供科学依据,还为该地区咖啡病害的有效防控提供理论支持,对当地的咖啡种植业具有重要的指导意义,也为其他地区的咖啡病害防控提供参考。

, authors=

王娜(1996—),女,硕士研究生,研究方向:咖啡植物病理学。

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* 娄予强(LOU Yuqiang),E-mail:
李亚男(LI Yanan),E-mail:
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王娜(1996—),女,硕士研究生,研究方向:咖啡植物病理学。

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王娜(1996—),女,硕士研究生,研究方向:咖啡植物病理学。

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Identification and biological characteristics of the pathogen causing Neopestalotiopsis leaf spot disease on Areca catechu[J]. Chinese Journal of Tropical Crops, 2023, 44(8): 1671-1678. 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Plant Pathology, 2019, 68(6): 1146-1156., articleTitle=Characteristics and distribution of Colletotrichum species in coffee plantations in Hainan, China, refAbstract=null)], funds=[Fund(id=1276238162615734866, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, awardId=2022YFF1302404, language=CN, fundingSource=国家重点研发计划项目(2022YFF1302404), fundOrder=null, country=null), Fund(id=1276238162708009555, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, awardId=202304BI090032-37, language=CN, fundingSource=云南省科技计划项目(202304BI090032-37), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1276238145876267521, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, xref=null, ext=[AuthorCompanyExt(id=1276238145884656130, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, companyId=1276238145876267521, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=Institute of Tropical and Subtropical Cash Crops, Yunnan Academy of Agricultural Sciences, Baoshan, Yunnan 678000, China), AuthorCompanyExt(id=1276238145888850435, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, companyId=1276238145876267521, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=云南省农业科学院热带亚热带经济作物研究所,云南保山 678000)])], figs=[ArticleFig(id=1276238159000244801, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, language=EN, label=Fig. 1, caption=Symptoms of coffee leaf diseases in fields, figureFileSmall=qqN9veMS3O+nNCIOujszmQ==, figureFileBig=nyZ9wcccFCDK4He7OnV32g==, tableContent=null), ArticleFig(id=1276238159067353666, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, language=CN, label=图1, caption=咖啡叶部病害田间症状, figureFileSmall=qqN9veMS3O+nNCIOujszmQ==, figureFileBig=nyZ9wcccFCDK4He7OnV32g==, tableContent=null), ArticleFig(id=1276238159767802435, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, language=EN, label=Fig. 2, caption=Leaf spots at 5 days post-inoculation, figureFileSmall=pZ0IPm4D+JqkI2t29Qo5VQ==, figureFileBig=vXKLFjsEAF1m7MYS7a4FhQ==, tableContent=null), ArticleFig(id=1276238159851688516, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, language=CN, label=图2, caption=接种5 d后叶片病斑, figureFileSmall=pZ0IPm4D+JqkI2t29Qo5VQ==, figureFileBig=vXKLFjsEAF1m7MYS7a4FhQ==, tableContent=null), ArticleFig(id=1276238160220787269, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, language=EN, label=Fig. 3, caption=Colony morphology (A) and colony diameter (B) of each strain on PDA medium, figureFileSmall=JbPnoJLzDx4I76I0SrXpFQ==, figureFileBig=XnHXu5x5ViwQ3ms0oNsDDA==, tableContent=null), ArticleFig(id=1276238160296284742, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, language=CN, label=图3, caption=各菌株在PDA培养基上的菌落形态(A)及菌落直径(B), figureFileSmall=JbPnoJLzDx4I76I0SrXpFQ==, figureFileBig=XnHXu5x5ViwQ3ms0oNsDDA==, tableContent=null), ArticleFig(id=1276238160623440455, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, language=EN, label=Fig. 4, caption=Conidia and mycelium morphology of each strain, figureFileSmall=YoS7lx3sOLi4TxZ7eHsyXg==, figureFileBig=Io1FiUHkXWkb5K/z7GP9fA==, tableContent=null), ArticleFig(id=1276238162099835466, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, language=CN, label=图4, caption=各菌株的分生孢子及菌丝形态, figureFileSmall=YoS7lx3sOLi4TxZ7eHsyXg==, figureFileBig=Io1FiUHkXWkb5K/z7GP9fA==, tableContent=null), ArticleFig(id=1276238162158555723, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, language=EN, label=Fig. 5, caption=Phylogenetic tree based on ITS gene sequences, figureFileSmall=xUOu+DJhCwtDg9f5lkUzrA==, figureFileBig=mXfgYGE0dHua2KWGh/VNUg==, tableContent=null), ArticleFig(id=1276238162238247501, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, language=CN, label=图5, caption=基于ITS构建的系统发育进化树, figureFileSmall=xUOu+DJhCwtDg9f5lkUzrA==, figureFileBig=mXfgYGE0dHua2KWGh/VNUg==, tableContent=null), ArticleFig(id=1276238162309550670, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, language=EN, label=Tab. 1, caption=

Incidence rates and lesion area of seven strains

, figureFileSmall=null, figureFileBig=null, tableContent=
菌株Strain发病率Incidence rate/%病斑面积Lesion area/cm2
Lu 11-252.780.12±0.05c
Lu 9-177.780.25±0.01c
Xin 5-2100.001.47±0.08a
Xin 7-572.220.30±0.07c
Xin 8-258.330.13±0.05c
Xin 15-280.560.79±0.23b
Lu 23-169.440.53±0.34bc
), ArticleFig(id=1276238162385048143, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, language=CN, label=表1, caption=

7个菌株发病率及病斑面积

, figureFileSmall=null, figureFileBig=null, tableContent=
菌株Strain发病率Incidence rate/%病斑面积Lesion area/cm2
Lu 11-252.780.12±0.05c
Lu 9-177.780.25±0.01c
Xin 5-2100.001.47±0.08a
Xin 7-572.220.30±0.07c
Xin 8-258.330.13±0.05c
Xin 15-280.560.79±0.23b
Lu 23-169.440.53±0.34bc
), ArticleFig(id=1276238162447962704, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, language=EN, label=Tab. 2, caption=

Colony morphology, colony diameter and growth rate of each strain

, figureFileSmall=null, figureFileBig=null, tableContent=
菌株Strain菌落形态Colony morphology菌落直径Colony diameter/cm生长速率Growth rate/(cm·d–1)
Lu 11-2菌落呈同心圆,正面内层菌落深褐色,微隆起,菌丝絮状,外层白色,菌丝较紧密,平板背面菌落中间黑褐色,边缘白色4.90±0.32c1.03±0.12c
Lu 9-1菌落近圆形,中部微隆起,正面内层菌落底部浅褐色,菌丝絮状,边缘白色,菌丝紧密,平板背面菌落中间为不规则的浅褐色,边缘白色4.10±0.15d0.74±0.04d
Xin 5-2菌落呈同心圆,正面内层菌落浅灰色,微隆起,菌丝絮状,外层深褐色,最外层白色,菌丝较紧密,气生菌丝稀疏,平板背面菌落呈同心圆,内层深褐色,外层褐色,最外层白色5.90±0.15b1.24±0.02b
Xin 7-5菌落呈同心圆,正面内层菌落深褐色,菌丝絮状,外层浅褐色,最外层白色,菌丝较紧密,平板背面菌落呈同心圆,内层深褐色,外层褐色,最外层白色4.93±0.02c0.78±0.04d
Xin 8-2菌落呈同心圆,正面内层菌落白色,菌丝絮状、稍微稀疏,外层白色,菌丝紧密,平板背面菌落呈同心圆,菌落白色,内层菌丝稍稀疏,外层菌丝紧密5.50±0.19b1.05±0.15c
Xin 15-2菌落灰白色,菌丝辐射状,气生菌丝稀疏,平板背面菌落灰白色7.82±0.31a1.65±0.09a
Lu 23-1菌落灰白色,气生菌丝稀疏,平板背面菌落灰白色5.75±0.11b1.25±0.00b
), ArticleFig(id=1276238162510877265, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276238142671819249, language=CN, label=表2, caption=

各菌株菌落形态、菌落直径及生长速率

, figureFileSmall=null, figureFileBig=null, tableContent=
菌株Strain菌落形态Colony morphology菌落直径Colony diameter/cm生长速率Growth rate/(cm·d–1)
Lu 11-2菌落呈同心圆,正面内层菌落深褐色,微隆起,菌丝絮状,外层白色,菌丝较紧密,平板背面菌落中间黑褐色,边缘白色4.90±0.32c1.03±0.12c
Lu 9-1菌落近圆形,中部微隆起,正面内层菌落底部浅褐色,菌丝絮状,边缘白色,菌丝紧密,平板背面菌落中间为不规则的浅褐色,边缘白色4.10±0.15d0.74±0.04d
Xin 5-2菌落呈同心圆,正面内层菌落浅灰色,微隆起,菌丝絮状,外层深褐色,最外层白色,菌丝较紧密,气生菌丝稀疏,平板背面菌落呈同心圆,内层深褐色,外层褐色,最外层白色5.90±0.15b1.24±0.02b
Xin 7-5菌落呈同心圆,正面内层菌落深褐色,菌丝絮状,外层浅褐色,最外层白色,菌丝较紧密,平板背面菌落呈同心圆,内层深褐色,外层褐色,最外层白色4.93±0.02c0.78±0.04d
Xin 8-2菌落呈同心圆,正面内层菌落白色,菌丝絮状、稍微稀疏,外层白色,菌丝紧密,平板背面菌落呈同心圆,菌落白色,内层菌丝稍稀疏,外层菌丝紧密5.50±0.19b1.05±0.15c
Xin 15-2菌落灰白色,菌丝辐射状,气生菌丝稀疏,平板背面菌落灰白色7.82±0.31a1.65±0.09a
Lu 23-1菌落灰白色,气生菌丝稀疏,平板背面菌落灰白色5.75±0.11b1.25±0.00b
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怒江干热河谷流域小粒咖啡叶部病害病原分离与鉴定
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王娜 , 严炜 , 马东晓 , 杨旸 , 胡发广 , 付兴飞 , 李贵平 , 喻好好 , 娄予强 * , 李亚男 *
热带作物学报 | 植物保护与生物安全 2025,46(2): 359-368
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热带作物学报 |植物保护与生物安全 2025 , 46 (2) : 359 -368
怒江干热河谷流域小粒咖啡叶部病害病原分离与鉴定
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王娜, 严炜, 马东晓, 杨旸, 胡发广, 付兴飞, 李贵平, 喻好好, 娄予强* , 李亚男*
作者信息
  • 云南省农业科学院热带亚热带经济作物研究所,云南保山 678000
通讯作者:
* 娄予强(LOU Yuqiang),E-mail:
李亚男(LI Yanan),E-mail:
Isolation and Identification of Pathogens Causing Foliar Diseases in Coffea arabica in the Nujiang Dry-hot Valley, Yunnan, China
Na WANG, Wei YAN, Dongxiao MA, Yang YANG, Faguang HU, Xingfei FU, Guiping LI, Haohao YU, Yuqiang LOU* , Yanan LI*
Affiliations
  • Institute of Tropical and Subtropical Cash Crops, Yunnan Academy of Agricultural Sciences, Baoshan, Yunnan 678000, China
出版时间: 2025-02-25 doi: 10.3969/j.issn.1000-2561.2025.02.011
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云南怒江干热河谷流域是小粒咖啡(Coffea arabica L.)种植的理想区域,但是咖啡叶部病害的持续发生,对咖啡树的健康、产量和品质造成了严重影响,给当地农业生产带来一定损失。为了有效防控这些病害,本研究采集怒江干热河谷流域不同咖啡种植地的典型病叶样本,对咖啡叶部病害进行病原菌分离与鉴定。通过病原分离、纯化、柯赫氏法则验证、形态学观察、ITS基因序列比对分析,鉴定到7个致病菌株,分别为Lu 11-2、Lu 9-1、Xin 5-2、Xin 7-5、Xin 8-2、Xin 15-2和Lu 23-1,其中Lu 11-2、Lu 9-1、Xin 5-2和Xin 7-5鉴定为链格孢菌(Alternaria sp.),Xin 15-2和Lu 23-1菌株鉴定为炭疽菌(Colletotrichum sp.),Xin 8-2菌株鉴定为拟盘多毛孢菌(Pestalotiopsis sp.)。该地区的咖啡叶部病害主要致病菌有3类:链格孢菌、拟盘多毛孢菌和炭疽菌,其中链格孢菌、拟盘多毛孢菌为咖啡褐斑病的主要致病菌,炭疽菌为咖啡炭疽病的致病菌。研究结果不仅为深入了解云南怒江干热河谷流域的咖啡叶部病害类型提供科学依据,还为该地区咖啡病害的有效防控提供理论支持,对当地的咖啡种植业具有重要的指导意义,也为其他地区的咖啡病害防控提供参考。

小粒咖啡  /  叶部病害  /  链格孢菌  /  拟盘多毛孢菌  /  炭疽菌

The Nujiang Dry-hot Valley region in Yunnan is an ideal area for the cultivation of Arabica coffee (Coffea arabica L.). However, the continuous occurrence of coffee leaf diseases has a significant impact on the health, yield and quality of coffee trees, causing considerable losses to local agricultural production. To effectively manage the diseases, this study focused on the isolation and identification of the pathogens associated with coffee leaf diseases in the Nujiang Dry-hot Valley region. Typical diseased leaf samples were collected from various coffee plantations, and through pathogen isolation, purification, verification using Koch’s postulates, morphological observation and ITS gene sequence analysis. Seven pathogenic strains, Lu 11-2, Lu 9-1, Xin 5-2, Xin 7-5, Xin 8-2, Xin 15-2 and Lu 23-1, were identified. Lu 11-2, Lu 9-1, Xin 5-2 and Xin 7-5 were identified as Alternaria sp., Xin 15-2 and Lu 23-1 were identified as Colletotrichum sp., and Xin 8-2 was identified as Pestalotiopsis sp. The primary pathogens causing coffee leaf diseases in this region could be categorized into three types, Alternaria sp., Pestalotiopsis sp. and Colletotrichum sp.. Specifically, Alternaria sp. and Pestalotiopsis sp. were the primary pathogens responsible for coffee leaf spot disease, while Colletotrichum sp. was the causative agent of coffee anthracnose disease. The findings of this study would not only provide a scientific basis for a deeper understanding of the types of coffee leaf diseases in the Nujiang Dry-hot Valley region of Yunnan but also offer theoretical support for effective disease management in the area. This research plays a significant guiding role for local coffee cultivation and serves as a reference for disease control in coffee-growing regions elsewhere.

Coffea arabica L.  /  leaf diseases  /  Alternaria sp.  /  Pestalotiopsis sp.  /  Colletotrichum sp.
王娜, 严炜, 马东晓, 杨旸, 胡发广, 付兴飞, 李贵平, 喻好好, 娄予强, 李亚男. 怒江干热河谷流域小粒咖啡叶部病害病原分离与鉴定. 热带作物学报, 2025 , 46 (2) : 359 -368 . DOI: 10.3969/j.issn.1000-2561.2025.02.011
Na WANG, Wei YAN, Dongxiao MA, Yang YANG, Faguang HU, Xingfei FU, Guiping LI, Haohao YU, Yuqiang LOU, Yanan LI. Isolation and Identification of Pathogens Causing Foliar Diseases in Coffea arabica in the Nujiang Dry-hot Valley, Yunnan, China[J]. Chinese Journal of Tropical Crops, 2025 , 46 (2) : 359 -368 . DOI: 10.3969/j.issn.1000-2561.2025.02.011
咖啡作为全球经济作物的重要支柱,影响着许多国家的农业经济和农民生计[1-2]。小粒咖啡(Coffea arabica L.)隶属于茜草科咖啡属,因其优良的品质和经济价值而受到广泛关注[2]。其生长对气候条件有较高的要求,尤其是温暖湿润的环境最为适宜[3],国际咖啡种植标准指出,咖啡适宜种植在海拔500~2000 m、温度15~25 ℃、年降雨量1500~2000 mm的具有火山灰质土壤的地区[4]。云南怒江干热河谷流域以其适宜的海拔、充足的阳光、适量的降水量和适宜的温度,为小粒咖啡的生长提供了优越的天然条件[5-6]。然而,干热河谷独特的气候条件也带来了病害发生及其扩散的复杂性和严重性。
云南怒江干热河谷流域多以山地、坡地为主,生物多样性丰富,咖啡叶部病害多发,叶部病害的发生不仅对咖啡树的健康造成严重威胁,还直接影响咖啡的产量和品质,导致咖啡产业的经济损失。常见的咖啡叶部病害主要有咖啡叶锈病、褐斑病、炭疽病、煤烟病等[7]。据报道,咖啡叶锈病由专性生物营养真菌Hemileia vastatrix引起,是一种世界性的毁灭性病害,导致咖啡种植园的产量严重下降和经济受损[8-9]。褐斑病可由多种病原菌引起,通常表现为叶片上出现褐色斑点,这些斑点扩大并发展成具有深棕色边缘的同心环,最后坏死斑点蔓延到整片咖啡叶,导致咖啡叶卷曲和脱落,使幼树大量死亡[10-11]。炭疽病主要感染咖啡的嫩叶、叶柄、枝条和果实,引发叶片脱落、枝条干枯及果实腐烂,严重损害咖啡的产量和品质[12-14]。煤烟病则是由昆虫分泌蜜露诱发的病害,导致咖啡叶面出现灰黑色的斑块或霉层[7],这些叶部病害的发生均不同程度地影响了咖啡植株的正常生长发育。
研究咖啡叶部病害病原菌种类及其生物学特性,以及它们与环境因素的关系是深入了解和应对咖啡病害的关键。本研究旨在对怒江干热河谷流域的小粒咖啡叶部病害进行系统地病原分离与鉴定,探究病原菌种类,明确致病菌种。以期为该地区咖啡病害的有效管理和防治提供科学依据,并提出切实可行的防控建议和对策,从而改善咖啡质量和提升咖啡生产效益。
于2023年7月采自云南省保山市潞江镇辛家山咖啡种植基地(海拔962.71 m,24°57′30″N,98°53′56″E)、泸水市咖啡种植区域(海拔1079.6 m,25°48′25″N,98°51′3″E)的小粒咖啡品种铁皮卡(Typica)病叶,分离纯化获得病原菌。
马铃薯葡萄糖琼脂(PDA)培养基干粉购自广东环凯微生物科技有限公司,按使用方法配制PDA培养基。
采用组织分离法分离病原菌[15],将采摘收集到的病叶置于超净工作台,用灭菌剪刀将病健交界处的组织剪成5 mm×5 mm的小块,用灭菌镊子将组织块置于75%医用酒精中浸泡1 min,后用灭菌离子水漂洗1 min,重复漂洗3次,取出置于灭菌吸水纸上吸干水分,然后将其接种到PDA平板上,每皿接种4块,于28 ℃培养箱中倒置培养。每天观察病原菌的生长情况,待组织周围长出菌丝后,挑取菌丝尖端进行纯化,重复纯化3次左右,待病原菌生长一致即可保存菌株,备用。
采用离体接菌的方法,采集健康无病的铁皮卡咖啡叶片,先用灭菌棉蘸取75%医用酒精擦拭咖啡叶面消毒,再用灭菌水擦拭一遍叶片,将叶片置于保鲜盒内。用无菌接种针刺伤叶脉一侧的叶片,另一侧不刺伤。将经纯化的病原菌菌饼接种到叶面,以菌丝面接触叶面,以PDA培养基作对照,每个菌株接种3片叶,3次重复。叶柄处用湿棉花包住,28 ℃保湿条件下培养7 d,并于接种后3、5、7 d对叶片进行拍照,使用Image J软件测量叶片病斑面积。待叶片发病后进行柯赫氏法则验证,并将发病组织进行分离纯化。
将纯化好的病原菌接种到PDA平板上,每个菌株接3皿,3次重复,置于28 ℃培养箱中倒置培养7 d,于接种后3、5、7 d观察病原菌的菌落生长情况,测量菌落直径并计算生长速率。采用德国Leica DM5000B显微镜观察菌丝和分生孢子形态,对病原菌进行形态鉴定,并随机选取50个分生孢子测量大小。
将上述病原菌送样,委托昆明擎科生物有限公司进行DNA测序,测序引物:ITS1(5′-TCCGTAGGTGAACCTGCGG-3′),ITS4(5′-TCCTCCGCTTATTGATATGC-3′)。将测序结果上传至NCBI进行BLAST同源分析,下载与分离病原菌亲缘关系相近的菌株序列,利用MEGA 11.0软件构建系统发育树。
利用IBM SPSS Statistics 25软件进行数据统计与分析,采用GraphPad Prism v8.0软件制图。
咖啡叶部病害一年四季均可发生,叶部病害的发生既会导致咖啡生长受阻,又会导致产量降低,从而影响咖啡品质。发现的咖啡叶部病害田间症状表现如图1,发病初期叶片出现小黄点,然后慢慢扩大为圆形或近圆形的褐色病斑,病斑边界明显,中心为灰白色,周围有黄色晕圈(图1A图1B);发病后期病斑连片形成大病斑,呈灰白色,病斑边缘有黑色点状物(图1C图1D)。
采用组织分离法对采集到的病叶病原进行分离纯化,得到45个菌株,根据菌落形态特征和生长情况从中选取9株进行离体叶片回接试验,每个菌株接种3片叶,3次重复,接种时分别做刺伤和不刺伤处理。结果如图2所示,接种4 d后,有7个菌株出现明显的发病症状,Xin 1-3和Xin 12-1菌株未发病。测量发病菌株的病斑面积及统计发病率(表1),发现Xin 5-2菌株的病斑面积最大,为(1.47±0.08)cm2,且发病率最高,为100%;Xin 15-2菌株次之,病斑面积为(0.79±0.23)cm2;而菌株Lu 11-2的病斑面积最小,仅为(0.13±0.05)cm2,且发病率最低,为52.78%。对发病的7个叶片进行病原分离纯化,得到与原接种菌株形态特征和培养性状一致的病原菌,符合柯赫氏法则,说明接种菌株是致病菌。
将符合柯赫氏法则的病原菌菌株接种到PDA培养基上,培养7 d后观察菌落形态特征及生长情况。结果发现这些菌株在某些特征上存在共性,同时也表现出一定的差异性。所有菌株的菌落均为同心圆形态,边缘菌丝为白色且菌丝形态相似;在菌落颜色和生长速率方面有差异,如Lu 11-2、Lu 9-1、Xin 5-2和Xin 7-5菌株的菌落颜色由内到外逐渐变淡,由深褐色到浅褐色再到白色,而Xin 8-2、Xin 15-2和Lu 23-1菌株的菌丝则呈现灰白色或白色。此外,7个菌株的菌落生长速率略有差异,其中Xin 15-2菌株的菌落生长速率显著高于其他菌株,其5 d的菌落直径达(7.82±0.31)cm,平均生长速率为(1.65±0.09)cm/d,而Lu 9-1菌株生长速率较慢,5 d的菌落直径为(4.10±0.15)cm,平均生长速率为(0.74±0.04)cm/d(图3表2)。
7个菌株的分生孢子及菌丝形态如图4,Lu 11-2菌株的分生孢子呈卵形,有隔,横隔1~3个,纵隔0~1个,分生孢子平均大小为(24.60±4.08)µm×(11.39±2.27)µm,长宽比为2.18±0.17,菌丝直径为(3.78±0.80)µm;Lu 9-1菌株的分生孢子呈梨形,有隔,横隔1~2个,纵隔0~1个,分生孢子平均大小为(24.87±6.07)µm×(11.17±1.21)µm,长宽比为2.23±0.46,菌丝直径为(4.10±0.77)µm;Xin 5-2的分生孢子呈卵形或梨形,有隔,横隔1~3个,纵隔0~2个,分生孢子平均大小为(26.44±2.71)µm×(12.51±3.27)µm,长宽比为2.24±0.53,菌丝直径为(5.31±0.83)µm;Xin 7-5的分生孢子呈倒梨形,有隔,横隔1~2个,纵隔0~2个,分生孢子平均大小(31.51±6.20)µm×(14.04±2.97)µm,长宽比为2.29±0.41,菌丝直径为(3.61±0.87)µm;Xin 8-2的分生孢子共5个细胞,4分隔,基部细胞颜色较淡,倒三角形或倒圆锥形,中间3个细胞颜色偏深,翁形,顶端细胞颜色偏淡,圆锥形或三角形,顶端具1根附属丝,基部具1~3根附属丝,分生孢子平均大小为(27.43±2.87)µm×(6.52±0.77)µm,长宽比为4.26±0.60,菌丝直径为(3.52±0.77)µm;Xin 15-2的分生孢子呈长圆柱形,单胞,分生孢子平均大小为(15.95±4.70)µm×(4.94±1.03)µm,长宽比为3.41±1.31,菌丝直径为(5.31±0.83)µm;Lu 23-1的分生孢子呈长圆柱形,单胞,分生孢子平均大小为(18.38±2.88)µm×(5.04±0.81)µm,长宽比为3.69±0.56,菌丝直径为(3.70±0.67)µm。
将7个菌株的ITS序列测序结果进行整理,分别得到不同大小的目标片段,Lu 11-2菌株的ITS片段大小为539 bp,Lu 9-1的为539 bp,Xin 5-2的为580 bp,Xin 7-5为575 bp,Xin 8-2为575 bp,Xin 15-2为583 bp,Lu 23-1为587 bp。将7个菌株的ITS序列分别在NCBI上进行BLAST比对,并下载同源性较高的代表菌株序列,以AB496639.1 Methanobacterium alcaliphilum作外群联合构建系统发育进化树(图5)。Lu 11-2、Lu 9-1、Xin 5-2、Xin 7-5菌株与链格孢菌(Alternaria sp.)以100%的支持率聚为一支,其中Lu 11-2和Lu 9-1鉴定为交链格孢(A. alternata),Xin 5-2鉴定为极细链格孢(A. tenuissima),Xin 7-5鉴定为链格孢属(Alternaria sp.);Xin 15-2和Lu 23-1菌株与炭疽菌(Colletotrichum sp.)聚为一支,其中Xin 15-2鉴定为果生炭疽菌(C. fructicola),Lu 23-1则鉴定为博宁炭疽菌(C. boninense);菌株Xin 8-2与拟盘多毛孢菌(Pestalotiopsis sp.)聚为一支,能明显与其他种区分。结合各菌株的形态学特征,将菌株Lu 11-2、Lu 9-1、Xin 5-2、Xin 7-5鉴定为链格孢菌,Xin 15-2和Lu 23-1则鉴定为炭疽菌,Xin 8-2鉴定为拟盘多毛孢菌。
在怒江干热河谷流域的咖啡种植区,咖啡叶部病害发生频繁且多样,为明确该区域咖啡叶部病害的种类及其病原,采集不同咖啡种植地的典型病叶样本,利用形态学和分子生物学方法,确定了咖啡叶部的病原菌分别是链格孢菌(Alternaria sp.)、拟盘多毛孢菌(Pestalotiopsis sp.)和炭疽菌(Colletotrichum sp.)。
链格孢菌(Alternaria sp.)是一种分布广泛的植物病原真菌,具有广泛的寄主范围,能够侵染多种植物,造成严重的经济损失。已有研究发现,交链格孢(A. alternata)可以引起苹果叶斑病,导致苹果叶片上形成棕色或褐色病斑,严重时导致叶片干枯脱落[16-17];交链格孢也是绣球花叶斑病的致病菌之一,在绣球花叶片上的症状表现为深褐色病斑,对植物的观赏价值造成严重影响[18];覃茜等[19]从苏铁叶片上分离出引起褐斑病的病原真菌巴斯基链格孢(A. burnsii),并证实该病原菌能引起多种苏铁褐斑病;此外,链格孢菌也是龙牙百合褐斑病的主要致病菌[20-21]。2024年,ZHANG等[10]报道了由交链格孢引起的咖啡叶斑病。本研究对采集到的咖啡病叶进行病原菌分离鉴定,鉴定到4个菌株属于链格孢菌,与ZHANG等[10]的研究结果相似,表明链格孢菌是咖啡褐斑病的致病菌之一,且咖啡褐斑病是怒江干热河谷流域咖啡种植区域的主要叶部病害之一。
拟盘多毛孢菌(Pestalotiopsis sp.)是一类广泛分布于自然界中的微生物,它们与多种植物病害有关,既可以作为病原菌导致植物病害,也可以作为内生菌或腐生菌存在于植物体内。拟盘多毛孢作为内生菌大多被研究者用来探究真菌的次级代谢产物等,如红树林内生拟盘多毛孢(P. microspora)可以通过生物合成途径产生抗肿瘤的化合物[22];耐铝树种千年桐的内生链格孢可吸收环境中的Al3+,提高宿主植物对铝胁迫的耐受性[23]。2020年,安小丽等[24]从贵州省茶树叶片上分离出茶拟盘多毛孢(P. theae),该病原菌可引起茶叶斑病;李晶等[25]报道了Neopestalotiopsis sonneratae会引起槟榔叶斑病,初期槟榔叶片出现黑褐色小病斑,随着病斑的扩展,最终导致叶片干枯倒垂;袁榕等[26]在华中五味子上鉴定出5种不同的拟盘多毛孢属真菌,并且这些菌株具有较强的致病性;还有其他研究表明,拟盘多毛孢还可引起草莓根腐病[27]、番石榴环斑病[28]、松树赤枯病[29]等。本研究对咖啡褐斑病进行病原菌分离鉴定,鉴定到1个菌株属于拟盘多毛孢菌,并且符合柯赫氏法则,说明拟盘多毛孢菌是咖啡褐斑病的致病菌之一。
炭疽菌(Colletotrichum sp.)作为一种重要的植物病原菌,引起了全球的广泛关注,它能够侵染多种农作物和经济作物,导致炭疽病的发生,对农业生产造成严重损失[30]。近年来,随着对炭疽菌的深入研究,研究者们在炭疽菌的病原鉴定、生物学特性以及病害防治方面取得了一系列进展。炭疽菌的研究涵盖了多个作物,包括砀山酥梨、咖啡、油茶等。研究发现,不同地区的炭疽菌种类存在差异,同一作物可能受到多种炭疽菌的侵染,如,砀山酥梨炭疽病的主要病原菌为果生炭疽菌(C. fructicola[31];而油茶炭疽病的病原菌则包括果生炭疽菌和胶孢炭疽菌(C. gloeosporioides[32];宁夏地区葡萄炭疽病的主要病原菌为毁灭炭疽菌(C. destructivum)、果生炭疽菌和暹罗炭疽菌(C. siamense[33];然而,导致重庆多花黄精炭疽病的病原菌有5种,分别为松针炭疽菌(C. fioriniae)、博宁炭疽菌(C. boninense)、胶孢炭疽菌(C. gloeosporioides)、果生炭疽菌和冬麦炭疽菌(C. liriopes[34]。国内外已经鉴定出多种可侵染咖啡的炭疽菌,CRISTÓBALMARTÍNEZ等[35]在发病的咖啡植株上鉴定到5种炭疽菌,2019年,CAO等[36]在海南咖啡种植区鉴定到8种炭疽菌,这8种炭疽菌中也包含了本研究中鉴定到的果生炭疽菌和博宁炭疽菌。这些研究表明咖啡炭疽病在全球分布广泛且危害严重。
本研究对怒江干热河谷流域的小粒咖啡叶部病害进行病原菌分离鉴定,鉴定到小粒咖啡叶部病害的主要致病菌为链格孢菌(Alternaria sp.)、拟盘多毛孢菌(Pestalotiopsis sp.)和炭疽菌(Colletotrichum sp.)。研究结果可为该地区咖啡病害的防控提供理论支持,对农业生产具有重要意义。随着研究的不断深入,期待未来能够开发出更多高效、环保的病害防控技术,为咖啡产业的可持续发展做出贡献。
  • 国家重点研发计划项目(2022YFF1302404)
  • 云南省科技计划项目(202304BI090032-37)
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2025年第46卷第2期
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doi: 10.3969/j.issn.1000-2561.2025.02.011
  • 接收时间:2024-08-19
  • 首发时间:2026-06-23
  • 出版时间:2025-02-25
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  • 收稿日期:2024-08-19
  • 修回日期:2024-10-16
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国家重点研发计划项目(2022YFF1302404)
云南省科技计划项目(202304BI090032-37)
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    云南省农业科学院热带亚热带经济作物研究所,云南保山 678000

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* 娄予强(LOU Yuqiang),E-mail:
李亚男(LI Yanan),E-mail:
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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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