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Article(id=1276213327835567092, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276213295170323272, articleNumber=null, orderNo=null, doi=10.3969/j.issn.1000-2561.2024.04.004, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1670342400000, receivedDateStr=2022-12-07, revisedDate=1672761600000, revisedDateStr=2023-01-04, acceptedDate=null, acceptedDateStr=null, onlineDate=1782202568252, onlineDateStr=2026-06-23, pubDate=1713974400000, pubDateStr=2024-04-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1782202568252, onlineIssueDateStr=2026-06-23, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1782202568252, creator=13701087609, updateTime=1782202568252, updator=13701087609, issue=Issue{id=1276213295170323272, tenantId=1146029695717560320, journalId=1235980609244409860, year='2024', volume='45', issue='4', pageStart='653', pageEnd='871', issueExtLink='null', onlineDate='null', pubDate='1713974400000', pubDateStr='2024-04-25', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1782202560465, creator='13701087609', updateTime=1782203706550, updator='13701087609', preIssue=null, nextIssue=null, articleTotal=null, ext={EN=IssueExt(id=1276218103419761358, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276213295170323272, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1276218103419761359, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1276213295170323272, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null, downloadFileDto=null}, startPage=682, endPage=690, ext={EN=ArticleExt(id=1276213328095613942, articleId=1276213327835567092, tenantId=1146029695717560320, journalId=1235980609244409860, language=EN, title=Resistance Gene Detection and Resistance Identification of PMMoV in Pepper Germplasm, columnId=1236256430337085821, journalTitle=Chinese Journal of Tropical Crops, columnName=Omics & Biotechnology, runingTitle=null, highlight=null, articleAbstract=

Pepper mild mottle virus (PMMoV) is one of the most serious pathogens worldwide. Currently, five pathogenic types of PMMoV have been reported, including P0, P1, P1,2, P1,2,3 and P1,2,3,4. The wide host range and strong pathogenicity of PMMoV seriously affect the yield and quality of pepper, and breeding for disease resistance is the most economical and effective means to control this virus. Our previous study determined that PMMoV infected with Danzhou pepper germplasm nursery in Hainan is the P1,2 pathogenic type. In order to screen PMMoV-resistant pepper germplasm resources, 110 germplasms were tested for PMMoV resistance using the reported L3-linked molecular markers. 46 pepper germplasms containing molecular markers of resistance were inoculated with PMMoV P1,2 pathogenic type by inoculation at seeding stage. Local symptoms were investigated 5 d after inoculation, and systemic symptoms were investigated 20 d after inoculation, and disease indices were calculated. The results showed that there were sig-nificant differences in disease resistance among different pepper germplasms, and the disease indexes of various germplasms ranged from 8.86-60.00. The germplasms showing high resistance (HR) to PMMoV P1,2 pathogenic type were CF19-34m and 19FB6-1. The germplasms showing resistance (R) wasCCJ93-1. The germplasms showing medium-resistance (MR) included 17YB32, 17SCa28m, 15SM39-2, 14SM555×14SM1, 14SM567-2, 14SM526-1, 14SM565-1, 14SM502-1, 14SM519-1, 14SM504m, 14SM503m, 14SM519-3, 14SM518-2, 14SM519-2, CCJ52-1, etc. The presenting susceptible (S) germplasms were 14SM514m, CCJ22-1, CCJ113-2, CCJ135-1, CCJ157-1, L537-1m, L545, CCJ87-2 and CCJ133-1, and the obtained disease-resistant germplasms accounted for 80.43% of the total identified materials. The results of this study would lay the foundation for creating germplasm resources resistant to pepper mild mottle virus and breeding resistant varieties.

, authors=null, authorsList=Jianfen CHEN, Zhenmu CAO, Yuling QIN, Longbin SHEN, Weixia LIU, Dan ZHU, Yiting WU, Ziji LIU, Xu WANG, authorCompany=null, correspAuthors=Ziji LIU, Xu WANG, 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=1276213329077081088, articleId=1276213327835567092, tenantId=1146029695717560320, journalId=1235980609244409860, language=CN, title=辣椒种质资源PMMoV抗性基因检测与抗性鉴定, columnId=1236256430517440904, journalTitle=热带作物学报, columnName=组学与生物技术, runingTitle=null, highlight=null, articleAbstract=

辣椒轻斑驳病毒（Pepper mild mottle virus，PMMoV）是世界范围内最严重的病原体之一，目前已报道的PMMoV包括P0、P1、P1,2、P1,2,3、P1,2,3,4五种致病型。辣椒轻斑驳病毒寄主范围广、致病力强，严重影响辣椒的产量和品质，开展抗病育种是防治该病毒最为经济有效的手段。本课题组前期研究已明确了感染海南儋州辣椒种质资源圃的PMMoV为P1,2致病型，为了筛选抗PMMoV的辣椒种质资源，以110份种质为材料，利用已报道的与L3连锁的分子标记对辣椒种质进行检测。结果表明：在110份材料中共检测到46份辣椒种质含有与L3连锁的分子标记。采用苗期人工接种鉴定方法，对含有抗性分子标记的46份辣椒种质接种PMMoV P1,2致病型进行抗性鉴定。接种5 d后调查局部症状，接种20 d后调查系统症状，计算病情指数。结果表明：不同的辣椒种质在抗病性上存在显著差异，各种质的病情指数在8.86~60.00之间，对PMMoV P1,2致病型表现高抗（HR）的种质为CF19-34m和19FB6-1；表现抗病（R）的种质为CCJ93-1；表现中抗（MR）的种质包括17YB32、17SCa28m、15SM39-2、14SM555×14SM1、14SM567-2、14SM526-1、14SM565-1、14SM502-1、14SM519-1、14SM504m、14SM503m、14SM519-3、14SM518-2、14SM519-2、CCJ52-1等34份；表现感病（S）的种质包括14SM514m、CCJ22-1、CCJ113-2、CCJ135-1、CCJ157-1、L537-1m、L545、CCJ87-2、CCJ133-1，获得的高抗、抗病、中抗种质占总鉴定材料的80.43%。本研究结果为创制抗辣椒轻斑驳病毒种质资源和培育抗性品种奠定基础。

, authors=

陈建分（1992—），女，硕士，研究方向：蔬菜抗病育种。

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* 刘子记（LIU Ziji），E-mail：
王旭（WANG Xu），E-mail：。
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2.海南大学园艺学院，海南海口　570208, bio={"content":"

陈建分（1992—），女，硕士，研究方向：蔬菜抗病育种。

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陈建分（1992—），女，硕士，研究方向：蔬菜抗病育种。

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Theoretical and Applied Genetics, 2008, 117(7): 1107-1118., articleTitle=Fine mapping and DNA fiber FISH analysis locates the tobamovirus resistance gene L3 of Capsicum chinense in a 400-kb region of R-like genes cluster embedded in highly repetitive sequences, refAbstract=null), Reference(id=1276466455784845784, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, doi=null, pmid=null, pmcid=null, year=2006, volume=null, issue=null, pageStart=66, pageEnd=75, url=null, language=null, rfNumber=[23], rfOrder=37, authorNames=李锡香, 张宝玺, journalName=辣椒种质资源描述规范和数据标准, refType=null, unstructuredReference=李锡香, 张宝玺. 辣椒种质资源描述规范和数据标准[M]. 北京: 中国农业出版社, 2006: 66-75., articleTitle=null, refAbstract=null), Reference(id=1276466455877120473, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, doi=null, pmid=null, pmcid=null, year=2006, volume=null, issue=null, pageStart=66, pageEnd=75, url=null, language=null, rfNumber=[23], rfOrder=38, authorNames=LI X X, ZHANG B X, journalName=Descriptive specifications and data standards of pepper germplasm resources, refType=null, unstructuredReference=LI X X, ZHANG B X. Descriptive specifications and data standards of pepper germplasm resources[M]. Beijing: China Agricultural Publishing House, 2006: 66-75. (in Chinese), articleTitle=null, refAbstract=null), Reference(id=1276466456183304666, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, doi=null, pmid=null, pmcid=null, year=2017, volume=null, issue=10, pageStart=44, pageEnd=50, url=null, language=null, rfNumber=[24], rfOrder=39, authorNames=秦蕾, 梁燕, 默宁, 张洋, 赵贵叶, journalName=中国蔬菜, refType=null, unstructuredReference=秦蕾, 梁燕, 默宁, 张洋, 赵贵叶. 辣椒种质资源TMV抗性的鉴定与评价[J]. 中国蔬菜, 2017(10): 44-50., articleTitle=辣椒种质资源TMV抗性的鉴定与评价, refAbstract=null), Reference(id=1276466456258802139, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, doi=null, pmid=null, pmcid=null, year=2017, volume=null, issue=10, pageStart=44, pageEnd=50, url=null, language=null, rfNumber=[24], rfOrder=40, authorNames=QIN L, LIANG Y, MO N, ZHANG Y, ZHAO G Y, journalName=China Vegetables, refType=null, unstructuredReference=QIN L, LIANG Y, MO N, ZHANG Y, ZHAO G Y. Identification and evaluation of resistance to TMV in pepper (Capsicum annuum L.) germplasm resources[J]. China Vegetables, 2017(10): 44-50. (in Chinese), articleTitle=Identification and evaluation of resistance to TMV in pepper (Capsicum annuum L.) germplasm resources, refAbstract=null), Reference(id=1276466456548209116, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, doi=null, pmid=null, pmcid=null, year=2012, volume=10, issue=3, pageStart=16, pageEnd=19, 23, url=null, language=null, rfNumber=[25], rfOrder=41, authorNames=姚明华, 焦春海, 王飞, 李宁, journalName=辣椒杂志, refType=null, unstructuredReference=姚明华, 焦春海, 王飞, 李宁. 非洲辣椒种质资源鉴定与评价[J]. 辣椒杂志, 2012, 10(3): 16-19, 23., articleTitle=非洲辣椒种质资源鉴定与评价, refAbstract=null), Reference(id=1276466456623706589, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, doi=null, pmid=null, pmcid=null, year=2012, volume=10, issue=3, pageStart=16, pageEnd=19, 23, url=null, language=null, rfNumber=[25], rfOrder=42, authorNames=YAO M H, JIAO C H, WANG F, LI N, journalName=Journal of China Capsicum, refType=null, unstructuredReference=YAO M H, JIAO C H, WANG F, LI N. Identification and evaluation of hot pepper germplasm resources from Africa[J]. Journal of China Capsicum, 2012, 10(3): 16-19, 23. (in Chinese), articleTitle=Identification and evaluation of hot pepper germplasm resources from Africa, refAbstract=null), Reference(id=1276466456988611038, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, doi=null, pmid=null, pmcid=null, year=2010, volume=49, issue=9, pageStart=2146, pageEnd=2148, url=null, language=null, rfNumber=[26], rfOrder=43, authorNames=王飞, 姚明华, 焦春海, journalName=湖北农业科学, refType=null, unstructuredReference=王飞, 姚明华, 焦春海. 泰国辣椒种质资源鉴定与评价[J]. 湖北农业科学, 2010, 49(9): 2146-2148., articleTitle=泰国辣椒种质资源鉴定与评价, refAbstract=null), Reference(id=1276466457072497119, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, doi=null, pmid=null, pmcid=null, year=2010, volume=49, issue=9, pageStart=2146, pageEnd=2148, url=null, language=null, rfNumber=[26], rfOrder=44, authorNames=WANG F, YAO M H, JIAO C H, journalName=Hubei Agricultural Sciences, refType=null, unstructuredReference=WANG F, YAO M H, JIAO C H. Identification and evaluation of pepper germplasm resources from Thailand[J]. Hubei Agricultural Sciences, 2010, 49(9): 2146-2148. (in Chinese), articleTitle=Identification and evaluation of pepper germplasm resources from Thailand, refAbstract=null), Reference(id=1276466457139605984, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, doi=null, pmid=null, pmcid=null, year=2020, volume=null, issue=9, pageStart=25, pageEnd=30, url=null, language=null, rfNumber=[27], rfOrder=45, authorNames=于海龙, 靳远, 刘婧, 张宝玺, 张正海, 曹亚从, 王立浩, journalName=中国蔬菜, refType=null, unstructuredReference=于海龙, 靳远, 刘婧, 张宝玺, 张正海, 曹亚从, 王立浩. 我国辣椒病毒病发生情况及发展趋势——基于2018年和2019年辣椒主产区的调查[J]. 中国蔬菜, 2020(9): 25-30., articleTitle=我国辣椒病毒病发生情况及发展趋势——基于2018年和2019年辣椒主产区的调查, refAbstract=null), Reference(id=1276466457441595873, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, doi=null, pmid=null, pmcid=null, year=2020, volume=null, issue=9, pageStart=25, pageEnd=30, url=null, language=null, rfNumber=[27], rfOrder=46, authorNames=YU H L, JIN Y, LIU J, ZHANG B X, ZHANG Z H, CAO Y C, WANG L H, journalName=China Vegetables, refType=null, unstructuredReference=YU H L, JIN Y, LIU J, ZHANG B X, ZHANG Z H, CAO Y C, WANG L H. Occurrence and development trend of pepper virus disease in China—based on main pepper producing areas investigation in 2018 and 2019[J]. China Vegetables, 2020(9): 25-30. (in Chinese), articleTitle=Occurrence and development trend of pepper virus disease in China—based on main pepper producing areas investigation in 2018 and 2019, refAbstract=null), Reference(id=1276466457542259170, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, doi=null, pmid=null, pmcid=null, year=2021, volume=34, issue=9, pageStart=1, pageEnd=6, url=null, language=null, rfNumber=[28], rfOrder=47, authorNames=杨中周, 邓竹根, 李曼, 戴祖云, journalName=中国瓜菜, refType=null, unstructuredReference=杨中周, 邓竹根, 李曼, 戴祖云. 辣椒轻斑驳病毒研究的回顾与展望[J]. 中国瓜菜, 2021, 34(9): 1-6., articleTitle=辣椒轻斑驳病毒研究的回顾与展望, refAbstract=null), Reference(id=1276466457819083235, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, doi=null, pmid=null, pmcid=null, year=2021, volume=34, issue=9, pageStart=1, pageEnd=6, url=null, language=null, rfNumber=[28], rfOrder=48, authorNames=YANG Z Z, DENG Z G, LI M, DAI Z Y, journalName=China Cucurbits and Vegetables, refType=null, unstructuredReference=YANG Z Z, DENG Z G, LI M, DAI Z Y. Review and prospect of research on Pepper mild mottle virus (PMMoV)[J]. China Cucurbits and Vegetables, 2021, 34(9): 1-6. (in Chinese), articleTitle=Review and prospect of research on Pepper mild mottle virus (PMMoV), refAbstract=null)], funds=[Fund(id=1276466448998461873, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, awardId=322MS132, language=CN, fundingSource=海南省自然科学基金面上项目(322MS132), fundOrder=null, country=null), Fund(id=1276466449073959346, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, awardId=1630032022009, language=CN, fundingSource=中央级公益性科研院所基本科研业务费专项(1630032022009), fundOrder=null, country=null)], companyList=[AuthorCompany(id=1276466427196469606, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, xref=1., ext=[AuthorCompanyExt(id=1276466427250995559, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, companyId=1276466427196469606, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China), AuthorCompanyExt(id=1276466427259384168, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, companyId=1276466427196469606, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=1.中国热带农业科学院热带作物品种资源研究所，海南海口　571101)]), AuthorCompany(id=1276466427330687337, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, xref=2., ext=[AuthorCompanyExt(id=1276466427339075946, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, companyId=1276466427330687337, language=EN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.School of Horticulture, Hainan University, Haikou, Hainan 570208, China), AuthorCompanyExt(id=1276466427603317099, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, companyId=1276466427330687337, language=CN, country=null, province=null, city=null, postcode=null, companyName=null, departmentName=null, remark=2.海南大学园艺学院，海南海口　570208)])], figs=[ArticleFig(id=1276466445680767397, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, language=EN, label=Fig. 1, caption=Detection of PMMoV, CMV and TSWV by RT-PCR, figureFileSmall=2+bWWiy/AiIogqTqAxxv4w==, figureFileBig=RyeAQEMac77YJIh6ew66AQ==, tableContent=null), ArticleFig(id=1276466446133752230, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, language=CN, label=图1, caption=PMMoV、CMV、TSWV RT-PCR检测

M: DL2000 DNA marker.

, figureFileSmall=2+bWWiy/AiIogqTqAxxv4w==, figureFileBig=RyeAQEMac77YJIh6ew66AQ==, tableContent=null), ArticleFig(id=1276466446565765543, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, language=EN, label=Fig. 2, caption=The detection of L3 resistance gene in pepper germplasm, figureFileSmall=elLvxCaflaLmniV9ZY/5ng==, figureFileBig=hTdEQfFNS5aAPAvjsMkEqg==, tableContent=null), ArticleFig(id=1276466446913892776, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, language=CN, label=图2, caption=辣椒种质L3抗性基因检测

M: DL2000 DNA marker.

, figureFileSmall=elLvxCaflaLmniV9ZY/5ng==, figureFileBig=hTdEQfFNS5aAPAvjsMkEqg==, tableContent=null), ArticleFig(id=1276466447010361769, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, language=EN, label=Fig. 3, caption=Resistance performance of some pepper germplasms to PMMoV, figureFileSmall=9rLa/QSzAcyv9hvT3qRdgQ==, figureFileBig=QGEHKU5qP2sEiVStfoIuhg==, tableContent=null), ArticleFig(id=1276466447320740266, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, language=CN, label=图3, caption=部分辣椒种质在苗期鉴定中的抗性表现, figureFileSmall=9rLa/QSzAcyv9hvT3qRdgQ==, figureFileBig=QGEHKU5qP2sEiVStfoIuhg==, tableContent=null), ArticleFig(id=1276466447438180779, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, language=EN, label=Tab. 1, caption=

Origin of pepper germplasm

, figureFileSmall=null, figureFileBig=null, tableContent=
编号No.名称Name来源Source编号No.名称Name来源Source编号No.名称Name来源Source
117YB32中国山西3814SM531S×L538m选育材料75CCJ135-1选育材料
217YB5选育材料3914SM528×14SM526选育材料76CCJ157-1英国伦敦
317SCa28m选育材料4014SM526-1选育材料77CCJ110-1法国
417YB82m中国广西贵港4114SM557-1选育材料78L505m选育材料
5CF19-13-1网络购买4214SM515m选育材料79L506m选育材料
6CF19-14-1网络购买4314SM520-1选育材料80L502-6-1m选育材料
7CF19-15-1网络购买4414SM527-1选育材料81L528m美国
8CF19-15-2网络购买4515SM8m选育材料82L542-1-1刚果
9CF19-16-1网络购买4614SM565-1选育材料83L531美国
10CF19-16-2网络购买4714SM514m选育材料84L533m美国
11CF19-19-1网络购买4814SM520m选育材料85L535-1美国
1219FB-2网络购买4914SM525×L501-1-1选育材料86L534m美国
13CF19-22-1网络购买5014SM502-1选育材料87L539美国
14CF19-22-2网络购买5114SM517m选育材料88L517m选育材料
15CF19-34m网络购买5214SM519-1选育材料89L537-1m非洲
16CF19-20-1网络购买5314SM504m选育材料90CCJ117-1选育材料
1719FB6-1泰国5414SM510-2选育材料91L518-1选育材料
1819FB-17-1泰国5514SM517-1选育材料9217YB14×17YB27杂交组合
1919FB8-2泰国5614SM510-1选育材料93L532-2美国
2019FB9-1泰国5714SM503m选育材料94L538m美国
2119FB11泰国5814SM519-3选育材料95L509选育材料
2219FB4m泰国5914SM518-2选育材料96L545墨西哥
23CF19-20-2网络购买6014SM519-2选育材料97L507-1-1m选育材料
24CCJ74-2中国台湾与法国材料杂交后代6114SM561×L502-7-1选育材料98L525-1m哥伦比亚
25CCJ121-1中国台湾与法国材料杂交后代6214SM562×L512-1-1选育材料99望都中国河北望都
26CCJ105-1中国台湾与法国材料杂交后代63CCJ52-1选育材料100CCJ64m中国台湾
2715SM39-2圭亚那64CCJ148-1英国伦敦101CCJ87-2泰国
28CCJ93-1种间杂交材料65CCJ149-1英国伦敦与中国琼海材料杂交后代102CCJ88-1泰国
2919FB9m泰国66CCJ158-1英国伦敦103CCJ126-1中国琼海
30樟树港中国湖南樟树港67CCJ1-1选育材料104CCJ137-1选育材料
31L522-1m-1巴西68CCJ2-1选育材料105CCJ133-1选育材料
3214SM508m选育材料69CCJ20-1选育材料106CCJ145m泰国
3314SM530×14SM549S选育材料70CCJ22-1选育材料107CCJ152-1英国伦敦
3414SM555×14SM1选育材料71CCJ112-1选育材料108热辣2号选育品种
3514SM521-1选育材料72CCJ113-2选育材料109热辣6号选育品种
3614SM521m选育材料73CCJ127-1选育材料1105×32选育杂交组合
3714SM567-2选育材料74CCJ122-1英国伦敦
), ArticleFig(id=1276466447803085228, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, language=CN, label=表1, caption=

参试辣椒种质来源

, figureFileSmall=null, figureFileBig=null, tableContent=
编号No.名称Name来源Source编号No.名称Name来源Source编号No.名称Name来源Source
117YB32中国山西3814SM531S×L538m选育材料75CCJ135-1选育材料
217YB5选育材料3914SM528×14SM526选育材料76CCJ157-1英国伦敦
317SCa28m选育材料4014SM526-1选育材料77CCJ110-1法国
417YB82m中国广西贵港4114SM557-1选育材料78L505m选育材料
5CF19-13-1网络购买4214SM515m选育材料79L506m选育材料
6CF19-14-1网络购买4314SM520-1选育材料80L502-6-1m选育材料
7CF19-15-1网络购买4414SM527-1选育材料81L528m美国
8CF19-15-2网络购买4515SM8m选育材料82L542-1-1刚果
9CF19-16-1网络购买4614SM565-1选育材料83L531美国
10CF19-16-2网络购买4714SM514m选育材料84L533m美国
11CF19-19-1网络购买4814SM520m选育材料85L535-1美国
1219FB-2网络购买4914SM525×L501-1-1选育材料86L534m美国
13CF19-22-1网络购买5014SM502-1选育材料87L539美国
14CF19-22-2网络购买5114SM517m选育材料88L517m选育材料
15CF19-34m网络购买5214SM519-1选育材料89L537-1m非洲
16CF19-20-1网络购买5314SM504m选育材料90CCJ117-1选育材料
1719FB6-1泰国5414SM510-2选育材料91L518-1选育材料
1819FB-17-1泰国5514SM517-1选育材料9217YB14×17YB27杂交组合
1919FB8-2泰国5614SM510-1选育材料93L532-2美国
2019FB9-1泰国5714SM503m选育材料94L538m美国
2119FB11泰国5814SM519-3选育材料95L509选育材料
2219FB4m泰国5914SM518-2选育材料96L545墨西哥
23CF19-20-2网络购买6014SM519-2选育材料97L507-1-1m选育材料
24CCJ74-2中国台湾与法国材料杂交后代6114SM561×L502-7-1选育材料98L525-1m哥伦比亚
25CCJ121-1中国台湾与法国材料杂交后代6214SM562×L512-1-1选育材料99望都中国河北望都
26CCJ105-1中国台湾与法国材料杂交后代63CCJ52-1选育材料100CCJ64m中国台湾
2715SM39-2圭亚那64CCJ148-1英国伦敦101CCJ87-2泰国
28CCJ93-1种间杂交材料65CCJ149-1英国伦敦与中国琼海材料杂交后代102CCJ88-1泰国
2919FB9m泰国66CCJ158-1英国伦敦103CCJ126-1中国琼海
30樟树港中国湖南樟树港67CCJ1-1选育材料104CCJ137-1选育材料
31L522-1m-1巴西68CCJ2-1选育材料105CCJ133-1选育材料
3214SM508m选育材料69CCJ20-1选育材料106CCJ145m泰国
3314SM530×14SM549S选育材料70CCJ22-1选育材料107CCJ152-1英国伦敦
3414SM555×14SM1选育材料71CCJ112-1选育材料108热辣2号选育品种
3514SM521-1选育材料72CCJ113-2选育材料109热辣6号选育品种
3614SM521m选育材料73CCJ127-1选育材料1105×32选育杂交组合
3714SM567-2选育材料74CCJ122-1英国伦敦
), ArticleFig(id=1276466447920525741, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, language=EN, label=Tab. 2, caption=

Primer sequence for detection virus

, figureFileSmall=null, figureFileBig=null, tableContent=
病毒名称Virus name引物名称Primer name序列（5ʹ-3ʹ）Sequence (5ʹ-3ʹ)片段大小Fragment size/bp
PMMOV[19]PMMoV-FGCCGTGCTAGATTCTCTAGTGTC220
PMMoV-RCGCTCTCGAACAGAGCTTG
CMV[20]CMV-FCGTTTAGTGACTTCAGACAG735
CMV-RATGGACAAATCTGAATCAAC
TSWV[21]TSWV-FATGTYTAAGGTTAAGCTCACTAAG750
TSWV-RTTAAGCAAGTTCTGTGAGTTTTGCC
), ArticleFig(id=1276466448155406766, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, language=CN, label=表2, caption=

病毒检测引物序列

, figureFileSmall=null, figureFileBig=null, tableContent=
病毒名称Virus name引物名称Primer name序列（5ʹ-3ʹ）Sequence (5ʹ-3ʹ)片段大小Fragment size/bp
PMMOV[19]PMMoV-FGCCGTGCTAGATTCTCTAGTGTC220
PMMoV-RCGCTCTCGAACAGAGCTTG
CMV[20]CMV-FCGTTTAGTGACTTCAGACAG735
CMV-RATGGACAAATCTGAATCAAC
TSWV[21]TSWV-FATGTYTAAGGTTAAGCTCACTAAG750
TSWV-RTTAAGCAAGTTCTGTGAGTTTTGCC
), ArticleFig(id=1276466448272847279, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, language=EN, label=Tab. 3, caption=

Identification results of PMMoV resistance in pepper germplasms

, figureFileSmall=null, figureFileBig=null, tableContent=
编号No.种质名称Germplasms name栽培种Cultivar病情指数Disease index抗性级别Resistance level编号No.种质名称Germplasms name栽培种Cultivar病情指数Disease index抗性级别Resistance level
117YB32C. chinense Jacquin37.78±1.11nmlMR25CCJ22-1C. chinense Jacquin55.56±2.63dS
217SCa28mC. annuum L.39.26±1.74ljkiMR26CCJ112-1C. chinense Jacquin38.94±2.55gfMR
3CF19-22-1C. annuum L.38.52±2.20nmlkMR27CCJ113-2C. chinense Jacquin60.00±1.39aS
4CF19-34mC. annuum L.8.86±1.35uHR28CCJ135-1C. chinense Jacquin60.00±2.12aS
519FB6-1C. annuum L.8.79±2.31uHR29CCJ157-1C. chinense Jacquin58.52±1.96bS
619FB11C. annuum L.37.04±1.51oMR30CCJ110-1C. chinense Jacquin36.74±2.28gfMR
715SM39-2C. chinense Jacquin36.30±2.04pMR31L506mC. chinense Jacquin38.58±2.33jihgMR
8CCJ93-1C. chinense Jacquin17.78±1.99sR32L502-6-1mC. chinense Jacquin37.04±1.59nmlMR
914SM555×14SM1C. chinense Jacquin29.63±2.65rMR33L528mC. chinense Jacquin39.44±2.18fMR
1014SM567-2C. chinense Jacquin39.26±2.60qMR34L531C. chinense Jacquin38.95±2.53ihgMR
1114SM526-1C. chinense Jacquin37.04±2.14nmlMR35L533mC. chinense Jacquin37.04±1.66oMR
1214SM565-1C. chinense Jacquin40.00±1.50jihgMR36L535-1C.chinense Jacquin39.93±1.35gfMR
1314SM514mC. chinense Jacquin58.52±2.77bS37L539C. chinense Jacquin37.04±1.13onmMR
1414SM502-1C. chinense Jacquin39.74±1.49gfMR38L537-1mC. chinense Jacquin54.81±1.04dS
1514SM519-1C. chinense Jacquin39.06±1.61mlkMR39L532-2C. chinense Jacquin38.14±1.66gfMR
1614SM504mC.chinense Jacquin39.37±2.47mlkjMR40L545C. chinense Jacquin57.04±1.54cS
1714SM503mC. chinense Jacquin39.26±1.46jihgMR41CCJ64mC. chinense Jacquin38.79±2.77hgfMR
1814SM519-3C.chinense Jacquin35.22±2.08gfMR42CCJ87-2C. chinense Jacquin40.74±2.29ihgS
1914SM518-2C. chinense Jacquin39.26±1.40kjihMR43CCJ137-1C.chinense Jacquin37.04±1.60onmMR
2014SM519-2C. chinense Jacquin37.78±1.77nmlMR44CCJ133-1C. chinense Jacquin58.52±2.53bS
21CCJ52-1C. chinense Jacquin34.30±1.49nmlMR45热辣2号C. chinense Jacquin37.04±1.76onMR
22CCJ148-1C. chinense Jacquin37.75±2.00feMR46热辣6号C. chinense Jacquin38.52±1.35mlkMR
23CCJ2-1C. chinense Jacquin38.15±1.46eMR4714SM517（CK–）C. chinense Jacquin55.56±1.65dS
24CCJ20-1C. chinense Jacquin37.44±1.35gfMR48中椒107（CK+）C. annuum L.11.11±0.82tR
), ArticleFig(id=1276466448583225776, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1276213327835567092, language=CN, label=表3, caption=

辣椒种质PMMoV抗性鉴定结果

, figureFileSmall=null, figureFileBig=null, tableContent=
编号No.种质名称Germplasms name栽培种Cultivar病情指数Disease index抗性级别Resistance level编号No.种质名称Germplasms name栽培种Cultivar病情指数Disease index抗性级别Resistance level
117YB32C. chinense Jacquin37.78±1.11nmlMR25CCJ22-1C. chinense Jacquin55.56±2.63dS
217SCa28mC. annuum L.39.26±1.74ljkiMR26CCJ112-1C. chinense Jacquin38.94±2.55gfMR
3CF19-22-1C. annuum L.38.52±2.20nmlkMR27CCJ113-2C. chinense Jacquin60.00±1.39aS
4CF19-34mC. annuum L.8.86±1.35uHR28CCJ135-1C. chinense Jacquin60.00±2.12aS
519FB6-1C. annuum L.8.79±2.31uHR29CCJ157-1C. chinense Jacquin58.52±1.96bS
619FB11C. annuum L.37.04±1.51oMR30CCJ110-1C. chinense Jacquin36.74±2.28gfMR
715SM39-2C. chinense Jacquin36.30±2.04pMR31L506mC. chinense Jacquin38.58±2.33jihgMR
8CCJ93-1C. chinense Jacquin17.78±1.99sR32L502-6-1mC. chinense Jacquin37.04±1.59nmlMR
914SM555×14SM1C. chinense Jacquin29.63±2.65rMR33L528mC. chinense Jacquin39.44±2.18fMR
1014SM567-2C. chinense Jacquin39.26±2.60qMR34L531C. chinense Jacquin38.95±2.53ihgMR
1114SM526-1C. chinense Jacquin37.04±2.14nmlMR35L533mC. chinense Jacquin37.04±1.66oMR
1214SM565-1C. chinense Jacquin40.00±1.50jihgMR36L535-1C.chinense Jacquin39.93±1.35gfMR
1314SM514mC. chinense Jacquin58.52±2.77bS37L539C. chinense Jacquin37.04±1.13onmMR
1414SM502-1C. chinense Jacquin39.74±1.49gfMR38L537-1mC. chinense Jacquin54.81±1.04dS
1514SM519-1C. chinense Jacquin39.06±1.61mlkMR39L532-2C. chinense Jacquin38.14±1.66gfMR
1614SM504mC.chinense Jacquin39.37±2.47mlkjMR40L545C. chinense Jacquin57.04±1.54cS
1714SM503mC. chinense Jacquin39.26±1.46jihgMR41CCJ64mC. chinense Jacquin38.79±2.77hgfMR
1814SM519-3C.chinense Jacquin35.22±2.08gfMR42CCJ87-2C. chinense Jacquin40.74±2.29ihgS
1914SM518-2C. chinense Jacquin39.26±1.40kjihMR43CCJ137-1C.chinense Jacquin37.04±1.60onmMR
2014SM519-2C. chinense Jacquin37.78±1.77nmlMR44CCJ133-1C. chinense Jacquin58.52±2.53bS
21CCJ52-1C. chinense Jacquin34.30±1.49nmlMR45热辣2号C. chinense Jacquin37.04±1.76onMR
22CCJ148-1C. chinense Jacquin37.75±2.00feMR46热辣6号C. chinense Jacquin38.52±1.35mlkMR
23CCJ2-1C. chinense Jacquin38.15±1.46eMR4714SM517（CK–）C. chinense Jacquin55.56±1.65dS
24CCJ20-1C. chinense Jacquin37.44±1.35gfMR48中椒107（CK+）C. annuum L.11.11±0.82tR
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辣椒种质资源PMMoV抗性基因检测与抗性鉴定
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陈建分 1, 2 , 曹振木 1 , 秦于玲 1 , 申龙斌 1 , 刘维侠 1 , 朱丹 1 , 吴怡婷 1 , 刘子记 1, * , 王旭 2, *
热带作物学报 | 组学与生物技术 2024,45(4): 682-690
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热带作物学报 |组学与生物技术 2024 , 45 (4) : 682 -690
辣椒种质资源PMMoV抗性基因检测与抗性鉴定
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陈建分（1992—），女，硕士，研究方向：蔬菜抗病育种。

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陈建分1, 2, 曹振木1, 秦于玲1, 申龙斌1, 刘维侠1, 朱丹1, 吴怡婷1, 刘子记1, * , 王旭2, *
作者信息
  • 1.中国热带农业科学院热带作物品种资源研究所，海南海口　571101
  • 2.海南大学园艺学院，海南海口　570208
通讯作者:
* 刘子记（LIU Ziji），E-mail：
王旭（WANG Xu），E-mail：。
Resistance Gene Detection and Resistance Identification of PMMoV in Pepper Germplasm
Jianfen CHEN1, 2, Zhenmu CAO1, Yuling QIN1, Longbin SHEN1, Weixia LIU1, Dan ZHU1, Yiting WU1, Ziji LIU1, * , Xu WANG2, *
Affiliations
  • 1.Tropical Crops Genetic Resources Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China
  • 2.School of Horticulture, Hainan University, Haikou, Hainan 570208, China
出版时间: 2024-04-25 doi: 10.3969/j.issn.1000-2561.2024.04.004
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摘要
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辣椒轻斑驳病毒（Pepper mild mottle virus，PMMoV）是世界范围内最严重的病原体之一，目前已报道的PMMoV包括P0、P1、P1,2、P1,2,3、P1,2,3,4五种致病型。辣椒轻斑驳病毒寄主范围广、致病力强，严重影响辣椒的产量和品质，开展抗病育种是防治该病毒最为经济有效的手段。本课题组前期研究已明确了感染海南儋州辣椒种质资源圃的PMMoV为P1,2致病型，为了筛选抗PMMoV的辣椒种质资源，以110份种质为材料，利用已报道的与L3连锁的分子标记对辣椒种质进行检测。结果表明：在110份材料中共检测到46份辣椒种质含有与L3连锁的分子标记。采用苗期人工接种鉴定方法，对含有抗性分子标记的46份辣椒种质接种PMMoV P1,2致病型进行抗性鉴定。接种5 d后调查局部症状，接种20 d后调查系统症状，计算病情指数。结果表明：不同的辣椒种质在抗病性上存在显著差异，各种质的病情指数在8.86~60.00之间，对PMMoV P1,2致病型表现高抗（HR）的种质为CF19-34m和19FB6-1；表现抗病（R）的种质为CCJ93-1；表现中抗（MR）的种质包括17YB32、17SCa28m、15SM39-2、14SM555×14SM1、14SM567-2、14SM526-1、14SM565-1、14SM502-1、14SM519-1、14SM504m、14SM503m、14SM519-3、14SM518-2、14SM519-2、CCJ52-1等34份；表现感病（S）的种质包括14SM514m、CCJ22-1、CCJ113-2、CCJ135-1、CCJ157-1、L537-1m、L545、CCJ87-2、CCJ133-1，获得的高抗、抗病、中抗种质占总鉴定材料的80.43%。本研究结果为创制抗辣椒轻斑驳病毒种质资源和培育抗性品种奠定基础。

辣椒  /  种质资源  /  PMMoV  /  分子标记  /  抗病性鉴定

Pepper mild mottle virus (PMMoV) is one of the most serious pathogens worldwide. Currently, five pathogenic types of PMMoV have been reported, including P0, P1, P1,2, P1,2,3 and P1,2,3,4. The wide host range and strong pathogenicity of PMMoV seriously affect the yield and quality of pepper, and breeding for disease resistance is the most economical and effective means to control this virus. Our previous study determined that PMMoV infected with Danzhou pepper germplasm nursery in Hainan is the P1,2 pathogenic type. In order to screen PMMoV-resistant pepper germplasm resources, 110 germplasms were tested for PMMoV resistance using the reported L3-linked molecular markers. 46 pepper germplasms containing molecular markers of resistance were inoculated with PMMoV P1,2 pathogenic type by inoculation at seeding stage. Local symptoms were investigated 5 d after inoculation, and systemic symptoms were investigated 20 d after inoculation, and disease indices were calculated. The results showed that there were sig-nificant differences in disease resistance among different pepper germplasms, and the disease indexes of various germplasms ranged from 8.86-60.00. The germplasms showing high resistance (HR) to PMMoV P1,2 pathogenic type were CF19-34m and 19FB6-1. The germplasms showing resistance (R) wasCCJ93-1. The germplasms showing medium-resistance (MR) included 17YB32, 17SCa28m, 15SM39-2, 14SM555×14SM1, 14SM567-2, 14SM526-1, 14SM565-1, 14SM502-1, 14SM519-1, 14SM504m, 14SM503m, 14SM519-3, 14SM518-2, 14SM519-2, CCJ52-1, etc. The presenting susceptible (S) germplasms were 14SM514m, CCJ22-1, CCJ113-2, CCJ135-1, CCJ157-1, L537-1m, L545, CCJ87-2 and CCJ133-1, and the obtained disease-resistant germplasms accounted for 80.43% of the total identified materials. The results of this study would lay the foundation for creating germplasm resources resistant to pepper mild mottle virus and breeding resistant varieties.

pepper (Capsicum annuum L.)  /  germplasm  /  PMMoV  /  molecular markers  /  disease resistance identification
陈建分, 曹振木, 秦于玲, 申龙斌, 刘维侠, 朱丹, 吴怡婷, 刘子记, 王旭. 辣椒种质资源PMMoV抗性基因检测与抗性鉴定. 热带作物学报, 2024 , 45 (4) : 682 -690 . DOI: 10.3969/j.issn.1000-2561.2024.04.004
Jianfen CHEN, Zhenmu CAO, Yuling QIN, Longbin SHEN, Weixia LIU, Dan ZHU, Yiting WU, Ziji LIU, Xu WANG. Resistance Gene Detection and Resistance Identification of PMMoV in Pepper Germplasm[J]. Chinese Journal of Tropical Crops, 2024 , 45 (4) : 682 -690 . DOI: 10.3969/j.issn.1000-2561.2024.04.004
正文
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辣椒（Capsicum annuum L.）为茄科辣椒属一年生或多年生二倍体作物，是全球重要的蔬菜作物之一。辣椒果实富含类胡萝卜素、蛋白质、维生素C等多种营养物质，既可直接食用又可用作调味品，辣椒提取物在医药和化工领域也具有广泛应用。据联合国粮食及农业组织（FAO）统计，2019年全球鲜食辣椒和加工辣椒总种植面积约为3.7190×106 hm2，总产量约为4.2283×107 t；我国2019年辣椒种植面积约为8.4700×105 hm2，总产量达1.9333×107 t，居全球首位[1]。
长期以来，病毒病一直是威胁辣椒生产的重要因素。目前，我国报道的病毒种类有30余种，近年来辣椒轻斑驳病毒（Pepper mild mottle viru，PMMoV）已逐渐成为海南[2]、广西[3]、广东[4]、四川[5]、安徽[6]、黑龙江[7]、山东[8-9]、西藏[10]等多个辣椒产区普遍发生的病害。PMMoV属于烟草花叶病毒属（Tobamovirus），主要通过种传、汁液传播[11-12]。辣椒感染PMMoV后叶片症状表现轻微皱缩、褪绿斑驳和花叶症状，果实被侵染后表现为扁平、变小、褪绿斑驳、凹凸斑点、畸形与坏死等现象。
L1、L2、L3、L4系列等位基因是辣（甜）椒抗烟草花叶病毒属病毒的主要抗性基因[13-15]，这些基因被引入辣椒栽培种中保护植株免受烟草花叶病毒的侵染。目前我国辣椒生产上的PMMoV致病型多以P1,2株系为主[16-17]，P1,2致病型病毒株系能够系统侵染带有L1、L2抗性基因的辣（甜）椒，而带有L3抗性基因的辣（甜）椒对该病毒株系表现为抗性，L3基因能够应对我国辣椒生产上的主要问题。张宝玺等[18]通过常规育种技术和分子标记相结合，创制出含有抗PMMoV（P0,1,2）L3基因新品种中椒105号、中椒106号、中椒107号、中椒108号。TOMITA等[15]通过图位克隆获得L3基因，并利用同位克隆的方法获得了L的6个等位基因：L1、L1a、L1c、L2、Lb和L4。鉴定抗性种质是选育抗病品种不可缺少的步骤，目前，高抗PMMoV的辣椒种质鲜有报道。由于辣椒的栽培种遗传多样性比较丰富、L基因组结构较为复杂，目前尚未利用基因序列开发功能标记。因此，本研究利用已报道的L3连锁标记，遗传距离为0.00015 cM，以110份辣椒种质资源为研究对象，结合抗病基因检测和苗期抗性鉴定，精准筛选抗辣椒轻斑驳病毒的种质资源，以期为选育抗PMMoV辣椒品种提供科学依据。
1　材料与方法
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1.1　材料
参试辣椒种质资源共110份，材料名称和来源地详见表1。PMMoV鉴定毒原P1,2株系由本课题组从田间发病植株上分离提纯保存。
1.2　方法
1.2.1　毒原扩繁及检测
将PMMoV毒源人工接种在本氏烟草（Nicotiana benthamiana）上进行扩繁，待发病后选取感病叶片，利用RT-PCR方法进行分子检测，引物由生工生物工程（上海）股份有限公司合成，引物序列见表2。
1.2.2　 L3抗性基因检测
与L3连锁的分子标记参照TOMITA等[22]的方法。根据Bioteke DNA试剂盒（北京百泰克生物技术有限公司）提取gDNA，采用1%琼脂糖凝胶电泳和紫外分光光度计测定DNA的质量和浓度，并将浓度稀释至100 ng/μL，保存于–20 ℃冰箱备用。PCR反应体系为20 μL：PCR-mix（2×Es Taq Master Mix）10 μL，模板DNA 1.0 μL，正向引物和反向引物（10 μmol/L）各1 μL，ddH2O补齐至20 μL。PCR扩增程序：94 ℃预变性5 min；94 ℃变性45 s，60 ℃退火45 s，72 ℃延伸1.5 min，35个循环；最后72 ℃延伸10 min，4 ℃保存。
1.2.3　PMMoV苗期人工接种抗性鉴定
将筛选出含有抗性基因分子标记的辣椒种质进行苗期人工接种鉴定，以14SM517m材料为感病对照（CK–），中椒107为抗病对照（CK+）。2022年8月3日，将所有辣椒种子经10%磷酸三钠溶液浸种35 min，用蒸馏水清洗除去辣味，在55 ℃下温汤浸种4 h，置于28 ℃恒温培养箱中培养。出芽后播种于50孔塑料穴盘内育苗，每份参试材料3次重复，每重复15株。幼苗长至两叶一心期时，移栽至15.2 cm×15.6 cm塑料盆中，育苗基质为高温灭菌基质（蛭石∶营养土=1∶2）。
待辣椒幼苗长至4~5叶期时，取感染PMMoV的烟草新叶，加入10倍于鲜叶质量的0.01 mol/L磷酸缓冲液（PBS，pH 7.0）中，在冰上研磨匀浆，离心去渣取上清液，接种液现配现用。在叶面抹薄层600目金刚砂，用手指蘸取少量接种液轻轻摩擦叶面。接种30 min后用清水冲去叶面多余汁液，为避免病毒接不上的情况，3 d后再接种1次。将幼苗置于28 ℃光照培养箱中培养，接种后5 d调查局部症状，20 d调查系统症状，计算病情指数，并进行抗性分级。
1.2.4　病情调查与统计
PMMoV抗性调查的病情指数计算及抗性级别的划分均参考《辣椒种质资源描述规范和数据标准》[23]。
按上述标准划分单株病情级别。0：无任何症状；1：心叶明脉，或接种叶少量急性小枯斑；3：少数叶片呈花叶，或接种叶脱落，茎部产生坏死斑；5：多数叶片花叶，少数叶片畸形，皱缩，或茎部产生坏死条斑；7：多数叶片畸形，皱缩，植株矮化，或茎、枝和叶脉系统坏死；9：植株严重矮化，停止生长；或植株严重系统坏死，甚至死亡。病情指数计算公式为：
品系群体抗病性划分标准：免疫（I），DI=0；高抗（HR），0<DI≤10；抗病（R），10<DI≤20；中抗（MR），20<DI≤40；感病（S），40<DI ≤60；高感（HS），60<DI≤100。
1.3　数据处理
试验数据通过Excel软件进行分析、计算平均值和标准差，使用SPSS 25软件进行单因素方差分析，采用LSD或Duncan’s法进行多重比较分析。
2　结果与分析
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2.1　毒原扩繁检测结果
RT-PCR鉴定结果表明，PMMoV特异性引物能扩增出220 bp左右的条带，与目的条带一致，而CMV和TSWV检测引物均未扩增出条带（图1），初步证明此毒原为PMMoV单一感染毒原，可用于接种鉴定。
2.2　辣椒种质L3抗性基因检测
L3抗性分子标记鉴定结果表明，17YB32、17SCa28m、CF19-34m、19FB6-1、15SM39-2、CCJ93-1、14SM555×14SM1、14SM567-2、14SM526-1、14SM 565-1、14SM514m、14SM502-1、14SM519-1、14SM 504m、14SM503m、14SM519-3、14SM518-2、14SM 519-2等46份种质含有与抗性基因L3连锁的分子标记（图2），其余64份种质均未扩增出目标条带，说明这些种质不含L3抗性基因。
2.3　辣椒种质PMMoV抗性鉴定
辣椒种质PMMoV抗性鉴定结果表明（图3、表3），46份供试种质存在显著性差异，未发现对PMMoV免疫的种质，各种质的病情指数在8.86~60.00之间。病情指数能够反映辣椒种质抗PMMoV能力的强弱，数值越高，表明抗性越低，反之，表明抗性越高。其中，高抗种质为CF19-34m和19FB6-1，病情指数分别为8.86和8.79，抗病性显著强于感病种质，叶片几乎无感染症状；抗病种质CCJ93-1次之，叶片仅有轻微的花叶症状；中抗材料17YB32、17SCa28m、15SM39-2、14SM555×14SM1、14SM567-2、14SM526-1、14SM565-1、14SM502-1、14SM519-1、14SM504m、14SM503m、14SM519-3、14SM518-2、14SM519-2、CCJ52-1等34份，植株少数叶片出现花叶、皱缩；感病种质14SM514m、CCJ22-1、CCJ113-2、CCJ135-1、CCJ157-1、CCJ537-1m、L545、CCJ87-2、CCJ133-1，这9份材料中CCJ113-2和CCJ135-1的病情指数最大，病情指数为60.00。感病种质的抗性能力弱，植株叶片多数皱缩和心叶畸形，植株矮化。总体来看，高抗、抗病和中抗材料占总鉴定材料的80.43%。
3　讨论
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本研究结合L3抗性基因检测和苗期人工接种抗性鉴定方法，在110份辣椒种质中筛选出抗PMMoV P1,2种质37份，这些抗病种质可为辣椒的抗病育种提供优良的种质资源。近年来，辣椒轻斑驳病毒在国内快速蔓延，研究者正加快筛选对其具较大抗性的辣椒种质。秦蕾等[24]对51份辣椒种质资源进行了TMV抗性鉴定，获得抗病材料15份，其中高抗材料2份，占总鉴定材料的3.92%；姚明华等[25]对引进的42份非洲辣椒材料进行了TMV抗性鉴定，获得抗性材料9份；王飞等[26]在74份泰国辣椒种质中筛选出7份TMV抗性材料。已有研究筛选到的抗性材料相对较少，本研究鉴定结果表明，不同的辣椒种质表现的抗性有所不同，其中多数抗病材料表现为中抗，仅获得2份高抗材料和1份抗病材料。这可能是由于抗病性受到多种因素的影响，包括外在的环境因子、内在基因表达及辣椒与病毒的相互作用等。其次，可能高效L基因主要存在于辣椒野生种当中。
目前，辣椒抗PMMoV的L基因已被克隆。本研究利用已报道的与L3连锁的分子标记对110份辣椒种质进行分子鉴定，结果显示有46份种质含有抗性分子标记，包括一年生辣椒栽培种（C. annuum L）和中国辣椒栽培种（C. chinense Jacquin）。在接种PMMoV P1,2菌株后，其中有9份含有抗性分子标记的种质表现为感病，这种现象可能与品种的遗传背景有关。本研究采用的是与L3连锁的标记而不是功能标记，因此存在假阳性的可能，为了准确鉴定辣椒种质对PMMoV的抗性，需要结合抗性鉴定结果进行综合判断。
病毒病一直是威胁辣椒生产的主要病害，对辣椒生产造成严重的经济损失[27]。由于PMMoV具有种传的特性，使用化学杀菌剂和传统的农业防治措施无法控制病毒病的为害。与传统育种方式相比，利用分子标记技术可以快速对已知抗性基因进行检测，缩短了育种年限，有利于寻找新抗源，拓宽辣椒品种抗性遗传多样性，对PMMoV抗病品种的选育尤为重要。
本课题组前期研究已经明确了海南儋州辣椒种质资源圃感染的PMMoV为P1,2致病型，应用L3抗性基因可以有效克服PMMoV P1,2致病型的为害。然而，病毒毒株会克服抗性基因而变异，L3基因对PMMoV P1,2,3,4致病型不具有抗性[28]。因此，未来的工作需要挖掘鉴定其他PMMoV抗性基因用于辣椒抗性种质资源的创制，为辣椒抗PMMoV育种提供更高效的抗源材料。
基金
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  • 海南省自然科学基金面上项目(322MS132)
  • 中央级公益性科研院所基本科研业务费专项(1630032022009)
文献
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2024年第45卷第4期
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doi: 10.3969/j.issn.1000-2561.2024.04.004
  • 接收时间：2022-12-07
  • 首发时间：2026-06-23
  • 出版时间：2024-04-25
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  • 收稿日期：2022-12-07
  • 修回日期：2023-01-04
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海南省自然科学基金面上项目(322MS132)
中央级公益性科研院所基本科研业务费专项(1630032022009)
作者信息
    1.中国热带农业科学院热带作物品种资源研究所，海南海口　571101
    2.海南大学园艺学院，海南海口　570208

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* 刘子记（LIU Ziji），E-mail：
王旭（WANG Xu），E-mail：。
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2种不同金属材料的力学参数
科
Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
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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