黄麻双生病毒 CoYVV 的分子鉴定和抗性种质筛选  

Molecular identification of a geminivirus CoYVV and screening of resistant germplasms in jute

作　　者：杨昕 林文忠 陈思远 杜振国 林杰 祁建民 方平平[1,2] 陶爱芬 张立武[1,2,3] YANG Xin;LIN Wen-Zhong;CHEN Si-Yuan;DU Zhen-Guo;LIN Jie;QI Jian-Min;FANG Ping-Ping;TAO Ai-Fen;ZHANG Li-Wu(Key Laboratory of Ministry of Education for Genetics,Breeding and Multiple Utilization of Crops/Fujian Key Laboratory for Crop Breeding by Design/College of Agriculture,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China;Experiment Station of Ministry of Ag-riculture and Rural Affairs for Jute and Kenaf in Southeast China/Fujian Public Platform for Germplasm Resources of Bast Fiber Crops/Fujian International Science and Technology Cooperation Base for Genetics,Breeding and Multiple Utilization Development of Southern Economic Crops,Fujian Agriculture and Forestry University,Fuzhou 350002,Fujian,China;State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops,Fuzhou 350002,Fujian,China)

机构地区：[1]福建农林大学农学院/作物遗传育种与综合利用教育部重点实验室/福建省作物设计育种重点实验室,福建福州350002 [2]福建农林大学农业农村部东南黄红麻实验观测站/福建省麻类种质资源共享平台/福建省南方经济作物遗传育种与多用途开发国际科技合作基地,福建福州350002 [3]闽台作物有害生物生态防控国家重点实验室,福建福州350002

出　　处：《作物学报》2022年第3期624-634,共11页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(31771369);国家现代农业产业技术体系建设专项(CARS-19-E06)资助。

摘　　要：越南黄麻黄脉病毒(Corchorus yellow vein Vietnam virus,CoYVV)是影响黄麻纤维品质和产量的双生病毒,其基因组含2个环状单链DNA(DNA-A和DNA-B)。鉴定黄麻双生病毒CoYVV并筛选抗病毒种质,不仅丰富对双生病毒分布和进化的认识,而且为黄麻抗性基因克隆奠定了基础。2018年福建省三明市福建农林大学洋中科教基地发现黄麻种质资源圃长果种黄麻福农5号疑似感染黄色花叶病。利用CTAB法提取该品种感病叶片DNA,通过PCR、滚环扩增及测序技术获得病毒基因组序列。通过同源序列比对和进化树构建明确该病毒与新旧世界双生病毒的关系。含有CoYVV DNA-A和CoYVV DNA-B侵染性克隆载体的农杆菌等比例混合,子叶注射法侵染具有代表性的7份长果种和7份圆果种黄麻品种(系),定期观察发病株数(病株率)并PCR检测接种黄麻的侵染率。PCR检测结果证实,福农5号病样感染了CoYVV,命名为CoYVV-FS。其DNA-A为2724 bp,DNA-B为2691 bp。进化分析表明,相较于旧世界菜豆金黄花叶病毒(begomoviruses),CoYVV-FS与新世界begomoviruses更近。农杆菌接种14份黄麻品种43 d时,统计病株率和侵染率。结果表明该病毒可以成功侵染13份黄麻品种,病株率和侵染率分别在0~60%和0~100%之间,说明该病毒在黄麻中具有较高侵染性。807元引马里的病株率和侵染率皆为0,表现为抗性种质;而巴长4号和印度墨绿子在接种30 d时产生黄脉、卷曲等典型的表型,表现出对CoYVV具有高度感病性。黄麻种植资源圃发现的CoYVV-FS与前期已报道的CoYVV属于同一分离物,它能侵染多数黄麻种质,在长果种黄麻上引起严重症状。Corchorus yellow vein Vietnam virus(CoYVV)is a geminivirus that affects yield and quality in jute.Its genome is consisted of two circular single-stranded DNAs,named DNA-A and DNA-B,respectively.Detection of CoYVV and screening of resistant germplasm will not only increase our understanding of geminivirus distribution and evolution but also lay the foundation for resistance gene cloning.Funong 5(C.olitorius.)was suspected to be infected with leaf mosaic symptoms in jute germplasm resource nursery of Yangzhong Science and Education Base of Fujian Agriculture and Forestry University(Sanming,Fujian,China)in 2018.The diseased samples were collected and subjected to total DNA extraction by CTAB.The virus in the sample was identified by PCR,RCA,and sequencing.Then the mixture of Agrobacterium tumefaciens containing two plasmids(infectious clone vector plasmids of CoYVV DNA-A and DNA-B)in 1:1 ratio was injected to 7 representative C.capsularis varieties and 7 representative C.olitorius ones.After inoculation,we investigated symptom development(disease incidence)and PCR was used to detect the rate of infection.The results showed that the symptomatic samples in the field were infected by an isolate of CoYVV(CoYVV-FS).The size of the DNA-A and DNA-B of CoYVV-FS were 2724 bp and 2691 bp,respectively.Phylogenetic analysis revealed that CoYVV-FS was more related to New World begomoviruses than to Old World begomoviruses.Moreover,the inocu-lation of 14 representative jute varieties showed that CoYVV was infectious to jute.The disease incidence was 0 in the 7 C.cap-sularis varieties and 0-60%in the 7 C.olitorius varieties.The infection rates for the 14 jute varieties was from 0 to 100%.Both disease incidence and infection rates of 807 yuanyinmali(C.olitorius)were 0,indicating that it was the resistant germplasm;while Bachang4(C.olitorius)and Yindumolvzi(C.olitorius)produced yellow vein and curly-leaf phenotypes after 30 days of inoculation,showing that it was the sensitive germplasm.The CoYVV-FS in jute germplasm resource nurser

关 键 词：黄麻 越南黄麻黄脉病毒 CoYVV 侵染性克隆 抗性鉴定 

分 类 号：S435.63[农业科学—农业昆虫与害虫防治]