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Acta Agronomica Sinica ›› 2022, Vol. 48 ›› Issue (3): 624-634.doi: 10.3724/SP.J.1006.2022.14017

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

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

YANG Xin1,2,3(), LIN Wen-Zhong3, CHEN Si-Yuan1,2, DU Zhen-Guo3, LIN Jie3, QI Jian-Min1,2, FANG Ping-Ping1,2, TAO Ai-Fen1,2, ZHANG Li-Wu1,2,3,*()   

  1. 1Key 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
    2Experiment Station of Ministry of Agriculture 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
    3State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops, Fuzhou 350002, Fujian, China
  • Received:2021-01-28 Accepted:2021-06-16 Online:2022-03-12 Published:2021-07-22
  • Contact: ZHANG Li-Wu E-mail:295548772@qq.com;lwzhang@fafu.edu.cn;zhang_liwu@hotmail.com
  • Supported by:
    National Natural Science Foundation of China(31771369);China Agriculture Research System(CARS-19-E06)

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Abstract:

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 inoculation of 14 representative jute varieties showed that CoYVV was infectious to jute. The disease incidence was 0 in the 7 C. capsularis 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 nursery belonged to the same isolate as the CoYVV previously reported, which could infect most jute varieties (lines) and cause serious symptoms in C. olitorius.

Key words: jute, Corchorus yellow vein Vietnam virus, CoYVV, infectious clone, resistance evaluation

Table 1

Primers for detection and full-length genome cloning of CoYVV in this study"

引物Primer 序列Sequence (5°-3°)
DeP3 GANGSATGHGTRCADGCCATATA
AV494 GCCYATRTAYAGRAAGCCMAG
CoVV-A-CP-F-149 CCATGTATAGGAAGCCCAGGATC
CoVV-A-CP-R-542 ATGACCTGATAACGATCACGGAG
CoYVV-AF TTCGTGCATCTGTACTTCTTCTGG
CoYVV-AR CTGCAGATTGTTCGATCTATCCAA

Fig. 1

Virus-infected symptoms of jute Funong 5 in the field"

Fig. 2

Genomic structure of DNA-A and DNA-B in CoYVV-FS in jute"

Table 2

Open reading frames encoded in DNA-A and DNA-B of CoYVV in jute"

开放阅读框
Open reading frame		 序列位置
Position
(nt)		 序列长度
Sequence length (bp)		 氨基酸数目
Amino acid residues (aa)		 蛋白分子量
Protein molecular weight (kD)		 预测功能
Function prediction		 参考文献
Reference
AC1 1449-2528 1080 359 40.6 复制相关蛋白
Replication-associated protein		 [26]
AC2 1139-1564 426 141 16.0 转录激活蛋白
Transcriptional activator protein		 [26]
AC3 1009-1401 393 130 15.8 增强病毒DNA复制蛋白
AC3 protein enhances viral DNA replication		 [27]
AC4 2087-2377 291 96 11.0 与病毒的移动相关蛋白
AC4 protein involved in virus movement		 [28]
AV1/V1 236-1012 777 258 29.9 外壳蛋白
Coat protein		 [26]
BC1 1246-2124 879 292 33.3 移动蛋白
Movement protein		 [29]
BV1 423-1184 762 253 29.1 核穿梭蛋白
Nuclear shuttle protein		 [30]

Table 3

24 geminiviruses for homologous comparison with CoYVV-FS used in the study"

病毒名称
Name of viruses		 缩写名
Abbreviation		 GenBank登录号
GenBank accession No.		 属
Genus		 病毒类型
Type of viruses
DNA-A DNA-B
南瓜曲叶病毒
Squash leaf curl virus		 SMLCV DQ285016 DQ285017 菜豆金黄花叶病毒属Begomovirus 来自新世界的双组分双生病毒
It is a geminivirus from NW consist
of two DNA components
菜豆矮花叶病毒
Bean dwarf mosaic virus		 BDMV NC_001931 NC_001930 菜豆金黄花叶病毒属Begomovirus 来自新世界的双组分双生病毒
It is a geminivirus from NW consist
of two DNA components
鹿藿金花叶病毒
Rhynchosia golden mosaic virus		 RhGMV AF239671 — 菜豆金黄花叶病毒属Begomovirus 来自新世界的单组分双生病毒
It is a geminivirus from NW consist
of one DNA components
黄花捻斑点病毒
Sida mottle virus		 SiMoV AY090555 — 菜豆金黄花叶病毒属Begomovirus 来自新世界的单组分双生病毒
It is a geminivirus from NW consist
of one DNA components
胜红蓟耳突病毒
Ageratum enation virus		 AEV JX436472 — 菜豆金黄花叶病毒属Begomovirus 来自旧世界的单组分双生病毒
It is a geminivirus from OW consist
of one DNA components
泽兰黄脉病毒
Eupatorium yellow vein virus		 EpYVV AB007990 — 菜豆金黄花叶病毒属Begomovirus 来自旧世界的单组分双生病毒
It is a geminivirus from OW consist
of one DNA components
印度木薯花叶病毒
Indian cassava mosaic virus		 ICMV AJ314739 AJ314740 菜豆金黄花叶病毒属Begomovirus 来自旧世界的双组分双生病毒
It is a geminivirus from OW consist
of two DNA components
南非木薯花叶病毒
South African cassava mosaic virus		 SACMV AF155806 AF155807 菜豆金黄花叶病毒属Begomovirus 来自旧世界的双组分双生病毒
It is a geminivirus from OW consist
of two DNA components
病毒名称
Name of viruses		 缩写名
Abbreviation		 GenBank登录号
GenBank accession No.		 属
Genus		 病毒类型
Type of viruses
DNA-A DNA-B
中国南瓜曲叶病毒
Squash leaf curl China virus		 SLCCNV AF509743 AF509742 菜豆金黄花叶病毒属Begomovirus 来自中国(旧世界)的双组分双生病毒
It is a geminivirus from China (OW)
consist of two DNA components
苎麻花叶病毒
Ramie mosaic virus		 RaMoV KC171650 KC171651 菜豆金黄花叶病毒属Begomovirus 来自中国(旧世界)的双组分双生病毒
It is a geminivirus from China (OW)
consist of two DNA components
中国大青花叶病毒
Clerodendrum golden mosaic China virus		 ClGMCNV FJ011668 FJ011669 菜豆金黄花叶病毒属Begomovirus 来自中国(旧世界)的双组分双生病毒
It is a geminivirus from China (OW)
consist of two DNA components
番茄黄曲叶病毒
Tomato yellow leaf curl virus		 TYLCV AJ489258 — 菜豆金黄花叶病毒属Begomovirus 来自中国(旧世界)的单组分双生病毒
It is a geminivirus from China (OW)
consist of one DNA components
番茄曲叶病毒
Tomato leaf curl virus		 ToLCV S53251 — 菜豆金黄花叶病毒属Begomovirus 来自中国(旧世界)的单组分双生病毒
It is a geminivirus from China (OW)
consist of one DNA components
胜红蓟黄脉病毒
Ageratum yellow vein virus		 AYVV AM940137 — 菜豆金黄花叶病毒属Begomovirus 来自中国(旧世界)的单组分双生病毒
It is a geminivirus from China (OW)
consist of one DNA components
黄麻金色花叶病毒
Corchorus golden mosaic virus		 CoGMV AB971842 AB971843 菜豆金黄花叶病毒属Begomovirus 来自旧世界双组分双生病毒
It is a geminivirus from OW consist
of two DNA components
越南黄麻黄脉病毒
Corchorus yellow vein Vietnam virus		 CoYVV AY727903 AY727904 菜豆金黄花叶病毒属Begomovirus 越南(旧世界)报道的CoYVV分离物
The isolate of CoYVV reported in
Vietnam (OW)
玉米坏死线条病毒
Maize streak reunion virus		 MSRV JQ624879 — 玉米线条病毒属
Mastrevirus		 代表玉米线条病毒属的病毒
It represents Mastrevirus
甜菜曲顶病毒Beet curly top virus BCTV AY548948 — 曲顶病毒属
Curtovirus		 代表曲顶病毒属的病毒
It represents Curtovirus
番茄伪曲顶病毒
Tomato pseudo-curly top virus		 TPCTV X84735 — 番茄伪曲顶病毒属Topocuvirus 代表番茄伪曲顶病毒属的病毒
It represents Topocuvirus
亚利桑那州菠菜曲顶病毒
Spinach curly top Arizona virus		 SCTAV HQ443515 — 甜菜曲顶病毒属
Becurtovirus		 代表甜菜曲顶病毒属的病毒
It represents Becurtovirus
弯叶画眉草线条病毒
Eragrostis curvula streak virus		 ECSV FJ665629 — 画眉草病毒属
Eragrovirus		 代表画眉草病毒属的病毒
It represents Eragrovirus
芜菁曲顶病毒
Turnip curly top virus		 TCTV JQ742019 — 芜菁曲顶病毒属Turncurtovirus 代表芜菁曲顶病毒属的病毒
It represents Turncurtovirus
苜蓿曲叶病毒
Alfalfa leaf curl virus		 ALCV MH603810 — 大戟小刺潜伏病毒属Capulavirus 代表大戟小刺潜伏病毒属的病毒
It represents Capulavirus
葡萄藤红斑病毒
Grapevine red blotch virus		 GRBV JX559642 — 葡萄斑点病毒属
Grablovirus		 代表葡萄斑点病毒属的病毒
It represents Grablovirus

Table 4

Similarity analysis of CoYVV-FS and other geminiviruses"

序号
No.		 缩写名
Abbreviation		 DNA-Aa DNA-Ba AC1b AC2 b AC3 b AC4 b AV1 b BC1 b BV1 b
1 CoYVV (the isolates in Vietnam) 100.00 99.93 100.00 100.00 100.00 100.00 100.00 100.00 99.60
2 CoGMV 79.80 71.73 78.93 65.96 65.38 51.06 83.72 86.30 63.60
3 SiMoV 77.09 — 49.72 63.72 48.55 26.67 84.50 — —
4 BDMV 83.09 77.74 55.03 53.44 50.38 — 83.72 75.00 51.56
5 RhGMV 82.45 — 66.29 56.34 47.06 — 84.50 — —
6 SMLCV 81.15 73.89 51.45 56.06 51.47 0 86.05 70.89 51.17
7 AYVV 77.88 — 68.17 31.88 46.84 37.23 74.90 — —
8 AEV 76.05 — 64.51 49.24 48.51 35.38 72.59 — —
9 RaMoV 76.02 0 65.92 51.16 50.00 32.14 73.36 0 0
10 ToLCV 73.93 — 67.64 45.80 46.67 38.10 71.54 — —
11 EpYVV 77.35 — 63.90 0 44.78 0 67.18 — —

Fig. 3

Phylogenetic tree of DNA-A of geminiviruses OW: Old World begomoviruses (eastern hemisphere, Europe, Africa, Asia); NW: New World begomoviruses (western hemisphere, the Americas); China: the begomoviruses reported in China (Old World); Vietnam: the isolates of CoYVV reported in Vietnam (Old World)."

Fig. 4

Phylogenetic tree of DNA-B of geminiviruses OW: Old World begomoviruses (eastern hemisphere, Europe, Africa, Asia); NW: New World begomoviruses (western hemisphere, the Americas); China: the begomoviruses reported in China; Vietnam: the isolates of CoYVV reported in Vietnam (Old World)."

Fig. 5

Phenotype of jute plants after inoculation with Agrobacterium infecting clones A: negative control, Bachang 4; B: Bachang 4 after inoculation; C: Yindumolyuzi after inoculation; D: negative control Yindumolyuzi."

Table 5

Disease statistics of 14 jute representative varieties after inoculation"

品种名称
Name of accessions		 种类
Species		 病株率
Disease incidence (%)		 侵染率
Infection rates (%)
爱店野生种Aidianyeshengzhong C. capsularis 0 100
粤圆5号Yueyuan 5 C. capsularis 0 60
黄麻179 Huangma 179 C. capsularis 0 100
新选1号Xinxuan 1 C. capsularis 0 90
梅峰2号Meifeng 2 C. capsularis 0 100
C46 C. capsularis 0 90
闽麻5号Minma 5 C. capsularis 0 90
奠边青麻Dianbianqingma C. olitorius 0 100
宽叶长果Kuanyechangguo C. olitorius 0 100
807元引马里807 Yuanyinmali C. olitorius 0 0
西贡长荚Xigongchangjia C. olitorius 0 20
福农5号Funong 5 C. olitorius 0 100
印度墨绿子Yindumolyuzi C. olitorius 40 80
巴长4号Bachang 4 C. olitorius 60 80
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