First Report of Broad Bean Wilt Virus 2 Infecting Perilla frutescens in China

Vigna marina (Burm.) Merr., the dune bean or notched cowpea, is a tropical creeping vine that grows on sand dunes along the coastal regions of Taiwan. Although V. marina is a weed, some varieties are also grown for fodder and food. This legume is a natural host of Bean common mosaic virus in the Solomon Islands (1) and Alfalfa mosaic virus or Beet western yellows virus in Australia (2). In April 2009, plants of V. marina showing severe mosaic and chlorotic ringspots on the foliage were found in the coastal region of Hualien County in eastern Taiwan. Indirect ELISA on a single diseased plant showed positive results with antibodies against the cucumber isolate of Cucumber mosaic virus (CMV) but negative to Broad bean wilt virus-1, Broad bean wilt virus-2, and some potyviruses (Agdia Inc., Elkhart, IN). A pure isolate of CMV was obtained from V. marina through three successive passages of single lesion isolation in sap-inoculated Chenopodium quinoa. Results of mechanical inoculations showed that the CMV-V. marina isolate was successfully transmitted to C. amaranticolor, C. murale, C. quinoa, Chrysanthemum coronarium, Gomphrena globosa, Nicotiana benthamiana, N. tabacum cv. Vam-Hicks, Phaseolus limensis, P. lunatus, P. vulgaris, Tetragonia tetragonioides, V. marina, V. radiata, and V. unguiculata subsp. sesquipedalis. These results of artificial inoculations were confirmed by ELISA. Homologous reactions of the CMV-V. marina isolate with a stock polyclonal antiserum against the CMV-cucumber isolate (4) were observed in sodium dodecyl sulfate-immunodiffusion. To determine the specific CMV subgroup, total RNA was extracted from inoculated leaves of C. quinoa using the Total Plant RNA Extraction Miniprep System (Viogene, Sunnyvale, CA). A DNA fragment of 940 bp covering the 3′ end of the coat protein gene and C-terminal noncoding region of RNA-3 was amplified using the Cucumovirus-specific primers (3) after reverse transcription (RT)-PCR with AccuPower RT/PCR PreMix Kit (Bioneer, Daejeon, Korea). The product was gel purified by Micro-Elute DNA/Clean Extraction Kit (GeneMark Technology Co., Tainan, Taiwan) and cloned in yT&A Cloning Vector System (Yeastern Biotech Co., Taipei, Taiwan) for sequencing (Mission Biotech Co., Taipei, Taiwan) and the sequence was submitted to GenBank (No. HM015286). Pairwise comparisons of the sequence of CMV-V. marina isolate with corresponding sequences of other CMV isolates revealed the maximum (95 to 96%) nucleotide identities with CMV subgroup IB isolates (strains Nt9 and Tfn) compared with 94 to 95% identities with subgroup IA isolates (strains Y and Fny) or 77 to 78% identities with subgroup II (strains LS and Q). These results suggest that CMV is the causal agent for the mosaic disease of V. marina in Taiwan and the isolate belongs to subgroup I. To our knowledge, this is the first report of V. marina as a natural host of CMV. This strain of CMV with specific pathogenicity could threaten crop production in the coastal zones. In addition, V. marina associated with native coastal vegetation was injured by CMV infection, which might lead to ecological impacts on shoreline fading. References: (1) A. A. Brunt. Surveys for Plant Viruses and Virus Diseases in Solomon Islands. FAO, Rome, 1987. (2) C. Büchen-Osmond, ed. Viruses of Plants in Australia. Retrieved from http://www.ictvdb.rothamsted.ac.uk/Aussi/aussi.htm . September, 2002. (3) S. K. Choi et al. J. Virol. Methods 83:67, 1999. (4) S. H. Hseu et al. Plant Prot. Bull. (Taiwan) 29:233, 1987.

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First Report of Broad bean wilt virus 2 and Ranunculus mild mosaic virus in Ranunculus asiaticus in Southern Italy

Plant Disease ◽

10.1094/pdis-05-15-0563-pdn ◽

2016 ◽

Vol 100 (1) ◽

pp. 235-235 ◽

Cited By ~ 2

Author(s):

M. Minutolo ◽

R. Sorrentino ◽

V. Masenga ◽

D. Alioto

Keyword(s):

Mosaic Virus ◽

Southern Italy ◽

Broad Bean ◽

Mild Mosaic ◽

First Report ◽

Ranunculus Asiaticus ◽

Mild Mosaic Virus ◽

Wilt Virus ◽

Broad Bean Wilt Virus

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First Report of Broad bean wilt virus 2 in Gynura procumbens in Korea

Plant Disease ◽

10.1094/pdis-10-16-1413-pdn ◽

2017 ◽

Vol 101 (3) ◽

pp. 514-514 ◽

Cited By ~ 3

Author(s):

H.-R. Kwak ◽

W.-R. Go ◽

M. Kim ◽

C.-S. Kim ◽

H.-S. Choi ◽

...

Keyword(s):

Broad Bean ◽

First Report ◽

Wilt Virus ◽

Broad Bean Wilt Virus

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First Report of Broad Bean Wilt Virus 2 in Achyranthes bidentata in Korea

Plant Disease ◽

10.1094/pdis-06-18-0967-pdn ◽

2019 ◽

Vol 103 (1) ◽

pp. 165 ◽

Cited By ~ 1

Author(s):

S.-J. Kwon ◽

J.-K. Seo

Keyword(s):

Broad Bean ◽

Achyranthes Bidentata ◽

First Report ◽

Wilt Virus ◽

Broad Bean Wilt Virus

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First Report of Broad bean wilt virus 2 in Leonurus sibiricus in Korea

Plant Disease ◽

10.1094/pdis-07-14-0695-pdn ◽

2014 ◽

Vol 98 (12) ◽

pp. 1748-1748 ◽

Cited By ~ 3

Author(s):

J.-K. Seo ◽

O. J. Shin ◽

H.-R. Kwak ◽

M.-K. Kim ◽

H.-S. Choi ◽

...

Keyword(s):

Host Range ◽

Broad Bean ◽

Rt Pcr ◽

Total Rna ◽

First Report ◽

Leonurus Sibiricus ◽

Plant Pathol ◽

Wilt Virus ◽

Broad Bean Wilt Virus ◽

Test Plants

Leonurus sibiricus L. (family Lamiaceae) has been used as a traditional herbal remedy to treat various gynecologic diseases. Although it is a widely distributed subtropical weed in Southeast Asia, L. sibiricus have been commercially cultivated on a small scale in many geographic areas of Korea. In August 2012, field-grown L. sibiricus plants showing mosaic, yellowing, and stunting symptoms were collected near a pepper field in Andong, Korea. Since L. sibiricus is only consumed as a raw material of traditional medicine in Korea, symptomatic plants lose commercial value entirely. To identify the causal agent(s) of the virus-like symptoms, total RNA was extracted from the symptomatic leaves, and a transcriptome library was generated using the TruSeq Stranded Total RNA with Ribo-Zero plant kit (Illumina, San Diego, CA) according to the standard protocol. Next-generation sequencing (NGS) was performed using an Illumina HiSeq2000 sequencer. De novo assembly of the quality filtered NGS reads (101-bp paired-end reads) were performed using the Trinity pipeline and the assembled contigs (92,329 contigs) were analyzed against the viral reference genome database in GenBank by BLASTn and BLASTx searches (3). The entire NGS procedure was performed by Macrogen Inc. (Seoul, South Korea). Among the analyzed contigs, only two large contigs were clearly of viral origin. Nucleotide blast searches showed that the first and second contigs (5,914 and 3,534 bp, respectively) have maximum identities of 91 and 95% to RNA1 of the isolate RP3 (GenBank Accession No. JX183225) and RNA2 of the isolate RP7 (JX183234) of Broad bean wilt virus 2 (BBWV-2), which were isolated from pepper in Korea. The NGS results were confirmed by analyzing the sequences of the fragments covering the entire BBWV-2 genome amplified by RT-PCR using specific primers for BBWV-2 as described previously (1). To obtain the complete genome sequence, terminal sequences of both RNA segments were analyzed by the 5′ and 3′ rapid amplification of cDNA ends (RACE) method as described previously (1). The assembled full-length sequences of BBWV-2 RNA1 and RNA2 isolated from L. sibiricus were 5,951 and 3,575 nucleotides in length, respectively, and deposited in GenBank under the accessions KM076648 and KM076649, respectively. BBWV-2 belongs to the genus Fabavirus in the family Secoviridae and it is known to have a wide host range. To investigate the host range of the BBWV-2 isolated from L. sibiricus, sap from the symptomatic leaves of L. sibiricus was inoculated to the test plants including Nicotiana benthamiana, Capsicum annuum (red pepper), and C. annuum var. gulosum (Paprika). RT-PCR detection and sequencing of the amplicons showed that all the inoculated test plants were infected with the BBWV-2 isolated from L. sibiricus. Currently, BBWV-2 is epidemic in pepper fields in Korea (1,2). Because BBWV-2 is easily transmitted by various aphids, and L. sibiricus is widely distributed in both wild and cultivated fields in Korea, this host might serve as a potential source of BBWV-2 to other crops such as pepper. To the best of our knowledge, this is the first report of BBWV-2 in L. sibiricus. References: (1) H.-R. Kwak et al. Plant Pathol. J. 29:274, 2013. (2) H.-R. Kwak et al. Plant Pathol. J. 29:397, 2013. (3) S.-E. Schelhorn et al. PLoS Comput. Biol. 9:e1003228, 2013.

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First Report of Broad bean wilt virus on Thyme (Thymus vulgaris L.)

Plant Disease ◽

10.1094/pdis.2001.85.1.99b ◽

2001 ◽

Vol 85 (1) ◽

pp. 99-99 ◽

Cited By ~ 3

Author(s):

M. G. Bellardi ◽

C. Rubies-Autonell

Keyword(s):

Broad Bean ◽

Protein A ◽

Enzyme Linked Immunosorbent Assay ◽

Thymus Vulgaris ◽

Chenopodium Amaranticolor ◽

Borago Officinalis ◽

First Report ◽

Chlorotic Mottle ◽

Wilt Virus ◽

Broad Bean Wilt Virus

In the spring and summer of 1998, a severe virus-like disease consisting of chlorotic mottle on leaves, yellowing, and stunting was observed at the Giardino delle Erbe of Casola-Valsenio (Emila-Romagna region, northern Italy). Most of the symptomatic plants were infected with a filamentous virus that was not identified. Moreover, one thyme plant showing yellow leaves was also found infected by an isometric virus, the identity of which was established by the following host reactions and serological assay. It was mechanically transmitted to 33 species belonging to 11 botanical families. All the Chenopodiaceae (Chenopodium amaranticolor Coste et Reyn., C. murale L., C. foliosum Ash., and C. quinoa Willd.) tested showed local and systemic symptoms in 3 to 4 days; among Leguminosae, broad bean (Vicia faba L.) showed necrotic local lesions and wilt. Virus particles reacted in protein A sandwich-enzyme-linked immunosorbent assay with the antiserum to Broad bean wilt virus (BBWV), serotype I (supplied from the Istituto di Fitovirologia Applicata, CNR, Turin, Italy). BBWV in the field was in all probability transmitted to thyme by aphids from weeds and/or other medicinal and aromatic species cultivated in the same herb garden, all recently shown to be hosts for BBWV: Polygonum fagopyrum L., Hedisarum coronarium L., Borago officinalis L., Phytolacca Americana L., Digitalis lanata Ehrh., and D. purpurea L. (1). This is the first report of BBWV in T. vulgaris and demonstrates that it is more prevalent in Italy than previously reported. References: (1) C. Rubies-Autonell and M. G. Bellardi. 1999. 7th International Plant Virus Epidemiology Symposium, Aguadulce, Spain.

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