• Korean Journal of Environmental Biology
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• v.41 no.1
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• pp.1-13
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• 2023

Broad bean wilt virus 2 (BBWV2) is a species in the genus Fabavirus and family Secoviridae, which is transmitted by aphids and has a wide host range. The BBWV2 genome is composed of two single-stranded, positive-sense RNAs, RNA-1 and RNA-2. The representative symptoms of BBWV2 are mosaic, mottle, vein clearing, wilt, and stunting on leaves, and these symptoms cause economic damage to various crops. In 2019, Perilla fructescens leaves with mosaic and yellowing symptoms were found in Geumsan, South Korea. Reverse-transcription polymerase chain reaction (RT-PCR) was performed with specific primers for 10 reported viruses, including BBWV2, to identify the causal virus, and the results were positive for BBWV2. To characterize a BBWV2 isolate (BBWV2-GS-PF) from symptomatic P. fructescens, genetic analysis and pathogenicity tests were performed. The complete genomic sequences of RNA-1 and RNA-2 of BBWV2-GS-PF were phylogenetically distant to the previously reported BBWV2 isolates, with relatively low nucleotide sequence similarities of 76-80%. In the pathogenicity test, unlike most BBWV2 isolates with mild mosaic or mosaic symptoms in peppers, the BBWV2-GS-PF isolate showed typical ring spot symptoms. Considering these results, the BBWV2-GS-PF isolate from P. fructescens could be classified as a new strain of BBWV2.

• The Plant Pathology Journal
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• v.29 no.3
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• pp.274-284
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• 2013

The incidence of Broad bean wilt virus 2 (BBWV2) on red pepper was investigated using the samples obtained from 24 areas of 8 provinces in Korea. Two hundred and five samples (79%) out of 260 collected samples were found to be infected with BBWV2. While the single infection rate of BBWV2 was 21.5%, the co-infection rate of BBWV2 with Cucumber mosaic virus, Pepper mottle virus, Pepper mild mottle virus and/or Potato virus Y was 78.5%. To characterize the genetic diversity of BBWV2 Korean isolates, 7 isolates were fully sequenced and analyzed. Phylogenetic analyses revealed that BBWV2 isolates could be divided largely into two groups as Group I and Group II. Based on the partial sequence analyses, 153 selected BBWV2 isolates were subgrouped into GS-I (21.6%), GS-II (3.9%) and GS-III (56.9%). BBWV2 GS-III, which was predominant in Korea, appears to be a new combination between Group I RNA-1 and Group II RNA-2. Viral disease incidence of BBWV2 on red pepper was under 2% before 2004. However, the incidence was increased abruptly to 41.3% in 2005, 58.2% in 2006 and 79% in 2007. These rapid increases might be related with the emergence of new combinations between BBWV2 groups.

• Research in Plant Disease
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• v.13 no.2
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• pp.75-81
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• 2007

The incidence of virus disease on peppers was investigated at the 52 areas in the whole country in 2002, 2004 to 2006. Among the six viruses, Cucumber mosaic virus (CMV), Pepper mottle virus (PepMoV), Pepper mild mottle virus (PMMoV), Broad bean wilt virus 2 (BBWV2), Tobacco mild green mosaic virus (TMGMV) and Tomato spotted wilt virus (TSWV), infecting peppers in Korea, the incidence of CMV, PepMoV, PMMoV and BBWV2 was 29.4%, 13.6%, 14.3%, 25.6%, orderly. TMGMV and TSWV had the same low infection rate of 1.0%. The infection rate of CMV was higher as 53.3% and 34.2% in 2002 and 2004, but it was decreased to 18.2% and 11.9% in 2005 and 2006, respectively. The infection rate of BBWV2 was lower as 1.3% in 2002 and 1.8% in 2004, but it was increased abruptly to 41.3% in 2005 and 58.2% in 2006. For the types of mixed infections of pepper viruses, the incidence of CMV+PepMoV was 62.6% in 2002 and 50.0% in 2004, and that of CMV+BBWV2 was increased suddenly from 33.3% in 2005 to 83.2% in 2006. The triplex infection of CMV+BBWV2+PepMoV was 6.4% in average. CMV caused severe mosaic and BBWV2 induced ring spots, and the two mixed virions caused chlorosis on the leaves of red peppers. TSWV induced the typical symptoms of multiple ring spots on the leaves and fruits of red peppers.

• Research in Plant Disease
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• v.18 no.4
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• pp.268-276
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• 2012

Broad bean wilt virus 2 (BBWV2), genus Fabavirus, subfamily Comovirinae, family Secoviridae, causes damage in many economically important horticultural and ornamental crops. Sequence alignments showed several conserved sequences in 5' non-coding regions (5' NCRs) of RNA 1 and RNA 2 in all BBWV2 strains characterized so far. Based on this observation, we generated a hpRNA construct (pIR-BBWV2) harboring an inverted repeat containing a 210 bp cDNA fragment homologous to 5' NCR portion of BBWV2 RNA 1 to investigate the silencing potential for its ability to interfere with a rapidly replicating BBWV2. Agrobacterium-mediated transient expression of the IR-BBWV2 had a detrimental effect on BBWV2 infection, showing no distinct symptoms in non-inoculated leaves of the agroinfiltrated Nicotiana benthamiana plants. BBWV2 genomic RNAs were not detected by RT-PCR from tissues of both the inoculated leaves and upper leaves of the agroinfiltrated plants. Accumulation of virus-derived small interfering RNAs was detected in the inoculated leaf tissues of N. benthamiana plants elicited by transient expression of IR-BBWV2 indicating that RNA silencing is responsible for the resistance to BBWV2.

• Research in Plant Disease
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• v.23 no.3
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• pp.234-240
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• 2017

Incidence of virus diseases in red pepper of open field in Yeongyang-Gun, Gyeongbuk Province was investigated by reverse transcription-polymerase chain reaction in 2012-2016. The infection rate of Cucumber mosaic virus (CMV), Broad bean wilt virus 2 (BBWV2), Pepper mottle virus, Potato virus Y, Pepper mild mottle virus and Tomato spotted wilt virus was 46.1%, 41.5%, 2.0%, 2.0%, 4.4% and 0.1% respectively. Incidence rate of single and mixed infection was 31.2% and 62.6%. Most of single infections were CMV and BBWV2. Among mixed infections, the incidence rate of CMV+BBWV2 mixed infection was the highest as 49.3% and most of mixed infections of triplex and tetraplex included CMV+BBWV2 mixed infection. CMV single infection caused mosaic, chlorosis, yellowing and vein necrosis and BBWV2 single infection induced cholosis and mosaic. CMV+BBWV2 mixed infection caused severe mosaic with chlorosis or malformation.

• The Plant Pathology Journal
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• v.29 no.4
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• pp.397-409
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• 2013

The full-genome sequences of fourteen isolates of Broad bean wilt virus 2 (BBWV2), collected from broad bean, pea, spinach, bell pepper and paprika plants in Korea during the years 2006-2012, were determined and analyzed comparatively along with fifteen previously reported BBWV2 genome sequences. Sequence analyses showed that RNA-1 and RNA-2 sequences of BBWV2 Korean isolates consisted of 5950-5956 and 3568-3604 nucleotides, respectively. Full-length genome sequence-based phylogenetic analyses revealed that the BBWV2 Korean isolates could be divided into three major groups comprising GS-I (isolates BB2 and RP7) along with isolate IP, GS-II (isolates BB5, P2, P3 and RP3) along with isolate B935, and GS-III including 16 BBWV2 Korean isolates. Interestingly, GS-III appears to be newly emerged and predominant in Korea. Recombination analyses identified two recombination events in the analyzed BBWV2 population: one in the RNA-1 of isolate K and another one in the RNA-2 of isolate XJ14-3. However, no recombination events were detected in the other 21 Korean isolates. On the other hand, out of 29 BBWV2 isolates, 16 isolates were found to be re-assortants, of which each RNA segment (i.e. RNA1 and RNA2) was originated from different parental isolates. Our findings suggested that reassortment rather than recombination is a major evolutionary force in the genetic diversification of BBWV population in Korea.

• Korean Journal Plant Pathology
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• v.3 no.3
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• pp.223-235
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• 1987

Samples showing yellow mosaic symptom of Lilium spp. with necrotic fleck, stunting, malformation, and colour breaking were collected from lily-growing areas in the southern part of Korea. Two viruses were distinguished under a electron microscope and their host range, serological reaction, stability in sap, type of aphid transmission, and relations with cells and tissues were examined. Broad bean wilt virus (BBWV) was transmitted by sap-inoculation to 23 plant species in 8 families and by the aphid, Myzus persicae. This virus was inactivated after 10 min at 70C, at dilution of $10^{-3}$, and after 6 days at about 20C. Electron microscopic examination of purified preparation showed that the virus is spherical particle of 28nm in diameter. The virus reacted positively with BBWV-antiserum in agar gel diffusion test. In ultrathin sections of BBWV infected tissues, large aggregates or crystalline array of virus particles and vesicular body were found in the cytoplasm, vacuole, and nucleus of mesophyll cells. Cucumber mosaic virus (CMV) was transmitted by sap-inoculation. Electron microscopic examination of its purified preparation showed spherical particles of 30nm in diameter. The virus reacted positively with CMV-Y strain-antiserum in agar gel diffusion test. In ultrathin sections of CMV infected tissues, crystalline array of virus particles were found in the vacuole and a large number 0f virus particles were found in the cytoplasm and the plasmodesmata of mesophyll cells. When each of these viruses was retransmitted to Lilium tigrinum. L. concolor, and L. auratum, BBWV induced slight symtoms and colour breaking, but CMV induced yellowing mosaic or necrotic fleck.

• Korean Journal Plant Pathology
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• v.14 no.1
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• pp.83-91
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• 1998

Gladioli (Gladiolus hybridus) showing flower colour breaking leaf mosaics, notched leaf, and dwarfing or lack of visible symptoms were collected from gladioli growing areas in Taegu and Kyungpook province, Korea. The three viruses isolated from the naturally infected gladioli were identified as broad bean wilt virus (BBWV), cucumber mosaic virus (CMV), and tobacco rattle virus (TRV) by their host range, immunosorbent electron microscopy (ISEM), enzyme-linked immunosorbent assay (ELISA), direct tissue blotting immunoassay (DTBIA), and intracellural symptoms. By DTBIA and ISEM, TRV was detected in gladiolus showing notched leaf, while CMV was frequently detected in gladioli with dwarfing, color breaking and malformation of flowers. BBWV was also often detected in many symptomless gladiolus plants, but TRV was detected in notched-leaf of gladiolus. Electron microcopic examination of negatively stained preparations showed that BBWV and CMV are spherical particles of 28 nm and 30 nm in diameter, and TRV is rigid rod-shaped particles of 40∼200 nm in length. The rigid rodshaped virus particles reacted positively with TRV antiserum in ISEM and DTBIA, respectively.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2002.11b
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• pp.54-54
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• 2002

The nucleotide sequence of a Korea isolate of broad bean wilt favavirus from Rehmannia glutinosa Lib., designated BBWV-RE, was determined. Direct amino acid sequencing of the virus coat proteins suggests that a comparison of several favaviruses in terms of nucleotide sequence and amino acid sequences showed that BBWV-2 isolates display high sequence identity. The small coat protein genes of RNA-2 were also determined for three other Japanese isolates(E, L, and 1-2) and two ATCC isolates(PV132 and PV176) of BBWV. the CP sequence suggested distinct evolution lineages. Serotype 2 favaviruses are more prevalent in Asia, Australia and North America, Wheres serotype 1 is more prevalent in europe.(중략)

• The Plant Pathology Journal
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• v.21 no.3
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• pp.258-261
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• 2005

We conducted a survey on pepper virus diseases in 31 regions in Korea from November 2001 to December 2004. Using electron microscopy, test plant reaction, rapid immuno-filter paper assay (RIPA), reverse transcription-polymerase chain reaction (RT-PCR) and/or analysis of viral nucleotide sequences, we found a number of viruses from 1,056 samples that we collected. These included Cucumber mosaic virus (CMV), Pepper mottle virus (PepMoV), Pepper mild mottle virus (PMMoV), Broad bean wilt virus 2 (BBWV2), Tobacco mild green mosaic virus (TMGMV), and Tomato spotted wilt virus (TSWV). Of the samples analyzed, $343(32.5\%)$ were infected with CMV, $209(19.8\%)$ with PepMoV, $141(13.4\%)$ with PMMoV, $12(1.1\%)$ with BBWV2, $40(3.8\%)$ with TMGMV, $5(0.5\%)$ with TSWV, $153(14.5\%)$ with CMV and PepMoV, $54 (5.1\%)$ with CMV and PMMoV, $31(2.9\%)$ with PepMoV and PMMoV, $3(0.3\%)$ with CMV and BBWV2, $1(0.1\%)$ with CMV, PepMoV and BBWV2, $8(0.8\%)$ with CMV, PepMoV and PMMoV, and $30 (2.8\%)$ samples were infected with viruses which were not identified. CMV was the most predominant virus in all inspected fields and the number of the samples infected with PMMoV was relatively low as compared PepMoV infection level in pepper. TMGMV was only found in the southern part of Korea, while TSWV was isolated in Anyang and Yesan. However, we did not encounter in this survey the Alfalfa mosaic virus (AMV), Potato virus Y (PVY), Tobacco mosaic virus (TMV), and Pepper vein chlorosis virus (PVCV).