Jang-Kyung Choi

Infection of soybean by cucumber mosaic virus as determined by viral movement protein

Summary.To characterize the host range determinant of the soybean strain of Cucumber mosaic virus (CMV) we analyzed a series of pseudorecombinants and chimeric viruses between infectious transcripts from two soybean strains (CMV-SC and CMV-SD) and an ordinary strain (CMV-Y). CMV-Y could not infect soybeans, even locally. Systemic infection of the two soybean-adapted soybean isolates on soybean plants mapped to RNA3. Chimeric RNA3s from between CMV-SC and CMV-Y, and chimeric RNA3s from between CMV-SC and CMV-SD, were made and inoculated onto wild soybean Iwate and soybean cv. Tsurunoko. The 3a region determined the viral systemic movement in the plants. In the wild soybean ecotype Hyougo, cell-to-cell movement of two different CMV soybean strains, one of which infects systemically while the other does not, in the inoculated leaves were almost the same, suggesting that the resistance of soybean operates at the level of long-distance movement. Our results clearly suggest that movement protein is a host determinant of CMV soybean strains.

The complete genome sequence of pepper severe mosaic virus and comparison with other potyviruses.

Summary.The complete nucleotide sequence of pepper severe mosaic virus (PepSMV) was determined. The viral genome consisted of 9890 nucleotides, excluding a poly (A) tract at the 3′ end of the genome. The PepSMV RNA genome encoded a single polyprotein of 3085 amino acid residues, resulting in ten functionally distinct potyviral proteins. The lengths of the 5′ nontranslated region (NTR) and the 3′ NTR were 164 and 468 nucleotides, respectively. The genome organization of the virus was typical for members of the genus Potyvirus in the family Potyviridae. The coat protein amino acid sequence identity between PepSMV and the other 45 potyviruses ranged from 53.4 to 79.7%. Sequence alignments and phylogenetic analyses of the potyviral polyprotein sequences revealed that PepSMV was the closest to potato virus Y (PVY) and closely related to members of the PVY subgroup. Our genome sequence data clearly confirmed that PepSMV belongs to a separate species in the genus Potyvirus.

Variation in the Pathogenicity of Lily Isolates of Cucumber mosaic virus

Two isolates of Cucumber mosaic virus (CMV) originated from lily plants, named Ly2-CMV and Ly8-CMV, were compared with their pathological features in several host plants. Ly2-CMV and Ly8-CMV could induce systemic mosaic symptom in Nicotiana benthamiana, but Ly2-CMV could not systemically infect tomato and cucumber plants that have been used for CMV-propagative hosts. While Fny-CMV used as a control infected systemically the same host plants, producing typical CMV symptoms. Ly8-CMV could infect systemically two species of tobacco (N. tabacum cv. Xanthi-nc and N. glutinosa) and zucchini squash (Curcubita pepo), but Ly2 failed systemic infection on these plants. As resulted from tissue-print immunoblot assay, different kinetics of systemic movement between Ly2-CMV and Ly8-CMV were crucial for systemic infection in tobacco (cv. Xanthi-nc). Sequence analysis of full-length genome of two lily isolates showed Ly2 and Ly8 belonged to subgroup IA of CMV. The lily isolates shared overall 98 % sequence identity in their genomes. Coat protein, 3a protein, and 2b protein involved in virus movement was highly conserved in genomes of the isolates Ly2 and Ly8. Although there is the low frequency of recombinants and reassortants in natural CMV population, phylogenetic analysis of each viral protein among a number of CMV isolates suggested that genetic variation in a defined population of CMV lily isolates was stochastically produced.

Molecular Characterization and Infectious cDNA Clone of a Korean Isolate of Pepper mild mottle virus from Pepper

A Korean isolate of Pepper mild mottle virus (PMMoV-Kr) was isolated from a diseased hot pepper plant and its biological and molecular properties were compared to that of PMMoV-J and PMMo V -So The genomic RNA of PMMoV-Kr consists of 6,356 nucleotides. The nucleotide and amino acid sequences identities of four viral proteins and two noncoding regions among PMMoV-Kr, PMMoV-S and PMMoV-J were , respectively. Full-length cDNA amplicon of PMMoV-Kr was directly amplified by RT-PCR with a set of 5`-end primer anchoring T7 RNA promoter sequence and 3`-end virus-specific primer. Capped transcript RNAs from the full-length cDNA clone were highly infectious and caused characteristic symptoms of wild type PMMoV when mechanically inoculated to systemic host plants such as Nicotiana benthamiana and pepper plants.

Patterns in disease progress and the influence of single and multiple viral infections on pepper ( Capsicum annuum L.) growth

The patterns and progress of disease caused by multiple infections of Cucumber mosaic virus (CMV), Pepper mild mottle virus (PMMoV) and Pepper mottle virus (PepMoV) and their effects on growth of pepper plants (Capsicum annuum L.) were investigated in this study. Each virus induced distinct symptoms, but more severe symptoms, including reduced growth rates, were observed when pepper plants were simultaneously infected by more than one virus. When CMV was included in multiple viral inoculations, co-inoculations and sequential inoculations, PepMoV and PMMoV symptoms were observed but the symptoms characteristic of CMV were not masked, even though CMV titre did not increase greatly. In multiple viral infections, PepMoV titre and CMV did not increase significantly, but PMMoV titre gradually increased in most cases. Growth rates of pepper plants were greatly reduced during the 30 to 40-day post-inoculation period under both single-infection and multiple-infection conditions, but multiple viral infections of CMV pre-inoculated peppers were affected to a greater extent. A significant reduction in fruit size and fruit number was observed in single and multiple viral inoculations, and fruit malformation rates were high in CMV single-infection and multiple viral infections with CMV.

First Report of the Peanut Stripe Strain of Bean common mosaic virus (BCMVPSt) Infecting Mungbean in Korea

A virus causing chlorotic ringspot, yellow mosaic and vein clearing symptoms was prevalent on mungbean plants around Taean, Korea. The isolate caused mosaic on Chenopodium quinoa, Nicotiana benthamiana, Phaseolus vulgaris and Vida laba but no symptoms on peanut plants. Inclusion bodies such as scroll, pinwheel and laminated aggregates induced by the virus in the host cells were similar to those produced by members of the Potyvirus subdivision III. Multiple alignment as well as cluster dendrograms of the 709 nucleotide region comprising part of the coat protein gene and 3`untranslated region (UTR) showed that the isolate belongs to the BCMV-PSt subgroup. Altogether, these results support the identification of the causal virus as peanut stripe strain of Bean common mosaic virus (BCMV-PSt).

Characterization of an Isolate of Cucumber mosaic virus Isolated from Chinese aster (Callistephus chinensis)

An isolate of Cucumber mosaic virus (CMV), designated as Cas-CMV, was isolated from Chinese aster (Callistephus chinensis) showing severe mosaic symptom, and its properties was compared to the well-characterized Fny-CMV (subgroup IA) and As-CMV (subgroup IB) by host reaction in several indicator plants, dsRNA analysis, RT-PCR analysis, and restriction enzyme profile of the PCR products. Cas-CMV differed markedly in their host reaction to Fny-CMV or As-CMV in Cucurbita pepo cv. Black beauty. In the zucchini squash, all strains induced chlorotic spot on inoculated leaves and mosaic symptoms on upper leaves. However, symptoms induced by Cas-CMV were developed lethal necrosis on the young plants 15 to 20 days after inoculation. In experiments of dsRNA analysis and RT-PCR analysis, properties of Cas-CMV was come within subgroup I CMV. Moreover, restriction enzyme analysis using HindIII of the RT-PCR products showed that Cas-CMV belong to a member of CMV subgroup IA.

Occurrence of Bean common mosaic virus (BCMV) Infecting Peanut in Korea

A virus causing vein banding, sometimes yellow mosaic and rugose symptoms on peanut was prevalent around Suwon, Korea. A survey conducted in the area found disease incidence, depending on cultivar, to range from 79 to . The virus was found to be seed-transmissible in all the five peanut cultivars tested with transmission rates ranging from 2 to . Host range analysis failed to differentiate 9 field isolates collected from different peanuts cultivars showing various symptoms. Inclusion bodies such as scroll, pinwheel and long laminated aggregates induced by the virus in host plant cells were similar to those induced by members of the Potyvirus subdivision III. The virus showed homology with Bean common mosaic virus (BCMV), BCMV-BICMV/AzMV strains and only with Desmodium mosaic virus. Based on biological characterization, electron microscopy and molecular analyses of a Korean isolate (Daewon 1), the virus was identified as peanut stripe strain of BCMV.

Characterisation of a satellite RNA of Cucumber mosaic virus that induces chlorosis in Capsicum annuum

The presence of Cucumber mosaic virus (CMV) satellite RNA dramatically changes symptoms on some hosts. A satellite RNA present in a strain of CMV (PepY-CMV) that induced chlorosis in pepper (Capsicumannuum) was shown to induce chlorosis in pepper in combination with another strain (Fny-CMV) that by itself induced a green mosaic symptom. The location of sequences within the PepY satellite RNA (PepY-satRNA) of CMV that conferred the ability to induce chlorosis on pepper plants were analyzed by exchanging sequence domains between cDNA clones of PepY-satRNA and an attenuated mosaic satellite RNA (Paf-satRNA), as well as site-directed mutagenesis of various clusters of the 22-nt sequence differences between the two satellite RNAs in the delimited central domain. The symptoms induced by site-directed mutants of PepY-satRNA and Paf-satRNA in the presence of Fny-CMV demonstrated an insertion within PepY-satRNA of 11 nt at positions 86–96 relative to Paf-satRNA determined the chlorosis-inducing phenotype. Within the chlorosis-inducing domain, deletion of nucleotides did not affect the satRNA replication but abolished the ability of PepY-satRNA to elicit chlorosis symptom. Conversely, a mutant satellite RNA derived from Paf-satRNA in which eleven nucleotides were inserted indicated that sequences of 11 nucleotides were found to be sufficient for chlorosis induction in pepper.

Identification and Characterization of Three Isolates of Cucumber mosaic virus Isolated from Weed Hosts

Three isolates of Cucumber mosaic virus (CMV) were isolated from weed hosts showing typical mosaic symptoms, and some properties of the viruses were investigated. CMV isolates, designated as Is-CMV, Jd-CMV and Pla-CMV from Isodon inflexus, Jeffersonia dubia and Phryma leptostachya var. asiatica, respectively, were identified and characterized by biological reaction in several host plants, serological property, dsRNA analysis, reverse transcription-polymerase chain reaction (RT-PCR), restriction fragment-length polymorphism (RFLP). All isolates systemically infected in Nicotiana benthamiana, Cucurbita pepo cv. Black beauty and Cucumis sativus, and did not reveal any differences in these host plants between the isolates. However, remarkable difference in the symptoms was found between the CMVs in Capsicum annuum. Is-CMV induced an asymptomatic symptoms, while Jd-CMV and Pla-CMV produced severe mosaic symptoms in C. annuum plants. In dsRNA analysis, all isolates revealed four major bands with estimated molecular size of 3.4, 3.2, 2.1 and 1.0 kbp. The cDNAs of coat protein gene of the isolates were amplified by RT-PCR using a genus-specific single pair primers that designed to amplify a DNA fragment of approximately ranging from 938 to 966 bp. By restriction mapping analysis using RFLP of the RT-PCR products as well as by serological properties of gel diffusion test, the CMV isolates belong to a typical members of CMV subgroup IA. This is the first report on the occurrence of CMV in the three weed hosts.