• Journal of Crop Science and Biotechnology
• /
• 제11권2호
• /
• pp.127-134
• /
• 2008

Tobamoviruses are the major viral pathogens of tomato and bell pepper. The preliminary results showed that Acibenzolar-Smethyl(ASM; S-methylbenzo(1,2,3) thiadiazole-7-carbothiate) pre-treatment to tomato and tobacco plants reduces the concentration of Tomato mosaic tobamovirus(ToMV) and Tobacco mosaic tobamovirus(TMV) in tomato and bell pepper seedlings, respectively. Pre-treatment of the indicator plant(Nicotiana glutinosa) with the ASM followed by challenge inoculation with tobamoviruses produced a reduced number and size of local lesions(67 and 79% protection over control to TMV and ToMV inoculation, respectively). In order to understand the mechanism of resistance the gene expression profiles of antiviral genes was examined. RT-PCR products showed higher expression of two viral resistance genes viz., alternative oxidase(AOX) and RNA dependent RNA polymerase(RdRp) in the upper leaves of the ASM-treated tomato plants challenge inoculation with ToMV. Further, the viral concentration was also quantified in the upper leaves by reverse transcription PCR using specific primer for movement protein of ToMV, as well as ELISA by using antisera against tobamoviruses. The results provided additional evidence that ASM pre-treatment reduced the viral movement to upper leaves. The results suggest that expressions of viral resistance genes in the host are the key component in the resistance against ToMV in the inducer-treated tomato plants.

• The Plant Pathology Journal
• /
• 제17권2호
• /
• pp.106-109
• /
• 2001

A multi-rapid immunofilter paper assay (multi-RIPA) system was prepared for simultaneous diagnosis of three Tobamoviruses, Cucumber green mottle mosaic virus (CGMMV), Kyuri green mottle mosaic virus (KGMMV), and Zucchini green mottle mosaic virus (ZGMMV) in cucurbitaceous crops. Each of these viruses was specifically detected by the multi-RIPA from cucumber, watermelon, zucchini, and bottle gourd inoculated with the three Tobamoviruses singly or in combination. The three viruses could infect cucumber, watermelon, and bottle gourd ; however, CGMMV could not infect zucchini as the latex-coated CGMMV antibody showed a negative reaction in the multi-RIPA of the virus-infected plant extract. When the minimum detection level of multi-RIPA was compared with that of double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) using CGMMV, the latter was 10 times more sensitive than the former. The detection limit of the multi-RIPA for the purified CGMMV was 50 ng/ml. In a survey of the threeviruses in cucurbits growing in commercial fields in 1999 and 2000, CGMMV was detected in watermelon and cucumber, and ZGMMV was detected only in zucchini growing in plastic houses at the suburbs of Chonju, Korea. However, KGMMV was not found in the commercially growing cucurbit crops in our study, The results suggest that the multi-RIPA can be a simple, rapid, specific and convenient tool to detect simultaneously the Tobamoviruses.

• The Plant Pathology Journal
• /
• 제17권5호
• /
• pp.243-248
• /
• 2001

Two Tobamoviruses, Cucumber green mottle mosaic virus (CGMMV) and Zucchini green mottle mosaic virus (ZGMMV), occurred in Korea in 463 ha in 1998, 33.9 ha in 1999, and 44.2 ha in 2000. CGMMV was detected in watermelon, cucumber, oriental melon, and melon, whereas ZGMMV was mainly detected in zucchini squash. Thirty-six CGMMV isolates wee classified into three types by analysis of single strand cDNA conformational polymorphism (SSCP) of the coat protein gene. In a comparison of serological relationships among CGMMV, ZGMMV, and Kyuri green mottle mosaic virus (KGMMV), the three tobamoviruses specifically reacted with each homologous antibody in the double-antibody sandwich enzyme-linked immunosorbent assay and rapid imunofilter paper assay (RIPA), although ZGMMV and KGMMV were slightly biologcially similar. In a survey of the three tobamoviruses in cucurbitgrowing field in Korea by RIPA, CGMMV and ZGMMV were detected but KGMMV was not found in commercially growing cucurbit crops so far. Seed contamination ratio of CGMMV in bottle gourd seeds tested was 84%, while seed trasmission ratio from the virus-contaminated seeds was 2.0%. Soil transmission ratio was 0-3.5% in fields naturally infested with CGMMV or ZGMMV. Control measures of the virus diseases are roguing and sanitation. These suggest that it is important to rogue the first infected crops, which include the seed and soil, especially early in the season. This may be practicable to control the diseases because CGMMV and ZGMMV have a narrow host range restricted to cucurbitaceous crops.

• The Plant Pathology Journal
• /
• 제22권2호
• /
• pp.164-167
• /
• 2006

An immunocapture reverse transcription-polymerase chain reaction (IC/RT-PCR) was developed for simultaneous detection of two pepper-infecting RNA viruses, Pepper mud mottle virus (PMMoV) and Tobacco mild green mosaic virus (TMGMV). The assay could be performed in a single tube for simultaneous and sensitive detection of these tobamoviruses. This detection system revealed thousand-fold increase in detection sensitivity compare to ELISA. This method could save time and reagent cost compare to common RT-PCR which needs several reactions and several procedures of viral RNA extractions for the same number of samples.

• 식물병연구
• /
• 제7권3호
• /
• pp.164-169
• /
• 2001

Nicotiana glutinosa를 이용한 단병반분리를 통해 국내에 시판되고 있는 상용고추종자로부터 괴사국부병반의 크기가 다른 두 종류의 Tobamovirus를 분리하고 이들을 각각 Tobamovirus-6(T-6)과 Tobamovirus-19(T-19)로 명명하였다. 기주범위와 혈청학적 연관성 조사에서 T-6은 pepper mild mottle vitus(PMMoV)와, T-19는 tomato mosaic virus (ToMV)와 유사한 특성을 보였다. 또한 외피단백질 염기서열 분석에서 T-6은 PMMoV와 T-19는 ToMV와 매우 유사하였다.

• 한국식물병리학회:학술대회논문집
• /
• 한국식물병리학회 2003년도 정기총회 및 추계학술발표회
• /
• pp.125.3-126
• /
• 2003

A near full-length sequence of a new tobamovirus infecting Hibiscus rosa-sinensis L. was determined. The genome consists of 58 nucleotides (nt) 5' UTR, followed by a 4.9 kb ORF which methyl transferase helicase domain (128 kDa), readthrough protein RNA dependent RNA polymerase (RdRp) 185 kDa and a 52 kDa protein. The 128 kDa protein had a maximum homology of 51.4 ％ to TMGMV and amino acids (an) were 54.3 ％ identical to TMV- vulgare strain. The 185 kDa RdRp had a maximum homology of 53.5％ to TMV-Ob and KGMMV-Y and a 59.6％ homology at the an level to CGMMV-SH. The MP gene encodes 282 aa and its theoretical molecular weight is 30.4 kDa. The nt and an sequence identities of MP ranged from 38.8％ to 43.9％ and 30.9％ to 37.9％, respectively. The CP gene encodes 163 residues and with a theoretical molecular weight of 18.2 kDa The (nt) and aa sequences of the CP were 46.9 ％ to 51.6％ and 45.3％ to 57.1％ identical to other tobamoviruses, respectively. The predicted virion origin of assembly (OAS) was located in the CP gene. Phylogenetic trees generated based on the nt and as sequences of RdRp, MP and CP genes indicated that this new virus clustered with subgroup II tobamoviruses. Although the CP ORF of this virus shared a high nt and aa sequence identity with Sunn-hemp mosaic virus (SHMV), Western analysis showed that it is serologically unrelated to SHMV. We propose the name Hibiscus virus S (HVS) for this Singapore isolate. This is the first report on a near full-length sequence of a Tobamovirus that infects hibiscus.

• Molecules and Cells
• /
• 제25권2호
• /
• pp.205-210
• /
• 2008

To develop molecular markers linked to the $L^4$ locus conferring resistance to tobamovirus pathotypes in pepper plants, we performed AFLP with 512 primer combinations for susceptible (S pool) and resistant (R pool) DNA bulks against pathotype 1.2 of pepper mild mottle virus. Each bulk was made by pooling the DNA of five homozygous individuals from a T10 population, which was a near-isogenic $BC_4F_2$ generation for the $L^4$ locus. A total of 19 primer pairs produced scorable bands in the R pool. Further screening with these primer pairs was done on DNA bulks from T102, a $BC_{10}F_2$ derived from T10 by back crossing. Three AFLP markers were finally selected and designated L4-a, L4-b and L4-c. L4-a and L4-c each underwent one recombination event, whereas no recombination for L4-b was seen in 20 individuals of each DNA bulk. Linkage analysis of these markers in 112 $F_2$ T102 individuals showed that they were each within 2.5 cM of the $L^4$ locus. L4-b was successfully converted into a simple 340-bp SCAR marker, designated L4SC340, which mapped 1.8 cM from the $L^4$ locus in T102 and 0.9 cM in another $BC_{10}F_2$ population, T101. We believe that this newly characterized marker will improve selection of tobamovirus resistance in pepper plants by reducing breeding cost and time.

• The Plant Pathology Journal
• /
• 제16권2호
• /
• pp.118-124
• /
• 2000

The coat protein (CP) gene of kyuri green mottle mosaic virus zucchini strain (KGMMV-Z) isolated from zucchini (Cucurbita pepo) in Chonfu, Korea in 1999 was sequenced by the reverse transcription and polymerase chain reaction with degenerate and generate primers originated from tobamoviruses. The degenerate primers were very effective in amplification of KGMMV-Z CP region. The KGMMV-Z CP gene consisted of 486 nucleotides and had the same nucleotide length compared with those of cucurbit-infecting tobamoviruses. KGMMV-Z CP gene shared 43.8, 44.2, and 44.4% nucleotide sequence similarity with the CP gene of cucumber green mottle mosaic virus watermelon strain (CGMMZ-W), CGMMV-KW1, and CGMMV-SH, respectively, whereas three CGMMV strains among themselves showed 98.6-99.6% nucleotide similarity. The deduced amino acids of KGMMV-Z CP gene were 161 amino acid residues with the molecular weight of 17,181 daltons. The first 24 codons of KGMMV-Z CP gene corresponded to the sequences of the N-terminal amino acid of the viral capsid protein. The amino acid sequences of KGMMV-Z CP had 45.3% similarity compared with those of three CGMMV strains. However, the amino acid sequences of CGMMV strains were identical. These results showed that two cucurbit-infecting tobamovirus members, KGMMV-Z and CGMMV were genetically distantly related.

• The Plant Pathology Journal
• /
• 제22권2호
• /
• pp.147-151
• /
• 2006

Disease incidences of cucumber virus diseases in Jeonnam province were 52.5%, 16.1%, 35.2%, and 50.9% in 1999, 2000, 2001, and 2002, respectively. Rod- and flexuous rod-shaped virus particles were observed with the frequencies of 63.2% and 10.5%, respectively from the samples collected in 1999 under EM observation. Rod-shaped virus particles are considered as tobamovirus while flexuous rod shaped particles are considered as potyviruses. To further confirm their nature, total of 312 diseased virus samples were collected from 2000 to 2002, and tested by RT-PCR. Disease incidences of tobamoviruses including Cucumber green mottle mosaic virus and Kyuri green mottle mosaic virus were 48.7% and 3.8%, respectively while those of potyviruses including Zucchini yellow mosaic virus, Papaya ringspot virus, and Watermelon mosaic virus were 15.7%, 9.3%, and 5.1%, respectively. Interestingly, Cucumber mosaic virus was hardly detected. About 5.8% of tested samples were infected with more than one virus. Tobamovirus infection was consistently observed from September to December regardless of planting time, whereas infection of potyviruses was observed in many cucumber cultivating areas where it was planted in September and October.

• 식물병연구
• /
• 제13권2호
• /
• pp.82-87
• /
• 2007

박과 작물에 발생하는 종자전염성 막대형 바이러스인 CGMMV, KGMMV, ZGMMV 3종의 단독 및 동시 VC/RT-PCR 유전자 진단법을 개발하였다. CGMMV 등 3종의 바이러스에 대한 동시진단용 조합선발을 위하여 12조합 중에서 동시 진단용 프라이머 조합 CGMMV-C724, KGMMV-K513, ZGMMV-Z407A를 선발하였다. CGMMV등 3종에 대한 triplex VC/RT-PCR 유전자 진단법은 박, 수박, 오이, 멜론, 쥬키니 호박, 애호박, 담배(N. benthamiana) 작물의 식물체 즙액과 프라이머 간에 간섭현상 없이 특이적으로 진단되었다.