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Generation and Retention of Defective RNA3 from Cucumber Mosaic Virus and Relevance of the 2b Protein to Generation of the Subviral RNA

  • Emi Ota;Chikara Masuta;Minoru Takeshita
    • The Plant Pathology Journal
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    • v.39 no.6
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    • pp.592-599
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    • 2023
  • A defective RNA3 (D3Yα) of strain Y of cucumber mosaic virus (CMV-Y) was examined on host-specific maintenance, experimental conditions, and a viral factor required for its generation in plants. D3Yα was stably maintained in cucumber but not in tomato plants for 28 days post inoculation (dpi). D3Yα was generated in Nicotiana tabacum or N. benthamiana after prolonged infection in the second and the third passages, but not in plants of N. benthamiana grown at low temperature at 28 dpi or infected with CMV-Y mutant that had the 2b gene deleted. Collectively, we suggest that generation and retention of D3Yα depends on potential host plants and experimental conditions, and that the 2b protein has a role for facilitation of generation of D3Yα.

Symptoms of Cucumber Virus Diseases Occurred in Sangju and Gurye in 2006 and 2007 (2006년과 2007년 상주와 구례에서 발생한 오이 바이러스병의 병징 특성)

  • Cho, Jeom-Deog;Lee, Jung-Hwan;Ko, Sug-Ju;Choi, Hong-Soo;Lee, Su-Heon;Choi, Gug-Seoun;Kim, Jeong-Soo
    • Research in Plant Disease
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    • v.17 no.2
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    • pp.196-204
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    • 2011
  • Virus diseases occurring on cucumber was surveyed at main plantation areas of 'Sangju' and 'Gurye' in 2006 and 2007. Viral infection rate on cucumber was ranged from 14% to 90% in fields and the average infection rate was 46% at Sangju area. Cucumber cultivated at Gurye area had viral incidence ranged 9-100% and averaged 48%. The majorly infected viruses were Cucumber green mottle mosaic virus, Zucchini yellow mosaic virus, Papaya ring spot virus and Watermelon mosaic virus-2, and their infection rates were 23.5%, 13.0%, 9.0% and 2.0%, respectively in 2006 and 2007. Mixed infection rate of duplex, triplex and tetraplex was 31.5%, 7.5% and 2.5%, respectively. The rate of infection type was 47.7% by single infection, 31.5% by double infection and 88.7% by both infection types. Important viruses involved in mixed infection types were Zucchini yellow mosaic virus, Cucumber green mottle mosaic virus, Papaya ring spot virus and Broad been wilt virus2, orderly. Symptom in single infection was almost same on 'Dadagi' line and 'Chicheong' line of cucumber cultivars. Cucumber green mottle mosaic virus produced various symptoms of chlorotic spot, vein chlorosis and vein wrinkle and so on. Main symptoms of vein chlorosis, and severe mosaic and malformation were induced by Zucchini yellow mosaic virus. Mild symptoms were occurred relatively by Papaya ring spot virus and Watermelon mosaic virus2.

Cross-Protection Effectiveness of Cucumber mosaic virus (CMV) Isolates Associated with Satellite RNA for Prevention of CMV Disease in Pepper Plants (Satellite RNA 보유 Cucumber mosaic virus(CMV)의 고추 CMV병에 대한 교차방어 효과)

  • 최장경;성미영;정혜진;홍진성;이상용
    • Research in Plant Disease
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    • v.7 no.3
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    • pp.155-163
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    • 2001
  • Two attenuated Cucumber mosaic virus (CMV) isolates, Paf-CMV and Rs2-CMV that had been selected from CMV isolates associated with satellite RNA (satRNA) were tested for cross-protection effect in pepper plants. The viruses selected as attenuated strains appeared to be identical serologically and physically to the challenge virus (Mf-CMV), but they were lower in the dilution end-point of infectivity of crude sap than Mf-CMV When symptoms were observed in several indicator plants after inoculation, Paf-CMV and Rs2-CMV were symptomless or showed mild mosaic symptoms while another satRNA isolate Ap-CMV developed severe mosaic symptoms on the leaves as Mf-CMV. The nucleotide sequences of the satRNAs were determined by sequencing full-length cDNA clones. Paf-, Rs2- and Ap-satRNAs were 386, 335, and 347 nucleotides long, respectively, The sequences were then compared with the other known Y-satRNA, revealing that nucleotide sequences of the satRNAs consisted of 5'- and 3'-terminal conserved regions. However variations occurred on the middle regions of the sequences, especially those related to symptom interference, showing significant differences between Paf-satRNA and other isolates. Infectious transcripts of Paf-satRNA and Rs2-satRNA induced mild mosaic symptoms in pepper plants when supported by genomic RNAs of Mf-CMV. Under greenhouse conditions, Paf-CMV and Rs2-CMV were tested for cross-protection effect in pepper and tobacco (Nicotiana tabacum cv, Xanthi nc) plants against Mf-CMV. No symptoms were developed on the plants vaccinated with Paf-CMV until 3 weeks after inoculation with the virulent strain; however another attenuated isolate, Rs2-CMV, showed less effectiveness in cross-protection. Depending on the concentration of the challenged virus, symptoms sometimes appeared later in the upper leaves. However, in plants challenged with low concentrations (below 0.2 mg/ml) of the challenge inoculum, symptoms caused by the virulent strain did not develop on the plants vaccinated with Paf-CMV. In the field experiments, the number of pepper plants with severe mosaic symptoms in the control plots was progressively increased after transplanting and reached approximately 50% after 50 days. On the other hand, the incidence of mosaic disease appeared very low on the plants that had received the protective inoculation with Paf-CMV.

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Identification and Sequence Analysis of RNA3 of a Resistance-Breaking Cucumber mosaic virus Isolate on Capsicum annuum

  • Lee Mi-Yeon;Lee Jang-Ha;Ahn Hong-Il;Yoon Ju-Yeon;Her Nam-Han;Choi Jang-Kyung;Choi Gug-Seon;Kim Do-Sun;Harn Chee-Hark;Ryu Ki-Hyun
    • The Plant Pathology Journal
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    • v.22 no.3
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    • pp.265-270
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    • 2006
  • Cultivated hot pepper crops showing severe mosaic symptom were found in Korea in 2004 and their causal agent was identified as Cucumber mosaic virus (CMV). These pepper crops was resistant to the virus in the filled, and they belonged to pathotype 0 (P0) resistant pepper. Resistance screening of selected pepper plants showed that a pepper isolate of CMV was the P0 resistance-breaking virus. This P0 resistance-breaking isolate of CMV, named as Ca-P1, was isolated from leaves of the virus-infected Capsicum annuum cv. Manidda that showed systemic severe mosaic symptom. Ca-P1-CMV could induce systemic mosaic symptoms on P0-susceptible (P0-S) and P0-resistant (P0-R) cultivars whereas an ordinary strain (Fny-CMV) could not infect P0-R. This result suggests that Ca-P1-CMV can overcome P0 resistant pepper cultivars. To analyze its genome sequence, the complete nucleotide sequence of RNA3 of Ca-P1-CMV was determined from the infectious full-length cDNA clone of the virus. RNA3 of Ca-P1-CMV consisted of 2,219 nucleotides. Overall sequence homology of RNA3-encoded two viral proteins (movement protein and coat protein) revealed high similarity (75.2-97.2%) with the known CMV strains. By sequence analysis with known representative strains of CMV, Ca-P1-CMV belongs to a typical member of CMV subgroup IB. The resistance and resistance-breaking mechanisms of pepper and counterpart CMV, respectively, remain to be investigated, which will enrich the genetic resources and accelerate CMV-resistant pepper breeding programs.

Ultrastructural Aspects of Mixed Infections with Turnip mosaic virus (TuMV- ACl8 and -C5) and Ribgrass mosaic virus (RMV-CA1) in Oriental Cabbage

  • Cho, Jeom-Deog;Park, Hong-Soo;Kim, Jeong-Soo;La, Yong-Jun;Kim, Kyung-Soo
    • The Plant Pathology Journal
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    • v.18 no.4
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    • pp.192-198
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    • 2002
  • Mixed infections of two economically important viruses, Turnip mosaic virus(TuMV) in the family Potyviridae and Ribgrass mosaic virus(RMV) in the genus Tobamo-virus, were studied ultrastructurally on oriental cabbage. TuMV-ACl8 (alpine isolate in Korea) induced chlorotic spots on inoculated leaves of both ‘SSD63’ inbred line known as susceptible to TuMV, and ‘Tambok’ commercial cultivar, known as resistant to the virus, in the early stages of infection. TuMV-C5 (Taiwan isolate) caused severe mosaic and malformation on the upper leaves of ‘SSD63’, and necrotic spots in both inoculated and upper leaves of ‘Tambok’. RMV-CA1 (oriental cabbage isolate from alpine in Korea) induced vein chlorosis, leaf malformation, and midrib necrotic streak in the upper leaves of both ‘SSD63’ and ‘Tambok’. Both oriental cabbages infected with a combination of TuMV-ACl8 and RMV-CA1 showed synergistic symptoms of severe yellowing, severe mosaic, and necrotic spot or vein necrosis on their leaves. A combination of TuMV-C5 and RMV-CA1 produced synergistic symptoms only in ‘SSD63’. In ‘Tambok’ infected with the combination of TuMV-C5 and RMV-CA1, the number of necrotic spots on the inoculated leaves was one half lesser than that on singly infected with TuMV-C5. A few necrotic spots progressed systemically. In cells infected with a combination of TuMV-ACl8 and RMV-CA1 or TuMV-C5 and RMV-CA1, the particles of the two viruses made nonagon-like rings(NLR); one TuMV particle was surrounded loosely by nine RMV particles. Two unrelated viruses of TuMV and RMV were compacted in the central part of the spiral aggregates(SA) that was induced strikingly in cells by the mixed infections. The SA showed NLR in its center of the cross-sectioned side. Many particles of RMV of Tobamovirus were closely associated with Potyvirus-characteristic cylindrical inclusions. The SAs in the mixed infections were formed easily by the Potyvirus of TuMV-ACl8 or -C5 isolates.

Evaluation of Resistance in Pepper Germplasm to Cucumber mosaic virus by High Resolution Melting Analysis (High Resolution Melting Analysis을 이용한 고추 유전자원의 Cucumber mosaic virus 저항성 평가)

  • Ro, Na Young;Hur, On Sook;Ko, Ho Cheol;Kim, Sang Gyu;Rhee, Ju Hee;Gwag, Jae-Gyun;Kwon, Jin-Kyung;Kang, Byoung-Cheorl
    • Research in Plant Disease
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    • v.18 no.4
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    • pp.290-297
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    • 2012
  • In this study, total number of 1941 Capsicum accessions conserved at RDA Genebank was evaluated for their response to Cucumber mosaic virus (CMV). These accessions were composed with 9 species originated from 89 countries, included 839 Capsicum annuum, 277 C. baccatum, 395 C. chinense, 343 C. frutescens, 49 C. pubescens, and other 38 wild pepper species (C. chacoense, C. galapagoense, etc.). Resistant to CMV was screened with the 240H02SP6 SNP marker related to the Cmr1 (Cucumber mosaic resistance 1). Eighty nine accessions of pepper germplasm were resistant to CMV based on the marker. One hundred sixty two accessions showed heterozygosity. One thousand two hundred seventy accessions were susceptible to CMV. Four hundred twenty accessions did not show distinction by 240H02SP6 marker. These 89 resistant pepper germplasm can be used in a pepper breeding program against CMV.

Study on X bodies in epidermal cells of Carina generalis infected with a mosaic virus (칸나 모자이크병의 X체에 관한 연구)

  • Lee Chang Un
    • Korean journal of applied entomology
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    • v.15 no.4 s.29
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    • pp.199-204
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    • 1976
  • Canna nosaic X bodies, which do not exist in tissues of the healthy plant and are originating in cells of virus infected Canna (Canna generalis BAILEY) with mosaic symptom, are easily observed under microscope through application of vital staining for 2-3 minutes with $1\%$ eosin of $H_2O$ solution added with slight amount of $CH_3COOH$ and distinguishing with N/5HCl followed by washing to inspect. The result of this experiment is summarized as following: 1) The X bodies are observed not only in epiermal cells of leaf of the mosaic virus infected Canna but in those of leaf sheath, stem, and root also, and it is expected that the X bodies are to exist in the flower cells of the disease infected Canna which were missed in this experiment. 2) Shape and nature of X bodies are not constant; in early stage of the disease development, the X bodies have equal contents and vague contour with their small size and round shape, but along with progress of the disease development they attain granular contents and clear contour with their increasing sige and defining shape in cytoplasm. In case of same individual pant, fully developed X. bodies. are increasing in cytoplasm in propoition to severity of mosaic and nettling of the diseased leaf. 3) The staining character of X bodies to eosin is more dense than that of nuclei; Xbodies are stained light red or red while nuclei are stained yellowish brown or light red. 1) It is assumed to be a result of cytoplasmic concentration around nucleus that X bodies are usunlly developed adjacent to nucleus and they are considered to be a cytoplasmic prodct. 5) Thus, I confirm that X bodies originsting in canna plant cells infected with mosaic virus aye multipling in the alive cells.

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Detection of Soybean mosaic virus by Reverse Transcription Loop-mediated Isothermal Amplification (Reverse transcription Loop-mediated isothermal amplification을 이용한 Soybean mosaic virus의 진단)

  • Lee, Yeong-Hoon;Bae, Dae-Hyeon;Kim, Bong-Sub;Yoon, Young-Nam;Bae, Soon-Do;Kim, Hyun-Joo;Mainali, Bishwo P.;Park, In-Hee;Lee, Su-Heon;Kang, Hang-Won
    • Research in Plant Disease
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    • v.21 no.4
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    • pp.315-320
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    • 2015
  • Soybean mosaic virus (SMV) is a prevalent pathogen that causes significant yield reduction in soybean production worldwide. SMV belongs to potyvirus and causes typical symptoms such as mild mosaic, mosaic and necrosis. SMV is seed-borne and also transmitted by aphid. Eleven SMV strains, G1 to G7, G5H, G6H, G7H, and G7a were reported in soybean varieties in Korea. A reverse transcription loop-mediated isothermal amplification (RT-LAMP) method allowed one-step detection of gene amplification by simple procedure and needed only a simple incubator for isothermal template. This RT-LAMP method allowed direct detection of RNA from virus-infected plants without thermal cycling and gel electrophoresis. In this study, we designed RT-LAMP primers named SML-F3/B3/FIP/BIP from coat protein gene sequence of SMV. After the reaction of RT-LAMP, products were identified by electrophoresis and with the detective fluorescent dye, SYBR Green I under daylight and UV light. Optimal reaction condition was at $58^{\circ}C$ for 60 min and the primers of RT-LAMP showed the specificity for nine SMV strains tested in this study.

On Improvement of Garlic Productivity by Inactivation of Virus in Garlics (마늘 Virus 불활성화에 의한 생산성 향상에 관하여)

  • LEE Chang Un
    • Korean journal of applied entomology
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    • v.20 no.1 s.46
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    • pp.6-14
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    • 1981
  • The effect of heat or chemotherapeutant treatment on the mosaic virus infected garlic(Allium sativum L.) scales and that of chemotherapeutant added to the culture medium were summarized as following. The treatment of the virus infected garlic scales at $37\~57^{\circ}C$ for 35 days to one hour in water or in air shelved no effort of inactivating the virus. Although treatment of the garlic scales at $62\~72^{\circ}C$ for 90 to five minutes reduced the mosaic symptom on the leaves of the garlic plants grown after the heat treatment, it reduced the growth vigor of the plants so greatly that complete inactivation of the virus in garlics was not feasible. The mosaic symptom on the leaves of garlic plant was reduced when the infected garlic scales were grown after 24 hours soaking in $10\~50\;ppm$ Malachite Green, 2,4-Dichlorophnoxy Acetic Acid, or in $20\~100\;ppm$ Quinhydron. These chemotherapeutants, however, inhibited the growth of garlic plant at the high concentration. Garlic scales soaked in $10\~50\;ppm$ Naphthyl Acetic Acid showed the least mosaic symptom without its complete extiction on the garlic leaves. When incorporated into the modified Murashige-Skoog's medium, $0.5\~l.5\;ppm$ Naphthyl Acetic Acid could inactivate the mosaic virus in newy developed garlic plants showing no mosaic symptom on the leaves, no inclusion bodies and intact nuclei in the leaf tissue cells.

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Serological Investigation of Virus Diseases of Tobacco Plant (Nicotiana tabaccum L.) In Korea (혈청학적 방법에 의한 잎담배 바이러스병의 감염상 조사)

  • Park Eun Kyung;La Yong Joon;Heu Il;Lee Yong Deuk
    • Korean journal of applied entomology
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    • v.14 no.2 s.23
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    • pp.59-63
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    • 1975
  • A total of 40 virus infected tobacco plants (Nicotiana tabaccum L.) with various symptom types Were collected from Bucheon and Jeonju area by its symptoms were investigated on the incidence of tobacco mosaic virus (TMV), cucumber mosaic virus (CMV), alfalfa mosaic virus (AMV), potato virus X (PVX) and potato virus Y (PVY) by serological methods. van Slogteren's microprecipitin test was applied for the testing of PVX and PVY from infected plants and Ouchterlony agar double diffusion test was used for CMV, TMV and AMV. Results obtained are as follows: 1. TMV, CMV, AMV, PVX and PVY wcre found to occur on the tobacco plants growing in Korea. 2. The prevalence of each of these viruses among the 40 tobacco plants investigated was in the order of AMV: $(67.5\%)>CMV:(60.0\%)>TMY:(47.5\%)>PVY:(17.5\%)>PVX: (10.0\%).$ 3. In Burley variety, the percentage of infection by TMV was $15\%$, whereas it was as high as $80\%$ in Hicks variety. 4. Among the 40 tobacco plants investigated, $37.5\%$ showed infection with one kind of virus whereas the remaining $62.5\%$, revealed mixed infection with more than two different viruses.

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