• Title/Summary/Keyword: soybean virus

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Characterization of the in vitro Activities of the P1 and Helper Component Proteases of Soybean mosaic virus Strain G2 and Tobacco vein mottling virus

  • Lim, Hyoun-Sub;Jang, Chan-Yong;Nam, Ji-Ryun;Li, Meijia;Hong, Jin-Sung;Bae, Han-Hong;Ju, Ho-Jong;Kim, Hong-Gi;Ford, Richard E.;Domier, Leslie L.
    • The Plant Pathology Journal
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    • v.28 no.2
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    • pp.197-201
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    • 2012
  • Potyviruses express their RNA genomes through the production of polyproteins that are processed in host cells by three virus-encoded proteases. Soybean plants produce large amounts of protease inhibitors during seed development and in response to wounding that could affect the activities of these proteases. The in vitro activities of two of the proteases of Soybean mosaic virus (SMV) and Tobacco vein mottling virus (TVMV) were compared in the rabbit reticulocyte lysate in vitro translation system using synthetic RNA transcripts. Transcripts produced from SMV and TVMV cDNAs that included the P1 and helper component-protease (HC-Pro) coding regions directed synthesis of protein products that were only partially processed. Unprocessed poly-proteins were not detected from transcripts that included all of the P1, HC-Pro, P3 and portions of the cylindrical inclusion protein coding regions of either virus. Addition of soybean trypsin inhibitor to in vitro translation reactions increased the accumulation of the unprocessed polyprotein from TVMV transcripts, but did not alter the patterns of proteins produced from SMV. These experiments suggest that SMV-and TVMV-encoded proteases are differentially sensitive to protease inhibitors.

Interaction Study of Soybean mosaic virus Proteins with Soybean Proteins using the Yeast-Two Hybrid System

  • Seo, Jang-Kyun;Hwang, Sung-Hyun;Kang, Sung-Hwan;Choi, Hong-Soo;Lee, Su-Heon;Sohn, Seong-Han;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.23 no.4
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    • pp.281-286
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    • 2007
  • Interactions between viral proteins and host proteins are essential for virus replication. Especially, translation of viral genes completely depends on the host machinery. In potyviruses, interactions of genome-linked viral protein (VPg) with host translation factors including eIF4E, eIF(iso)4E, and poly(A)-binding protein (PABP) has previously been characterized. In this study, we investigated interactions between Soybean mosaic virus (SMV) viral proteins and host translation factors by yeast two-hybrid system. SMV VPg interacted with eIF4E, eIF(iso)4E, and PABP in yeast two-hybrid system, while SMV helper component proteinase (HC-pro) interacted with neither of those proteins. The interaction between SMV NIb and PABP was also detected. These results are consistent with those reported previously in other potyviruses. Interestingly, we found reproducible and specific interactions between SMV coat protein (CP) and PABP. Deletion analysis showed that the region of CP comprising amino acids 116 to 206 and the region of PABP comprising amino acids 520 to 580 are involved in CP/PABP interactions. Soybean library screening with SMV NIb by yeast two-hybrid assay also identified several soybean proteins including chlorophyll a/b binding preprotein, photo-system I-N subunit, ribulose 1,5-biphosphate carboxylase, ST-LSI protein, translation initiation factor 1, TIR-NBS type R protein, RNA binding protein, ubiquitin, and LRR protein kinase. Altogether, these results suggest that potyviral replicase may comprise a multi-protein complex with PABP, CP, and other host factors.

Immunochemical Detection of Soybean Mosaic Virus Infections in the Seeds of Soybean Cultivars in Korea (면역이중확산법에 의한 콩 종자의 모자이크 바이러스(SMV) 감염상 조사)

  • La Yong-Joon;Bak Won-Chull;Oh Jeung-Haing
    • Korean journal of applied entomology
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    • v.22 no.1 s.54
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    • pp.26-30
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    • 1983
  • Soybean mosaic virus (SMV) infection in seeds wits investigated by an immunochemical method Seven soybean cultivars or lines Bughae No.1, KAS 662-7, Chungbugbaeg, Gwanggyo Clark, Bongeui, and Gangrim were tested using hypocotyls of germinated seeds and presence of SMV was detected in six soybean cultivars but Gangrim. The level of SMV infection in the assayed cultivars varied from 2.1 to $12.5\%$. It seemed that seed coat mottling had no correlation with seeds. SMV infection of the seeds since virus has not always been detected from the mot tled seeds SMV has not been detected in the seeds of variety Gwanggyo which showed necrotic symptoms.

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Serological Detection of Plant Viruses with latex-test (Latex-Test에 의한 식물 바이러스의 검정)

  • 박은경;김정화;이영근
    • Journal of the Korean Society of Tobacco Science
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    • v.1 no.1
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    • pp.33-38
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    • 1979
  • A simple test using latex-agglutination was developed to detect serologically tobacco mosaic virus (TMV), soybean mosaic virus (SoyMV), and barley stripe mosaic virus (BSMV) from infected Plants. Latex spheres ( 0.81 $\mu$, Difco) were adsorbed with immuno globulin purified by electrophoresis from crude antiserum against viruses. The antibody- sensitized latex suspension was mixed with sap from virus -infected leaves in a glass capillary tube (inner diam. 1mm $\times$ 100 mm length) The mixture, after agitation, was observed under a stereo microscope at low magnification (X20 - X4O), to examine the reaction between antigen (Virus) and its antibody. Flocculation occurred when the reaction was positive.

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Optimization of a Virus-Induced Gene Silencing System with Soybean yellow common mosaic virus for Gene Function Studies in Soybeans

  • Kim, Kil Hyun;Lim, Seungmo;Kang, Yang Jae;Yoon, Min Young;Nam, Moon;Jun, Tae Hwan;Seo, Min-Jung;Baek, Seong-Bum;Lee, Jeom-Ho;Moon, Jung-Kyung;Lee, Suk-Ha;Lee, Su-Heon;Lim, Hyoun-Sub;Moon, Jae Sun;Park, Chang-Hwan
    • The Plant Pathology Journal
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    • v.32 no.2
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    • pp.112-122
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    • 2016
  • Virus-induced gene silencing (VIGS) is an effective tool for the study of soybean gene function. Successful VIGS depends on the interaction between virus spread and plant growth, which can be influenced by environmental conditions. Recently, we developed a new VIGS system derived from the Soybean yellow common mosaic virus (SYCMV). Here, we investigated several environmental and developmental factors to improve the efficiency of a SYCMV-based VIGS system to optimize the functional analysis of the soybean. Following SYCMV: Glycine max-phytoene desaturase (GmPDS) infiltration, we investigated the effect of photoperiod, inoculation time, concentration of Agrobacterium inoculm, and growth temperature on VIGS efficiency. In addition, the relative expression of GmPDS between non-silenced and silenced plants was measured by qRT-PCR. We found that gene silencing efficiency was highest at a photoperiod of 16/8 h (light/dark) at a growth temperature of approximately $27^{\circ}C$ following syringe infiltration to unrolled unifoliolate leaves in cotyledon stage with a final SYCMV:GmPDS optimal density $(OD)_{600}$ of 2.0. Using this optimized protocol, we achieved high efficiency of GmPDS-silencing in various soybean germplasms including cultivated and wild soybeans. We also confirmed that VIGS occurred in the entire plant, including the root, stem, leaves, and flowers, and could transmit GmPDS to other soybean germplasms via mechanical inoculation. This optimized protocol using a SYCMV-based VIGS system in the soybean should provide a fast and effective method to elucidate gene functions and for use in large-scale screening experiments.

Complete Genome Sequencing and Infectious cDNA Clone Construction of Soybean Mosaic Virus Isolated from Shanxi

  • Wang, Defu;Cui, Liyan;Zhang, Li;Ma, Zhennan;Niu, Yanbing
    • The Plant Pathology Journal
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    • v.37 no.2
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    • pp.162-172
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    • 2021
  • Soybean mosaic virus (SMV) is the predominant viral pathogen that affects the yield and quality of soybean. The natural host range for SMV is very narrow, and generally limited to Leguminosae. However, we found that SMV can naturally infect Pinellia ternata and Atractylodes macrocephala. In order to clarify the molecular mechanisms underlying the cross-family infection of SMV, we used double-stranded RNA extraction, rapid amplification of cDNA ends polymerase chain reaction and Gibson assembly techniques to carry out SMV full-length genome amplification from susceptible soybeans and constructed an infectious cDNA clone for SMV. The genome of the SMV Shanxi isolate (SMV-SX) consists of 9,587 nt and encodes a polyprotein consisting of 3,067 aa. SMV-SX and SMV-XFQ008 had the highest nucleotide and amino acid sequence identities of 97.03% and 98.50%, respectively. A phylogenetic tree indicated that SMV-SX and SMV-XFQ018 were clustered together, sharing the closest relationship. We then constructed a pSMV-SX infectious cDNA clone by Gibson assembly technology and used this clone to inoculate soybean and Ailanthus altissima; the symptoms of these hosts were similar to those caused by the virus isolated from natural infected plant tissue. This method of construction not only makes up for the time-consuming and laborious defect of traditional methods used to construct infectious cDNA clones, but also avoids the toxicity of the Potyvirus special sequence to Escherichia coli, thus providing a useful cloning strategy for the construction of infectious cDNA clones for other viruses and laying down a foundation for the further investigation of SMV cross-family infection mechanisms.

Molecular and Biological Characterization of an Isolate of Cucumber mosaic virus from Glycine soja by Generating its Infectious Full-genome cDNA Clones

  • Phan, Mi Sa Vo;Seo, Jang-Kyun;Choi, Hong-Soo;Lee, Su-Heon;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.30 no.2
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    • pp.159-167
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    • 2014
  • Molecular and biological characteristics of an isolate of Cucumber mosaic virus (CMV) from Glycine soja (wild soybean), named as CMV-209, was examined in this study. Comparison of nucleotide sequences and phylogenetic analyses of CMV-209 with the other CMV strains revealed that CMV-209 belonged to CMV subgroup I. However, CMV-209 showed some genetic distance from the CMV strains assigned to subgroup IA or subgroup IB. Infectious full-genome cDNA clones of CMV-209 were generated under the control of the Cauliflower mosaic virus 35S promoter. Infectivity of the CMV-209 clones was evaluated in Nicotiana benthamiana and various legume species. Our assays revealed that CMV-209 could systemically infect Glycine soja (wild soybean) and Pisum sativum (pea) as well as N. benthamiana, but not the other legume species.

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.

Affection of SMV -G5H Strain on Plant Growth and Seed Chemical Composition of Soybean Variety, Danyeobkong (콩 모자이크 바이러스 SMV-G5H 나병이 단엽콩 생육 및 종실 성분에 미치는 영향)

  • 김율호;노재환;김명기;임대준;이봉호;이상철
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.41 no.3
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    • pp.340-347
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    • 1996
  • Soybean [Glycjne max Merr. L.] variety 'Danyeobkong' was inoculated with soybean mosaic virus SMV-G5H strain in field and green house at Crop Experiment Station in 1995. The effects of this virus strain on plant characteristics and seed chemical composition of the soybean variety Danyeobkong were evaluated. Yield, the number of pod and 100 seed weight were significantly reduced by SMV-G5H strain infection. Inoculation of SMV-G5H strain of soybean mosaic virus caused the increment of seed protein content and the decrement of seed oil content in the soybean variety Danyeobkong. Changing pattern of fatty acid composition by SMV-G5H strain infection was varied according to the experimental place. In field trial, the percentage of linoleic and linolenic acids decreased and stearic and oleic acids increased in the seed oil from virus infected plants. In green house trial, however, linolenic and oleic acids were increased, whereas palmitic and linoleic acids were reduced.

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