• Journal of Microbiology and Biotechnology
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• v.31 no.8
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• pp.1088-1097
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• 2021

Grouper nervous necrosis virus (GNNV) infection causes mass grouper mortality, leading to substantial economic loss in Taiwan. Traditional methods of controlling GNNV infections involve the challenge of controlling disinfectant doses; low doses are ineffective, whereas high doses may cause environmental damage. Identifying potential methods to safely control GNNV infection to prevent viral outbreaks is essential. We engineered a virus-binding bacterium expressing a myxovirus resistance (Mx) protein on its surface for GNNV removal from phosphate-buffered saline (PBS), thus increasing the survival of grouper fin (GF-1) cells. We fused the grouper Mx protein (which recognizes and binds to the coat protein of GNNV) to the C-terminus of outer membrane lipoprotein A (lpp-Mx) and to the N-terminus of a bacterial autotransporter adhesin (Mx-AIDA); these constructs were expressed on the surfaces of Escherichia coli BL21 (BL21/lpp-Mx and BL21/Mx-AIDA). We examined bacterial surface expression capacity and GNNV binding activity through enzyme-linked immunosorbent assay; we also evaluated the GNNV removal efficacy of the bacteria and viral cytotoxicity after bacterial adsorption treatment. Although both constructs were successfully expressed, only BL21/lpp-Mx exhibited GNNV binding activity; BL21/lpp-Mx cells removed GNNV and protected GF-1 cells from GNNV infection more efficiently. Moreover, salinity affected the GNNV removal efficacy of BL21/lpp-Mx. Thus, our GNNV-binding bacterium is an efficient microparticle for removing GNNV from 10‰ brackish water and for preventing GNNV infection in groupers.

• Research in Plant Disease
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• v.7 no.1
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• pp.1-7
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• 2001

An isolate of Cucumber mosaic cucumovirus(CMV) was isolated from Hydrangea macrophylla for. otaksa(Sieb. et Zucc. ) Wils. showing mosaic symptoms, and designated as Hm-CMV. Hm-CMV was characterized by the tests of host range, physical properties, serological properties, RNA and coat protein compositions, and reverse transcription and polymerase chain reaction (RT-PCR) analysis. Twelve species in 4 families were used in the host range test of Hm-CMV and could be differentiated from Y-CMV used as a control CMV by the ringspot and line pattern on inoculated leaves of several tobacco plants. Thevirus produced local lesions on inoculated leaves of Chenopodium amarticolor, C. quinoa and Vigna unguiculata. The physical properties of the virus were as follows; thermal inactivation point(TIP) was 60$\^{C}$, dilution end point (DEP) was 10$\^$-3/, and longevity in vitro (LIP) was 3∼4 days. Hm-CMV was serologically identical to Y-CMV. SDS-polyaciylamide gel electrophoresis(SDS-PAGE) showed one major protein band of about 28 kDa. In RNA or dsRNA analysis, Hm-CMV consisted of four RNA or dsRNA species, but satellite RNA was not detected. In RT-PCR using CMV-common primer and CMV subgroup I-specific primer, bothe amplified expected size of about 490 bp and 200 bp DNA fragments from Hm-CMV, respectively. Restriction enzyme analysis of the 490 bp RT-PCR products using EcoR I and Msp I showed that Hm-CMV belonged to CMV subgroup I. However, Hm-CMV could be differentiated from other CMV subgroup I isolates by RNA fingerprinting by arbitrarily primed polymerase chain reaction (RAP-PCR).

• Proceedings of the Korean Society of Crop Science Conference
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• 2022.10a
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• pp.316-316
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• 2022

Azuki bean is the important bean crop in Korea, and is traditionally used as a filling material for red bean porridge, rice cakes, and bread, as well as for sediment. So far, the National Insititute of Crop Science has developed azuki bean varieties for various uses, such as for sediment, tea, and azuki bean sprouts. Among azuki bean elite lines, 25 strains were used to analyze the correlation between red bean quality characteristics and sediment yield. The crude protein of red beans was 25.0-28.9%, ash content was 3.8-5.3%, and fat was 0.4-1.0%. As for the appearance quality characteristics of azuki bean, one hundred seed weight was 11.1-19.5 g, the length of seed was 6.45-8.49 mm, the width seed was 4.84-6.45 mm, and the withd/lenght ratio was 0.72-0.89. When the azuki bean were boiled, the thickness of the seed coat was 0.14-0.27 mm and the length ratio of seed was 0.65-0.76 mm, showing that the length of seed was slightly larger than the width. The yield of azuki bean sediment was 239-284% for whole red bean paste, and 144-203% for fine azuki bean paste. As a result of analyzing the correlation between the quality characteristics of azuki beans, such as protein, husk and seed weight, and the yield of azuki bean sediment, the yield of whole azuki bean sediment showed a negative correlation with the seed coat thickness and the length ratio of the original grain at -0.62 and -0.45, respectively. The yield of fine sediment showed a negative correlation with the length ratio of whole azuki beans at -0.49, and a positive correlation with that of white beans at 0.41. However, protein and ash content did not show any correlation with sediment yield. In view of the above results, it is thought that it would be better to have a high seed weight or a low width/length ratio of seed to develope azuki bean cultivar for sediment.

• Proceedings of the Korean Society of Plant Pathology Conference
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• 2003.10a
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• pp.139.2-140
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• 2003

Ornithogalum showing mosaic symptoms were collected from the isolated field of National Plant Quarantine Service in Sengrimmyon of Kyungnam province. Electron microscopic examination of negatively strained preparation was filamentous particle of 740nm in lenght. Indirect-ELISA determined that the virus was serologically related to potyvirus. A single major protein band of Mr 30,000 was observed after sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Indicator plant test showed masaic, necrotic local lesion and sunken areas in leaves of Nicotiana clevelandii and Tetragonia expansa, while the others of indicator plants did not infect. An enzyme-aided purification protocal was used, which eliminated a highly viscous mucilage from extracts of the Omithogalum. Total RNA extracted from infected Omithogalum leaves were amplified of 411b.p fragment in reverse transcription (RT)-PCR when primers specilic for the coat protein gene. An isolate of Omithogalum mosaic virus (OrMV) of the genus Potyvirus was identified as the casual agent of the disease on the basis of electron microscopic, biological and serological reaction.

• Asian-Australasian Journal of Animal Sciences
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• v.26 no.11
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• pp.1518-1522
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• 2013

TYR (Tyrosinase) and TYRP1 (Tyrosinase-related protein 1) play crucial roles in determining the coat color of birds. In this paper, we aimed to characterize the relationship of TYR and TYRP1 genes with plumage colors in Korean quails. The SNPs were searched by cDNA sequencing and PCR-SSCP in three plumage color Korean quails (maroon, white and black plumage). Two SNPs ($367T{\rightarrow}C$ and $1153C{\rightarrow}T$) were found in the coding region of TYRP1 gene, but had no significant association with plumage phenotype in Korean quails. The expression of TYR was higher in black plumage quails than that in maroon plumage quails. In contrast, the expression of TYRP1 was lower in black plumage quails than that in maroon plumage quails. This study suggested that the melanic plumage color in Korean quails may be associated with either increased production of TYR or decreased production of TYRP1.

• KSBB Journal
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• v.26 no.3
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• pp.243-247
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• 2011

Using Bacillus subtilis spore display system, with cotG as an anchoring motif, levansucrase from Zymomonas mobilis, was displayed on the outer surface of Bacillus subtilis spore. Flow cytometry of DB104 (pSDJH-cotG-levU) spore, proved the surface localization of CotG-LevU fusion protein on the spore compared to that of DB104. Enzymatic activity of DB104 (pSDJH-cotG-levU) spore showed more than 1.5 times higher levansucrase specific activity compared to that of the host spore, which is a remarkable increase of enzymatic activity considering the existence of sacA (sucrase) and sacB (levansucrase) in the Bacillus subtilis chromosome. The spore integrity, revealed by sporulation frequency test after heat and lysozyme treatment of spore, did not changed at all in spite of the CotG-LevU fusion protein incorporation into the spore coat layer during spore formation process. These data prove again that Bacillus subtilis spore could be considered as good live immobilization vehicle for efficient bioconversion process.

• BMB Reports
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• v.43 no.4
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• pp.279-283
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• 2010

Polydnavirus genome is segmented and dispersed on host wasp chromosome. After replication, the segments form double- stranded circular DNAs and embedded in viral coat proteins. These viral particles are delivered into a parasitized host along with parasitoid eggs. A serine-rich protein (SRP) is predicted in a polydnavirus, Cotesia plutellae bracovirus (CpBV), genome in its segment no. 33 (CpBV-S33), creating CpBV-SRP1. This study explored its expression and physiological function in the diamondback moth, Plutella xylostella, larvae parasitized by C. plutellae. CpBV-SRP1 encodes 122 amino acids with 26 serines and several predicted phosphorylation sites. It is persistently expressed in all tested tissues of parasitized P. xylostella including hemocyte, fat body, and gut. Its physiological function was analyzed by injecting CpBV-S33 and inducing its expression in nonparasitized P. xylostella by a technique called SERI (segment expression and RNA interference). The expression of CpBV-SRP1 significantly impaired the spreading behavior and total cell count of hemocytes of treated larvae. Subsequent RNA interference of CpBV-SRP1 rescued the immunosuppressive response. This study reports the persistent expression of CpBV-SRP1 in a parasitized host and its parasitic role in suppressing the host immune response by altering hemocyte behavior and survival.

• The Plant Pathology Journal
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• v.24 no.1
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• pp.93-96
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• 2008

In August 2006, a severe disease incidence showing mosaic and/or necrotic symptoms on two bell pepper varieties including red-colored 'Special' and yellow-colored 'Fiesta' was observed in a greenhouse located in Gwangyang, Jeonnam province, Korea. To identify causal viruses, total RNAs were extracted from 11 fruit samples with and without symptoms. Specific oligonucleotide primers for Cucumber mosaic virus (CMV), Pepper mottle virus (PepMoV), Tomato spotted wilt virus (TSWV) and Pepper mild mottle virus (PMMoV) were designed based on the sequences available on GenBank. Database comparisons of the deduced amino acid sequences of each sequence produced 100% and 98% matches with nucleocapsid protein gene of TSWV (Acc. No. ABE11605) and coat protein gene of CMV (Acc. No. DQ018289), respectively, suggesting that the symptoms on bell pepper fruits might be caused by the infection of CMV and TSWV. To our knowledge this is the first report of necrotic as well as mosaic virus disease on bell pepper fruits by the infection of CMV and TSWV in Jeonnam province, Korea.

• The Plant Pathology Journal
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• v.28 no.4
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• pp.381-389
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• 2012

Molecular characterization of Cucumber mosaic virus (CMV) was done by using samples from tomato and cucurbitaceous plants collected from different locations in the northwest region of Iran. After screening by enzyme-linked immunosorbent assay, 91 CMV-infected samples were identified. Biological properties of eight representative isolates were compared with each other revealing two distinct phenotypes on squash and tomato plants. Phylogenetic analyses based on nucleotide sequences of the coat protein (CP), movement protein (MP) and 2b of the new isolates, together with that of previously reported isolates, led to the placement of the Iranian isolates in subgroups IA and IB according to CP and MP genes, but in subgroup IA according to the 2b gene. These data suggest that reassortment may have been a major event in the evolution of CMV in Iran, and that the Iranian isolates are derived from a common recent ancestor that had passed through a bottleneck event.

• The Plant Pathology Journal
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• v.32 no.4
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• pp.371-376
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• 2016

Grapevine Algerian latent virus (GALV) is a member of the genus Tombusvirus in the Tombusviridae and infects not only woody perennial grapevine plant but also herbaceous Nicotiana benthamiana plant. In this study, we developed GALV-based gene expression and virus-induced gene silencing (VIGS) vectors in N. benthamiana. The GALV coat protein deletion vector, pGMG, was applied to express the reporter gene, green fluorescence protein (GFP), but the expression of GFP was not detected due to the necrotic cell death on the infiltrated leaves. The p19 silencing suppressor of GALV was engineered to inactivate its expression and GFP was successfully expressed with unrelated silencing suppressor, HC-Pro, from soybean mosaic virus. The pGMG vector was used to knock down magnesium chelatase (ChlH) gene in N. benthamaina and the silencing phenotype was clearly observed on systemic leaves. Altogether, the GALV-derived vector is expected to be an attractive tool for useful gene expression and VIGS vectors in grapevine as well as N. benthamiana.