• The Plant Pathology Journal
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• 제19권3호
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• pp.166-170
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• 2003

Ninety samples showing mosaic symptoms on soybean (Glycine max) cv. Sukryangputkong were collected from the Cheongsongkun area, Kyungbuk province in Korea. Initially, DAS-ELISA was conducted far detection of Soybean mosaic virus (SMV). Negative samples were chosen at random and mechanically inoculated on soybean cv. Buffalo, which reported not to produce mosaic symptoms when mechanically inoculated with SMV. An isolate of SMV, designated as B-1, from Buffalo showing mosaic and mottle symptoms was used for identification and biological characterization of the causal vim. The purified B-1 isolate had spherical particles of approximately 24nm. It positively reacted with the antiserum against Cowpea mosaic virus (CPMV) but not with Cucumber mosaic virus (CMV) and SMV antisera. CPMV was newly isolated from soybean and had been characterized by host range and by serological and electron microscopic methods. Results of this study suggest that CPMV is the possible cause of mosaic disease in vegetable soybean and that based on sympto-matology, a difference between the typical mosaic and rugose symptoms caused by SMV and CPMV was observed. This is first report of CPMV from soybean in Korea.

• The Plant Pathology Journal
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• 제24권3호
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• pp.328-336
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• 2008

Out of various fungi and bacteria tested for inhibition of Tobacco mosaic virus(TMV) infection using Nicotiana tabacum cv. Xanthi-nc, a bacterial isolate JB-l, identified as Pseudomonas putida had a strong direct inhibitory activity against the TMV infection. Its systemic or indirect activity was also noted at more than a half level of the direct control efficacy. Disease severity was reduced significantly in the susceptible tobacco N. tabacum cv. NC 82 by the treatment of the bacterial culture filtrate, somewhat more by the pretreatment than by simultaneous treatment, probably by inhibiting the TMV transmission and translocation in the plants, showing negative serological, which responses in the viral detection by DAS-ELISA. TMV-inhibitory substances from P. putida JB-1 were water-soluble, stable to high temperature(even boiling), and to a wide range of pH. As proteinase K nullified their antiviral activity, the TMV inhibition activity of P. putida may be derived from proteinaceous materials. In electron microscopy, TMV particles treated with the JB-1 culture were shown to be shrunken with granule-like particles attached on them. All of these aspects suggest that P. putida JB-1 may be developed as a potential agent for the control of TMV.

• 한국동물위생학회지
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• 제33권1호
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• pp.1-6
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• 2010

Monoclonal antibody (mAb)-based enzyme immune-slide assay (EISA) was used for the detection of Peste des petits ruminants (PPR) virus from field samples collected from a natural outbreak. The clinicopathological study was undertaken to diagnose the case primarily of PPR. Antigen was detected from discharges and faeces of infected goats and swabs of postmortem lesions prepared on glass slide or glass plate using acetone fixation. Nasal discharge collected at the early stage of disease course or lung is an appropriate ante- or postmortem sample for this technique, respectively. Convalescent polyclonal sera collected from recovered animals which were diagnosed as PPR by EISA showed high antibody titer against PPR by C-ELISA, demonstrating the satisfactory specificity of the test. Therefore, EISA is a sensitive and specific assay to confirm PPR infection both in field and laboratory conditions and especially suitable for developing country.

• 한국가금학회지
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• 제37권2호
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• pp.167-172
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• 2010

닭 전염성 F낭병 (IBD)은 닭에서 전염성이 강하고 발병으로 인하여 양계 산업에 막대한 경제적 피해를 입히는 닭의 바이러스성 전염병이다. 이 연구에서는 수 분 이내에 검사 시료로부터 닭 전염성 F낭병 바이러스를 검출할 수 있는 시판용 면역크로마토그래피법 검사 킷트를 이용하여 IBD 진단에 있어서의 유용성을 조사하였다. 사용한 면역크로마토그래피법 검사 킷트는 닭 전염성 F낭병 바이러스 VP2에 특이적인 단클론 항체를 이용하여 닭 전염성 F낭병 바이러스를 검출하도록 고안되었다. 바이러스 감염 역가를 알고 있는 IBDV를 사용하여 조사한 결과, IC 검사 킷트의 검출 한계는 $10^{3.1}$ 내지 $10^{3.9}$ $EID_{50}$/mL이었다. 이 검사 킷트는 닭의 다른 전염성 바이러스인 뉴캣슬병 바이러스, 닭 전염성 기관지염 바이러스, 조류인플루엔자 바이러스 및 전염성 후두기관염 바이러스에 대하여 비특이 반응을 나타내지 않았다. 고병원성의 IBDV를 실험적으로 감염시킨 후 3일 내지 4일에 폐사한 닭의 장기별로 조사한 결과, 모든 폐사 닭의 F낭, 장편도, 비장, 신장 시료들은 면역크로마토그래피법 검사 킷트에서 강한 양성반응을 나타내었다. 검사 시료 중 F낭 시료가 IC 검사 킷트에서 가장 강한 양성반응을 나타내었다. 폐사 닭의 간, 흉선, 선위의 경우, 각각 검사시료의 87.5%, 37.5% and 0%가 양성 반응을 나타내었다. 면역크로마토그래피법 검사 킷트에서 음성이었던 시료 중 흉선 시료 한 점을 제외한 모든 시료는 DAS-ELISA와 동일한 검사 결과를 나타내었으나, 검사 시료 중 흉선과 선위 일부에서 RT-PCR 검사에서 양성 반응을 나타내었다. 야외 IBD 발생 농장과 비발생 농장에서 수거한 폐사닭 231수의 조직을 면봉으로 도말하여 채취한 시료를 조사하여 RT-PCR법과 비교한 결과, 상대적 민감도와 특이도는 각각 100% (109/109) 및 97.5% (119/122)를 나타났으며, 두 검사 방법간 kappa value는 0.97이었다. 우리의 연구 결과는 IC 검사 킷트는 야외 양계 농장에서 폐사 닭을 대상으로 IBD를 진단하는 데 적용하기에 매우 유용하다는 것을 말해준다.

• 대한약침학회지
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• 제19권1호
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• pp.37-44
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• 2016

Objectives: This study examined the relative efficacies of a derivative of betulinic acid (dBA) and its poly (lactide-co-glycolide) (PLGA) nano-encapsulated form in A549 lung cancer cells in vivo and in co-mutagen [sodium arsenite (SA) + benzo[a]pyrene (BaP)]-induced lung cancer in mice in vivo. Methods: dBA was loaded with PLGA nanoparticles by using the standard solvent displacement method. The sizes and morphologies of nano-dBA (NdBA) were determined by using transmission electron microscopy (TEM), and their intracellular localization was verified by using confocal microscopy. The binding and interaction of NdBA with calf thymus deoxyribonucleic acid (CT-DNA) as a target were analyzed by using conventional circular dichroism (CD) and melting temperature (Tm) profile data. Apoptotic signalling cascades in vitro and in vivo were studied by using an enzyme-linked immunosorbent assay (ELISA); the ability of NdBA to cross the blood-brain barrier (BBB) was also examined. The stage of cell cycle arrest was confirmed by using a fluorescence-activated cell-sorting (FACS) data analysis. Results: The average size of the nanoparticles was ~ 110 nm. Confocal microscopy images confirmed the presence of NdBA in the cellular cytoplasm. The bio-physical properties of dBA and NdBA ascertained from the CD and the Tm profiles revealed that NdBA had greater interaction with the target DNA than dBA did. Both dBA and NdBA arrested cell proliferation at G0/G1, NdBA showing the greater effect. NdBA also induced a greater degree of cytotoxicity in A549 cells, but it had an insignificant cytotoxic effect in normal L6 cells. The results of flow cytometric, cytogenetial and histopathological studies in mice revealed that NdBA caused less nuclear condensation and DNA damage than dBA did. TEM images showed the presence of NdBA in brain samples of NdBA fed mice, indicating its ability to cross the BBB. Conclusion: Thus, compared to dBA, NdBA appears to have greater chemoprotective potential against lung cancer.

• The Plant Pathology Journal
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• 제33권1호
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• pp.43-52
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• 2017

A survey was conducted to determine the status of Lucerne transient streak virus (LTSV) in three high-yielding alfalfa regions in central Saudi Arabia (Riyadh, Qassim, and Hail) during 2014. Three hundred and eight symptomatic alfalfa, and seven Sonchus oleraceus samples were collected. DAS-ELISA indicated that 59 of these samples were positive to LTSV. Two isolates of LTSV from each region were selected for molecular studies. RT-PCR confirmed the presence of LTSV in the selected samples using a specific primer pair. Percentage identity and homology tree comparisons revealed that all Saudi isolates were more closely related to each other but also closely related to the Canadian isolate-JQ782213 (97.1-97.6%) and the New Zealand isolate-U31286 (95.8-97.1%). Comparing Saudi isolates of LTSV with ten other sobemoviruses based on the coat protein gene sequences confirmed the distant relationship between them. Eleven out of fourteen plant species used in host range study were positive to LTSV. This is the first time to document that Trifolium alexandrinum, Nicotiana occidentalis, Chenopodium glaucum, and Lathyrus sativus are new host plant species for LTSV and that N. occidentalis being a good propagative host for it.

• Food Science and Biotechnology
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• 제17권5호
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• pp.1092-1096
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• 2008

Commercial food products from major cities of Coahuila, Mexico were screened to identify residues of transgenic deoxyribonucleic acid (DNA) and/or proteins. After performed, an inventory on all products that contained a soybean-based ingredient in a commercial grocery store in the city of Saltillo, Coahuila, Mexico, 245 food products were identified and grouped in 15 classes according to the soybean ingredient as well as the manufacturing process used for their elaboration. Similar sampling was made for the different food classes in the cities of Monclova, Piedras Negras, and Torreon. A total of 88 samples were analyzed and DNA was extracted by the hexadecyltrimethyl-ammonium bromide (CTAB) technique with slight modification to obtain better DNA quality (1). In addition, segments of the transgenic genes one that codifies for 5-enolpyruvylshikimate-3-phosphate synthase (epsps), cry 1A, and the cauliflower mosaic virus (CaMV) promoter were amplified using polymerase chain reaction (PCR). The transgenic proteins 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) and insecticidal crystal protein (Cry 1Ab/Ac) were identified using double antibody sandwich-enzymatic linked immunoassay analysis (DAS-ELISA). Presence of transgenic genes and/or proteins was identified in 35.3% of the commercial products samples.

• The Plant Pathology Journal
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• 제33권5호
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• pp.508-513
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• 2017

The complete genome sequence of a Slovak SL-1 isolate of Tomato mosaic virus (ToMV) was determined from the next generation sequencing (NGS) data, further confirming a limited sequence divergence in this tobamovirus species. Tomato genotypes Monalbo, Mobaci and Moperou, respectively carrying the susceptible tm-2 allele or the Tm-1 and Tm-2 resistant alleles, were tested for their susceptibility to ToMV SL-1. Although the three tomato genotypes accumulated ToMV SL-1 to similar amounts as judged by semiquantitative DAS-ELISA, they showed variations in the rate of infection and symptomatology. Possible differences in the intra-isolate variability and polymorphism between viral populations propagating in these tomato genotypes were evaluated by analysis of the capsid protein (CP) encoding region. Irrespective of genotype infected, the intra-isolate haplotype structure showed the presence of the same highly dominant CP sequence and the low level of population diversity (0.08-0.19%). Our results suggest that ToMV CP encoding sequence is relatively stable in the viral population during its replication in vivo and provides further demonstration that RNA viruses may show high sequence stability, probably as a result of purifying selection.

• The Plant Pathology Journal
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• 제25권4호
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• pp.369-375
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• 2009

The potential antiviral effects of the culture filtrates (CF) from Serratia marcescens strain Gsm01 against yellow strain of Cucumber mosaic virus (CMV-Y) were investigated. The culture filtrate of S. marcescens strain Gsm01 applied on Chenopodium amaranticolor showed high inhibitory activity, likewise no necrosis appeared when applied on the tobacco plants 2 days before CMV-Y inoculation. When plants were challenge inoculated with CMV-Y for eighteen days, the disease incidence in plants with culture filtrate of S. marcescens Gsm01 did not exceed 59%, whereas 100% of control plants were severely infected. The results of double antibody sandwich-enzyme linked immunosorbent assay (DAS-ELISA), reverse transcriptase polymerase chain reaction (RT-PCR), dot blotting, and western blotting showed that culture filtrate treatment highly affected the accumulation of CMV-Y or its CP protein gene in the treated plant leaves. It was also observed that the culture filtrate had no RNase activity on genomic RNAs of CMV-Y, suggesting that culture filtrate may not contain ribosome inactivating proteins (RIPs) or proteins with RNase activity. These data shows that culture filtrate of S. marcescens strain Gsm01 seems to be a promising source of antiviral substance for the practical use.

• Journal of Plant Biotechnology
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• 제6권4호
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• pp.221-227
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• 2004

Protocol was established for production of virus free healthy seeds using meristem ($0.3-0.5\;\cal{mm}$ in size) culture and field management under net house condition in tomato. The isolated meristem was found well established in MS liquid medium containing $0.1\;\cal{mg}\;1^{-1}\;of\;GA_3$. For shoot and root development either from primary meristem or from nodal segment of meristem derived plants, semisolid MS medium having $0.5\;\cal{mg}\;1^{-1}$ of IBA was found most effective. The elimination of the studied viruses (ToMV, CMV, ToLCV) in meristem-derived plants was confirmed by DAS-ELISA test. For field management of the virus eradicated meristem-derived plants, use of net house was found very effective measures to check viral vector visit and eventually infection. The meristem-derived plants were vigor and high yielder than the native seed derived plants and produced healthy seeds. Due to stop vector visit, no viral symptoms were observed in both $R_1\;and\;R_2$ plants cultivated in net house condition. Starting of viral infestation was observed in $R_2$ generation when they were planted in open house condition without control of vector visit. Therefore, for management of viral diseases, use of virus free meristem derived plantlets and their subsequent cultivation in soil under net house condition without using any vector killing insecticide can be recommended for producing healthy seeds in tomato. The developed protocol for environmentally healthy tomato seed production in Bangladesh may be used in the countries having similar tropical like environment conducive for viral vector visit.