• The Plant Pathology Journal
• /
• v.35 no.3
• /
• pp.257-273
• /
• 2019

Tomato (Solanum lycopersicum) is one of the most widely grown and economically important vegetable crops in the world. Tomato chlorosis virus (ToCV) is one of the recently emerged viruses of tomato distributed worldwide. ToCV-tomato interaction was investigated at the molecular level for determining changes in the expression of tomato genes in response to ToCV infection in this study. A cDNA library enriched with genes induced in response to ToCV infection were constructed and 240 cDNAs were sequenced from this library. The macroarray analysis of 108 cDNAs revealed that the expression of 92 non-redundant tomato genes was induced by 1.5-fold or greater in response to ToCV infection. The majority of ToCV-induced genes identified in this study were associated with a variety of cellular functions including transcription, defense and defense signaling, metabolism, energy, transport facilitation, protein synthesis and fate and cellular biogenesis. Twenty ToCV-induced genes from different functional groups were selected and induction of 19 of these genes in response to ToCV infection was validated by RT-qPCR assay. Finally, the expression of 6 selected genes was analyzed in different stages of ToCV infection from 0 to 45 dpi. While the expression of three of these genes was only induced by ToCV infection, others were induced both by ToCV infection and wounding. The result showed that ToCV induced the basic defense response and activated the defense signaling in tomato plants at different stages of the infection. Functions of these defense related genes and their potential roles in disease development and resistance to ToCV are also discussed.

• Journal of Apiculture
• /
• v.32 no.4
• /
• pp.281-293
• /
• 2017

Sacbrood virus (SBV) is one of the main pathogenic RNA viruses of honeybee. SBV is found worldwide and many local strains have been reported, such as kSBV, cSBV, and wSBV. In this study, SBV-specific DNA fragments were cloned and sequenced by reverse-transcription PCR from 4 populations of A. mellifera, 4 sequences from 1 population belonged to the 2134D51 genotype (349 nucleotides, nt) and 12 sequences from 3 populations belonged to the 2100D0 genotype (400 nt) among the 16 determined sequences. A total of 87 points of mismatches were found by comparison with the most similar sequences in GenBank. Seventeen single-nucleotide polymorphisms (SNP) were detected, and 6 SNP-patterns in the 2100D0 genotype and 2 SNP-patterns in the 2134D51 genotype were identified based on SNP positions. In SNP-pattern 2, 10 SNPs were detected, but only 2 SNPs were found in SNP-pattern7. Meanwhile, one SNP-pattern was found from one RNA-sample, multi SNP-patterns were detected from other RNA-samples. Large numbers of SNP variants indicate that vast numbers of point-mutations on SBV have occurred since SBV invaded Korea and that SNP smay have been introduced individually over time. Thorough analysis of SNP variants will not only define the local infection-route, but also the relationships between SNP-pattern and SBV-pathogenic abilities.

• Journal of Microbiology and Biotechnology
• /
• v.31 no.12
• /
• pp.1709-1715
• /
• 2021

Outbreaks of food poisoning due to the consumption of norovirus-contaminated shellfish continue to occur. Male-specific (F+) coliphage has been suggested as an indicator of viral species due to the association with animal and human wastes. Here, we compared two methods, the double agar overlay and the quantitative real-time PCR (RT-PCR)-based method, for evaluating the applicability of F+ coliphage-based detection technique in microbial contamination tracking of shellfish samples. The RT-PCR-based method showed 1.6-39 times higher coliphage PFU values from spiked shellfish samples, in relation to the double agar overlay method. These differences indicated that the RT-PCR-based technique can detect both intact viruses and non-particle-protected viral DNA/RNA, suggesting that the RT-PCR based method could be a more efficient tool for tracking microbial contamination in shellfish. However, the virome information on F+ coliphage-contaminated oyster samples revealed that the high specificity of the RT-PCR- based method has a limitation in microbial contamination tracking due to the genomic diversity of F+ coliphages. Further research on the development of appropriate primer sets for microbial contamination tracking is therefore necessary. This study provides preliminary insight that should be examined in the search for suitable microbial contamination tracking methods to control the sanitation of shellfish and related seawater.

• The Plant Pathology Journal
• /
• v.39 no.3
• /
• pp.255-264
• /
• 2023

Sweet potato symptomless virus 1 (SPSMV-1) is a single-stranded circular DNA virus, belonging to the genus Mastrevirus (family Geminiviridae) that was first identified on sweet potato plants in South Korea in 2012. Although SPSMV-1 does not induce distinct symptoms in sweet potato plants, its co-infection with different sweet potato viruses is highly prevalent, and thus threatens sweet potato production in South Korea. In this study, the complete genome sequence of a Korean isolate of SPSMV-1 was obtained by Sanger sequencing of polymerase chain reaction (PCR) amplicons from sweet potato plants collected in the field (Suwon). An infectious clone of SPSMV-1 (1.1-mer) was constructed, cloned into the plant expression vector pCAMBIA1303, and agro-inoculated into Nicotiana benthamiana using three Agrobacterium tumefaciens strains (GV3101, LBA4404, and EHA105). Although no visual differences were observed between the mock and infected groups, SPSMV-1 accumulation was detected in the roots, stems, and newly produced leaves through PCR. The A. tumefaciens strain LBA4404 was the most effective at transferring the SPSMV-1 genome to N. benthamiana. We confirmed the viral replication in N. benthamiana samples through strand-specific amplification using virion-sense- and complementary-sense-specific primer sets.

• Korean Journal of Veterinary Service
• /
• v.46 no.2
• /
• pp.115-122
• /
• 2023

This study was performed to investigate the distribution of mosquito vectors related to the zoonotic disease in Daejeon. Samples were taken using a blacklight trap once a month from March to November 2021 at the slaughterhouse in Daejeon. A total of 820 mosquitoes were captured and classified into 5 genera and 8 species. Among the collected mosquitoes, 319 (38.9%) and 295 (35.93%) were Aedes vexans nipponii and Culex pipiens pallens, respectively, making them the dominant species. The overall number of mosquitoes collected started to increase from May and reached the largest value of 329 (40.12%) in June. Trapped mosquitoes are created 72 pools by environmental condition and by species. The pools were tested by PCR methods for 7 zoonotic pathogens. Flavivirus-positive products were confirmed by DNA sequencing. Japanese encephalitis viruses were detected in 3 pools collected from cow lairage (Culex pipiens pallens) in May, cow by-product processing room (Aedes vexans nipponii) in June and cow lairage (Mansonia uniformis) in June. Culex flavivirus were detected in 4 pools. Based on the results of this study, it is considered that continous surveillence of mosquitoes in livestock assembly facilities (slaughterhouse) should be performed for controlling mosquito populations and mediating disease spread by mosquitoes.

• Korean journal of applied entomology
• /
• v.58 no.2
• /
• pp.89-99
• /
• 2019

In total, 654,362 adult mosquitoes were captured using black light traps in Gangwon-do Province of the Republic of Korea from 2012 to 2017. The collected mosquitoes were identified to the species level, placed in pools of up to 50 mosquitoes each, by species and date of collection, and screened for flaviviruses using a reverse transcription-polymerase chain reaction assay. A total of 276,224 adult mosquitoes were grouped in 7,721 pools for virus testing, and 68 flavivirus positive pools (0.9%) were detected. Flavivirus-positive products were confirmed by DNA sequencing. Japanese encephalitis viruses were detected in single pools collected from Chuncheon (2012, 2017: Culex pipiens, 2,728 and 1,111 mosquitoes, respectively), Hoengseong (2013: Culex orientalis, 19), and Gangneung (2017: C. pipiens, 724). All the Japanese encephalitis viruses detected were revealed as genotype V. Chaoyang viruses were detected in 63 pools of 5,055 Aedes vexans nipponii and a single pool of 585 C. pipiens collected in Gangwon-do Province from 2012 to 2017. Chuncheon was the region with the highest minimum infection rates (MIR, 0.32) and maximum likehood estimate (MLE, 0.33; confidence interval (CI) 95%, 0.23-0.46) of A. vexans nipponii for Chaoyang virus, followed by Hoengseong (MIR 0.30, MLE 0.30, CI 0.16-0.52) and Gangneung (MIR 0.21, MLE 0.21, CI 0.13-0.31). Monthly MIR and MLE values of A. vexans nipponii for Chaoyang virus were the highest in October (MIR 0.38, MLE 0.38, CI 0.07-1.25).

• Korean Journal of Clinical Laboratory Science
• /
• v.49 no.4
• /
• pp.446-454
• /
• 2017

Human papillomaviruses (HPVs) are major causes of cervical cancer. Sixteen high risk HPVs, including HPV 16, HPV 18, HPV31, HPV 33, HPV 35, HPV 39, HPV 45, HPV 51, HPV 52, HPV 53, HPV 56, HPV 58, HPV 59, HPV 66, HPV 68, and HPV 69 are found in cervical cancer. HPVs 16 and 18 are mainly presented in 70% of cervical cancer. Therefore, identifying the presence of these high-risk HPVs is crucial. The objective of this study is to establish the HPV ViroCheck for detecting 16 HR-HPVs and genotypes of HPVs 16 and 18, as well as to analyze the analytical performance of HPV ViroCheck. We performed the analytical sensitivity of HPV E6 / E7 genes of 16 high risk HPVs to confirm the limit of detection. Then, a cross reactivity of HPV ViroCheck with microorganisms and viruses related to the cervix were analyzed for analytical specificity. Analytical sensitivity of high risk HPV genotypes ranged from 1 to 100 copies when using cloned DNAs. The limit of detection was 10 cells for both SiHa and HeLa cells. Cervical-related microorganisms and viruses did not show cross-reactivity to HPV DNA. Moreover, the intra- and inter-assay coefficient variations (CVs) were below 5%. In conclusion, HPV Virocheck will be useful for the detection of 16 HR HPVs, as well as HPV 16 and HPV 18 genotypes rapidly.

• Korean journal of applied entomology
• /
• v.56 no.1
• /
• pp.19-28
• /
• 2017

Polydnaviruses (PDVs) are a group of double-stranded DNA viruses symbiotic to some endoparasitoid wasps. Cotesia plutellae bracovirus (CpBV) is a PDV symbiotic to an endoparasitoid wasp, C. plutellae, parasitizing young larvae of Plutella xylostella. An early expressed gene, CpBV-ELP1, plays an important role in the parasitism by suppressing host cellular immunity by its cytotoxic activity against hemocytes. This study aimed to test its oral toxicity against insect pest by expressing it in a recombinant tobacco plant. A recombinant CpBV-ELP1 protein was produced using a baculovirus expression system and secreted to cell culture medium. The cell cultured media were used to purify CpBV-ELP1 by a sequential array of purification steps: ammonium sulfate fractionation, size exclusion chromatography, and ion exchange chromatography. Purified rCpBV-ELP1 exhibited a significant cytotoxicity against Spodoptera exigua hemocytes. CpBV-ELP1 was highly toxic to the fifth instar larvae of S. exigua by injection to hemocoel. It also showed a significant oral toxicity to fifth instar larvae of S. exigua by a leaf-dipping assay. CpBV-ELP1 was cloned into pBI121 vector under CaMV 35S promoter with opaline synthase terminator. Resulting recombinant vector (pBI121-ELP1) was used to transform Agrobacterium tumefaciens LBA4404. The recombinant bacteria were then used to induce callus of a tobacco (Nicotiana tabacum Xanthi) leaves and subsequent generation (T1) plants were selected. T1 generation tobacco plants expressing CpBV-ELP1 gave significant insecticidal activities against S. exigua larvae. These results suggest that CpBV-ELP1 gene can be used to control insect pests by constructing transgenic crops.

• Journal of Life Science
• /
• v.21 no.8
• /
• pp.1156-1162
• /
• 2011

Equine coital exanthema caused by equine herpesvirus type 3 (EHV-3) is a venereal disease which seriously drops horse reproduction rates. Here, we isolated EHV-3 from infected horses and investigated their biological characteristics. Initial cytopathic effects such as rounding of cells were detected 48 hours post infection of the virus into RK-13 cells. The infected cells were going to detach from the surface of culture flasks 72 hours post infection. The type of isolated viruses from swabbed samples was EHV-3 by PCR analysis. Glycoprotein G (gG) of isolated EHV-3 has a 99.25 percent similarity rate to that of EHV-3 334/74 control strain. The isolated EHV-3 was named Georo strain. Georo strain consisted of four major proteins including 145 kD, 60 kD, 45 kD and 40 kD, as shown by SDS-PAGE analysis. We hope the newly isolated Georo strain of EHV-3 can be used for studying various aspects of Korean equine coital exanthema.

• Asian Pacific Journal of Cancer Prevention
• /
• v.14 no.9
• /
• pp.5287-5291
• /
• 2013

The human papilloma virus (HPV) causes skin and mucous membrane infections. It crosses from one person to another by skin-to-skin contact, such as sexual contact. There are more than 100 types of HPV that can influence different parts of the body. Some types of HPV can cause cancer (such as cervical or anal cancer) and others can cause warts (such as genital or plantar warts). HPV infection is one of the most common sexually transmitted infections (STIs) in Iran and around the world. Considerable molecular evidence suggests a role for human papilloma virus (HPV) in the pathogenesis of carcinoma. Epidemiological studies on human papilloma viruses (HPVs) infections in general population are critical for the performing of health policy guidelines for developing the strategies to hinder the primary and secondary different cancer. In different parts of Iran, there is a lack of population-based studies to determine the prevalence of HPV in the general population. The aim of this population-based study was therefore to report the prevalence ratse of HPV types among Iranian patients. To study the risk of human papilloma virus (HPV) infection, we managed a retrospective study in Kerman province, southeast of Iran. For this purpose, 410 patients tested for the presence of HPV DNA using PCR and INNo-Lipa assays. HPV DNA was detected in 108 out of 410 patients (26.34%), while it was not detected in any of the control group samples. Patients included 23 (21.1%) males and 86 (78.8%) females. HPV type 6 was the most common (49%) followed by HPV type 16 (10.1%), and also HPV type11 (9.2%). The prevalence of HPV in Iran is comparable to those reported in other regions of the world. In a similar manner, it seems that HPV types 6, 16 and11 are the most common types in Kerman. Additional studies on larger group of patients, particularly in those with pre-invasive forms of disease, are needed to explain the roles of different HPV types in this location of Iran.