• Animal cells and systems
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• 제3권1호
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• pp.89-96
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• 1999

Nucleotide sequences of a 501 base-pair (bp) fragment in the mitochondrial cytochrome b (cyt b) gene were analyzed for 12 populations of Rana rugosa from Korea and Japan using a polymerase chain reaction (PCR) and direct silver sequencing. Two genetically distinct groups (type-A and type-B) were found in Korea. Type-A was found throughout most of South Korea and type-B was restricted to the mid-southeastern regions (Samchok, Yongdok, Chongsong and Pohang). But in the Tonghae population, both types were found. The level of mitochondrial DNA (mtDNA) sequence differences ranged from 0% to .0.8% among six populations of type-A, and 0 to 1.0% among 4 populations of type-B. However, sequence differences between type-A and type-B ranged from 5.4% to 6.6%, Using Kimura's two-parameter distance, the level of genetic sequence divergence between type-A and type-B was 6.7%. The Japanese R. rugosa was clustered very far from the Korean R. rugosa with 14.7%. In the neighbor-joining and UPGMA tree, all Korean samples were grouped, but subdivided into two types in 99% of the bootstrap iteration.

• 한국동물위생학회지
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• 제32권1호
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• pp.11-18
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• 2009

Porcine epidemic diarrhea virus (PEDV), an enveloped single stranded RNA virus in the family Coronaviridae, causes acute viral enteric disease in piglets. Recently outbreaks of porcine epidemic diarrhea (PED) have been rare in Europe but frequent in Asia. In Korea, the increase of PED prevalence is showing specially in postweaning pigs. The purpose of this study was to investigate nucleotide sequence of nucleocapsid protein gene of PEDV field isolates from postweaning pigs in Korea and get more information about the viruses. A total of 15 postweaing pigs clinically suspected of PEDV infection by severe watery diarrhea and dehydration were used in this study. Viral RNA was extracted from small intestines and stools of the pigs. The N gene was amplified by nested RT-PCR, purificated, sequenced, analyzed and then compared with published sequences of other PEDV strains. Three PEDVs were isolated from the suspected postweaning pigs. The N gene of three PEDV field isolates consisted of 483 nucleotides. These PEDV field isolates showed nucleotide sequence homology range from 99.6% to 95% with Chinese strains, from 99.8% to 95.2% with Korean strains, from 97.3% to 95.7% with Japanese strains and from 96.5% to 95.7% with Belgium and British strains. The encoded pritein shared range from 98.8% to 95.6% with Chinese strains, from 99.4% to 95% with Korean strains, from 97.5% to 96.3% with Japanese strains, from 95.6% to 95% with Belgium and British strains. By phylogenetic tree analysis based on nucleotide sequence, three PEDV field isolates were clustered into two groups which were Chinese isolate groups and other Korean isolate groups. These results indicated that some of PEDV field isolates prevailing in Korean postweaning pigs may be associated with those of Chinese strains and other Korean strains.

• 한국동물위생학회지
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• 제39권4호
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• pp.259-266
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• 2016

Porcine epidemic diarrhea virus (PEDV) causes an acute and lethal watery diarrhea in piglets that is great economic losses to the swine country worldwide. The spike (S) glycoprotein is an important determinant for PEDV biological properties. In the present study, we determined the full-length S gene sequences of five Chung-nam PEDV field isolates collected in 2016. The S gene was amplified by RT-PCR, purificated, sequenced, analyzed and then compared with published sequences of other PEDV strains. 5 field strains share 98.5%~99.9% homologies with each other at the nucleotide sequence level and 96.7%~99.9% homologies with each other at the amino acids sequence level. Most field strains have nucleotide insertions, deletions and mutation regions, and show lower homologies (93.1~93.8%) with classical and vaccine strains, however higher homologies (99.1%~99.5%) with US PEDV isolates in 2013. By phylogenetic tree analysis based on nucleotide sequence, five PEDV field isolates were clustered into Genogroup 2b but differ genetically from the vaccine strains (SM-98 and DR-13).

• The Plant Pathology Journal
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• 제35권1호
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• pp.41-50
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• 2019

Sugarcane bacilliform viruses (SCBV), which belong to the genus Badnavirus, family Caulimoviridae, are an important DNA virus complex that infects sugarcane. To explore the genetic diversity of the sugarcane-infecting badnavirus complex in China, we tested 392 sugarcane leaf samples collected from Fujian, Yunnan, and Hainan provinces for the occurrence of SCBV by polymerase chain reaction (PCR) assays using published primers SCBV-F and SCBV-R that target the reverse transcriptase/ribonuclease H (RT/RNase H) regions of the viral genome. A total of 111 PCR-amplified fragments (726 bp) from 63 SCBV-positive samples were cloned and sequenced. A neighbor-joining phylogenetic tree was constructed based on the SCBV sequences from this study and 34 published sequences representing 18 different phylogroups or genotypes (SCBV-A to -R). All SCBV-tested isolates could be classified into 20 SCBV phylogenetic groups from SCBV-A to -T. Of nine SCBV phylogroups reported in this study, two novel phylogroups, SCBV-S and SCBV-T, that share 90.0-93.2% sequence identity and show 0.07-0.11 genetic distance with each other in the RT/RNase H region, are proposed. SCBV-S had 57.6-92.2% sequence identity and 0.09-0.66 genetic distance, while SCBV-T had 58.4-90.0% sequence identity and 0.11-0.63 genetic distance compared with the published SCBV phylogroups. Additionally, two other Badnavirus species, Sugarcane bacilliform MO virus (SCBMOV) and Sugarcane bacilliform IM virus (SCBIMV), which originally clustered in phylogenetic groups SCBV-E and SCBV-F, respectively, are first reported in China. Our findings will help to understand the level of genetic heterogeneity present in the complex of Badnavirus species that infect sugarcane.

• 한국균학회지
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• 제47권1호
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• pp.89-94
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• 2019

중국에서 범부채의 녹병균이 Puccinia iridis로 동정됨에 따라 우리나라에서도 범부채의 녹병균을 재검토하였다. 저자들이 채집한 2점의 시료를 형태적으로 검토한 결과 모두 P. iridis의 특징과 일치하였다. 또한 유전분석한 결과 ITS 및 LSU rDNA 영역의 염기서열이 기존에 기록된 P. iridis와 각각 100% 및 99%의 상동성을 나타냈다. 이를 Neighbor-joining 분석법으로 계통수를 작성하였을 때도 P. iridis 계통군에 속하였다. 따라서 우리나라에서 범부채의 녹병균으로 P. iridis의 존재가 확인되었다. 한편, 우리나라에서 2003년에 범부채의 녹병균으로 기록된 Puccinia belamcandae에 대한 재검토는 향후 숙제로 남게 되었다.

• 한국미생물·생명공학회지
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• 제47권1호
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• pp.25-33
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• 2019

Lactobacillus rhamnosus BFE5264, isolated from a Maasai fermented milk product ("kule naoto"), was previously shown to exhibit bile acid resistance, cholesterol assimilation, and adhesion to HT29-MTX cells in vitro. In this study, we re-annotated and analyzed the previously reported complete genome sequence of strain BFE5264. The genome consists of a circular chromosome of 3,086,152 bp and a putative plasmid, which is the largest one identified among L. rhamnosus strains. Among the 2,883 predicted protein-coding genes, those with carbohydrate-related functions were the most abundant. Genome analysis of strain BFE5264 revealed two consecutive CRISPR regions and no known virulence factors or antimicrobial resistance genes. In addition, previously known highly variable regions in the genomes of L. rhamnosus strains were also evident in strain BFE5264. Pairwise comparison with the most studied probiotic strain L. rhamnosus GG revealed strain BFE5264-specific deletions, probably due to insertion sequence-mediated recombination. The latter was associated with loss of the spaCBA pilin gene cluster and exopolysaccharide biosynthetic genes. Comparative genomic analysis of the sequences from all available L. rhamnosus strains revealed that they were clustered into two groups, being within the same species boundary based on the average nucleotide identities. Strain BFE5264 had a sister group relationship with the group that contained strain GG, but neither ANI-based hierarchical clustering nor core-gene-based phylogenetic tree construction showed a clear distinctive pattern associated with the isolation source, implying that the genotype alone cannot account for their ecological niches. These results provide insights into the probiotic mechanisms of strain BFE5264 at the genomic level.

• The Plant Pathology Journal
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• 제37권2호
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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.

• 한국자원식물학회지
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• 제35권1호
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• pp.100-108
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• 2022

한반도 미기록 외래식물인 서양쐐기풀(쐐기풀과)를 한강 수계에서 발견하였다. 서양쐐기풀은 유럽, 온대 아시아, 서북아프리카 원산으로 알려져 있으며, 세계의 여러 지역에 도입되어 있는데 특히, 동아시아(일본), 오세아니아, 남아메리카에서 외래종으로 보고되었다. 본 분류군은 단성화이며, 자웅이주를 가지는 특징에 의해 근연분류군과 구분된다. Neighbor-Joining 계통수에서 국내 난지한강공원과 양재천에서 채집된 서양쐐기풀은 U. dioica s.s.군으로 유집되었다. 주요 형질에 대한 기재문과 화상자료 및 검색표를 제시하였다.

• Journal of Veterinary Science
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• 제23권5호
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• pp.73.1-73.8
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• 2022

Background: Lumpy skin disease (LSD), a disease transmitted by direct and indirect contact with infected cattle, is caused by the Lumpy skin disease virus (LSDV). The disease affects cattle herds in Africa, Europe, and Asia. The clinical signs of LSD range from mild to the appearance of nodules and lesions in the skin leading to severe symptoms that are sometimes fatal with significant livestock economic losses. Objectives: This study aimed to characterize LSDV strains in the blood of infected cattle in Thailand based on the GPCR gene and determine the phylogenetic relationship of LSDV Thailand isolates with published sequences available in the database. Methods: In total, the blood samples of 120 cattle were collected from different farms in four provinces in the northeastern part of Thailand, and the occurrence of LSDV was examined by PCR based on the P32 antigen gene. The genetic diversity of LSDV based on the GPCR gene was analyzed. Results: Polymerase chain reaction assays based on the P32 antigen gene showed that 4.17% (5/120) were positive for LSDV. All positive blood samples were amplified successfully for the GPCR gene. Phylogenetic analysis showed that LSDV Thailand isolates clustered together with LSDVs from China and Russia. Conclusions: The LSD outbreak in Thailand was confirmed, and a phylogenetic tree was constructed to infer the branching pattern of the GPCR gene from the presence of LSDV in Thailand. This is the first report on the molecular characterization of LSDV in cattle in Thailand.

• 식물병연구
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• 제28권2호
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• pp.98-107
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• 2022

The incidence of Tomato brown rugose fruit virus (ToBRFV) and biological and molecular characterization of the Palestinian isolates of ToBRFV are described in this study. Symptomatic leaf samples obtained from Solanum lycopersicum L. (tomatoes) and Nicotiana tabacum L. (cultivated tobacco) plants were tested for tobamoviruses infection by reverse transcription polymerase chain reaction. Tomato leaf samples collected from Tulkarm and Qalqilia are infected with ToBRFV-PAL with an infection rate of 76% and 72.5%, respectively. Leaf samples collected from Jenin and Nablus were found to be mixed infected with ToBRFV-PAL and Tobacco mosaic virus (TMV) (100%). Sequence analysis of the ToBRFV-PAL genome showed that the net average nucleotide divergence between ToBRFV/F48-PAL strain and the Israeli and Turkish strains was 0.0026398±0.0006638 (±standard error of mean), while it was 0.0033066±0.0007433 between ToBRFV/F42-PAL and these two isolates. In the phylogenetic tree constructed with the complete genomic sequence, all the ToBRFV isolates were clustered together and formed a sister branch with the TMV. The sequenced Palestinian isolates of ToBRFV-PAL shared the highest nucleotide identity with the Israeli ToBRFV isolate suggesting that the virus was introduced to Palestine from Israel. The findings of this study enhance our understanding of the biological and molecular characteristics of ToBRFV which would help in the management of the disease.