• Title/Summary/Keyword: Molecular phylogenetic tree

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First Report of Charcoal Rot Caused by Macrophomina phaseolina on Peanut Plants in Korea (땅콩에서 Macrophomina phaseolina에 의한 균핵마름병 발생 보고)

  • Soo Yeon Choi;You Kyoung Lee;Chang Ok Geum;Shinhwa Kim;Hyunjung Chung;Sang-Min Kim;Yong Hoon Lee
    • The Korean Journal of Mycology
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    • v.51 no.4
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    • pp.383-387
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    • 2023
  • Peanut plants showing mild wilt were found in fields of Iksan, Korea, in August 2021. The diseased peanut plants were collected, and the causal pathogens were isolated using potato dextrose agar (PDA) medium. The isolated IS-1 strain formed white mycelia on PDA, which turned black with age. Sclerotia were produced on the PDA and barley leaves laid on water agar 7 d after incubation at 30℃. The sequences of both the internal transcribed spacer (ITS) region and calmodulin gene of IS-1 showed a 100% similarity with that of Macrophomina phaseolina. A phylogenetic tree constructed using the ITS regions of fungal pathogens causing disease in peanut plants indicated that the IS-1 stain belongs to M. phaseolina. The inoculation of IS-1 sclerotia into peanut seedlings resulted in yellowing and wilt symptoms in aboveground plants and brown to dark rots in roots 35-40 d after inoculation. Overall, the morphological characteristics, molecular identification, and pathogenicity of IS-1 indicate that the causal pathogen is M. phaseolina. This is the first report of charcoal rot caused by M. phaseolina on peanut plants in Korea. Further study is needed to develop the control measures for charcoal rot in peanut plants.

Molecular Characterization and Phylogenetic Analysis of Season Influenza Virus Isolated in Busan during the 2006-2008 Seasons (부산지역에서 유행한 계절인플루엔자바이러스의 유전자 특성 및 계통분석('06-'08 절기))

  • Park, Yon-Koung;Kim, Nam-Ho;Choi, Seung-Hwa;Lee, Mi-Oak;Min, Sang-Kee;Kim, Seong-Joon;Cho, Kyung-Soon;Na, Young-Nan
    • Journal of Life Science
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    • v.20 no.3
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    • pp.365-373
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    • 2010
  • To monitor newly emerged influenza virus variants and to investigate the prevalence pattern, our laboratory performed isolation of the viruses from surveillance sentinel hospitals. In the present study, we analysed influenza A/H1N1, A/H3N2, B viruses isolated in Busan during the 2006/07 and 2007/08 seasons by sequence analysis of the hemagglutinin (HA1 subunit) and neuraminidase (NA) genes. The isolates studied here were selected by the stratified random sample method from a total of 277 isolates, in which 15 were A/H1N1, 16 were A/H3N2 and 29 were B. Based on the phylogenetic tree, the HA1 gene showed that A/H1N1 isolates had a 96.7% to 97.7% homology with the A/Brisbane/59/2007, A/H3N2 isolates had a 98.4% to 99.7% homology with the A/Brisbane/10/2007, and B isolates had a 96.5% to 99.7% homology with the B/Florida/4/2006(Yamagata lineage), which are all the vaccine strains for the Northern Hemisphere in 2008~2009 season. In the case of the NA gene, A/H1N1 isolates had 97.8% to 98.5% homologies, A/H3N2 isolates had 98.9% to 99.4% homologies, and B isolates had 98.9% to 100% homologies with each vaccine strain in the 2008~2009 season, respectively. Characterization of the hemagglutinin gene revealed that amino acids at the receptor-binding site and N-linked glycosylation site were highly conserved. These results provide useful information for the control of influenza viruses in Busan and for a better understanding of vaccine strain selection.

Analysis of Microsatellite Loci for Swimming Crab Portunus trituberculatus Populations in the Korean Side of the Yellow Sea (서해안에서 채집된 꽃게(Portunus trituberculatus) 집단에 대한 microsatellite 좌위의 분석)

  • Lee, Hye Jin;Yoon, Seong Jong;Hyun, Young Se;Kim, Hye Jin;Hwang, Sung-Il;Bae, Joo-Seung;Chung, Ki Wha
    • Journal of Life Science
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    • v.23 no.9
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    • pp.1088-1095
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    • 2013
  • The swimming crab, Portunus trituberculatus, inhabits seafloor habitats containing sand or pebbles and is widely distributed throughout the world. The present study investigated genetic polymorphisms of 10 microsatellites in 281 samples of P. trituberculatus collected from four locations along the coastal water of the Korean side of the Yellow Sea (Yeonggwang, Taean, Sorea, and Yeonpyeong-do Island). The number of alleles per locus ranged from 50 to 129, with a mean of 69.5. The observed and expected hetrozygosity varied from 0.111 to 1.000 and from 0.609 to 0.979, respectively. The inbreeding coefficients (Fis) varied among the loci from -0.0207 to 0.8175. The genetic differentiation (Fst) was less than 0.05 (range 0.0020-0.0124). Therefore, the four groups of P. trituberculatus appeared to exhibit little genetic differentiation. The lack of differentiation was confirmed in a phylogenetic tree constructed by the unweighted pair group method with the arithmetic average (UPGMA). The hypervariation between the populations and the lack of genetic differentiation may reflect active gene flow among the Yellow Sea populations and the absence of geographical boundaries. The highly polymorphic microsatellite loci will be useful for molecular and phylogenetic studies, as well as stock management, of swimming crab, which is an important fishery resource.

Evaluate of SSRs for Heat Tolerance using Korean Major Wheat Cultivars and Heat Resistant Turkey Resources (주요 국산밀 품종과 내고온성 터키 유전자원을 이용한 내고온성 관련 SSR 마커 평가)

  • Son, Jae-Han;Kim, Kyeung-Hoon;Cheong, Young-Keun;Park, Jong-Chul;Kim, Kyong-Ho;Kim, Yang-Kil;Oh, Young-Jin;Song, Tae-Hwa;Kim, Bo-kyeong;Kang, Chon-Sik
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.60 no.3
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    • pp.293-299
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    • 2015
  • High temperature is one of major environmental stress. Heat tolerance managing is difficult through the phenotypic selection, so marker assistant selection (MAS) using molecular markers like as RAPD, SSR etc. was tried to select useful traits for heat tolerance. Fourteen SSR markers reported by previous research were selected for this research. We tried to evaluate 14 SSR markers for MAS using 31 useful wheat resources including 24 crossing line from Turkey, six Korean wheat cultivars and Chinese spring. The average of the number of alleles and PIC values in this study were 6.14 and 0.64, respectively. Two major clades and four sub clades were grouped by phylogenetic tree using UPGMA. Four Korean wheat cultivars were distinct from other Turkey resources in the phylogenetic dendrogram. From the results, we expected that these markers were able to adapt to screening wheat genotyping for heat tolerance.

Molecular Detection and Characterization of Orf Virus from Outbreak of Contagious Pustular Dermatitis in Korean Indigenous Goats (한국 재래 산양의 전염성 농피성 피부병에서 orf virus의 검출과 B2L 유전자를 통한 계통발생학적 분석)

  • Park, Jin-Ho;Kim, Guk-Jong;Choi, Wook;Kim, Eun-Ha;Han, Jae-Chul;Ou, Sung-Guk;Lee, Jon-Hwa;Cho, Mae-Rim;Song, Hee-Jong;Chae, Jun-Seok
    • Journal of Veterinary Clinics
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    • v.21 no.2
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    • pp.102-108
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    • 2004
  • Orf virus (ORFV), a member of genus Parapoxvirus (family-Poxviridae), a causative agent of contagious ecthyma in sheep and goat leading to a condition commonly known as vesicular dermatitis. Recently, twelve goats from Iksan in Jeonbuk province were observed with clinical signs like necrotic vesicular lesions around the mucosa of mouth, nasal cavity, eye, ear, teats, abdomen and groin. Based on these clinical symptoms, contagious ecthyma infection was suspected. The skin scrapping was collected from lesions for isolation of DNA and subsequent PCR amplification of ORFV specific 235 bp region of B2L gene. All of the samples were found positive by PCR analysis. Sequencing and further phylogenetic analysis of the PCR product revealed 100% identity to Japan isolate of ORFV (Okinawa, GenBank accession number AB080769), and showed 99.6% of similarity to New Zealand strain (NZ-2, GenBank accession number U06671). It was concluded that ORFV strain detected in the present study is homologous to Japan isolate and New Zealand strain. The PCR test based on amplification of B2L gene is a highly useful tools for rapid and specific diagnosis of contagious ecthyma.

Genetic diversity and population structure of indigenous chicken of Bangladesh using microsatellite markers

  • Rashid, Muhammad Abdur;Manjula, Prabuddha;Faruque, Shakila;Bhuiyan, A.K. Fazlul Haque;Seo, Dongwon;Alam, Jahangir;Lee, Jun Heon;Bhuiyan, Mohammad Shamsul Alam
    • Asian-Australasian Journal of Animal Sciences
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    • v.33 no.11
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    • pp.1732-1740
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    • 2020
  • Objective: The objectives of this study were to investigate the genetic diversity, population structure and relatedness among the five chicken populations of Bangladesh using microsatellite markers. Methods: A total of 161 individuals representing 5 chicken populations (non-descript Deshi [ND], naked neck [NN], hilly [HI], Aseel [AS], and red jungle fowl [JF]) were included in this study to investigate genetic diversity measures, population structure, genetic distance and phylogenetic relationships. Genotyping was performed using 16 selected polymorphic microsatellite markers distributed across 10 chromosomes. Results: The average observed and expected heterozygosity, mean number of alleles and polymorphic information content were found to be 0.67±0.01, 0.70±0.01, 10.7 and 0.748, respectively in the studied populations. The estimated overall fixation index across the loci (F), heterozygote deficiency within (FIS) and among (FIT) chicken populations were 0.04±0.02, 0.05 and 0.16, respectively. Analysis of molecular variance analysis revealed 88.07% of the total genetic diversity was accounted for within population variation and the rest 11.93% was incurred with population differentiation (FST). The highest pairwise genetic distance (0.154) was found between ND and AS while the lowest distance was between JF and AS (0.084). Structure analysis depicted that the studied samples can be categorized into four distinct types or varieties (ΔK = 3.74) such as ND, NN, and HI where AS and JF clustered together as an admixed population. The Neighbor-Joining phylogenetic tree and discriminant analysis of principal component also showed close relatedness among three chicken varieties namely AS, HI, and JF. Conclusion: The results reflected that indigenous chicken of Bangladesh still possess rich genetic diversity but weak differentiation among the studied populations. This finding provides some important insight on genetic diversity measures that could support the designing and implementing of future breeding plans for indigenous chickens of Bangladesh.

Characterization and Functional Study of PyrR Orthologues from Genome Sequences of Bacteria (세균 게놈 유래성 PyrR Orthologue의 기능 분석)

  • 김사열;조현수;설경조;박승환
    • Microbiology and Biotechnology Letters
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    • v.31 no.2
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    • pp.103-110
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    • 2003
  • The regulation of pyrimidine nucleotide synthesis has been proved to be controlled by a regulatory protein PyrR-mediated attenuation in the Gram-positive bacteria. After several bacterial genome sequencing projects, we have discovered the PyrR orthologues in the databases for Haemophilus influenzae and Synechocystis and sp. PCC6803 genome sequences. To investigate whether these PyrR orthologue proteins regulate pyrimidine nucleotide synthesis as well as the cases of Bacillus, the PyrR regions of each strains were amplified by PCR and cloned with pUC19 or T-vector in Escherichia coli and with a shuttle vector pHPS9 for E. coli and B. subtilis. For the regulation test of the PyrR orthologues, the aspartate-transcarbamylase (ATCase) assay was carried out. From the results of the ATCase assay, it was confirmed that Synechocystis sp. PCC6803 could not restore by pyrimidines to a B. subtilis, PyrR but H. influenzae PyrR could. For Purification of PyrR orthologue proteins, PyrR orthologue genes were cloned into the expression vector (pET14b). Over-expressed product of PyrR orthologue genes was purified and analyzed by the SDS-PACE. The purified PyrR orthologue proteins from H. influenzae and Synechocystis sp. PCC6803 turned out to be molecular mass of 18 kDa and 21 kDa, respectively. The result of uracil phosphoribosyl transferase (UPRTase) assay with purified PyrR orthologue proteins showed that H. influenzae PyrR protein only has UPRTase activity. In addition, we could predict several regulatory mechanisms that PyrR orthologue proteins regulate pyrimidine de novo synthesis in bacteria, through phylogenetic analysis for PyrR orthologue protein sequences.

Genetic signature of strong recent positive selection at interleukin-32 gene in goat

  • Asif, Akhtar Rasool;Qadri, Sumayyah;Ijaz, Nabeel;Javed, Ruheena;Ansari, Abdur Rahman;Awais, Muhammd;Younus, Muhammad;Riaz, Hasan;Du, Xiaoyong
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.7
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    • pp.912-919
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    • 2017
  • Objective: Identification of the candidate genes that play key roles in phenotypic variations can provide new information about evolution and positive selection. Interleukin (IL)-32 is involved in many biological processes, however, its role for the immune response against various diseases in mammals is poorly understood. Therefore, the current investigation was performed for the better understanding of the molecular evolution and the positive selection of single nucleotide polymorphisms in IL-32 gene. Methods: By using fixation index ($F_{ST}$) based method, IL-32 (9375) gene was found to be outlier and under significant positive selection with the provisional combined allocation of mean heterozygosity and $F_{ST}$. Using nucleotide sequences of 11 mammalian species from National Center for Biotechnology Information database, the evolutionary selection of IL-32 gene was determined using Maximum likelihood model method, through four models (M1a, M2a, M7, and M8) in Codeml program of phylogenetic analysis by maximum liklihood. Results: IL-32 is detected under positive selection using the $F_{ST}$ simulations method. The phylogenetic tree revealed that goat IL-32 was in close resemblance with sheep IL-32. The coding nucleotide sequences were compared among 11 species and it was found that the goat IL-32 gene shared identity with sheep (96.54%), bison (91.97%), camel (58.39%), cat (56.59%), buffalo (56.50%), human (56.13%), dog (50.97%), horse (54.04%), and rabbit (53.41%) respectively. Conclusion: This study provides evidence for IL-32 gene as under significant positive selection in goat.

Qualitative and Quantitative Analysis of Microbial Community Structure in the Sequencing Batch Reactor for Enriching ANAMMOX Consortium (연속회분식 반응기를 이용한 혐기성 암모늄 산화균 농후배양에서의 정성 및 정량적 미생물 군집구조 분석)

  • Bae, Hyo-Kwan;Jung, Jin-Young
    • Journal of Korean Society of Environmental Engineers
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    • v.31 no.10
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    • pp.919-926
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    • 2009
  • Enrichment of anaerobic ammonium oxidation (ANAMMOX) bacteria is the essential step for operating full-scale ANAMMOX bioreactor because adding a significant amount of seeding sludge is required to stabilize the ANAMMOX reactor. In this study, the enrichment of ANAMMOX bacteria from an activated sludge using sequencing batch reactor was conducted and verified by analyzing changes in the microbial community structure. ANAMMOX bacteria were successfully enriched for 70 days and the substrate removal efficiencies showed 98.5% and 90.7% for $NH_4\;^+$ and $NO_2\;^-$ in the activity test, respectively. The phylogenetic trees of Planctomycetes phylum showed that the diverse microbial community structure of an activated sludge was remarkably simplified after the enrichment. All 36 clones, obtained after the enrichment, were affiliated with ANAMMOX bacteria of Candidatus Brocadia (36%) and Candidatus Anammoxoglobus (64%) genera. The quantification using real-time quantitative PCR (RTQ-PCR) revea ed that the 16S rDNA concentration of ANAMMOX bacteria was 74.8% compared to the granular ANAMMOX sludge obtained from an upflow ANAMMOX sludge bed reactor which had been operated for more than one year. The results of molecular analysis supported that the enriched sludge could be used as a seeding sludge for a full-scale ANAMMOX bioreactor.

The classification and comparison of genetic diversity of genus Malus using RAPD (RAPD를 이용한 능금속 식물종의 계통관계와 유전적 다양성)

  • Huh, Man-Kyu
    • Journal of Life Science
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    • v.17 no.6 s.86
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    • pp.756-761
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    • 2007
  • Cenus Malus is a long-lived woody species primarily distributed throughout Asia. Many species of this genus are regarded as agriculturally and ecologically important. The phynetics and genetic diversity among eight species of genus Malus were reconstructed using the random amplified polymorphic DNA (RAPD) markers. In a simple measure of intraspecies variability by the percentage of polymorphic bands, the M. micromalus exhibited the lowest variation (34.7%). The M. pumila showed the highest (50.0%). Mean number of alleles per locus (A) ranged from 1.347 to 1.500 with a mean of 1.437. The phenotypic frequency of each band was calculated and used in estimating genetic diversify (H) within species. The mean of H was 0.190 across species, varying from 0.155 to 0.220. In particular, two cultivated species, M. pumila and M. asiatica, had high expected diversity, 0.314 and 0.307, respectively. On a per locus basis, the proportion of total genetic variation due to differences among species ranged from 0.388 to 0.472 with a mean of 0.423, indicating that 42.3% of the total variation was found among species. The phylogenetic tree showed three distinct elates. One includes M. sieversii, M. pumila, and M. asiatica. Another includes three M. baccata taxa. The other includes M. sieboldii, M. floribunsa, and M. micromalus. One variety and one form of M. sieboldii were well separated each other. RAPD markers are useful in germ-plasm classification of genus Malus and evolutionary studies.