• Title/Summary/Keyword: Neighbor-Joining tree

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Genetic Diversity and Molecular Markers in Introduced and Thai Native Apple Snails (Pomacea and Pila)

  • Thaewnon-Ngiw, Bungorn;Klinbunga, Sirawut;Phanwichien, Kantimanee;Sangduen, Nitsri;Lauhachinda, Nitaya;Menasveta, Piamsak
    • BMB Reports
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    • 제37권4호
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    • pp.493-502
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    • 2004
  • The genetic diversity and species-diagnostic markers in the introduced apple snail, Pomacea canaliculata and in the native Thai apple snails; Pila ampullacea, P. angelica, P. pesmei, and P. polita, were investigated by restriction analysis of COI and are reported for the first time. Twenty-one composite haplotypes showing non-overlapping distributions among species were found. Genetic heterogeneity analysis indicated significant differences between species (P < 0.0001) and within P. pesmei (P < 0.0001) and P. angelica (P < 0.0004). No such heterogeneity was observed in Pomacea canaliculata (P > 0.0036 as modified by the Bonferroni procedure), P. ampullacea (P = 0.0824-1.000) and P. polita (P = 1.0000). A neighbor-joining tree based on genetic distance between pairs of composite haplotypes differentiated all species and indicated that P. angelica and P. pesmei are closely related phylogenetically. In addition, the 16S rDNA of these species was cloned and sequenced. A species-specific PCR for P. canaliculata was successfully developed with a sensitivity of detection of approximately 50 pg of the target DNA template. The amplification of genomic DNA (50 pg and 25 ng) isolated from the fertilized eggs, and juveniles (1, 7, and 15 d after hatching) of Pomacea canaliculata was also successful, and suggested that Pomacea canaliculata and Pila species can be discriminated from the early stages of development.

Genetic Variation of the Mitochondrial Cytochrome b Sequence in Korean Rana rugosa (Amphibia; Ranidae)

  • Hyun Ick Lee;Dong Eun Yang;Yu Ri Kim;Hyuk Lee;Jung Eun Lee;Suh Yung Yang;Hei Yung Lee
    • 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.

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형태적 특징 및 다좌위 염기서열 분석에 의한 산철쭉 모무늬병균 Sphaerulina azaleae 동정 (Identification of Sphaerulina azaleae on Korean Azalea in Korea Based on Morphological Characteristics and Multilocus Sequence Typing)

  • 최인영;최영준;이귀재;주호종;조성완;신현동
    • 한국균학회지
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    • 제48권3호
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    • pp.329-335
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    • 2020
  • 2008년부터 2017년도에 제주, 홍천 등에서 산철쭉에서 모무늬 증상을 나타내는 잎을 채집하였다. 산철쭉모무늬 증상은 빈번하게 잎에만 발생하여 식물의 관상가치를 떨어트리고 조기낙엽을 유발하였다. 변색부는 잎의 윗면에 작고 담갈색 내지 흑자색 점무늬가 먼저 나타나며, 잎의 세맥으로 경계가 구분되어 모무늬 또는 부정형의 증상을 나타냈다. 산철쭉에서 분리한 균주를 동정하고자 형태적 특징과 actin (Act), translation elongation factor 1-alpha (EF), internal transcribed spacer (ITS), 28S nrDNA (LSU), RNA polymerase II second largest subunit (RPB2) 염기서열을 분석하였다. 형태적 특징을 재확인한 염기서열 분석결과 Sp. azaleae와 99~100%의 상동성을 나타냈으며, 계통수를 작성하였을 때도 Sp. azaleae 계통군에 속하였다. 따라서 산철쭉에 모무늬 증상에 관여하는 곰팡이는 Sp. azaleae로 동정되었다.

Phylogenetic relationships of Iranian Allium species using the matK (cpDNA gene) region

  • Zarei, Hemadollah;Fakheri, Barat Ali;Naghavi, Mohammad Reza;Mahdinezhad, Nafiseh
    • Journal of Plant Biotechnology
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    • 제47권1호
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    • pp.15-25
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    • 2020
  • Allium L. is one of the largest genera of the Amaryllidaceae family, with more than 920 species including many economically important species used as vegetables, spices, medicines, or ornamental plants. Currently, DNA barcoding tools are being successfully used for the molecular taxonomy of Allium. A total of 46 Allium species were collected from their native areas, and DNA was extracted using the IBRC DNA extraction kit. We used specific primers to PCR amplify matK. DNA sequences were edited and aligned for homology, and a phylogenetic tree was constructed using the neighbor-joining method. The results show thymine (38.5%) was the most frequent and guanine (13.9%) the least frequent nucleotide. The matK regions of the populations were quite highly conserved, and the amount of C and CT was calculated at 0.162 and 0.26, respectively. Analysis of the nucleotide substitution showed C-T (26.22%) and A-G (8.08%) to have the highest and lowest percent, respectively. The natural selection process dN/dS was 1.16, and the naturality test results were -1.5 for Tajima's D and -1.19 for Fu's Fs. The NJ dendrogram generated three distinct clades: the first contained Allium austroiranicum and A. ampeloprasum; the second contained A. iranshahrii, A. bisotunense, and A. cf assadi; and the third contained A. rubellum and other species. In this study, we tested the utility of the matK region as a DNA barcode for discriminating Allium. species.

A report on 53 unrecorded bacteria species in Korea in the class Gammaproteobacteria

  • Kanjanasuntree, Rungravee;Cha, Chang-Jun;Cho, Jang-Cheon;Im, Wan-Taek;Kim, Myung Kyum;Jeon, Che-Ok;Joh, Kiseong;Kim, Seung-Bum;Seong, Chi-Nam;Yi, Hana;Lee, Soon Dong;Bae, Jin-Woo;Kim, Wonyong
    • Journal of Species Research
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    • 제8권4호
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    • pp.319-336
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    • 2019
  • During an investigation of unrecorded prokaryotic species in Republic of Korea, a total of 53 bacterial strains belonging to the class Gammaproteobacteria were isolated from soil, seawater, tidal flats, rhizosphere, salt ponds, beach sand, urine, manure, sediment, and animal intestine (Russian grayling butterfly [Hipparchia autonoe], mouse [Mus musculus], and sea bass [Lateolabrax japonicus]). Strains were identified to species using the 16S rRNA gene sequence, showing high similarity (>98.7%) with the closest bacterial species and forming a robust clade in the neighbor-joining phylogenetic tree. The 53 strains of Gammaproteobacteria in this study have not been report previously in Korea. Therefore, we describe 27 genera of 16 families in 7 orders: 13 strains in the order Alteromonadales, 1 strain in the order Chromatiales, 11 strains in the order Enterobacterales, 7 strains in the order Oceanospirillales, 10 strains in the order Pseudomonadales, 8 strains in the order Vibrionales, and 3 strains in the order Xanthomonadales. Gram reaction, strain ID, isolation source, and morphological and basic biochemical characteristics are described for each species.

Genetic characteristics of Korean Jeju Black cattle with high density single nucleotide polymorphisms

  • Alam, M. Zahangir;Lee, Yun-Mi;Son, Hyo-Jung;Hanna, Lauren H.;Riley, David G.;Mannen, Hideyuki;Sasazaki, Shinji;Park, Se Pill;Kim, Jong-Joo
    • Animal Bioscience
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    • 제34권5호
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    • pp.789-800
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    • 2021
  • Objective: Conservation and genetic improvement of cattle breeds require information about genetic diversity and population structure of the cattle. In this study, we investigated the genetic diversity and population structure of the three cattle breeds in the Korean peninsula. Methods: Jeju Black, Hanwoo, Holstein cattle in Korea, together with six foreign breeds were examined. Genetic diversity within the cattle breeds was analyzed with minor allele frequency (MAF), observed and expected heterozygosity (HO and HE), inbreeding coefficient (FIS) and past effective population size. Molecular variance and population structure between the nine breeds were analyzed using a model-based clustering method. Genetic distances between breeds were evaluated with Nei's genetic distance and Weir and Cockerham's FST. Results: Our results revealed that Jeju Black cattle had lowest level of heterozygosity (HE = 0.21) among the studied taurine breeds, and an average MAF of 0.16. The level of inbreeding was -0.076 for Jeju Black, while -0.018 to -0.118 for the other breeds. Principle component analysis and neighbor-joining tree showed a clear separation of Jeju Black cattle from other local (Hanwoo and Japanese cattle) and taurine/indicine cattle breeds in evolutionary process, and a distinct pattern of admixture of Jeju Black cattle having no clustering with other studied populations. The FST value between Jeju Black cattle and Hanwoo was 0.106, which was lowest across the pair of breeds ranging from 0.161 to 0.274, indicating some degree of genetic closeness of Jeju Black cattle with Hanwoo. The past effective population size of Jeju Black cattle was very small, i.e. 38 in 13 generation ago, whereas 209 for Hanwoo. Conclusion: This study indicates genetic uniqueness of Jeju Black cattle. However, a small effective population size of Jeju Black cattle indicates the requirement for an implementation of a sustainable breeding policy to increase the population for genetic improvement and future conservation.

A New Distinct Clade for Iranian Tomato spotted wilt virus Isolates Based on the Polymerase, Nucleocapsid, and Non-structural Genes

  • Abadkhah, Mahsa;Koolivand, Davoud;Eini, Omid
    • The Plant Pathology Journal
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    • 제34권6호
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    • pp.514-531
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    • 2018
  • Tomato spotted wilt virus (TSWV; Genus Orthotospovirus: Family Tospoviridae) is one of the most destructive viruses affecting a wide range of horticultural crops on a worldwide basis. In 2015 and 2016, 171 leaf and fruit samples from tomato (Solanum lycopersicum) plants with viral symptoms were collected from the fields in various regions of Iran. ELISA test revealed that the samples were infected by TSWV. The results of RT-PCR showed that the expected DNA fragments of about 819 bp in length were amplified using a pair of universal primer corresponding to the RNA polymerase gene and DNA fragments of ca 777 bp and 724 bp in length were amplified using specific primers that have been designed based on the nucleocapsid (N) and non-structural (NSs) genes, respectively. The amplified fragments were cloned into pTG19-T and sequenced. Sequence comparisons with those available in the GenBank showed that the sequences belong to TSWV. The high nucleotide identity and similarities of new sequences based on the L, N, and NSs genes showed that minor evolutionary differences exist amongst the isolates. The phylogenetic tree grouped all isolates six clades based on N and NSs genes. Phylogenetic analysis showed that the Iranian isolates were composed a new distinct clade based on a part of polymerase, N and NSs genes. To our knowledge, this is the first detailed study on molecular characterization and genetic diversity of TSWV isolates from tomato in Iran that could be known as new clade of TSWV isolates.

Morphological and Molecular Identification of Stellantchasmus dermogenysi n. sp. (Digenea: Heterophyidae) in Thailand

  • Wongsawad, Chalobol;Nantarat, Nattawadee;Wongsawad, Pheravut;Butboonchoo, Preeyaporn;Chai, Jong-Yil
    • Parasites, Hosts and Diseases
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    • 제57권3호
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    • pp.257-264
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    • 2019
  • We tried a series of morphological and molecular approaches to identify a new species of Stellantchasmus (Digenea: Heterophyidae) originating from the wrestling half-beaked fish, Dermogenys pusillus of Thailand. Adult worm samples of the new species were recovered from hamsters experimentally infected with the metacercariae from D. pusillus in Thailand. Two isolates (Thai and Korean) of Stellantchasmus falcatus were used as comparative control groups. Worm samples of 3 Stellantchasmus groups were morphologically observed and molecularly analyzed with the mitochondrial cytochrome c oxidase 1 gene. The morphological characteristics of S. dermogenysi n. sp. are similar to S. falcatus originating from brackish water fish, but minor difference was noted including the absence of the prepharynx, position of the ovary near the ceca end, smaller body size, and shorter esophageal length. A phylogenetic tree derived from neighbor-joining and maximum-likelihood methods suggests that S. dermogenysi n. sp. is separated from S. falcatus supported by high bootstrap values. The relative divergences persist between these host-specific trematodes, which we suggest should be recognized as 2 distinct species. Comparisons of S. dermogenysi n. sp. with S. falcatus isolated from mullets in Thailand and Korea indicate a genetic divergence of mitochondrial DNA of 19.4% and 21.7%, respectively. By the present study, a new species, Stellantchasmus dermogenysi n. sp. (Digenea: Heterophyidae), is proposed in Thailand based on molecular evidences, in addition to minor morphological differences between S. falcatus and the new species.

Presence of Diverse Sugarcane Bacilliform Viruses Infecting Sugarcane in China Revealed by Pairwise Sequence Comparisons and Phylogenetic Analysis

  • Ahmad, Kashif;Sun, Sheng-Ren;Chen, Jun-Lu;Huang, Mei-Ting;Fu, Hua-Ying;Gao, San-Ji
    • 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.

Transferability of EST SSR-Markers from Foxtail Millet to Barnyard Millet (Echinochloa esculenta)

  • Myung Chul Lee;Yu-Mi Choi;Myoung-Jae Shin;Hyemyeong Yoon;Seong-Hoon Kim
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2020년도 춘계학술대회
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    • pp.45-45
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    • 2020
  • A large number of expressed sequence tags (ESTs) in public databases have provided an opportunity for the systematic development of simple sequence repeat (SSR) markers. EST-SSRs derived from conserved coding sequences show considerable cross-species transferability in related species. In the present study, we assessed the utility of foxtail millet EST-SSRs in barnyard millet. A total of 312 EST-SSRs of foxtail millet were tested using 84 Echinochloa crus-galli germplasm accessions; a high rate of transferability (62%) and 46 primer sets (13%) were shown the polymorphism in barnyard millet. The 13% of functional EST-SSRs) was demonstrated between cereals and barnyard millet. SSR marker profile data were scored for the computation of pairwise distances as well as a Neighbor Joining (NJ) tree of all the genotypes. The averaged values of gene diversity (HE) and polymorphism information content (PIC) were 0.213 and 0.179 within populations, respectively. The 84 barnyard millet germplasm accessions were divided into five different groups, which agreed well with their geographical origins. The exotic 12 accessions of India type barnyard millet (E. frumentacea) were all separated form Korean local collection genotype. The present results provide evidence of divergence between cultured and wild type barnyard, as a millet and grass. The polymorphic SSR markers indicated in this study were of great value in analysis of genetic diversity that can be further used for crop improvement through breeding.

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