• Title/Summary/Keyword: Genetic diversity ITS

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Development of EST-SSR markers for the Korean endemic species Chrysosplenium aureobracteatum (Saxifragaceae)

  • SHIN, Jae-Seo;KIM, Bo-Yun;KIM, Yong-In;LEE, Jung-Hoon;KIM, Young-Dong
    • Korean Journal of Plant Taxonomy
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    • v.50 no.1
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    • pp.22-26
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    • 2020
  • Chrysosplenium aureobracteatum Y. I. Kim & Y. D. Kim (Saxifragaceae) is a recently described endemic species growing in the central part of the Korean peninsula. It requires constant monitoring for conservation due to its limited distributions. There is also a need for molecular markers for proper assessments of the genetic differentiation of C. aureobracteatum from species morphologically similar to it. In this study, we developed microsatellite markers that can be used to evaluate the genetic diversity of this species, representing fundamental data with which to conserve the natural populations of the species. A total of 17 expressed sequence tag-simple sequence repeat (EST-SSR) markers were developed by the Illumina pair-end sequencing of the transcriptomes of C. aureobracteatum. These markers were successfully applied to populations of C. aureobracteatum and to its most closely related species, C. barbatum, revealing high polymorphism in both species. The EST-SSR markers developed in this study were proven to be useful not only to monitor the population genetic structure of C. aureobracteatum for conservation purposes but also to study the genetic delimitation of the species from species closely related to it.

Genetic Diversity of Xanthomonas arboricola pv. pruni Isolated in Korea (우리나라에서 분리한 Xanthomonas arboricola pv. pruni의 유전적 다양성)

  • Park, So-Yeon;Lee, Young-Sun;Shin, Jong-Sub;Koh, Young-Jin;Jung, Jae-Sung
    • Journal of Life Science
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    • v.19 no.5
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    • pp.684-687
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    • 2009
  • Xanthomonas arboricola pv. pruni, the causal agent of bacterial shot holes in stone fruits, was known to have a low population diversity. To investigate the genetic characteristics of X. arboricola pv. pruni isolated in Korea, three strains which have identical 16S rDNA sequences - including type strain (LMG852), Japanese isolate (MAFF301420) and Korean isolate (XWD1) - were analysed based on the nucleotide sequences of three DNA regions and RAPD pattern. No sequence diversity among the three strains was found within the ITS, glnA and atpD gene sequences. However, five of 756 nucleotides of the atpD gene determined (accession number FJ429319) were different from those of the French strain available from the Genbank database. RAPD analyses performed with 40 different arbitrary primers revealed that two strains isolated from Korea and Japan showed similarity in their band patterns distinguished by type strain. These results suggest that Korean and Japanese strains are very close and belong to a population with a low genetic diversity, and might have a different origin from strains found in West Europe.

Phylogenetic Relationships Using ITS2 Sequence and RAPD-PCR Data from Four Species of Korean Pseudo-nitzschia (Bacillariophyceae) (ITS2 부위의 염기서열 및 RAPC-PCR에 의한 Pseudo-nitzschia 4종의 유연관계)

  • Cho, Eun-Seob;Lee, Young-Sik
    • Journal of Life Science
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    • v.14 no.1
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    • pp.32-37
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    • 2004
  • A portion of ribosomal internal transcribed spacer (ITS) 2 was sequenced from the samples of Pseudo nitzschia (P. deticatissima, P. multiseries, P. pungens and P. subfraudulenta) to investigate the genetic characteristics by measuring tile magnitude of genetic diversity and the degree of similarity coefficient using random amplified polymorphic DNAs (RAPD)-PCR patterns. The phylogenetic trees inferred from the genetic distance analyses showed the placement of P. delicatissima formed a quite long distance from p. P. multiseries, P. pungens, and even P. subfraudulenta. The phylogenetic tree from RAPD patterns showed that P. multiseries and P. pungens had dissimilarity coefficient of 0.31, while P. delicatissima and three species of Pseudo-nitzschia had that of 0.81. It is likely thought that the genetic position of P. delicatissima formed far from P. multiseries, P. punges, and P. subfraudulenta. These results imply that ITS2 region is expected to support a useful molecular characters for recognizing at the species level and for even discriminating P. multiseries from P. pungens. RAPD method also will be used to differentiate the species of Pseudo-nitzschia in a short time.

Genetic Diversity and Phylogenetic Analysis of the Iranian Leishmania Parasites Based on HSP70 Gene PCR-RFLP and Sequence Analysis

  • Nemati, Sara;Fazaeli, Asghar;Hajjaran, Homa;Khamesipour, Ali;Anbaran, Mohsen Falahati;Bozorgomid, Arezoo;Zarei, Fatah
    • Parasites, Hosts and Diseases
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    • v.55 no.4
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    • pp.367-374
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    • 2017
  • Despite the broad distribution of leishmaniasis among Iranians and animals across the country, little is known about the genetic characteristics of the causative agents. Applying both HSP70 PCR-RFLP and sequence analyses, this study aimed to evaluate the genetic diversity and phylogenetic relationships among Leishmania spp. isolated from Iranian endemic foci and available reference strains. A total of 36 Leishmania isolates from almost all districts across the country were genetically analyzed for the HSP70 gene using both PCR-RFLP and sequence analysis. The original HSP70 gene sequences were aligned along with homologous Leishmania sequences retrieved from NCBI, and subjected to the phylogenetic analysis. Basic parameters of genetic diversity were also estimated. The HSP70 PCR-RFLP presented 3 different electrophoretic patterns, with no further intraspecific variation, corresponding to 3 Leishmania species available in the country, L. tropica, L. major, and L. infantum. Phylogenetic analyses presented 5 major clades, corresponding to 5 species complexes. Iranian lineages, including L. major, L. tropica, and L. infantum, were distributed among 3 complexes L. major, L. tropica, and L. donovani. However, within the L. major and L. donovani species complexes, the HSP70 phylogeny was not able to distinguish clearly between the L. major and L. turanica isolates, and between the L. infantum, L. donovani, and L. chagasi isolates, respectively. Our results indicated that both HSP70 PCR-RFLP and sequence analyses are medically applicable tools for identification of Leishmania species in Iranian patients. However, the reduced genetic diversity of the target gene makes it inevitable that its phylogeny only resolves the major groups, namely, the species complexes.

Development of Repetitive DNA Probes for Genetic Analysis of Phytophthora capsici (Phytophthora capsici의 유전적 특성 분석을 위한 Repetitive DNA Probe의 개발)

  • Song, Jeong-Young;Kim, Hong-Gi
    • The Korean Journal of Mycology
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    • v.30 no.1
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    • pp.66-72
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    • 2002
  • To develop DNA markers for analysis of genetic characteristics of Phytophthora capsici population, randomly selected clones from HindIII-digested genomic DNA library of P. capsici 95CY3119 were surveyed by hybridizing to Southern blots of HindIII-digested total genomic DNA of P. capsici. Probe DNAs inserted into selected individual clones strongly hybridized with HindIII digests of P. capsici. Among probes examined, PC9 revealed the repetitive and highly polymorphic bands to HindIII digests of inter-and intra-field P. capsici isolates. Genetic diversity of individual isolates was also clearly revealed in cluster analysis based on its band patterns. The other probe, PC22, was hybridized only to DNA from P. capsici and this was highly repetitive. However, there was no response to other Phytophthora species and Pythium sp. These DNA probes could be used as very useful markers in analysing genetic diversity and identification for P. capsici population throughout the world.

Development of EST-SSRs and Assessment of Genetic Diversity in Germplasm of the Finger Millet, Eleusine coracana (L.) Gaertn.

  • Wang, Xiaohan;Lee, Myung Chul;Choi, Yu-Mi;Kim, Seong-Hoon;Han, Seahee;Desta, Kebede Taye;Yoon, Hye-myeong;Lee, Yoonjung;Oh, Miae;Yi, Jung Yoon;Shin, Myoung-Jae;Kim, Kyung-Min
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.66 no.4
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    • pp.443-451
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    • 2021
  • Finger millet (Eleusine coracana) is widely cultivated in tropical regions worldwide owing to its high nutritional value. Finger millet is more tolerant against biotic and abiotic stresses such as pests, drought, and salt than other millet crops; therefore, it was proposed as a candidate crop to adapt to climate change in Korea. In 2019, we used expressed sequence tag simple sequence repeat (EST-SSR) markers to evaluate the genetic diversity and structure of 102 finger millet accessions from two geographical regions (Africa and South Asia) to identify appropriate accessions and enhance crop diversity in Korea. In total, 40 primers produced 116 alleles, ranging in size from 135 to 457 bp, with a mean polymorphism information content (PIC) of 0.18225. Polymorphism was detected among the 40 primers, and 13 primers were found to have PIC values > 0.3. Principal coordinate and phylogenetic analyses, based on the combined data of both markers, grouped the finger millet accessions according to their respective collection areas.Therefore, the 102 accessions were classified into two groups, one from Asia and the other from Africa. We have conducted an in-depth study on the finger millet landrace pedigree. By sorting out and using the molecular characteristics of each pedigree, it will be useful for the management and accession identification of the plant resource. The novel SSR markers developed in this study will aid in future genetic analyses of E. coracana.

Studies on the Construction of Mutant Diversity Pool (MDP) lines, and their Genomic Characterization in Soybean

  • Dong-Gun Kim;Sang Hoon Kim;Chang-Hyu Bae;Soon-Jae Kwon
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2021.04a
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    • pp.9-9
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    • 2021
  • Mutation breeding is useful for improving agronomic characteristics of various crops. In this study, we constructed soybean Mutant Diversity Pool (MDP) from 1,695 gamma-irradiated mutants through two selection phases over M1 to M12 generations; we selected 523 mutant lines exhibiting at least 30% superior agricultural characteristics, and, second, we eliminated redundant morphological phenotypes in the M12 generation. Finally, we constructed 208 MDP lines and investigated 11 agronomic traits. We then assessed the genetic diversity and inter-relationships of these MDP lines using target region amplification polymorphism (TRAP) markers. Among the different TRAP primer combinations, polymorphism levels and PIC values averaged 59.71% and 0.15, respectively. Dendrogram and population structure analyses divided the MDP lines into four major groups. According to an analysis of AMOVA, the percentage of inter-population variation among mutants was 11.320 (20.6%), whereas mutant inter-population variation ranged from 0.231 (0.4%) to 14.324 (26.1%). Overall, the genetic similarity of each cultivar and its mutants were higher than within other mutant populations. In an analysis of the genome-wide association study (GWAS) using based on the genotyping-by-sequencing (GBS), we detected 66 SNPs located on 13 different chromosomes were found to be highly associated with four agronomic traits: days of flowering (33 SNPs), flower color (16 SNPs), node number (6 SNPs), and seed coat color (11 SNPs). These results are consistent with those previously reported for other genetic resource populations, including natural accessions and recombinant inbred line. Our observations suggest that genomic changes in mutant individuals induced by gamma rays occurred at the same loci as those of natural soybean population. This study has demonstrated that the integration of GBS and GWAS can serve as a powerful complementary approach to gamma-ray mutation for the dissection of complex traits in soybean.

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Development of PCR-Based Sequence Characterized DNA Markers for the Identification and Detection, Genetic Diversity of Didymella bryoniae with Random Amplified polymorphic DNA(RAPD)

  • Kyo, Seo-Il;Shim, Chang-Ki;Kim, Dong-Kil;Baep, Dong-Won;Lee, Seon-Chul;Kim, Hee-Kyu
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.130-130
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    • 2003
  • Gummy stem blight pathogen is very difficult not only to monitor the inoculum levels prior to host infection, and also it is destructive and hard to control in field condition. We have applied RAPD technique to elucidate the genetic diversity of the genomic DNA of Didymella bryoniae and also to generate specific diagnostic DNA probe useful for identification and detection. The 40 primers produced clear bands consistently from the genomic DNA of twenty isolates of Didymella bryoniae, and two hundred seventy-three amplified fragments were produced with 40 primers. The combined data from 273 bands was analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYS-PC (Version 1.80) to generate a dendrogram. At the distance level of 0.7, two major RAPD groups were differentiated among 20 strains. RAPD group (RG) I included 8 isolates from watermelon except one isolate from melon. RAPD group (RG) IV included 12 isolates from squash, cucumber, watermelon and melon.. In amplification experiment with SCAR specific primer RG1F-RG1R resulted in a single band of 650bp fragment only for 8 isolates out of 20 isolates that should be designated as RAPD Group 1. However, same set of experiment done with RGIIF-RGIIR did not result in any amplified product.. Our attempts to detect intraspecific diversity of ITS region of rDNA by amplifying ITS region and 17s rDNA region for 20 isolates and restriction digestion of amplified fragment with 12 enzymes did not reveal polymorphic band. In order to develop RAPD markers for RGIV specific primer, a candidate PCR fragment( ≒1.4kb) was purified and Southern hybridized to the amplified fragment RGIV isolates. This promising candidate probe recognized only RGIV isolates

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Genetic Diversity of Penicillium isolates Isolated from Pears with Postharvest Decay in Storage (수확 후 저장 배에서 분리한 Penicillium속 균의 유전적 다양성)

  • Han, Do-Suk;Hong, Sung-Kee;Kang, Hee-Wan
    • The Korean Journal of Mycology
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    • v.40 no.1
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    • pp.11-18
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    • 2012
  • This study was carried out to identify the genetic diversity of Penicillium isolates that were isolated from pears with postharvest decay in storage. URP-PCR was used to detect DNA diversity of 84 Penicillium isolates. Based on URP-PCR profiles, 18 Penicillium isolates were selected and their PCR polymorphic bands were produced by additional primers URP1F, URP2R, URP2F, and URP4R. UPGMA cluster analysis using the polymorphic bands showed four clustered groups and futhermore cultural and morphological features characterized the 18 Penicillium isolates. Group 1 was dominant, which occupies 70% in the four clustered groups and identified as P. expansum based on ITS sequence and morphological features.

Breeding System and Allozyme Genetic Diversity of Deutzia paniculata Nakai, an Endemic Shrub in Korea (고유종 꼬리말발도리의 생식특성과 동위효소 유전다양성)

  • Chang, Chin-Sung;Kim, Hui
    • Journal of Korean Society of Forest Science
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    • v.103 no.4
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    • pp.519-527
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    • 2014
  • Deutzia paniculata is an endemic species, which is geographically restricted within southern part of Korea. Four populations of D. paniculata were sampled across its natural range, from the smallest population, Mt. Dalum, which held less than 100 individuals, to the largest, Mt. Unmum, over 3,500 individuals. Artificial pollination study showed that D. paniculata had an obligate outcross breeding system. Major pollinators were two bee species, Lasioglossum exiliceps and Allograpta balteata (de Geer). The breeding system and patterns of allozyme variation of D. paniculata were investigated to understand the population biology and to explain on reserve designs and management proposals relevant to this species. D. paniculata held relatively low genetic variation at the eight allozyme loci surveyed. Measures of genetic variation in this species alleles per locus ($A_s=1.33$), proportion of polymorphic loci (P=23.85%), and expected heterozygosity ($H_{es}=0.110$) were similar to values reported for endemic species. Mt. Dalum population (DAL) was composed with one clone based on allozyme data. Individuals of D. paniculata were frequently included in root connected clusters. Population genetic structure between and within four populations was probably the result of shrinking effective population size and the extinctions of intervening populations. For the conservation of genetic diversity, maximum number of different genotype need to be protected based on genetic structure and mating system.