• Title/Summary/Keyword: intraspecific groups

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Usability of DNA Sequence Data: from Taxonomy over Barcoding to Field Detection. A Case Study of Oomycete Pathogens

  • Choi, Young-Joon;Thines, Marco
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.41-41
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    • 2015
  • Oomycetes belong to the kingdom Straminipila, a remarkably diverse group which includes brown algae and planktonic diatoms, although they have previously been classified under the kingdom Fungi. These organisms have evolved both saprophytic and pathogenic lifestyles, and more than 60% of the known species are pathogens on plants, the majority of which are classified into the order Peronosporales (includes downy mildews, Phytophthora, and Pythium). Recent phylogenetic investigations based on DNA sequences have revealed that the diversity of oomycetes has been largely underestimated. Although morphology is the most valuable criterion for their identification and diversity, morphological species identification is time-consuming and in some groups very difficult, especially for non-taxonomists. DNA barcoding is a fast and reliable tool for identification of species, enabling us to unravel the diversity and distribution of oomycetes. Accurate species determination of plant pathogens is a prerequisite for their control and quarantine, and further for assessing their potential threat to crops. The mitochondrial cox2 gene has been widely used for identification, taxonomy and phylogeny of various oomycete groups. However, recently the cox1 gene was proposed as a DNA barcode marker instead, together with ITS rDNA. To determine which out of cox1 or cox2 is best suited as universal oomycete barcode, we compared these two genes in terms of (1) PCR efficiency for 31 representative genera, as well as for historic herbarium specimens, and (2) in terms of sequence polymorphism, intra- and interspecific divergence. The primer sets for cox2 successfully amplified all oomycete genera tested, while cox1 failed to amplify three genera. In addition, cox2 exhibited higher PCR efficiency for historic herbarium specimens, providing easier access to barcoding type material. In addition, cox2 yielded higher species identification success, with higher interspecific and lower intraspecific divergences than cox1. Therefore, cox2 is suggested as a partner DNA barcode along with ITS rDNA instead of cox1. Including the two barcoding markers, ITS rDNA and cox2 mtDNA, the multi-locus phylogenetic analyses were performed to resolve two complex clades, Bremia lactucae (lettuce downy mildew) and Peronospora effuse (spinach downy mildew) at the species level and to infer evolutionary relationships within them. The approaches discriminated all currently accepted species and revealed several previously unrecognized lineages, which are specific to a host genus or species. The sequence polymorphisms were useful to develop a real-time quantitative PCR (qPCR) assay for detection of airborne inoculum of B. lactucae and P. effusa. Specificity tests revealed that the qPCR assay is specific for detection of each species. This assay is sensitive, enabling detection of very low levels of inoculum that may be present in the field. Early detection of the pathogen, coupled with knowledge of other factors that favor downy mildew outbreaks, may enable disease forecasting for judicious timing of fungicide applications.

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A checklist of vascular plants in limestone areas on the Korean Peninsula (한반도 석회암지대의 관속식물 목록)

  • KIM, Jung-Hyun;NAM, Gi-Heum;LEE, Seung-bae;SHIN, Sookyung;KIM, Jin-Seok
    • Korean Journal of Plant Taxonomy
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    • v.51 no.3
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    • pp.250-293
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    • 2021
  • Limestone areas are sedimentary rock outcrops consisting of calcium carbonate created several hundreds of millions of years ago by calcium-secreting marine organisms and subsequently lifted above sea level by tectonic movement. Limestone areas support very high levels of endemic species of plants and are recognized as biodiversity areas with much biological information. The purpose of this study is to devise a strategy for the comprehensive conservation of the vegetation of limestone areas through analyses of the floristics and plant species compositions in ten limestone areas on the Korean Peninsula. The results of 153 field surveys from April of 2010 to October of 2016 identified 1,202 taxa in total, representing 1,096 species, 18 subspecies, 84 varieties, 2 forms, and 2 hybrids in 530 genera and 133 families. Among them, 55 taxa were endemic plants to Korea, and 38 taxa were red data plants. The floristic target plants amounted to 102 taxa, specifically 27 taxa of grade V and 75 taxa of grade IV. In all, 121 alien plants were recorded in the investigated area. Calciphilous plants amounted to 102 taxa, specifically 14 taxa of calciphilous indicator plants, 30 taxa of superlative most calciphilous plants, and 58 taxa of comparative more calciphilous plants. A cluster analysis showed a high degree of similarity between sites that are geographically adjacent with similar habitat environments. Limestone areas also supported groups distinct from those in non-limestone areas, demonstrating the specificity of limestone flora. Plant geography approaches therefore appear to be crucial to gain a better understanding of the level of biodiversity in limestone areas, not only at the interspecific but also at the intraspecific level. These results highlight the importance of protecting limestone habitats to preserve not only their interspecific but also the intraspecific diversity, which is highly threatened.

Analysis of phylogenetic relationship among Korean Pinellia Tenore (Araceae) using RAPD markers (RAPD markers에 의한 한국산 반하속 식물의 유연관계 분석)

  • Tae, Kyoung-Hwan;Kim, Dong-Kap;Kim, Joo-Hwan
    • Korean Journal of Plant Taxonomy
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    • v.35 no.3
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    • pp.161-174
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    • 2005
  • In order to presume the relationships between two species of P. ternata and P. tripartia, and their populations of the Korean Pinellia, RAPD analysis was performed. The length of the amplified DNA fragments ranged from 300 to 2,500bp. Seventy scorable RAPD makers were found from the PCR reactions with 7 random oligoprimers and were analyzed by Nei-Li's genetic coefficient. Also, some regional groups instead of same taxa were clustered from the phenogram of UPGMA analysis and NJ tree. Populations within each species were clustered at low genetic distance, there had the closed relationship. According to the regional individuals, Pinellia ternata was showed the variation pattern of morphological (leaf shape and flower color) and cytological characters(somatic chromosome numbers). So we suggested to difference of characteristic variety based on variety of habitat. According to this study, new species (Pinellia sp.) was affiliated with Pinellia and had the closest relationship with Hallasan and Japan population. The RAPD data was very useful to define the genetic variation and to discuss the relationships among the intraspecific taxa and their populations of the Korean Pinellia.

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.

Interspecific Relationship of Polygonatum Species Collected from Gyeongnam Area Using Cluster Analysis (경남지역 둥굴레속의 Cluster 분석에 의한 종간 유연관계)

  • Shim, Jae-Suk;Park, Jeong-Min;Jeon, Byong-Sam;Kang, Jin-Ho
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.1
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    • pp.30-34
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    • 2005
  • Polygonatum species are a useful medical crop in Korea but basic study in the species was not well conducted. The study was carried out to analyse genetic diversity and intraspecific relationship of 47 Polygonatum species collected from Gyeongnam province. Their analysis was done through principle component analysis and average linkage cluster analysis with their twelve morphological traits. The result of principle component analysis showed the Prin 1, Prin 2 and Prin 3 represented 79% of total variation. By the 0.7 average distance of the cluster analysis and the calculated Euclidian distance, the 47 collected species were grouped into five groups. Group I included 22 collected species representing P. ordoratum var. pluriflorum, group II did 5 ones representing P. involucratum, group III was divided into two subclasses, 2 species including P. inflatum and 7 species including P. thunbergii, group IV also consisted of 2 subclasses, a species similar to P. thunbergii and P.involucratum, respectively, and finally group V included 8 species representing P.lasianthum var. coreanum. meaning that the useful germplasms can be collected from relatively small area.

Systematic Study of the Subfamily Leuciscinae (Cyprinidae) from Korea (한국산 황어아과어류의 계통분류학적 연구)

  • KIM Ik Soo;LEE Geum Yeong;YANG Suh Y.
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.18 no.4
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    • pp.381-400
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    • 1985
  • Twelve species of subfamily Leuciscinae from Korea are reviewed and keys to species and genera are provided. The species include Tribolodon hakonensis, T. branti, Phoxinus phoxinus, Moroco oxycephalus, M. steindachneri, M. keumgang, M. semotilus, Zacco platypus, Z. temminckii, Opsariichthys uncirostris amurensis, Aphyocypris chinensis and Squaliobarbus curriculus. In the present study M. lagowskii and O. bidens recorded previously in Korea have to be classified to M. steindachneri and O.u.amurensis based on several characters such as caudal peduncle depth, their distributions and lateral line scales, etc. A. chinensis from Korea is easily distinguished from that of mainland China in head length and caudal peduncle length. The skull and weberian apparatus of M. keumgang, endemir to Korea are described and compared with the other species of this subfamily. The diploid chromosome number of 9 species has three patterns: 2n=50 (T. hakonensis, P. phoxinus, M. oxycephalus, M. steindachneri and M. keumgang), 2n=48(Z. platypus, Z. temminckii and A. chinensis), 2n=76 (O. u. amurensis). Polyploidization or intraspecific polymorphism of chromosome was not observed in any species studied. Both species of P. phoxinus and A. chinensis are shown to be electrophoretically distinct from other taxa examined. The genera of subfamily Leuciscinae in Korea are represented by three possible groups: 1) Zacco-Opsariichthys-Tribolodon-Phoxinus-Morco, 2) Aphyocypris, and 3) Squaliobarbus.

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Analyses of Inter-cultivar Variation for Salinity Tolerance in Six Korean Rapeseed Cultivars

  • Lee, Yong-Hwa;Lee, Tae-Sung;Kim, Kwang-Soo;Jang, Young-Seok;Nam, Sang-Sik;Park, Kwang-Geun
    • Horticultural Science & Technology
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    • v.30 no.4
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    • pp.417-425
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    • 2012
  • Salinity stress is one of the most serious factors limiting the productivity of agricultural crops. The aim of this study was to assess inter-cultivar (intraspecific) variation for salinity tolerance in six Korean rapeseed (Brassica napus L.) cultivars at the seedling stage. The effect of three different salinity stress levels (EC 4, 8, and 16 $dS{\cdot}m^{-1}$) on seedlings of six cultivars was investigated through leaf size, leaf dry weight, and leaf chlorosis. At the highest salinity level (16 $dS{\cdot}m^{-1}$), the mean decrease of leaf dry weight in 'Sunmang', 'Tammi', 'Tamla', 'Naehan', 'Youngsan', and 'Halla' was about 56.2, 56.9, 78.4, 79.3, 77.4, and 80.9%, respectively. 'Tammi' and 'Sunmang' showed much less reduction in leaf dry weight than all the other cultivars. In addition, diluted seawater treatments increased the occurrence of leaf chlorosis in six cultivars. At EC 8 and 16 $dS{\cdot}m^{-1}$, 'Naehan', 'Youngsan', and 'Halla' showed a higher level of leaf chlorosis than 'Tammi' 'Sunmang', and 'Tamla'. On the basis of these results, six cultivars were placed into salinity-tolerant and sensitive groups. 'Tammi' and 'Sunmang' were the salinity-tolerant cultivars, while 'Naehan', 'Halla', 'Youngsan', and 'Tamla' were the salinity-sensitive cultivars. 'Tammi' and 'Naehan' rated as the most tolerant and most sensitive cultivar, respectively. To further analyze protein expression profiles in 'Tammi' and 'Naehan', 2-D proteomic analysis was performed using the plants grown under diluted seawater treatments. We identified eight differentially displayed proteins that participate in photosynthesis, carbon assimilation, starch and sucrose metabolism, amino acid metabolism, cold and oxidative stress, and calcium signaling. The differential protein expressions in 'Tammi' and 'Naehan' are likely to correlate with the differential growth responses of both cultivars to salinity stress. These data suggest that 'Tammi' is better adapted to salinity stressed environments than 'Naehan'.

Identification of Korean Mountain Cultivated Ginseng by RAPD (RAPD(Random Amplified Polymorphic DNA)를 이용한 장뇌삼의 지역별 품종 구분)

  • Choi, Ji-Young;Lee, Ju-Hee;Lee, Su-Gwang;Kang, Ho-Duck
    • Journal of agriculture & life science
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    • v.43 no.6
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    • pp.35-43
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    • 2009
  • This study was conducted to examine the genetic variations and intraspecific relationships between 9 individuals of Panax ginseng C.A Meyer by using RAPD (Randomly Amplified Polymorphic DNA) analysis. The 34 primers out of 40 random primers were amplified for all tested plants. The 48 (40%) among 244 bands derived from 34 primers shown polymorphism, and the 72 (64%) rest of bands showed similar forms. By regional groups Sangju and Andong samples located in Kyungsang buk-do showed a high similarity. However, Punggi located in Kyungsang buk-do showed higher similarity with Jinan's of Junla buk-do. In this way, it did not show that Panax ginseng from the same area has similarities. In future study we need to more specific molecular phylogenetic analysis such as AFLP technology and gene sequencing with nuclear chloroplast DNA in all samples.