• Title/Summary/Keyword: random amplified polymorphic DNA (RAPD-PCR)

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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.

Intergeneric Transfer of Isolated Nuclei from Lentinus edodes into Protoplasts of Pleurotus florida (사철느타리버섯 원형질체내에 표고 핵의 전이)

  • Yoo, Young-Bok;Shin, Pyung-Gyun
    • The Korean Journal of Mycology
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    • v.24 no.2 s.77
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    • pp.111-126
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    • 1996
  • Transfer of the isolated nuclei from Lentinus edodes into protoplasts of Pleurotus florida was induced with polyethlene glycol (PEG) and $CaCl_2$. The intergeneric transfer products were classified into nuclear hybrid, heterokaryon or synkaryon, and reconstituted cell. These progenies except nuclear hybrids formed mature fruiting bodies on sawdust rice bran medium. Formation of fruit bodies was influenced by several factors such as light, temperature, nutrition and physic state of the culture media. Most of fruiting body characters were similar to those of P. florida in synkaryon and L. edodes in reconstituted cell, respectively. All these basidiocarps had clamp connections though initial heterokaryon colonies were lacking. Isozyme patterns of intergeneric progenies were quite different from those of parents. DNA polymorphisms of transfer products were also compared by random amplified polymorphic DNAs (RAPD) analysis based on polymerase chain reaction. The RAPD patterns were different from those of donor and recipient. DNA fingerprints ranged in size from 0.25 to 4.0 Kb. On the basis of RAPD, the transfer products were classified into five groups. Two synkaryon were analysed with distribution of progenies and segregation of genetic markers by random spore analyses. The genetic markers were segregated into wild type and riboflavine requiring auxotrophs.

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An analysis of the genetic diversity of a riparian marginal species, Aristolochia contorta (수변 경계종인 쥐방울덩굴의 유전적 다양성 분석)

  • Nam, Bo Eun;Park, Hyun Jun;Son, Ga Yeon;Kim, Jae Geun
    • Journal of Wetlands Research
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    • v.22 no.2
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    • pp.100-105
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    • 2020
  • Northern pipevine (Aristolochia contorta) commonly inhabits marginal areas between waterside and terrestrial vegetation. In particular, A. contorta is ecologically important in the marginal areas as a food plant of dragon swallowtail butterfly (Sericinus montela), which is designated as vulnerable species in the Republic of Korea. For long-term sustainability of the plant population, assessment of the genetic diversity of exist populations should be conducted. Genomic DNA of A. contorta leaf samples were extracted from four populations where the vigorous growth were observed in the South Korea. Intra-population genetic diversity and inter-population genetic distance were assessed using randomly amplified polymorphic DNA (RAPD) with five polymorphic random primers. Overall genetic diversity was lower, compared to other wetland species (h: 0.0607 ~ 0.1401; I: 0.0819 ~ 0.1759), while GP showed the highest intra-population genetic diversity. Despite of the geographical distance, GP showed the larger genetic distance from other populations. This result seemed to be caused by the fragmented habitat and lower sexual reproduction of A. controta. Mixture of the different source populations and construction of the proper environmental condition such as shade and physical support for sexual reproduction should be considered for conservation of A. contorta population.

Studies on Microbial Ecology of Actinomycetes in Tideland Soils. (서해 아암도 갯벌토양 미생물의 개체군 분석 및 RAPD 분석에 의한 방선균의 생태학적 연구)

  • 조영주;김정한;전은수;이상미;박동진;이재찬;이향범;김창진
    • Microbiology and Biotechnology Letters
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    • v.30 no.1
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    • pp.79-85
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    • 2002
  • Ecological characteristics of microorganisms in tideland soils were studied by investigation of microbial diversity and population. Twenty soil samples were taken at surface, 10, 20 and 30 cm depth each. Bacteria, actinomycetes and fungi were isolated on each selective isolation medium containing different concentration of NaCl. Actinomycetes were the most isolated from soil samples taken at 10 cm depth and isolated by humic acid-vitamin (HV) medium without sea water or salt. Twenty nine strains of actinomycetes were isolated at surface soil and 74, 39, 37 strains were at 10, 20, and 30 cm depth, respectively. All these isolates were analysed and grouped by random amplified polymorphic DNA (RAPD)-PCR analysis. Many of the isolates were clustered into Microtetraspora and Pseudonocardia. Fungal isolates were highly distributed at the surface soil and isolated well on potato dextrose agar (PDA) medium with sea water. Bacterial isolates were higly distributed at surface soil and isolated well by nutrient medium without sea water or salt. Soil samples taken at 10 cm depth showed the highest microbial diversity and population.

Genetic Characteristics and Virulence Factors of Pandemic Vibrio parahaemolyticus Isolated in South Korea and Japan (한국과 일본에서 유행하는 장염비브리오의 병원성 인자와 유전자의 특성)

  • Hong, Seuk-Won;Moon, Ji-Young;Lee, Bok-Kwon;Kim, Yung-Bu
    • Journal of Life Science
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    • v.17 no.3 s.83
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    • pp.386-395
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    • 2007
  • A total of 27 strains of Vibrio parahaemolyticus (18 strains isolated from Korea and 9 strains from Japan) were serotyped and examined for biochemical characteristics, antimicrobial susceptibility patterns, cytotoxicity assay, thermostable direct hemolysin (TDH) production and molecular epidemiology. Using polymerase chain reaction (PCR) method and DNA probe hybridization method, the strains were tested for toxR, tdh, trh and ORF 8 genes. The V. parahaemolyticus isolated from patients were belonged to 8 different serotypes : O3:K6, O1:K38, O3:K57, O4:K9, O4:Kl2, O4:K68, O5:Kl5 and O6:K46. Urease-positive strain possessed the trh gene, and conversely, urease-negative strains lacked the gene, indicating that urease production by V. parahemolyticus strains strongly correlates with the possession of the trh gene. Most strains showed multiple resistant to more than three antibiotics and the antibiogram could be classified into 6 group (I to VI). All of the O3:K6 strains isolated in South Korea and Japan producted TDH at high levels. The TDH titers ranged between 256 and 2.048, and the average titer was 1009. To distinguish the new and increasingly common V. parahaemolyticus strains from clinical isolates, ORF 8 is a useful genetic marker. After Southern hybridization, the HindIII restriction fragment patterns of the tdh gene were grouped one type, respectively. One type showed two bands one of which was 4.3kb and the other was 11.5kb in size. Variation between the O3:K6 serotype are minor when compared to the differences seen with the non O3:K6 strains. The migration patterns of Not I -digested of the total DNA of the O3:K6 strains were similar, and only slight variations were observed between the serotypes. By contrast, the O3:K6 strains and non O3:K6 had markedly different profiles. In conclusion, Random amplified polymorphic DNA (RAPD) profile using appropriate primers was an effective epidemiological marker.

Reproductive Ecology and Genetic Variations in the Sea Urchins Anthocidaris crassispina and Hemicentrotus pulcherrimus in the Cheju Coast (제주연안에 서식하는 보라성게 Anthocidaris crassispina와 말똥성게 Hemicentrotus pulcherrimus의 지역별 번식생태학적 특성과 유전적 변이의 비교)

  • 이정재;김범규;강상균;정상철;이기완;최광식
    • Journal of Aquaculture
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    • v.13 no.2
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    • pp.129-135
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    • 2000
  • Genetic variabilities of local populations of Anthocidaris crassispina and Hemicentrotus pulcherrimus were analysed by random amplified polymorphic DNA (RAPD) technique with phenotypic variabilities in timing of gonadal maturation, reproductive output and size of individuals in Cheju. H, pulcherrimus, collected from 4 locations during March 1997, indicated that Daepo individuals were significantly smaller than that at Hamdok, Wimi site A and Wimi site B (ANOVA, p>0.001). Gonodal-somatic index (GSI) of the Wimi site B population was significantly higher than that of three other locations (ANOVA, P>0.0001). An ANOVA test conducted on test size of A. crassispina, harvested from six different locations of Cheju during June 1997, indicated that the size of individuals from Pophwan was significantly smaller (P>0.0001) than that from five other locations. GSIs of urchin m Wimi and Hanlim were significantly higher than that from Pohwan and Oedo (ANOVA, p>0.0001). Genetic similarity, calculated from k13 primer analysis of total DNA, among the six different populations of A. crassispina varied from 0.67 to 0.92, and the values for H. pulcherrimus from 0.60 to 0.73; thus there was no genetic variation among different populations of the same species. Therefore, the populations are genetically homologous and the observed phenotypic variabilities were possibly associated with water temperature and food.

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Fruiting body development and genetic analysis of somatic hybrids by protoplast fusion in edible fungi (식용버섯의 원형질체 융합체의 자실체 발생 및 유전분석)

  • Yoo, Young Bok;Kong, Won Sik;Oh, Se Jong;Jhune, Chang Sung;Shin, Pyung Gyun;Kim, Beom Gi;Kim, Gyu Hyun;Park, Minsun;Min, Byung Re
    • Journal of Mushroom
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    • v.2 no.3
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    • pp.115-126
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    • 2004
  • Somatic hybrids of inter-compatible and inter-incompatible strains were obtained by protoplast fusion. The fusion products between compatible strains, Pleurotus ostreatus and P. florida, formed heterokaryons, while fusants between incompatible strains such as P. cornucopiae + P. florida, P. ostreatus + Ganoderma applanatum, P. florida + Ganoderma lucidum, and P. ostreatus + Flammulina velutipes formed synkaryons that retained genes from both parents. The heterokaryons showed the same level of basidioma development. In contrast, the synkaryons showed unique characteristics including clamp connection formation at mitosis, either partner basidioma development, and abnormal segregation and recombination compared with inter-compatible strains. Synkaryons can be classified into homokaryoyic and heterokaryotic type. A comparison of somatic hybrids with compatible and incompatible strains was made using random amplified polymorphic DNA (RAPD) analysis. The heterokaryons between compatible species showed the same level of variability and contained both parental RAPD bands. In contrast, most of the synkaryons between incompatible species showed similarity to those of either parental bands and non-parental RAPD bands. Synkaryons can be classified into microgenome insertion type and macrogenome insertion type. A tetrapolar mating system was found among monospore isolates in somatic hybrids and wild type P. ostreatus. Homokaryons from each somatic hybrid combination were paired with tester homokaryons of the initial wild type of P. ostreatus. The changed mating types were identified in progenies. The pattern of mating type switching in somatic hybrids depends on compatibility of fusion partner. There are several factors related to the mechanism of clamp connection formation and fruiting body development of synkaryons. Of these,the major factor may be associated with self-fertility and mating type switching such as homokaryotic fruiting of wild type P. ostreatus. This review will discuss these aspects.

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Characterization and breeding of a new cultivar Pleurotus ostreatus 'Heuksol' (느타리버섯 신품종 '흑솔'의 육성 및 특성)

  • Oh, Min-Ji;Im, Ji-Hoon;Shin, Pyung-Gyun;Oh, Youn-Lee;Jang, Kab-Yeul;Kon, Won-Sik
    • Journal of Mushroom
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    • v.15 no.3
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    • pp.129-133
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    • 2017
  • Oyster mushroom is a type of mushroom that is commonly cultivated and consumed in Korea. P. ostreatus 'Suhan' is a preferred cultivar for many mushroom farmers because it has a dark pileus and thick stipe. However, as it is very sensitive to environmental conditions, farmers consistently demand an alternative cultivar. To develop a new cultivar, the parental strains KMCC01680 ('Suhan') and KMCC00478 ('Gosol') were selected from various collected P. ostreatus strains by cultivating genetic resources. P. ostreatus 'Heuksol' was developed by the method of Mon-Mon crossing between monokaryotic strains derived from 'Suhan' and 'Gosol'. Thirty strains of 174 crossed strains were initially selected by cultivation experiments. After bulk cultivation tests, 'Heuksol' was selected. The nuclear DNA profile of 'Heuksol' was similar to those of the parental strains, 'Suhan' and 'Gosol', when RAPD (random amplified polymorphic DNA) primers and UPF (Universal PCR Fingerprinting) 2, 3, and 4 were used. The optimum temperature for mycelial growth was $30^{\circ}C$ for 'Heuksol', but medium-high temperatures were also appropriate, especially $13-20^{\circ}C$. The fruiting body production per bottle (1,100 mL) was approximately 140.8 g. When compared to the control strain 'Suhan', the thickness of the stipe of 'Heuksol' was greater than that of 'Suhan' (13.5 mm vs 9.4 mm). The pileus diameter of 'Heuksol' was similar to that of 'Suhan' and the pileus thickness of 'Heuksol' and 'Suhan' was 19.7 mm and 21.8 mm, respectively. 'Heuksol' had more a productive stipe number than 'Suhan' and the pileus of 'Heuksol' was dark gray, even at high temperatures. Therefore, it was suggested that this new cultivar, 'Heuksol', could provide an alternative to 'Suhan' and contribute to the profit of oyster mushroom farms.