• Title/Summary/Keyword: RAPD 프라이머

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Evaluation of the Genetic Diversity of Biovar 3 Strains of Pseudomonas syringae pv. actinidiae Isolated in Korea (RAPD 지문을 통한 우리나라에서 분리된 Pseudomonas syringae pv. actinidiae biovar 3 균주의 유전적 다양성 평가)

  • Lee, Young Sun;Kim, Gyoung Hee;Koh, Young Jin;Jung, Jae Sung
    • Journal of Life Science
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    • v.30 no.1
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    • pp.1-9
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    • 2020
  • Pseudomonas syringae pv. actinidiae, the causal agent of a bacterial canker disease in kiwifruit, is subdivided into five genetically distinct populations, namely biovars 1, 2, 3, 5, and 6. Of these, strains belonging to biovar 3 are responsible for a pandemic bacterial canker of kiwifruits since 2008. This study aimed to characterize the structure of the biovar 3 population and investigate the origin of biovar 3 strains isolated in Korea. The genetic variability of fifteen biovar 3 strains, thirteen Korean and two Chinese, were evaluated through random amplified polymorphic DNA (RAPD)-PCR. The RAPD results revealed the presence of eight lineages, designated as subgroups I-VIII, across the biovar 3 strains used in this study. As the strains in subgroups II and III from China were not found in the Korean examples, we concluded that six genetically different biovar 3 subgroups (I, IV, V, VI, VII, and VIII) are present in Korea. In PCR analysis using primers specific to the strains of New Zealand and Europe, Korean strains in subgroups V and VI amplified the relevant DNA bands, suggesting that these were introduced from these two origins, respectively. PCR primers specific to subgroup VIII were developed to monitor the spread of the first biovar 3 strain in Korea, and investigations revealed that this strain was not found in Korea after its first occurrence.

Primer for the Potato Specific Internal Control DNA and Screening Method for the Genetically Modified Potatoes by Competitive Duplex-PCR (감자 특이 Internal Control DNA 증폭용 Primer와 이를 이용한 유전자 변형 감자의 경쟁적 이중 PCR 검정법)

  • Seo, Hyo-Won;Yi, Jung-Yoon;Cho, Hyun-Mook;Kim, Sung-Yeul
    • Journal of Plant Biotechnology
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    • v.29 no.4
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    • pp.235-240
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    • 2002
  • We report the new method for the screening of genetically modified potato by competitive duplex-PCR using the potato specific single oligomer primer for the internal control and CaMV 35S promoter or NOS terminator specific primers. The single oligomer primer (rAGU4A) amplify the potato specific internal control band from the homozygous potato genomic DNA in the RAPD profiles of all analyzed potato varieties. The 530 bp internal control DNA was amplified independently to CaMV 35S promoter or NOS terminator DNA and identified as repetitive or microsatellite DNA of potato (AF541972). With this new technique, the transgenic potatoes which were transformed with vectors contained the different foreign genes are analyzed. In case of the commercialized transgenic potato varieties, 'Hew Leafs', those two genetic factors are used for promoter and terminator respectively So, this new PCR technique should be a promising method of cost effective and accurate screening for the commercialized GM potatoes on market.

Genomic Fingerprinting Patterns of Bifidobacteria Isolated from Healthy Koreans Using ERIC-, TAP-, and BOX-PCR (건강한 한국인으로부터 분리된 비피도박테리아의 ERIC-, TAP-, BOX- 중합효소연쇄반응을 이용한 유전자 지문 분석)

  • Lee, Do-Kyung;Kang, Byung-Yong;Chung, Myung-Jun;Lee, Kang-Oh;Kim, Kyung-Jae;Ha, Nam-Joo
    • Environmental Analysis Health and Toxicology
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    • v.23 no.1
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    • pp.1-9
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    • 2008
  • 유산균인 비피도박테리아는 사람과 동물에서 유익한 프로바이오틱 미생물로 알려져 있다. 본 연구에서는 이러한 비피도박테리아 균주의 분류를 위한 repetitive DNA element PCR fingerprinting (ERIC-또는 TAP-PCR)의 사용을 평가하였다. 사람분변으로부터 분리한 알려지지 않은 비피도박테리움 균주와 한국생명공학연구원 생물자원센터로부터 분양받은 표준균주를 가지고 분류 및 동정에 ERIC-PCR과 TAP-PCR을 이용한 RAPD-fingerprinting을 수행하였다. 그 결과 비피도박테리움 균주에 대한 속과 종단위의 분류가 가능하였으며, 실험에 사용된 모든 비피도박테리움 균주는 RAPD-fingerprinting 분석을 통해 유전적 다양성을 확인하였다. 또한 ERIC2와 TAP1 프라이머를 이용한 실험에서는 Bifidobacterium adolescenits 특이 유전자 단편을 확인하였으며 이는 B. adolescenits 균주의 동정에 유용할 것으로 사료된다.

Morphological Characteristics of Medium-Leaf Type Zoysiagrasses (Zoysia spp.) and Their Classification Using RAPDs (중엽형 한국잔디(Zoysia spp.)류의 형태적 특성과 RAPDs를 이용한 분류)

  • Choi, Joon-Soo
    • Asian Journal of Turfgrass Science
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    • v.24 no.2
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    • pp.88-96
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    • 2010
  • Zoysiagrass, especially Jungji implicating medium-leaf type zoysiagrass has been widely used in South Korea recently. This study was carried out to classify 36 selected medium-leaf type zoysiagrass accessions compared to 5 basic zoysiagrass species using RAPDs. Morphological characteristics such as leaf width, leaf angle, leaf sheath length, existance of trichomes and stolon color were measured as useful characteristics for identification of species in Zoysia genus. Nineteen RAPD markers were identified using 8 selected random primers. The dissimilarity coefficient of variants ranged from 0 to 0.736. Three zoysiagrass groups were clustered by dissimilarity coefficient analysis. Group 1 consisted of Z. japonica and some US varieties including 'Zenith' and 'Meyer'. Group 2 consisted of Z. sinica, Z. macrostachya and Korean commercial varieties such as 'Anyang', 'Samdeock', and 'Pyeongdong' medium-leaf type grasses. Group 3 was genetically distinct from Group 1 and Group 2, and included Z. matrella and Z. tenuifolia. 'Anyang', 'Samdeock', and 'Pyeongdong' medium-leaf type zoysiagrasses showed very close genetic relationship with Z. sinica and Z. macrostachya.

Analysis of Gene Diversity and Phenetic Relationship of Water Dropwort Species in Korea Using RAPD (OPB Primers) Markers (한국 미나리 집단에 대한 RAPD (OPB 프라이머)에 의한 유전적 다양성과 표현형 관계)

  • Huh, Man Kyu
    • Journal of Life Science
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    • v.32 no.8
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    • pp.595-600
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    • 2022
  • Water dropworts, Oenanthe javanica and O. javanica var. japonica are called "minari" in Korea and are eaten as a vegetable. Cicuta virosa is common European water hemlock and has toxic properties, such as cicutoxin. Molecular variations of water dropwort species in Korea were investigated using random amplified polymorphic DNA (RAPD). The six populations were studied with 10 primers (Operon, OPB) for RAPD analysis. The 72 DNA fragments (bands) were found among six populations. Among these 72 bands, 61 (84.7%) bands were polymorphic. The typical populations of Cicuta virosa in Korea were small, isolated, and patchily distributed for natural populations and they maintained a high level of genetic diversity. However, when cultivated populations of O. javanica var. japonica were large and widely grown in rice paddies as vegetables, they maintained a lower genetic diversity than those of C. virosa and wild populations of O. javanica. Although the diversity indices of wild populations were shown to be higher than those of cultivated populations, no significant difference for measures of genetic variability was shown. Total genetic diversity value (HT) was 0.342. The interlocus variation in the within-population genetic diversity (HS) was 0.201. The proportion of total genetic variation due to differences among populations (GST) range was 0.414, indicating that 41.4% of the total variation was among populations. In conclusion, the RAPD technique was a useful method for discrimination between C. virosa and O. javanica. In addition, RAPD-OPB markers could further distinguish the strains from different food sources.

Hybridity Verification of Progenies Obtained from Ovule Culture by Using RAPD Markers in Reciprocal Crosses of Alstroemeria (알스트로메리아 배주배양을 통하여 획득한 정역교배 자손의 혼종성 분석)

  • Lee, Ja-Hyun;Joung, Youn-Hwa;Han, Tae-Ho
    • FLOWER RESEARCH JOURNAL
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    • v.19 no.4
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    • pp.231-237
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    • 2011
  • In this study, we performed ovule culture after reciprocal crosses of two Alstroemeria accessions and investigated genetic contribution of parents by using RAPD markers. The best method was half-ovule culture on MS medium supplemented with $60g{\cdot}L^{-1}$ sucrose and $2.2g{\cdot}L^{-1}$ gelrite at 14 days after pollination. Embryos began to germinate after 6 weeks of culture. The complete plantlets were formed after 4 months of culture. In eight progenies and two parental cultivars, 59 polymorphic bands were obtained out of 89 total bands by RAPD analysis using 7 primers. Eight $F_1$ progenies from the crosses between two accessions using reciprocal crosses showed 1:1 contribution of maternal and paternal parents. It is confirmed that $F_1$ progenies were obtained from parental accessions by using RAPD markers. We conclude this cross combination showed pre-fertilization barriers with incompatibility between stigma or style, and pollen because progeny number was different in each cross combination. Thereby, it warrants overcoming pre-fertilization barrier together with post-fertilization barrier in order to broaden the heterozygosity within progeny populations in Alstroemeria breeding program.

Analysis of Genetic Relationships of Grapevine Cultivars (Vitis ssp.) in Korea Using RAPD Markers (RAPD를 이용한 한국 포도 품종의 계통유연관계 분석)

  • Yoo, Ki Yeol;Cho, Kang-Hee;Shin, Il-Sheob;Kim, Jeong Hee;Heo, Seong;Noh, Jung Ho;Kim, Hyun Ran
    • Korean Journal of Breeding Science
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    • v.41 no.4
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    • pp.437-443
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    • 2009
  • In this study, we used the random amplified polymorphic DNA (RAPD) technique to evaluate the genetic relationships among 29 grapevine cultivars (Vitis spp.). Sixty selective primers detected a total of 558 polymorphic bands. By UPGMA (unweighted pair-group method arithmetic average) cluster analysis with 558 polymorphic bands, the 29 grapevine cultivars were divided into six major groups at 58.8% genetic similarity. The "Super Hamburg" was clustered in group I. Group II consisted of "Wonkyo RA-23", "Muscat Hamburg", "Tano Red", and "Tankeumchu". Group III consisted of "Alden", "Wonkyo RA -21", "Wonkyo RA-30", and "Dutchess". Group IV included 14 grapevine cultivars ("Heukgoosul", "Heukbosuk", "Suok", "Wonkyo RA-29", "Wonkyo RA-22", "Kyoho", "Pione", "Beniizu", "Golden Muscat", "Jinok", "Doonuri", "Campbell Early", "Delaware", and "Schuyler"). Group V consisted of "Hongdan", "Tamnara", "Hongisul", and "Himrod seedless". Group VI included 2 cultivars ("Cheongsoo", and "S. 9110").

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.

Discrimination of Korean Apple Cultivars Using Combination of RAPD-SCAR Markers (RAPD-SCAR 마커 조합을 이용한 국내 육성 사과 품종 판별)

  • Cho, Kang-Hee;Heo, Seong;Kim, Hyun-Ran;Kim, Jeong-Hee;Shin, Il-Sheob;Han, Sang-Eun;Kim, Se-Hee;Kim, Dae-Hyun
    • Horticultural Science & Technology
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    • v.28 no.5
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    • pp.828-835
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    • 2010
  • Conventional methods for identification of apple cultivars are based on the evaluation of sets of morphological characteristics, however, closely related cultivars often cannot be distinguished by morphological traits. This study was conducted to develop DNA markers for discrimination of the apple cultivars bred in Korea. Thirty random primers generated eighty-three random amplified polymorphic DNA (RAPD) markers from thirty-one Korean bred and introduced apple cultivars. Fifty-two RAPD fragments were cloned and sequenced for conversion into sequence characterized amplified region (SCAR) markers. Among them only seventeen SCAR markers resulted in the amplification of single major bands the same size as the RAPD fragment cloned. Several combinations of six (AN11_433, AN08_566, A408_592, AK17_653, AO04_711, AO04_779 or AW15_368, AN11_433, A408_592, AK17_653, AO04_711, AO04_779, or AL1_427, AN11_433, AN08_566, A408_592, AK17_653, AO04_779) to seven (AL1_427, AN11_433, AN08_566, A408_592, AK17_653, AM16_708, AO04_779 or A330_424, AN11_433, AG14_502, AN08_566, A408_592, AK17_653, AO04_779 or A330_424, AN11_433, AK14_564, A408_592, AK17_653, AM16_708, AT14_789) SCAR markers provided enough polymorphism to identify sixteen Korean apple cultivars among thirty-one tested cultivars. Therefore, application of the seventeen SCAR markers was sufficient to identify the thirty-one tested apple cultivars. These markers could be utilized as a reliable tool for cultivar discrimination of Korean apples.

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.