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

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Chinese Cabbage Club root Pathogen, Plasmodiophora brassicae, Is Genetically Stable

  • Heo, Seung-Hwan;Jang, Se-Jeong;Choi, Jin-Soo;Jang, Chang-Soon;Song, Jeong-Young;Kim, Hong-Gi
    • Mycobiology
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    • v.37 no.3
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    • pp.225-229
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    • 2009
  • Single spore isolates of Plasmodiophora brassicae e4 and e9 obtained from diseased Chinese cabbage were identified as race 4 and race 9, respectively, by the Williams' differential variety set. To confirm the possibility of variation in same generation and progeny of a single spore isolate of P. brassicae, random amplified polymorphic DNA (RAPD) analysis was conducted using the URP 3, 6 and OPA 7 primers. There was no difference in band type at each part of the gall of Chinese cabbage obtained by inoculation of e4 and e9 and amplification using the URP 3 and 6 primers when the same generation was analyzed. In addition, the progeny analysis, which was expanded to the third generation and conducted using the URP 3 and OPA 7 primers, revealed no differences in the band type of the e4 isolate. Based on these results, the single spore isolate of P. brassicae was genetically stable.

Identification of Phellinus linteus by Comparison of Colony Shapes and Using PCR techniques (목질진흙버섯(Phellinus linteus)의 균총형태 비교 및 PCR 기법을 이용한 동정)

  • Kong, Won-Sik;Kim, Dong-Hyun;You, Chang-Hyun;Kim, Young-Ho;Kim, Kyung-Soo;Kim, Kwang-Ho
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.466-477
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    • 1998
  • Twenty-two Phellinus strains were characterized using colony morphologies and polymerase chain reaction (PCR) to divide into Phellinus linteus. There were some differences in mycelial growth and colony shapes among the strains when they were grown on various media such as PDA, MCM, MEA and YM. Phellinus linteus was slowly growing, formed golden-yellow colony, and produced blue pigment on PDA media. When the regions of internal transcribed spacer (ITS) were amplified from ribosomal RNA (rRNA) coding genes of P. igniarius and P. linteus strains by means of PCR, two types of band (700 bp and 800 bp) were appeared, respectively. For the amplified intergenic region I (IGRI), P. igniarius strains showed a different band among 500, 600, 700 and 800 bp according to the strains, whereas P. linteus strains did one specific band of 700 bp. By polymorphism analysis after digesting the amplified products with 6 different restriction enzymes, a band specific to P. linteus was generated when the products for ITS region were digested with HaeIII, suggesting that the enzyme digestion could provide effective method to distinguish between P. igniarius and P. linteus. And also, the analysis of genetic relationship showed that the genetic similarities were 89% and 95% in P. igniarius and P. linteus strains, respectively. Random amplification polymorphic DNA (RAPD) analysis using multiple primer sets and arbitrarily primed PCR (AP-PCR) with ITS3 primer could also result in a reproducible way to identify P. linteus strains.

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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.

Classification of Safflower(Carthamus tinctorious L.) Collections by RAPD Analysis (RAPD 분석에 의한 홍화의 품종군 분류)

  • Bang, Kyong-Hwan;Kim, Young-Guk;Park, Hee-Woon;Seong, Nak-Sul;Cho, Joon-Hyeong;Kim, Hong-Sig;Cho, Yong-Gu
    • Korean Journal of Medicinal Crop Science
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    • v.9 no.3
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    • pp.225-231
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    • 2001
  • This studies were conducted to provide the basic information on safflower collections and to identify the variations which could be utilized in safflower breeding programs, The RAPDs was used to clarify the genetic relationships among safflower collections and to classify them into distint genetic groups. Among 30 of 10 mer primers in RAPD analysis, twenty were selected as the appropriate primers for identification of the genetic characters in safflower collections. Amplified PCR showed the highly reproducible bands at $3.0{\sim}0.2kb$. The number of bands amplified with the each primer showed the variations ranged from 2 to 11, with the average of 5.6. Total of 111 bands were identified among 20 selected primers used in PCR reaction and 84 bands (75.7%) showed polymorphism. Based on the similarity value of 0.14 in dendrogram derived from the cluster analysis using RAPD-PCR, the 30 safflower collections were classified into 11 groups. The two main groups, VII and VIII included 7 collections (23%) and 8 collections (27%), respectively. Most of the collections in group VII were the Korean collections (85%).

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Phylogenetic Analysis of the Genus Dendronephthya (Nephtheidae, Alcyonacea) Based on Internal Transcribed Spacer Sequences of Nuclear rDNA

  • Lee, Young-Ja;Song, Jun-Im
    • Animal cells and systems
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    • v.4 no.4
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    • pp.319-324
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    • 2000
  • Species boundaries among the Alcyonacean soft coral, the genus Dendronephthya, are often obscured by inter- and intraspecific morphological variations. In the present study, we attempted to infer the genetic relationships of eight dendronephthians based on their molecular characters, the internal transcribed spacer (ITS) regions of ribosomal DNA, and then compared this result together with the random amplified polymorphic DNA (RAPD) data from our previous investigation. Dendronephthya. putteri and D. suensoni formed a divaricate form - VI grade specific clade, whereas D. castanea, D. gigantea, D. aurea and D. spinifera, formed a umbellate and glomerate form - IV and III grade specific clade. Therefore, we confirmed that the main characters the growth form and the anthocodial grade and formula, are important in identification of the species in dendronephthians despite some problems. Also, the relationships of the growth form are clarified as the glomerate form is much closer to the umbellate form than to the divaricate form based on two sets of independent molecular data. However, we cannot determine the molecular markers which limit the species boundaries among this genus with ITS sequences.

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Genetic Variation in Among Cultivated Field Populations of Korean Ginseng(Panax ginseng C.A.Meyer) Using RAPD (RAPD marker를 이용한 고려인삼(Panax ginseng C.A.Meyer)의 유전적 변이 분석)

  • 차선경;김영창;최재을;최장선;강권규
    • Korean Journal of Plant Resources
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    • v.16 no.3
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    • pp.251-256
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    • 2003
  • Genetic variation in field grown Korean ginseng(Panax ginseng C.A.Meyer) was evaluated using random amplified polymorphic(RAPD) markers. (This experiment was carried to collect the local native from farm of Chungnam National University in Korea in order to investigate genetic variation.) Some morphological characters showed considerable variation ranging 22 to 68cm in plant hight, 10 to 38mm in root diameter, 16 to 86g in root weight, and culum color and flowering date, respectively. Ten RAPD primers out of the 32 which produced reproducible bands in 662 Korean ginseng plants were selected for the further study. The total number of bands generated by 10 primers were 108 and among them 103 were polymorphic among the 662 plants with the polymorphism ratio of 94.5%. A total of 662 plants were classified into 16 groups based on polymorphic data with an URP 05 primer.

Population Genetic Analyses of Gibberella fujikuroi Isolated from maize in Korea

  • Park, Sook-Young;Seo, Jeong-Ah;Lee, Yin-Won;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • v.17 no.5
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    • pp.281-289
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    • 2001
  • We analyzed 88 strains of Gibberella fujikuroi (Analmorph: Fusarium section Liseola) from maize in Korea for mating population, mating type, fumonisin production vegetative compatibility, and random amplified polymorphic DNA (RAPD) patterns. We found 50 strains that were MATA-2, 22 that were MATA-1, 1 that was MATD-1, and 15 that were not reproducibly fertile with any of the mating type testers. Of the 50 MATA-2, 15 were female fertile, while 10 of the 22 MATA-1 strains were female fertile. A total of 1,138 nitrate non-utilizing (nit) mutants were recovered from a total of 88 strains. These strains were grouped into 39 vegetative compatability groups (VCGs) by demonstrating heterokaryosis between nit mutants. A single maize ear could be infected by more than one VCG of F. moniliforme. RAPD analysis measured genetic diversity among 63 strains of F. moniliforme. Several VCGs were distinguished by RAPD fingerprinting patterns. Most strains produced significant levels of fumonisins. However, 6 MATA-2 strains from a single VCG produced higher levels of fumonisin $\textrm{B}_3$ than that of fumonisin $\textrm{B}_1$ or $\textrm{B}_2$. From these data, we concluded that most Korean strains of F. moniliforme associated with maize belonged to mating population A and produced significant levels of fumonisins. Futhermore, RAPD analysis could differentiate strains associated with different VCGs.

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Identification of Fusarium Species Associated with Corn Ear Rot (옥수수 이삭썩음병에 관여하는 Fusarium속균의 동정)

  • Choi, Hyo-Won;Kim, Jung-Mi;Kim, Jin-Hee;Hong, Sung-kee;Kim, Wan-Gyu;Chun, Se-Chul
    • The Korean Journal of Mycology
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    • v.37 no.2
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    • pp.121-129
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    • 2009
  • In 2007, a total of 77 isolates of Fusarium spp. were obtained from ear rot symptoms of corns collected from 5 locations in Gangwon Province, Korea. The fungal isolates were identified based on their morphological features. Out of the isolates, fifteen isolates were identified as Fusarium verticillioides which formed microconidia in long chains on monophialides. Four isolates were identified as F. subglutinans which formed microconida only on false heads. Six isolates were identified as F. graminearum which produced red pigment in PDA culture. Besides these Fusarium species, F. napiform, F. nygamai, and F. oxysporum were identified from the rest isolates. To assess for genetic diversity of the isolates, a random amplified polymorphic DNA(RAPD) technique was carried out using URP primers. The results from the RAPD analysis showed that the isolates from corn were divided into 6 groups. These RAPD groups of the Fusarium species corresponded to morphological characters of the Fusarium species. The phylogenetic analysis of most isolates by DNA sequencing of EF-1$\alpha$ gene corresponded to morphological characters of the Fusarium species. The results of pathogenicity tests by two inoculation methods revealed that F. verticillioides, F. graminearum and F. subglutinans are strongly pathogenic to corn stalks.

Characteristics of Dioscorea alata L. Introduced from Tropical and Subtropical Regions (도입 마(Dioscorea alata L.)의 특성 분석)

  • Chang, K.J.;Yoo, K.O.;Park, C.H.;Park, J.I.;Hong, K.H.;Park, J.H.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.3 no.1
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    • pp.48-69
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    • 2001
  • A lot of clones of the genus Dioscorea have been introduced from some tropical and subtropical regions since 1997. In 33 clones of water yams (Dioscorea alata L.), some morphological characteristics were investigated at the field. Variation ranges of the total weight and tuber number per stump were within the ranges from 90 to 2,147 g with an average of 610 g ; and 1.3-4.7 with an average of 2.8, respectively. The color tones observed on the tuber-flesh were sorted into 3 color-categories, i.e., white, pale brown and pale purple, and those on leaves were sorted into 3 color-categories, i.e., green, heavy green and purplish green. Intraspecific genetic relationship of 19 variation types of the Yam classified by their external morphological characteristics such as leaf and tuber shape was assessed by DNA using random and specific primers. Twenty two out of 113 primers (100 random[10-mer] primers, two 15 mer [M13 core sequence, and (GGAT)4 sequence]) had been used in PCR-amplification. Only 12 primers, however, were successful in DNA amplification in all of the analyzed plants, resulting in 93 randomly and specifically amplified DNA fragments. The analyzed taxa showed very high polymorphisms(69 bands, 71.0%), allowing individual taxon to be identified based on DNA fingerprinting. Monomorphic bands among total amplified DNA bands of each primer was low under the 50%. Similarity indices between accessions were computed from PCR(polymerase chain reaction) data, and genetic relationships among intraspecific variations were closely related at the levels ranging from 0.66 to 0.90.

Effects of Genotypes on In Vitro Maturation and Fertilization of Frozen-Thawed Porcine Oocytes

  • Jia Y. H.;Jin H. J.;Wee M. S.;Cheong H. T.;Yang B. K.;Park C. K.
    • Reproductive and Developmental Biology
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    • v.29 no.4
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    • pp.207-212
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    • 2005
  • In the present study, we investigated the effects of genotypes on in vitro maturation and fertilization in porcine fresh/frozen-thawed oocytes. The porcine cumulus-oocyte complexes (COCs) were divided into four groups according to whether they were: (1) in vitro matured; (2) cryopreserved and in vitro matured; (3) in vitro fertilized and (4) cryopreserved, and in vitro fertilized. Maturation of porcine COCs was accomplished by incubation in NCSU23 medium. Immature oocytes were cryopreserved by Open Pulled Straws (OPS) method according to Vajta et al., (1998). Oocytes stained by Acetic-Orcein method were observed under the microscope. DNA extracted from the ovaries was analyzed by RAPD (random amplified polymorphic DNA) and SSCP (single strand conformational polymorphisrrt) method. The rates of oocytes maturation and fertilization were significantly high in AA genotype. The results indicated that in vitro maturation and fertilization in porcine fresh/frozen-thawed oocytes may be affected by genotypes in pigs.