• Title/Summary/Keyword: Random amplified polymorphic DNA

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Molecular Typing of Staphylococcus aureus Strains from Domestic Animals and Humans by REP-PCR Analysis (REP-PCR을 이용한 국내 사람과 동물유래 Staphylococcus aureus 분리주의 Molecular Typing)

  • Woo Yong-Ku;Kim Shin
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.60-66
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    • 2005
  • To select the rapid and efficient molecular subtyping method for epidemiologic monitoring of Staphylococcus aureus (S. aureus) strains at clinical laboratory levels, a total 116 of S. aureus and MRSA (methicillin-resistant S. aureus) strains from diverse animal species [Korean cattle, goat, pig, dog, chicken, mouse] and also humans were analyzed. To evaluate the discriminatory ability (DA) of individual PCR methods, random amplified polymorphic of DNA [RAPD; 4M & RA primer], repetitive extragenic palindromic sequences PCR (REP-PCR), and enterobacterial repetitive intergenic consensus sequences PCR (ERIC-PCR) methods were conducted and then compared on their Simpson's index of diversity (SID) values based on the dendrogram patterns, which was produced by software program (BiolD2+ & GelCompar II). In first, RAPD using the 4M primer (SID 0.915) was expressed more higher SID value than that of RA primer (SID 0.874). 4M primer was expressed more powerful DA than RA. Both REP-PCR (SID 0.930) and ERIC-PCR (SID 0.929) methods showed much more higher DA than that of RAPD. According to the present results, both REP-PCR and ERIC-PCR among the tested analysis methods were found as the most reliable and discriminative molecular subtyping method, because they expressed the highest DA for the present S. aureus and MRSA strains.

Analysis of Genetic Diversity of Apple Cultivars Using RAPD and SSR Markers (RAPD와 SSR 마커를 이용한 사과 품종의 유전적 다양성 분석)

  • Cho, Kang-Hee;Heo, Seong;Kim, Jeong-Hee;Shin, Il Sheob;Han, Sang Eun;Kim, Se Hee;Kim, Dae-Hyun;Kim, Hyun Ran
    • Korean Journal of Breeding Science
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    • v.42 no.5
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    • pp.525-533
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    • 2010
  • In this study, random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) analyses were utilized for evaluation of genetic diversity of 34 Korean bred and introduced apple cultivars. Thirty-seven RAPD primers detected a total of 193 polymorphic bands (36.2%) with an average of 5.6. Twenty-six SSR markers generated a total of 112 alleles with an average 4.3 alleles per locus. Genetic diversity of 34 cultivars estimated by polymorphic information content (PIC) value ranged from 0.536 (CH03d12) to 0.952 (CH04c06) with an average of 0.843. By UPGMA (unweighted pair-group method arithmetic average) cluster analysis with 305 polymorphic bands, the apple cultivars were classified four groups by similarity index of 0.640. The 'Seokwang' was included in group I. Group II consisted of 12 cultivars which have 'Golden Delicious' in their pedigree, with the exception of 'Spur Earliblaze' and 'Jonathan'. Group III included 13 cultivars which have usually 'Fuji' in their ancestry and bud sport of 'Fuji' cultivars. Group IV consisted of 8 cultivars with 'Hongro', 'Gamhong', and 'Saenara'. Similarity values among the tested apple cultivars ranged from 0.529 to 0.987, and the average similarity value was 0.647. The similarity index was the highest (0.987) between 'Hwarang' and 'Danhong', and the lowest (0.529) between 'Seokwang' and 'Hwarang'. The genetic relationships among the 34 studied apple cultivars were basically consistent with the known pedigree.

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.

Phylogenetic Relationships and Genetic Diversity in Collected Resources of Carthamus tinctorius by Random Amplified Polymorphic DNA Markers (RAPD 마커에 의한 수집된 홍화자원에서 계통관계와 유전적 다양성)

  • Sung, Jung-Sook;Cho, Gyu-Taek;Lee, Gi-An;Baek, Hyung-Jin;Huh, Man-Kyu
    • Journal of Life Science
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    • v.20 no.12
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    • pp.1764-1771
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    • 2010
  • Carthamus tinctorius L. (Compositae) is an herb primarily distributed throughout in the world. The species is regarded as ecologically important in the world. Safflower was used for medicines, as well as making red (carthamin) and yellow dyes. We have used the RAPD technique to investigate the phylogenetic relationships and genetic diversity of C. tinctorius. We obtained 123 bands from all the 26 cultivars. The average number of bands was 9.5 per primer. The genetic diversity of safflower is found among cultivars and there is a high among-cultivar differentiation. The OPC18-01 band is the specific marker for Syria cultivar, whereas no products were detected in individuals from other country cultivars. We found seven phenetic bands for determining the specific marker of cultivars with SCAR markers. Though the number of individuals sampled for analysis was small and probably not fully representative of the total available diversity in C. tinctorius, this study demonstrates that the regions (Morocco, Syria, and Turkey) of the Mediterranean Sea were more variable than other regions with the exception of India. In this result, although only simple result of RAPD is difficult to assert the center of species diversity of C. tinctorius, the regions of the Mediterranean Sea may be the most probable candidate for the origin of safflower. India was also the candidate of the center or secondary center of species diversity of C. tinctorius. RAPD markers were effective in classifying cultivar levels of safflower.

A study of morphological characteristics and hybridization on Lepista nuda (민자주방망이버섯(Lepista nuda )의 형태적 특성 및 교배에 관한 연구)

  • Jung, Won Soon;Kim, Jong Bong
    • Journal of Mushroom
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    • v.11 no.1
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    • pp.1-8
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    • 2013
  • In this study, twelve of Lepista nuda were collected from various localities in Korea. Also thirteen exotic L. nuda species were collected from Japan, France, Switzerland and Portugal. Spores were isolated under optical microscope. These spores were placed on the surface of YM medium for inducing to germination. Eleven mating-groups were selected by morphological characters of fruit body such as size, color and stipe patterns. Intra-isolate crosses were made between two single-spore isolates derived from mating-groups. Also, dikaryotic crossing using the isolates from L. nuda were carried out to evaluated tetrakaryon formation. Cross-mating compatibility tests also verified its dikaryotic state by microscopic or molecular genetic observation of clamp connection and Random Amplified Polymorphic DNA (RAPD) band pattern. To analyze the growth rate of hybrids and parents mycelium in dikaryons obtained from compatible mating groups were placed on PDA medium. Intra-isolate crosses determined eleven mating-groups within L. nuda. The typical clamp connection were mostly observed in mating-groups of Korean L. nuda in $K1{\times}K2$, $K1{\times}K3$, $K1{\times}K4$, $K1{\times}K6$, $K1{\times}K5$, $K2{\times}K4$, $K2{\times}K3$, $K2{\times}K6$, $K3{\times}K4$, $K4{\times}K5$, and $K4{\times}K6$. Korean L. nuda type of dikaryon, shown to cross-incompatibility with L. sordida, it seemed that mating induce more rapidly than wild types in a view of growth rate. In conclusion, it would be useful to improve mass production with better morphological characteristics through a special mating of L. nuda.

Characterization of a new variety Auricularia polytricha (Mont.) Sacc.'Geoni' (털목이 신품종 '건이')

  • Kim, Kil-Ja;Lee, Sook-Jae;Kim, Myeong-Seok;An, Ho-Sub;Kim, Dong-Gwan;Kwon, Oh-Do;Park, Heung-Gyu
    • Journal of Mushroom
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    • v.14 no.4
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    • pp.211-214
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    • 2016
  • Geoni, a new variety of Auricularia polytricha, was bred in JARES(Jeollanamdo Agricultural Research and Extension Services) in 2012. Geoni was selected through the monospore hybridization with JNM21008 and JNM21014 in 2010. Based on a performance test conducted from 2011 to 2012, the Geoni was selected from a line showing an excellent light brown pileus and strong pest resistance. Geoni has a favorable chewiness, light brown and smooth pileus. In addition, Geoni was rich in dietary fiber. MCM(mushroom complete medium), Malt and PDA(Potato Dextrose Agar) mediums were suitable for cultivating the Geoni. The number of effective stipes was 39 ea/0.9 kg and minor axis and major axis of pileus were 6.9 cm and 8.7 cm respectively. The yield of Geoni was 291 g/0.9 kg in plastic bag. Geoni was required 40~54 days for culture at $20^{\circ}C$ and 24days for the primordia and growth period, which is longer than that of the control(Pung-un). Somatic incompatibility was formed between parental strains and Geoni. Analysis of the genetic diversity of the new variety "Geoni"revealed a different profile from the parental strains when RAPD(random Amplified Polymorphic DNA) primers were used.

Genotypic Variation of Helicobacter pylori Isolated from Gastric Antrum and Body in Korean Patients

  • Park, Seon-Mee;Kwon, Soon-Kil;Son, Bo-Ra;Shin, Kyeong-Seob;Woo, Chan-Won;Kim, Eung-Gook;Kim, Seok-Yong
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.19-29
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    • 2000
  • Although most persons infected with Helicobacter pylori harbor a single strain of the organism, multiple strain colonization in the same patient is also occasionally reported in developed countries. The aims of this study were to determine the prevalence of multiple strain colonization in Korean patients and to detect the cagA, iceA1, and babA status of H. pylori isolated from the antrum and body of the stomach. H. pylori was obtained from 35 patients from the antrum and body of the stomach. The genomic diversity of H. pylori was determined by random amplified polymorphic DNA analysis. The status of cagA, iceA1, and babA genes of H. pylori was assessed by polymerase chain reaction with appropriate primers. Clearly different diversity patterns were identified among the isolates from 35 individual patients. Eighteen (51.4%) patients had a single strain of H. pylori. Eight (22.9%) and nine (25.7%) patients had subtypically (one or two bands difference) and typically (clearly different pattern) different strains of H. pylori in the antrum and body, respectively. Among the 70 isolates of H. pylori from 35 patients, the positive rates of 349-bp and 208-bp cagA gene fragments and the iceA1 gene were 68/70 (97.1%), 68/70 (97.1%), and 58/70 (82.9%), respectively. However, the babA gene was found in 22/66 cases (31.4%). In five out of 18 patients with a single strain, the genetic status of cagA, iceA1, and babA varied between the isolates from the antrum and the body. In 8/17 patients with sub typically or typically different strains, the gene status differed between antrum and body isolates. The prevalence of co-colonization with typically or subtypically different strains is high in Korea, and sub-clones with different pathogenic gene status exist within strains of identical RAPD patterns.

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Application of RAPD Markers to Early Selection of Elite Individuals of Pinus Species for a Clonal Forest Tree Breeding Program (소나무류 육종에 있어 임의 증폭 다형 디엔에이(RAPD)지표를 이용한 우량 임목의 조기 선발)

  • Yi, Jae-Seon;Cheong, Eun-Ju;Moon, Heung-Kyu;Dale, Glenn T.;Teasdale, Robert D.
    • Journal of Forest and Environmental Science
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    • v.11 no.1
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    • pp.81-101
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    • 1995
  • Random amplified polymorphic DNA (RAPD) technology, a recent approach in molecular genetics, is much usable to select the elite trees and to maximize the genetic gain in forest tree breeding program, providing a clue to determine the genetic marker-trait correlation. This review intorduces research on bark thickness and breeding strategy in Pinus elliottii, Pinus caribaea and their hybrid by Queensland Forest Service and ForBio Research Pty Ltd, University of Queensland, which employ RAPD technology. Genetic linkage map of $F_1$ hybrids includes 186 RAPD markers and 16 linkage groups (1641 cM long in total) and 6 quantitative trait loci are located putatively for bark thickness. Following recent research results and experiences in pine breeding programs, the forseeable stages in the application and development are proposed for marker assisted selectin; stage 1-determination of species specific markers for genes controlling traits of commercial interest, and stage 2-determination of marker-allele association for specific allelic variants within pure species. As pines inherit their megagametophytes from the seed parent and zygotic embryos from both male and female parents, the determination of marker-trait correlation is possible even in embryo stage, eventually making ways for the early selection of elite individuals.

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Characterization of a new cultivar of Auricularia auricula-judae 'Yong-A' (목이 신품종 '용아'의 특성)

  • Kim, Kil-Ja;Kim, Dami;Lee, Sook-Jae;An, Ho-Sub;Kim, Dong-Kwan;Kwon, Oh-Do
    • Journal of Mushroom
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    • v.16 no.3
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    • pp.198-202
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    • 2018
  • 'Yong-A' was bred by Jeollanamdo Agricultural Research and Extension Services in 2015. It was primarily selected through monospore crossing between JNM21008 and JNM21013 in 2009. The strain is named as JNM-Mi-194 and 'Yong-A', showing major characteristics with good production yield. The cultivation characteristics, yield potential, and fruiting body characteristics of 'Yong-A' are as follows; The wrinkle of the fruiting body was of branching type its shape was wave type. MCM, YM, Malt, and PDA media were suitable for growth of this cultivar. The number of effective stipes was 13 ea/0.9 kg. The minor axis of pileus was 5 cm and the major axis was 9 cm. The yield was 291 g per plastic bag (0.9 kg). Yong-A required 60 days for mycelial running at $20^{\circ}C$. The growth and primordial period required 24 days, which was shorter than that of JNM-Mi-21002 (the conventional cultivar). Somatic incompatibility was observed between parental lines and Yong-A. Analysis of genetic diversity in the new variety 'Yong-A' showed a different profile compared to that of the parent strain when Random Amplified Polymorphic DNA primers were used.

Determination of paraquat-resistant biotype on Conyza canadensis and the resistant mechanism (Paraquat 저항성 생태형 망초의 선발과 저항성 기작)

  • Kim, Sung-Eun;Kim, Seung-Yong;Ahn, Sul-Hwa;Chun, Jae-Chul
    • The Korean Journal of Pesticide Science
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    • v.9 no.1
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    • pp.88-96
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    • 2005
  • Paraquat-resistant biotype of Conyza canadensis (L.) Cronq. was determined by chlorophyll loss and random amplified polymorphic DNA (RAPD) analysis and the resistant mechanism was investigated with respect to absorption, translocation, and binding constant. RAPD analysis for paraquat resistant (R) and susceptible (S) biotypes found in a pear orchard revealed that the biotypes possessed remote genetic relationship. Chlorophyll loss, as an indication of paraquat toxicity, of S biotype was 7.8-fold greater than that of R biotype. There were no differences in contents of epicuticular wax and cuticle and amounts of [14C]paraquat penetrating the cuticle between the two biotypes. Little translocation of the herbicide out of the treated leaf was observed in either biotype. Binding constants of paraquat to the cell wall and thylakoid membrane were 7.4-fold and 16.9-fold, respectively, higher in R biotype than in S biotype. The results suggest that the resistance mechanism of C. canadensis biotype is due partly to high binding affinity of paraquat to the cell wall and thylakoid membrane.