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

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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.

RAPD Loci for Seed Protein and Oil Content in Soybean (Glycine max)

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    • Korean Journal of Plant Resources
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    • v.10 no.3
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    • pp.247-249
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    • 1997
  • Seed protein and oil content is important trait in the soybean. Both seed protein and oil content in this plant species is inherited quantitatively. A 68-plant $F_2$ segregation population derived from a mating between Mercury and PI 467.468 was evaluated with random amplified polymorphic DNA (RAPD) markers to identify QTL related to seed protein and oil content. Marker OPB12 was found to be associated with differences in seed protein content. Four markers, OPA09b, OPM07b, OPC14, and OPN11b had highly significant effects on seed oil content. By interval mapping, the interval between marker OPK3c and OPQ1b on linkage group 13 contained a QTL that explained 25.7% variation for seed oil content.

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Efficiency of RAPD and ISSR Markers in Differentiation of Homo- and Heterokaryotic Protoclones of Agaricus bisporus

  • Mahmudul, Islam Nazrul;Bian, Yin-Bing
    • Journal of Microbiology and Biotechnology
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    • v.20 no.4
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    • pp.683-692
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    • 2010
  • Morphologically, nine different slow-growing protoclones were screened from regenerated protoplasts of heterokaryotic Agaricus bisporus. As such, the present study is the first report on differentiating homo- and heterokaryotic protoclones using random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers. Among 80 primers tested, the seven ISSR and seven RAPD primers selected for the analysis generated a total of 94 ISSR and 52 RAPD fragments, respectively. The ISSR fingerprinting also detected more polymorphic loci (38.29%) than the RAPD fingerprinting (34.61%). A principal coordinate analysis (PCA) was employed to evaluate the resolving power of the markers as regards differentiating protoclones. As a result, the mean polymorphism information content (PIC) for each marker system (i.e., 0.787 for RAPD and 0.916 for ISSR) suggested that ISSR is more effective for determining polymorphisms. The dendrograms constructed using RAPD, ISSR, and an integrated RAPD and ISSR marker system were highly correlated with one another as revealed by a high Mantel correlation (r= 0.98). The pairwise similarity index values also ranged from 0.64 to 0.95 (RAPD), 0.67 to 0.98 (ISSR), and 0.67 to 0.98 (RAPD and ISSR), whereas the mean similarity index values of 0.82, 0.81, and 0.84 were obtained for the RAPD, ISSR, and combined data, respectively. As there was a good correspondence between the RAPD and ISSR similarity matrices, ISSR would appear to be an effective alternative to RAPD in the genetic diversity assessment and accurate differentiation of homo- and heterokaryotic protoclones of A. bisporus.

Analysis of Migration of the Oriental Fruit Moth, Grapholita molesta, in Apple-Cultivating Areas Based on Population Monitoring Using Sex Pheromone and RAPD Molecular Marker (성페로몬과 RAPD 분자지표를 이용한 사과 재배지 복숭아순나방(Grapholita molesta)발생 모니터링과 집단 이동 분석)

  • Kim, Yong-Gyun;Bae, Sung-Woo;Son, Ye-Rim;Park, Jung-A
    • Korean journal of applied entomology
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    • v.48 no.2
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    • pp.211-219
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    • 2009
  • Local and seasonal populations of the oriental fruit moth, Grapholita molesta, were monitored with sex pheromone trapping and RAPD (random amplified polymorphic DNA) molecular marker to analyze their movement in apple orchards. To detect their movements among farms, pheromone traps were placed at regions between apple farms ('outside-farms') as well as within-farms ('inside-farms'). Four seasonal adult peaks were evident in apple-cultivating fields from April to October in both trappings of inside- or outside-farms. After overwintering generation, populations of inside-farms were significantly reduced with frequent insecticide applications, compared to populations of outside-farms. Within apple farms, G. molesta tended to be unevenly distributed because of significant sublocal preference. Active movements of local and seasonal populations of G. molesta were supported by gene flow analysis using RAPD marker. Monitoring data using sex pheromone and seasonal reduction in initial genetic differentiation detected in the overwintering populations suggest that there must be significant movement of G. molesta among different orchards in apple-cultivating areas.

Genetic Diversity Based on Morphology and RAPD Analysis in Vegetable Soybean

  • Srinives, P.;Chowdhury, A.K.;Tongpamnak, P.;Saksoong, P.
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.46 no.2
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    • pp.112-120
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    • 2001
  • Genetic diversity of 47 East-Asian vegetable soybean was characterized by means of agro-morphological traits and RAPD markers. A field trial was conducted to evaluate 14 agro-morphological traits. To study RAPD-based DNA analysis, a total of sixty 10-mer random primers were screened. Of these, 23 polymorphic markers in 16 varieties used for screening. Among 207 markers amplified, 48 were polymorphic for at least one pairwise comparison within the 47 varieties. A higher differentiation level between varieties was observed by using RAPD markers compared to morphological markers. Correspondence analysis using both types of marker showed that RAPD data could fully discriminate between all varieties, whereas morphological markers could not achieve a complete discrimination. Genetic distances between the varieties were estimated from simple matching coefficients, ranged from 0.0 to 0.640 with an average of 0.295$\pm$0.131 for morphological traits and 0.042 to 0.625 with an average of 0.336$\pm$0.099 for RAPD data, respectively. Cluster analysis based on genetic dissimilarity of these varieties gave rise to 4 distinct groups. The clustering results based on RAPDs did not match with those based on morphological traits. Geographical distribution of most varieties in each of the groups were not well defined. The results suggested that the level of genetic diversity within this group of East-Asian vegetable soybean varieties was sufficient for a breeding program and can be used to establish genetic relationships among them with unknown or unrelated pedigrees.

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Analysis of Genetic Diversity in Soybean Varieties Using RAPD Markers (사료작물로 이용이 가능한 한국 재배콩의 RAPD 표지인자에 의한 유전적 다양성 분석)

  • Lee, Sung-Kyu;Kim, Bum-Jun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.18 no.4
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    • pp.277-284
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    • 1998
  • Random amplified polymorphic DNA (RAPD) analysis was used to detect the genetic diversity of soybean (Glycine max (L.) Merr.) varieties and field bean (Glycine soza Sieb. and Zucc.) Five soybean varieties and one field bean were analysed with random primers using the polymerase chain reaction (PCR). Nine primers of a total twenty random primer were selected to amplify DNA segments. A total of 74 PCR products were amplified and 67.6% of which were polymorphic. The size of DNA molecule is ranged 0.13~2.0Kb and typically generated four to eight major bands. Specific genetic marker were revealed in primer sequence 5'-CAG GCC CIT C-3', 5'-TGC TCT GCC C-3' and 5'-GTC CAC ACG G-3', respectively. Genetic similarity between each of the varieties were calculated from the pair-wise comparisons of amplification products and a dendrogram was constructed by an unweighted pair-group method with arithmethical means (UPGMA). The results indicate that intervarietal relationships of soybean have a narrow genetic base and between the varieties, Hwanggum-kong and Seckryang-bootkong is more closely related than the rest of varieties, and field bean is related quite distant.

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Development of SCAR Markers for the Discrimination of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma based on the RAPD (RAPD 분석을 통한 대황(大黃)과 종대황(種大黃) 감별용 SCAR 유전자 마커 개발)

  • Moon, Byeong-Cheol;Lee, Young-Mi;Chun, Jin-Mi;Lee, A-Young;Yoon, Tae-Sook;Cheon, Myeong-Sook;Choo, Byung-Kil;Kim, Ho-Kyoung
    • The Korea Journal of Herbology
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    • v.24 no.4
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    • pp.115-120
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    • 2009
  • Objectives : Due to the morphological similarity and frequent occurrence of intermediate forms as well as morphological variations of aerial part, the correct identification between Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma is very difficult. To develop a reliable method for correct identification and improving the quality standards of Rhei Radix et Rhizoma and Rhei Undulatai Rhizoma, we analyzed RAPD and developed SCAR marker. Methods : To amplify target DNA at the genomic level, 32 Operon 10-mer random primers were applied with four Rheum species, R. officinale, R. palmatum, R. tanguticum and R. undulatum. The nucleotide sequences were determined and species-specific primers were prepared depending on the species-specific RAPD amplicons after subcloned into the pGEM-Teasy vector. To develop the SCAR markers, species-specific PCR amplification and multiplex-PCR were carried out using the single species-specific primer pairs and combinations of them, respectively. Results : We used RAPD analysis of four Rheum plant species to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed two SCAR markers that amplified 314 bp and 390 bp DNA fragments in only R. undulatum but not in R. officinale, R. palmatum, R. tanguticum and R. undulatum, for distinguishing Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. Conclusions : These genetic markers can be used for the efficient discrimination of plants species and commercial herbal medicines between Rhei Undulatai Rhizoma and Rhei Radix et Rhizoma, to ultimately prevent indiscriminate distribution and prescription of these herbal medicines.

Development of RAPD-Derived SCAR Markers and Multiplex-PCR for Authentication of the Schisandrae Fructus (오미자 (五味子) 종 감별을 위한 RAPD 유래 SCAR Marker 및 Multiplex-PCR 기법 개발)

  • Lee, Young Mi;Moon, Byeong Cheol;Ji, Yunui;Seo, Hyeong Seok;Kim, Ho Kyoung
    • Korean Journal of Medicinal Crop Science
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    • v.21 no.3
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    • pp.165-173
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    • 2013
  • The fruits of Schisandra chinensis have been used as an edible ingredient and traditional medicine in Korea. Due to morphological similarities of dried mature fruits, the correct identification of S. chinensis from other closely related Schisandrae species is very difficult. Therefore, molecular biological tools based on genetic analysis are required to identify authentic Schisandrae Fructus. Random amplifed polymorphic DNA (RAPD) and Sequence Characterized Amplified Region (SCAR) were used to develop an easy, reliable and reproducible method for the authentication of these four species. In this paper, we developed several RAPD-derived species specific SCAR markers and established a multiplex-PCR condition suitable to discriminate each species. These genetic markers will be useful to distinguish and authenticate Schisandrae Fructus and four medicinal plants, S. chinensis, S. sphenanthera, S. repanda and K. japonica, in species level.

Identification of a Causal Pathogen of Watermelon Powdery Mildew in Korea and Development of a Genetic Linkage Marker for Resistance in Watermelon (Citrullus lanatus)

  • Han, Bal-Kum;Rhee, Sun-Ju;Jang, Yoon Jeong;Sim, Tae Yong;Kim, Yong-Jae;Park, Tae-Sung;Lee, Gung Pyo
    • Horticultural Science & Technology
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    • v.34 no.6
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    • pp.912-923
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    • 2016
  • Watermelon production is often limited by powdery mildew in areas with a large daily temperature range. Development of resistant watermelon cultivars can protect against powdery mildew; however, little is known about the characteristics of its causal agents. Here, we identified the genus and race of a causal pathogen of powdery mildew in Ansung province of South Korea, and developed molecular markers for the generation of resistant watermelon cultivars. The causal pathogen was determined to be Podosphaera xanthii based on multiple sequence alignments of internal transcribed spacers (ITS) of rDNA. The physiological race was identified as 1W, and the Ansung isolate was named P. xanthii 1W-AN. Following inoculation with the identified P. xanthii 1W-AN, we found inheritance of the resistant gene fitting a single dominant Mendelian model in a segregated population ('SBA' ${\times}$ PI 254744). To develop molecular markers linked to fungus-resistant loci, random amplified polymorphic DNA (RAPD) was accomplished between DNA pooled from eight near-isogenic lines (NILs; $BC_4F_6$), originated from PI 254744 and susceptible 'SBB' watermelon. After sequencing bands from RAPD were identified in all eight NILs and PI254744, 42 sequence-characterized amplifiedregion (SCAR) markers were developed. Overall, 107 $F_2$ plants derived from $BC_4F_6$ NIL-1 ${\times}$ 'SBB' were tested, and one SCAR marker was selected. Sequence comparison between the SCAR marker and the reference watermelon genome identified three Nco I restriction enzyme sites harboring a single nucleotide polymorphism, and codominant cleavage-amplified polymorphic site markers were subsequently developed. A CAPS marker was converted to a high-resolution melt (HRM) marker, which can discriminate C/T SNP (254PMR-HRM3). The 254PMR-HRM3 marker was evaluated in 138 $F_{2:3}$ plants of a segregating population ('SBA' ${\times}$ PI254744) and was presumed to be 4.3 cM from the resistance locus. These results could ensure P. xanthii 1W-AN resistance in watermelon germplasm and aid watermelon cultivar development in marker-assist breeding programs.

Genetic Variations and Relationships of Persicaria thunbergii(Sieb. & Zucc.) H. Gross ex Nakai(Polygonaceae) by the RAPD Analysis (RAPD 분석에 의한 고마리(마디풀과)의 유전적 변이 및 유연관계)

  • Kim, Yong-Hyun;Tae, Kyoung-Hwan;Kim, Joo-Hwan
    • Korean Journal of Plant Resources
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    • v.21 no.1
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    • pp.66-72
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    • 2008
  • RAPD analyses were performed from twenty-four populations of Persicaria thunbergii(Sieb. & Zucc.) H. Gross ex Nakai. The length of amplified DNA fragments ranged from 200 to 1,900bp. 184 scorable RAPD markers were found from PCR reactions with sixteen random oligoprimers. Based on the results, populations of Persicaria thunbergii were classified into disturbance streams of urban and rural streams as well as natural streams. And the populations from natural streams showed having higher genetic similarites than those from highly disturbed streams, Also, the heterogenetic differences between the populations from natural and disturbed areas could be represented the results of the stream environmental changes.