• Title/Summary/Keyword: RAPD markers

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RAPD Analysis of Three Deer Species in Malaysia

  • El-Jaafari, Habiba A.A.;Panandam, Jothi M.;Idris, Ismail;Siraj, Siti Shapor
    • Asian-Australasian Journal of Animal Sciences
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    • v.21 no.9
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    • pp.1233-1237
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    • 2008
  • The genetic variability within and among three deer species in Malaysia, namely Cervus nippon (sika), Cervus timorensis (rusa) and Cervus unicolor (sambar), were evaluated using the RAPD technique. The DNA extracted from the buffy coat of 34 sika, 38 rusa and 9 sambar were analysed using ten primers that gave bands which showed good resolution. The primers generated 164 RAPD markers in total, and these ranged in size from 150 to 900 bp. The percent of polymorphism of the bands generated per primer ranged from 66.66-93.33% for rusa, 36.84-61.14% for sambar and 52.38-100% for sika. The overall percent polymorphism observed for the 164 RAPD markers was 99.39%. The results revealed five exclusive, monomorphic markers for sambar and one exclusive, monomorphic marker for sika; none was observed for rusa. However, these cannot be declared as markers for the identification of the species without analysis of more samples, populations and species. The means of within population genetic distances, based on Dice's and Jaccard's similarity indices, were similar for the rusa (0.383 and 0.542, respectively) and sika (0.397 and 0.558, respectively) populations with the sambar population being the least variable (0.194 and 0.323, respectively). The Dice based genetic distances within the species ranged from 0.194 to 0.397 and the genetic distances among the species were 0.791-0.911. The genetic distances based on Dice's and Jaccard's similarity indices between the rusa and sambar were 0.556 and 0.713, between the rusa and sika populations were 0.552 and 0.710, and between sambar and sika were 0.622 and 0.766, respectively.

Rapid molecular authentication of three medicinal plant species, Cynanchum wilfordii, Cynanchum auriculatum, and Polygonum multiflorum (Fallopia multiflorum), by the development of RAPD-derived SCAR markers and multiplex-PCR

  • Moon, Byeong-Cheol;Choo, Byung-Kil;Cheon, Myeong-Sook;Yoon, Tae-Sook;Ji, Yun-Ui;Kim, Bo-Bae;Lee, A-Young;Kim, Ho-Kyoung
    • Plant Biotechnology Reports
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    • v.4 no.1
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    • pp.1-7
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    • 2010
  • Definitive identification of original plant species is important for standardizing herbal medicine. The herbal medicines Cynanchi Wilfordii Radix (Baekshuoh in Korean and Beishuwu in Chinese) and Polygoni Multiflori Radix (Hashuoh in Korean and Heshuwu in Chinese) are often misidentified in the Korean herbal market due to morphological similarities and similar names. Therefore, we developed a reliable molecular marker for the identification of Cynanchi Wilfordii Radix and Polygoni Multiflori Radix. We used random amplified polymorphic DNA (RAPD) analysis of three plant species, Polygoni multiflorum, Cynanchum wilfordii, and Cynanchum auriculatum, to obtain several species-specific RAPD amplicons. From nucleotide sequences of these RAPD amplicons, we developed six sequence characterized amplification region (SCAR) markers for distinguishing Polygoni Multiflori Radix and Cynanchi Wilfordii Radix. Furthermore, we established SCAR markers for the simultaneous discrimination of the three species within a single reaction by using multiplex-PCR. These SCAR markers can be used for efficient and rapid authentication of these closely related species, and will be useful for preventing the distribution of adulterants.

Discrimination of Species Specific DNA Markers Using RAPD and AFLP Analysis between Atractylodes japonica Koidz. and Atractylodes macrocephala Koidz.

  • Bang, Kyong-Hwan;Park, Chun-Geon;Jin, Dong-Chun;Kim, Hong-Sig;Park, Hee-Woon;Park, Chung-Heon;Seong, Nak-Sul
    • Korean Journal of Medicinal Crop Science
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    • v.11 no.4
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    • pp.268-273
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    • 2003
  • To identify the variation of the RAPD patterns between two Atractylodes species, 52 kinds of random primers were applied to each eight of A japonica and A. macrocephala genomic DNA. Ten primers of 52 primers could be used to discriminate between the species and 18 polymorphisms among 67 scored DNA fragments (18 fragments are specific for A. japonica and A. macrocephala) were generated using these primers, 26.9% of which were polymorphic. RAPD data from the 10 primers was used for cluster analysis. The cluster analysis of RAPD markers showed that the two groups are genetically distinct. On the other hand, to identify the variation of the AFLP patterns and select the species specific AFLP markers, eight combinations of EcoRI/MseI primers were applied to the bulked A. japonica and A. macrocephala genomic DNA. Consequently, three combinations of EcoRI/MseI primers (EcoRI /Mse I ; AAC/CTA, AAC/CAA, AAG/CTA) used in this study revealed 176 reliable AFLP markers, 42.0% of which were polymorphic. 74 polymorphisms out of 176 scored DNA fragments were enough to clearly discriminate between two Atractylodes species.

Analysis of Genetic Diversity in Echinochloa Species Using Random Amplified Polymorphic DNAs(RAPDs) Markers (RAPD Marker를 이용한 피 수집종의 유연관계 분석)

  • Kim, Kil-Ung;Sohn, Jae-Keun;Shin, Dong-Hyun;Kim, Kyung-Min;Kim, Hak-Yoon;Lee, In-Jung
    • Korean Journal of Weed Science
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    • v.18 no.1
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    • pp.76-83
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    • 1998
  • Echinochloa species maintained by selling for more than 10 years were classified using random amplified polymorphic DNAs(RAPDs) analysis. Seventy-four decamer of randomly sequence markers were used to classify intraspecific variation irt Echinochloa species. The number of amplification products increased with increasing GC content of the primer in the range between 60% and 70% GC. Single-base substitutions of a primer altered amplification, providing new polymorphisms. The size of amplified DNA was mostly between 0.40kbp and 1.4kbp with the most common bands at 1.1kbp. Echinochloa species were detected with 6 primers which generated 26 polymorphic amplified DNAs. By hierarchical cluster analysis, Echinochloa species collected in Korea were divided into three groups. These results revealed that RAPD markers are useful tools for the determination of genetic variations in Echinochloa species.

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Screening of RAPD Markers for Fluoride Resistance in Bombyx mori L.

  • Chen, Keping;Yao, Qin;Li, Muwang;Wang, ong
    • International Journal of Industrial Entomology and Biomaterials
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    • v.7 no.1
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    • pp.11-14
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    • 2003
  • NF733xin, the near allele line was obtained by means of crossing and backcrossing the silkworm race T6, which contained fluoride resistance major gene, to race 733xin, which was highly susceptible to fluoride toxicity. Two hundred RAPD random primers were used in the RAPD analysis of these 3 strains. Two molecular markers, OPB-08850 and OPB-10917, were obtained. OPB-10917 was used to detect the backcross generations. It was found that all the fluoride resistant individuals in each backcross generation had the same special band. These results proved that this marker was reliable.

Identification of DNA polymorphisms in the field bean ( Glycine soza S. and Z. ) using RAPD markers (RAPD 표지인자를 이용한 돌콩 DNA 다형현상 분석)

  • 이성규
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.18 no.2
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    • pp.143-150
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    • 1998
  • Six field bean (GI-vcine soza S and Z ) plants were examined for their genetic polymorphisms and intraspecific variations using randomly amplified polymorphic DNA(RAPD) markers. In RAPD analysis of 5 random primers (Rp-1, Rp2, Rp-3, Rp-4, Rp-5), 30 of total 155 bands obtained kom 5 primers were polymorphic and sizes of polymirphic band ranged between 0.5 and 3.0 kb. Number of bands amplyfied per primer was varied from 2 to 11 and average number was 6.0. Genetic variation of intraspecies in the samples of six region was ranged behveen 11 to 25 percent, and genetic similarity among intraspecies was ranged from 0.69 to 0.78. In pairwise genetic similarity test of six field bean plants, Mun and Hoj showed highest coefficient of genetic similarity as 0.67, whereas Sin and Hoj was lowest as 0.45. According to the genetic similarity, the level of intraspecific variation is higher than that of regional distance in GI-vcine soza.

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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.

Genetic Diversity in Rauvolfia tetraphylla L.f using RAPD Markers

  • Padmalatha, K;Prasad, MNV
    • Journal of Plant Biotechnology
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    • v.33 no.2
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    • pp.139-145
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    • 2006
  • The present study is the first report of molecular variations in different accessions of Rauvolfia tetraphylla L.f, a medicinally important plant collected from seven locations of Andhra Pradesh, India. Molecular analysis was carried out using RAPD markers. Out of the 40 primers screened from OPA and OPC Kts, a total of 205 scorable polymorphic markers out of 397 total markers were generated. Polymorphism of 51.6% was found with 3 unique markers. Levels of genetic diversity within accessions i.e., the genetic distance ranged from 0.816-0.932. Cluster analysis based on Dice coefficient showed two major groups indicating that mostly in cross-pollinated plants, high levels of differentiation among accessions exists independent of geographical distance. Hence the results of the present study can be seen as a starting point for future researches on the population and evolutionary genetics of this species. Understanding such variation would also facilitate their use in various conservational management practices, rootstock breeding and hybridisation programmes.

Construction of a Genetic Linkage Map in Radish(Raphanus sativus L.) Using RAPD Markers (RAPD 마커를 이용한 무의 유전자지도 작성)

  • Ahn, Choon-Hee;Choi, Su-Ryun;Lim, Yong-Pyo;Chung, Hae-Joon;Yae, Byeong-Woo;Yoon, Wha-Mo
    • Journal of Plant Biotechnology
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    • v.29 no.3
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    • pp.151-159
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    • 2002
  • Genetic map and molecular marker have a great importance in improving and facilitating crop breeding program as well as in genome analysis and map-based cloning of genes representing desirable characters. This study aimed at developing RAPD markers and constructing a genetic linkage map using 82 BC$_1$F$_1$individuals originated from the cross between '835' and B$_2$in radish (Raphanus sativus L.). One of the parents for genetic linkage map construction, '835'(P$_1$) of egg type is susceptible to Fusarium wilt and have medium resistance to virus infection and the other parent, B$_2$(P$_2$) of round type, is susceptible to Fusarium wilt and virus, Screening of 394 RAPD primers in BC$_1$F$_1$) population resulted in selecting 128 polymorphic markers which displayed 1:1 segregation pattern. Two markers failed to display 1:1 segregation and showed the segregation ratio skewed to maternal genotype. Selected markers were categorized into 14 linkage group based on LOD score represented by MAPMAKER/EXP program. Five groups composed of single marker among them were excluded from the linkage map, and consequently, the remaining groups are well matched with the number of radish chromosome (n=9). The linkage map constructed with 128 markers covers 1,688.3 cM and the average distance between markers was 13.8 cM. For developing STS marker, we determined the partial nucleotide sequence of OPE10 marker at both ends and designed a oligonucleotide primer pair based on this sequence. STS PCR using the primer pair displayed a single, clear band of which segregation is perfectly matched with that of OPE10 marker. This implies that RAPD markers could readily convert into clear and reliable STS markers.

A taxonomic study of Abeliophyllum Nakai (O1eaceae) based on RAPD analysis (RAPD분석에 의한 미선나무속의 분류학적 연구)

  • 김동갑;박경량;김주환
    • Korean Journal of Plant Resources
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    • v.15 no.1
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    • pp.26-35
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    • 2002
  • Abeliophyllum distichum Nakai (O1eaceae) is a monotype of Korea and is distinguished from related genus Forsythia Vahl by the morphological characters such as fruit shape, flower color and etc. Even though several intraspecific taxa were reported according to the color of flowers and shape of fruits, there have been many controversals on the taxonomic indentity and status of rank on those taxa. In the present study, we performed the RAPD analysis to delimit the infraspecific taxa of Abeliophyllum distichum and to investigate the genetic polymorphism and relationships among 12 populations. 212 scorable RAPD markers with 70 common markers were found from the PCR reactions with 16 random oligoprimers and were analyzed by Nei's genetic distance. From 0.108 to 0.321 of genetic variations were showed among the taxa. Some regional groups instead of same taxa were clustered from the phonogram of UPGMA analysis. Also, we could not find distinct lineage among intraspecific taxa. The result from RAPD analysis supported that the infraspecific taxa of Abeliophyllum distichum might be the individual variations and treated as the same taxa. RAPD analysis was very useful to confirm the high gene pool with diverse genetic polymorphism among Abeliophyllum distichum populations.