• Title/Summary/Keyword: RAPD primer analysis

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Genetic Relationship among Ostericum koreanum Kitakawa Collections by RAPD Analysis (RAPD에 의한 강활 수집종간의 유연관계 분석)

  • Kim, Soo-Yong;Sim, Yong-Gu;Kwon, Soon-Tae;Oh, Sei-Myung
    • Korean Journal of Medicinal Crop Science
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    • v.13 no.3
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    • pp.109-113
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    • 2005
  • To analyze the genetic relationship 8 accessions of Ostericum koreanum Kitakawa, random amplified polymorphic DNA(RAPD) analysis was performed using 60 Operon primers. The 25 primers out of 60 random primers were amplified DNA by PCR using genomic DNA of O. koreanum. Eighty-five (49.1%) among 173 bands derived from 25 primers showed poly morphism, On the basis of similarity coefficient analysis by UPGMA, 8 accessions of O. koreanum Kitakawa could be classified into three groups at the similarity coefficient value of 0.71. Group I contained three accessions (Nam Gangwhal), Group II contained one accession (Nam Gangwhal) and Group III contained four accessions (Buk Gangwhal), The range of total genetic similarity coefficient value of 8 accessions of O. koreanum Kitakawa was $0.63{\sim}0.96$. Buk Gangwhal was flowered 18 to 26 days earlier than Nam Gangwhal, and Nam Gangwhal leaf stalk was thin and long as bolting rate high compared with Buk Gangwhal.

Phylogenetic relationships in different strains of Pholiota species based on PCR polymorphism (PCR 다형성 분석에 의한 비늘버섯 속 계통의 유연관계 분석)

  • Kwon, Woon-Hyuk;Park, Hyuk;Baek, Min-Jae;Cho, Woo-Jin;Choi, Woo-Jeong;Ahn, Chi-Beom;Shin, Do-Bin;Lee, Tae-Soo
    • Journal of Mushroom
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    • v.11 no.2
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    • pp.69-76
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    • 2013
  • Pholiota species were collected from different geographical regions of the world. Genetic diversity and phylogenetic relationships were analyzed by rDNA-ITS sequences and RAPD polymorphism. The sizes of rDNA-ITS PCR amplicons of Pholiota spp. varied from 233~271, 158~223 and 174~219 bp, respectively. A phylogenetic tree was constructed on the ITS region sequences and Pholiota strains were classified into 8 clusters. Twenty strains in seven Pholiota spp. were classified into seven clusters by RAPD polymorphism using 15 arbitrary primers. Our experimental results suggested that rDN-ITS and RAPD analysis are useful tool for classifying Pholiota spp. and strains.

Genetic Identity between Bhadawari and Murrah Breeds of Indian Buffaloes (Bubalus bubalis) Using RAPD-PCR

  • Saifi, H.W.;Bhushan, Bharat;Kumar, Sanjeev;Kumar, Pushpendra;Patra, B.N.;Sharma, Arjava
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.5
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    • pp.603-607
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    • 2004
  • Randomly Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) analysis was carried out with a battery of 11 random decamer primers to study band frequency (BF), genetic identity index (I) and mean average percentage difference (MAPD) between Bhadawari and Murrah breeds of buffalo. The primers OPA04 and BG15 resolved a band of 460 bp, which was present only in animals of Bhadawari breed. Whereas, the primers OPA14, BG27 and BG28 produced Murrah specific fragments of sizes 730 bp and 1,230 bp, respectively. The estimate of genetic identity index was highest (0.845) with the primer OPA01 and the lowest (0.479) with the primer BG27. The genetic identity index pooled over the primers was 0.596${\pm}$0.037 between these two breeds. The highest MAPD estimate (53.9) between the two breeds was obtained with the primer BG27 and the lowest (14.3) with the primer OPA01. It might be concluded that the genetic identity index between these two breeds calculated on the basis of BF showed moderate level of genetic identity with the primers employed. MAPD calculated on the basis of uncommon bands also demonstrated lower to medium level of genetic difference between Bhadawari and Murrah breeds of buffalo.

Assessment of Genetic Variability in Two North Indian Buffalo Breeds Using Random Amplified Polymorphic DNA (RAPD) Markers

  • Sodhi, M.;Mukesh, M.;Anand, A.;Bhatia, S.;Mishra, B.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.9
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    • pp.1234-1239
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    • 2006
  • Murrah and NiliRavi are the important North Indian buffalo breeds occupying the prominent position of being the highest milk producers. These breeds are more or less similar at morphological as well as physiological levels. The technique of RAPD-PCR was applied in the present study to identify a battery of suitable random primers to detect genetic polymorphism, elucidation of the genetic structure and rapid assessment of the differences in the genetic composition of these two breeds. A total of 50 random primers were screened in 24 animals each of Murrah and NiliRavi buffaloes to generate RAPD patterns. Of these, 26 (52%) primers amplified the buffalo genome generating 263 reproducible bands. The number of polymorphic bands for the 26 chosen RAPD primers varied from 3 (OPG 06 and B4) to 26 (OPJ 04) with an average of 10.1 bands per primer and size range of 0.2 to 3.2 kb. DNA was also pooled and analyzed to search for population specific markers. Two breed specific RAPD alleles were observed in each of Murrah (OPA02 and OPG16) and NiliRavi (OPG09) DNA pools. RAPD profiles revealed that 11 (4.2%) bands were common to all the 48 individuals of Murrah and NiliRavi buffaloes. Pair-wise band sharing calculated among the individual animals indicated considerable homogeneity of individuals within the breeds. Within breed, band sharing values were relatively greater than those of interbreed values. The low genetic distance (Nei's) value (0.109) estimated in this study is in accordance with the origin and geographical distribution of these breeds. The RAPD analysis indicated high level of genetic similarity between these two important North Indian buffalo breeds.

Parentage Identification of 'Daebong' Grape (Vitis spp.) Using RAPD Analysis

  • Kim, Seung-Heui;Jeong, Jae-Hun;Kim, Seon-Kyu;Paek, Kee-Yoeup
    • Journal of Plant Biotechnology
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    • v.4 no.2
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    • pp.67-70
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    • 2002
  • The RAPD data were used to assess genetic similarity among f grape cultivars. Of the 100 random primers tested on genomic DNA, 10 primers could be selected for Benetic analysis, and the selected primers generated a total of 115 distinct amplification fragments. A similarity matrix was constructed on the basis of the presence or absence of bands. The 7 grape cultivars analyzed with UPGMA were clustered into two groups of A and B. The similarity coefficient value of cultivars was high. The mean similarity index for all pairwise comparisons was 0.851, and ranged from 0.714 ('Rosaki' and 'Black Olympia') to 0.988 ('Kyoho' and 'Daebong'). After due consideration of differences in cultural and morphological characteristics of these two theoretically identical cultivars, it could be deduced that 'Daebong' is a bud sport of 'Kyoho' cultivar.

Development of Cleaved Amplified Polymorphic Sequence (CAPS) Marker for Selecting Powdery Mildew-Resistance Line in Strawberry (Fragaria×ananassa Duchesne) (딸기 흰가루병 저항성 계통 선발을 위한 분자마커 개발)

  • Je, Hee-Jeong;Ahn, Jae-Wook;Yoon, Hae-Suk;Kim, Min-Keun;Ryu, Jae-San;Hong, Kwang-Pyo;Lee, Sang-Dae;Park, Young-Hoon
    • Horticultural Science & Technology
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    • v.33 no.5
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    • pp.722-729
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    • 2015
  • Powdery mildew (PM) caused by Podosphaera aphanis is a major disease that can result in significant yield losses in strawberry (Fragaria ${\times}$ ananassa Duchesne). For preventing PM, pesticides are usually applied in strawberry. In this study, molecular markers were developed to increase breeding efficiency of PM-resistance cultivars by marker-assisted selection (MAS). An $F_2$ population derived from a cross between PM-resistance 'Seolhyang' and PM-susceptibility 'Akihime' was evaluated for disease resistance to PM and RAPD (random amplification of polymorphic DNA)-BSA (bulked segregant analysis). Among 200 RAPD primers tested, OPE10 primer amplified a 311bp-band present in with 331bp. Sequence alignment performed for searching polymorphisms and six single nucleotide polymorphism (SNP) were found in amplified regions. To develop polymorphic marker for distinguishing between resistant and susceptible, RAPD was converted to cleaved amplified polymorphic sequence (CAPS) marker. Among restriction enzymes associated with six SNPs, Eae I (Y/GGCCR) was successfully digested to 231bp in susceptible. The results suggest that the selected CAPS marker could be used for increasing efficiency of selecting powdery mildew resistant strawberry in breeding system.

RAPD Analysis on the Species of Pinelliae Tuber (RAPD 방법을 이용한 반하류 한약재의 감별 연구)

  • 배명효;김규열;정유헌;최호영
    • The Journal of Korean Medicine
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    • v.20 no.4
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    • pp.16-22
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    • 2000
  • This study intends to report the significance of several experimental results obtained from analysing the genes extracted from the plants and herbal medicine such as P. temalta (Thunb.) Breit, A. amurense var serratum Nakai, A. erubescens (Wall.) Schott, Pinelliae Tuber and Arisaematis Tuber, mainly by the method of RAPD(randomly amplified polymorphic DNA) and the method of RFLP(restriction fragment length polymorphism) on ITS(internal transcribed spacer) region. Genomic DNA could be extracted from both original plants and dried materials. DNA fragments of P. temata kind and A. amurense kind showed the same aspect separately within the same species under the method of RAPD using random primer, while various aspects(polymorphism) were discovered among different species. In RAPD analysis by uniprimer, common bands were extracted from all types of P. temata in the case of uniprimer #4, which were distinguished from the kind of A. amurense. Other polymorphic bands appeared in between different A. amurense species as well. In the case of uniprimer #11, particular band came out in the kind of P. temata. On the other hand, in the case of uniprimer #5, #6, and #8, various bands(polymorhism) were revealed in both kinds of P. temata and A. amurense. Although further study is needed to ascertain whether these results are due to the differences of species, kinds, or growing place, the results could be used as a scientific method of identifying the substitutes for A. amurense genus. The author believes that as if P. temata class of plants used in this experiment are different among themselves in terms of the shape, size and property, those are clearly a class of P. temata or belong to the same genus.

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Isolation and Identifieation of Entomopathogenic Nematodes from Soil and Insect (토양과 곤충 사체로부터 곤충병원성 선충의 분리 및 동정)

  • 한상미;한명세
    • Korean Journal of Environmental Biology
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    • v.17 no.3
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    • pp.321-330
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    • 1999
  • Nematodes were isolated using silkwom trap through the investigation of 100 soil samples from various biotopes in Korea. The 30 nematode strains from soil and dead insects by the pathogenicity aganinst silkworms (Bombyx mori mori) and insect pests of Calliphora vomitoria, Pseufazetia separata, Palomena angulosa, and Melolontha incana. Mortailty of the silkworm larvae and pupae were as high as 100% by nematode infection, those of insect of pests were varied from 20 to 100%. The 30 strains of entemopathogenic nematodes were classified into five groups of Rhabditidae, Diplogatroidae, Heterorhabitidae, Steinernematidae, and Tylenchida by morphological criteria. The genetic relationships among the 30 nematode strains were analyzed by various RAPD bands with twenty primers. The 30 nematode strains were classified into six major subgroups on the basis of the genetic similarity coefficient of 0.853. The grouping by RAPD was agree with those of morphological taxa in discrimination of the higher group, however, was not completely agree in the subgroup. The family Steinernematidae belong to Rhabditida was clarified as closer to the Tylenchida, rather than the other Rhabditida of Heterorhabitidae, Rhabditidae, and Diplogatroidae in genetic distance valule. From the result of the morphological classification and RAPD of the genomic DNA showed that genetic relationship analysis furnish infurmation on phylogenetic classification and relationships of entomopathogenic nematodes. The application of genetic similarity will overcome the limitation of taxonomy and classification of morphologically simple nematode. Several primers were confirmed those utility of identification for individual nematode strains, the methods of molecular genetics secured the simplicity, rapidity and accuracy on the selection of entomopathogenic nematodes.

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Molecular discrimination of Panax ginseng species

  • Um, Jae-Young;Chung, Hwan-Suck;Kim, Hyun-Ju;Kim, Dae-Ki;Shim, Kyung-Shik;Lee, Kang-Yong;Kim, Jeong-Sook;Choi, Tae-Jin;Kim, Nam-Song;An, Nyeon-Hyoung;Lee, Kang-Min;Lee, Young-Mi;Kim, Jeong-Joong
    • Advances in Traditional Medicine
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    • v.1 no.2
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    • pp.52-58
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    • 2000
  • In order to develop convenient and reproducible methods for identification of ginseng drugs at a DNA level, RAPD (randomly amplified polymorphic DNA) and PCR-RFLP (PCR-Restriction fragment length polymorphism) analysis were applied within Panax species. To authenticate Panax ginseng betvyeen Chinese and Korean ginseng population, RAPD analysis were carried out using 20 mer-random primer. The similarity coefficients among the DNA of ginseng plants analyzed were low, ranging from 0.197 to 0.491. In addition, using PCR-RFLP analysis, very different fingerprints were obtained within Korean ginseng plants. These results suggest that these methods are able to authenticate the concerned Panax species. Broader application of this approach to authenticate other morphologically similar medicinal materials is rationalized.

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Monitoring of Gentic Variability in Dicofol-susceptible, Dicofol-resistant, and its Reverse-selected Strains of Tetranychus urticae by RAPD-PCR

  • Song, Cheol;Park, Jin-Hee;Kim, Gil-Hah;Kwon, O-Yu;Cho, Kwang-Yun
    • Journal of Life Science
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    • v.9 no.1
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    • pp.14-16
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    • 1999
  • Genetic variability was monitored by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) in dicofol-susceptible (S), dicofol-resistant (R) and its reverse-selected (RS) strains of two-spotted spider mite, of Tetranychus urticae. Before the reverse-selection, RS strain, selected reversely from R strain, was 23-fold resistance ratio at {TEX}$LC_{50}${/TEX} to S strain. The resistance was started to in incline slowly to the resistance level of S strain after one year, and the resistance ratio was 4-fold in the 7 years after then. PCR-amplification of T. urticae DNA showed polymorphism in the amplifications with 12 primers in 100 kinds of arbitrary DNA sequences. RAPD amplification with primer OPR-12 (5`-ACAGGTGCGT-3`) showed amplified bands at 1,000 base pair in the S-and RS-strain, and at 350 base pair in R-strain. The results of polymorphism are genetic variabilities derived from development and selection of resistance in each strain. The peculiarly amplified fragments were guessed to participate in dicofol resistance. From the analysis of genetic similarity, it is inferred the gene composition of S-and RS-strain is much closer than that of R-strain.

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