• Title/Summary/Keyword: RAPD primer analysis

Search Result 230, Processing Time 0.027 seconds

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
    • /
    • v.18 no.4
    • /
    • pp.277-284
    • /
    • 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.

  • PDF

Molecular Discrimination of Dinoflagellates Cochlodinium Polykrikoides Margalef, Gyrodinium Impudicum Fraga et Bravo and Gymnodinium Catenatum Graham using RAPD-PCR Method (RAPD-PCR 방법을 이용한 Cochlodinium polykrikoides Gyrodinium impudicum, Gymnodinium catenatum의 분자생물학적 진단)

  • Cho, Eun-Seob
    • Journal of Life Science
    • /
    • v.13 no.5
    • /
    • pp.651-657
    • /
    • 2003
  • Randomly amplified polymorphic DNA (RAPD) analysis was used to study genetic relationships among C. polykrikoides, G. impudicum and G. catenatum, which possess similar morphological features. Four of 12 primers were selected and 59 amplification products ranged from 0.2 kb to 3.0 kb. The number of polymorphic products in C. polykrikoides, G. impudicum and G. catenatum was 16 (27.1%), 8 (13.5%), and 16 (27.1%), respectively, while 17 were monomorphic. Number of species-specific bounds was 26 (44.0%), 34 (57.6%), 26 (44.0%) in C. polykrikoides, G. impudicum and G. catenatum, respectively. The genetic similarity between C. polykrikoides and G. impudicum/G. catenatum was 0.83, whereas G. impudicum and G. catenatum was 0.78. Our results suggest that C. polykrikoides, G. impudicum and G. catenatum are extremely different on the basis of RAPD analysis, despite similarity based on their morphology. The RAPD technique appears to be efficient in detecting genetic variation in these dinoflagellates.

Analysis of antibiotic susceptibility of Salmonella Enteritidis isolated from Gyeongnam province and the bacterial genotyping by using RAPD-PCR (경남지역에서 분리한 Salmonella Enteritidis의 항생제 감수성 검사 및 random amplification polymorphic DNA (RAPD)-PCR을 이용한 유전형 분석)

  • Kim, Eun-Gyeong;Kim, Min-Kyung;Kwon, Hyun-Ae;Youn, Do-Kyung;Koo, Jeong-Heon;Park, So-Yeon;Lee, Hui-Geun;Jo, Myeong-Hui;Hah, Do-Yun;Kim, Cheol-Ho;Hwang, Bo-Won;Kim, Sang-Hyun
    • Korean Journal of Veterinary Service
    • /
    • v.41 no.3
    • /
    • pp.149-155
    • /
    • 2018
  • Salmonella Enteritidis (S. Enteritidis) are found in animals, humans, and environment. In addition, S. Enteritidis draws attention to the public health concerns due to carriage of antibiotic resistance traits. For these reasons, the prevalence and antibiotic resistance patterns of S. Enteritidis are significant issues with regard to public health. To address this issues, a total of 24 strains of S. Enteritidis from 164 samples collected from several slaughterhouses in Gyeong-Nam province in order for antibiotic resistance profiles. Subsequently, we characterized the genotyping by random amplification polymorphic DNA (RAPD)-PCR. As a result, very high level of resistance to protein synthesis inhibition antibiotics and most isolates were susceptible to others. Six random primers were used for RAPD-PCR to reveal genotypes of S. Enteritidis isolates. One of the primer, P1245, generated 147 distinct RAPD-PCR fragments ranging from 400~3000 bp. The number of RAPD-PCR products ranged from 4 to 8 for this primer. The RAPD-PCR fragments could be placed these strains into 3 subgroups and 2 classes by UPGMA cluster analysis. Interestingly, several S. Enteritidis that isolated from different slaughterhouses showed same genotype. These results showed only limited genetic variation among the isolates, those were grouped into a few different patterns of antibiotic resistance.

Mendelian Inheritance of Inter-Simple Sequence Repeats Markers in Abies Koreans Wilson (구상나무에 있어서 Inter-Simple Sequence Repeats Marker의 유전양식(遺傳樣式))

  • Hong, Yong-Pyo;Cho, Kyung-Jin;Kim, Yong-Yul;Shin, Eun-Kyeong
    • Journal of Korean Society of Forest Science
    • /
    • v.87 no.3
    • /
    • pp.422-428
    • /
    • 1998
  • Polymerase chain reaction(PCR)-based inter-simple sequence repeats(I-SSR) markers were analyzed in 48 megagametophytes of a single tree of Abies koreana $W_{ILS}$. Nineteen of the 35 primers, screened with 6 megagametophyte DNA and produced the clearest amplification products in the preliminary experiment, were used for PCR with 48 megagametophyte DNAs sampled from a single tree. On the basis of the chi-square test, a total of 51 amplicons, amplified by the 19 primers, were revealed to be segregated according to the Mendelian ratio(i.e., 1 : 1 segregation ratio) in the 48 megagametophytes at 5% significance level. Based on the linkage analysis, the observed 51 Mendelian loci turned out to be unlinked each other, which suggested that they are evenly distributed in the genome. However, majority of RAPD markers are known to belong to the independent linkage blocks, which frequently results in the amplification of RAPD markers from the restricted regions of the genome. Owing to the nature of even distribution of the 51 loci observed in this study, the I-SSR markers could give better resolution of estimating genetic diversity from the whole genome than RAPD markers. And I-SSR markers are also more suitable than RAPD markers for reconstructing phylogenetic relationship by a cladistic method which requires to fulfil the assumption of independent evolution of the different characters.

  • PDF

A Study of Optium Condition of RAPD for the Analysis of Genetic Characteristics by Autumn Leaf Color of Zelkova serrata (느티나무(Zelkova serrata)단풍의 유전적 특성분석을 위한 RAPD 적정 조건 구명에 관한 연구)

  • Choi, Byoung Kon;Bang, Kwang Ja
    • Journal of the Korean Society of Environmental Restoration Technology
    • /
    • v.7 no.5
    • /
    • pp.94-99
    • /
    • 2004
  • This study was carried out to find out what is the optimum conditions for RAPD of Zelkova serata. We changes the factors what affect to PCR band patterns, as a result, we established the optimum conditions as follows; template DNA 100mg, Primer 0.25uM, dNTP 100mM, Taq polymerase 1.0u, and total reaction volume was filled up to 10uL with distilled water. As the amount of primers went higher, PCR reaction rates were lowered. This reason was cause by exhaustion of primers during initial reaction. The amount of dNTP didn't showed noticable differtations between the range, but the optimum amount was 100mM for efficiency. Taq polymerase 1.0 unit was the best in the range. As the concentration of polymerase were increased, many non-specific bands were appeared, In primer selection, most Openron Random Primers are amplified in this experiment. The primers GC contents were 60, and set A, B, C, D, E, X were tested. Thermal cycler(ASTEC PC808, Japan) condition was, $95^{\circ}C$, 5min, initial denaturation, $94^{\circ}C$, 20sec, denaturation, $37^{\circ}C$, 40sec, annealing, $72^{\circ}C$, 1min, extention, 45cycle repeated and final extention $72^{\circ}C$10min.

Intraspecific Relationship Analysis of Eleutherococcus senticosus Max. by RAPD Markers (RAPD 분석에 의한 가시오갈피의 유연관계 분석)

  • 임정대;성은수;최강준;김승경;김명조;유창연
    • Korean Journal of Plant Resources
    • /
    • v.13 no.2
    • /
    • pp.104-110
    • /
    • 2000
  • To analyse the genetic relationship and intraspecific variations among the Eleutherococcus senticosus population, the polymerase chain reaction(PCR) was performed total genomic DNAs of 10 E. senticosus collections by random 10 primers. The genetic diversity and genetic distance among 10 collections of Eleutherococcus spp. were used to describe the dendrogram showing phylogenic relationship. Ten collections were classfied into two group(group I, II) at the similarity coefficient value of 0.50. Group I included E. senticosus of Bukhado(Japanese), youngwal(Korea), E. seoulense, and E. chiisanesis while group II included several internal and Russia collection. The range of polymorphism was from 66.7 to 90.9% in 87 amplified DNA fragments. The similarity value of all collections ranged from 0.41 to 0.92. The average of genetic distance was 0.61.

  • PDF

Specific and Sensitive Detection of Venturia nashicola, the Scab Fungus of Asian Pears, by Nested PCR

  • Koh, Hyun Seok;Sohn, San Ho;Lee, Young Sun;Koh, Young Jin;Song, Jang Hoon;Jung, Jae Sung
    • The Plant Pathology Journal
    • /
    • v.29 no.4
    • /
    • pp.357-363
    • /
    • 2013
  • The fungus Venturia nashicola is the causal agent of scab on Asian pears. For the rapid and reliable identification as well as sensitive detection of V. nashicola, a PCR-based technique was developed. DNA fingerprints of three closely related species, V. nashicola, V. pirina, and V. inaequalis, were obtained by random amplified polymorphic DNA (RAPD) analysis. Two RAPD markers specific to V. nashicola were identified by PCR, after which two pairs of sequence characterized amplified region (SCAR) primers were designed from the nucleotide sequences of the markers. The SCAR primer pairs, designated as D12F/D12R and E11F/E11R, amplified 535-bp and 525-bp DNA fragments, respectively, only from genomic DNA of V. nashicola. The specificity of the primer sets was tested on strains representing three species of Venturia and 20 fungal plant pathogens. The nested PCR primer pair specific to V. nashicola was developed based on the sequence of the species-specific 525-bp DNA fragment amplified by primer set E11F/E11R. The internal primer pair Na11F/Na11R amplified a 235-bp fragment from V. nashicola, but not from any other fungal species tested. The nested PCR assay was sensitive enough to detect the specific fragment in 50 fg of V. nashicola DNA.

Diversity of I-SSR Variants in Gingko biloba L. Planted in 6 Regions of Korea (국내(國內) 6개(個) 은행(銀杏)나무 식재지(植栽地)에 있어서 I-SSR 변이체(變異體)의 다양성(多樣性))

  • Hong, Yong-Pyo;Cho, Kyung-Jin;Hong, Kyung-Nak;Shin, Eun-Myeong
    • Journal of Korean Society of Forest Science
    • /
    • v.90 no.2
    • /
    • pp.169-175
    • /
    • 2001
  • Genomic DNAs were extracted from the leaves of 182 ginkgo trees (Ginkgo biloba L.) planted in 6 regions and subjected to the analysis of both I-SSR and RAPD markers. A total of 227 amplicon variants were generated by PCR using 15 I-SSR primers and 67 amplicons by PCR with 5 RAPD primers. Levels of genetic diversity within 6 populations were turned out to be similar (Shannon's Index, I-SSR : 0.35~0.40; mean of 0.38, RAPD : 0.31~0.38; mean of 0.35, combined : 0.35~0.40; mean of 0.37). Ranks of the level of genetic diversity estimated from I-SSR, RAPD, and combined data were not coincided each other. Majority of genetic diversity was allocated among individuals within populations (I-SSR : 94.31%, RAPD : 93.62%, combined : 93.57%), which resulted in pretty low level of population differentiation. Genetic differentiation between male and female groups was turned out to be quite low (I-SSR : 0.03, RAPD : 0.091, combined : 0.043), which slightly fluctuated when analysis was restricted to the data obtained from 3 regions where both male and female trees were sampled (I-SSR : 0.038, RAPD : 0.084, combined : 0.047). Genetic relationships among the populations, reconstructed by UPGMA, were not coincided with geographic affinity, which might be resulted from sharing of seed sources in some regions. Whereas independent cluster analyses with I-SSR data and RAPD data, respectively, reclassified by sexes revealed two sexual groups in which all the male and the female populations were clustered together, cluster analysis with combined data did not show clear sexual grouping.

  • PDF

Discrimation of the three Angelica species using the RADPs and Internal Root Structure (RAPD 분석과 뿌리의 내부구조 비교를 통한 당귀류의 감별)

  • Lee, Mi-Young;Im, Sung-Hee;Ju, Young-Seung;Han, Keong-Sik;Jeong, Ge-Jin;An, Deok-Gun;Kang, Heon-Cheol;Ko, Byong-Seob
    • Korean Journal of Medicinal Crop Science
    • /
    • v.8 no.3
    • /
    • pp.243-249
    • /
    • 2000
  • Analysis of random amplified polymorphic DNAs(RAPDs) and internal morphological features were performed using three species of medicinal plants in the genus of Angelica(A. gigas Nakai, A. sinensis(Oliv.) Diels., A. acutiloba Kitagawa) to distinguish between these three species. Fifty decarmer oligonucleotide primers were screened for the RAPDs of the herbal plant species. Five primers generated distinct RAPD markers specific to the species of Angelica, In analysis of the degree of similarity, A. sinensis(Oliv.) Diels is more closely related to A. acutiloba Kitagawa than to A. gigas Nakai. Furthermore, we proved the usefulness of RAPD analysis for the discrimination of the species using dry roots and commercial plant materials. In internal morphology of three species, A. sinensis(Oliv.) Diels seemed to be more specialized in systemic than A. acutiloba Kitagawa and A. gigas Nakai

  • PDF

Analysis of Relationship of the Korean Styracaceae by RAPD Markers (RAPD markers에 의한 한국산 때죽나무과(Styracaceae)식물의 유연관계)

  • Kim, Hyuk-Jin;Tae, Kyoung-Hwan;Kim, Joo-Hwan
    • Korean Journal of Plant Taxonomy
    • /
    • v.37 no.2
    • /
    • pp.143-153
    • /
    • 2007
  • In order to estimate the relationships among four taxa (Styrax japonicus, Styrax obassia, Styrax shiraianus and Pterostyrax hispidus) of two genus in the Styracaceae and their regional populations, RAPDs analysis was performed. Fifteen random primers generating 238 RAPD bands were enough to reveal differences in genotype bands. They were clustered into three groups by the UPGMA phenogram. The first group was Styrax japonicus populations, the second group was consisted of Styrax obassia populations and S. shiraianus populations, and third group was Pterostyrax hispidus population. The UPGMA phenogram based on RAPD analyses showed that Styrax obassia and S. shiraianus had the closest relationship among four taxa. Species as well as genera of the Korean Styracaceae were well discriminated by the RAPD analyses.