• Title/Summary/Keyword: PCR polymorphic

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Development of Sequence-Based DNA Markers for Evaluation of Phylogenetic Relationships in Korean Watermelon Varieties

  • Lee, Hee-Jeong;Cho, Hwa-Jin;Lee, Kyung-Ah;Lee, Min-Seon;Shin, Yoon-Seob;Harn, Chee-Hark;Yang, Seung-Gyun;Nahm, Seok-Hyeon
    • Journal of Crop Science and Biotechnology
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    • v.10 no.2
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    • pp.98-105
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    • 2007
  • Phylogenetic relationships in Korean watermelons were evaluated by genetic similarity coefficients using 15 SSR(simple sequence repeat), 14 SCAR(sequence characterized amplified region) and 14 CAPS(sequence characterized amplified region) markers. The SSR markers were selected from previously reported melon and watermelon SSRs through testing polymorphisms within a set of commercial $F_1$ varieties. The SCAR and CAPS markers were developed from polymorphic AFLP(amplified fragment length polymorphism) markers between inbred lines 'BN4001' and 'BN4002'. From the AFLP analysis, 105 polymorphic fragments were identified between the inbred lines using 1,440 primer combinations of EcoRI+CNNN and XbaI+ANNN. Based on the sequencing data of these polymorphic fragments, we synthesized sequence specific primer pairs and detected clear and reliable polymorphisms in 27 primer pairs by indels(insertion/deletion) or RFLP(restriction fragment length polymorphism). A total of 43 sequence-based PCR markers were obtained and polymorphic information content(PIC) was analyzed to measure the informativeness of each marker in watermelon varieties. The average PIC value of SCAR markers was 0.41, which was similar to that of SSR markers. Genetic diversity was also estimated by using these markers to assess the phylogenetic relationships among commercial varieties of watermelon. These markers differentiated 26 Korean watermelon varieties into two major phylogenetic groups, but this grouping was not significantly correlated with their morphological and physiological characteristics. The mean genetic similarity was 66% within the complete set of 26 commercial varieties. In addition, these sequence-based PCR markers were reliable and useful to identify cultivars and genotypes of watermelon.

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Use of RAPD Fingerprinting for Discriminating Two Populations of Hilsa shad (Tenualosa ilisha Ham.) from Inland Rivers of Bangladesh

  • Shifat, Rehnuma;Begum, Anwara;Khan, Haseena
    • BMB Reports
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    • v.36 no.5
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    • pp.462-467
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    • 2003
  • The Random Amplified Polymorphic DNA-Polymerase Chain Reaction (RAPD-PCR) was applied to analyze the genetic variation of the Hilsa shad, Tenualosa ilisha Ham., from the two major inland rivers (Padma and Meghna) in Bangladesh. Twenty-eight random 10-mer primers were primarily scored in 8 individuals from each of the two locations. Fifteen primers, which gave polymorphism, were selected and used in the final analysis of 34 individuals from the two sites. Using these primers, 480 scorable DNA fragments were found, of which 98 (20.41%) were polymorphic. By comparing the RAPD banding patterns, variations were found between and within the populations. A dendrogram was constructed with the polymorphic fragments to analyze the genetic distances between the Hilsa shad populations. The results show two major clusters of Padma and Meghna, assuming different spawning populations with different stocks or races of Hilsa shad in the major Bangladesh rivers.

Molecular Typing of Pseudomonas aeruginosa by Randomly Amplified Polymorphic DNA

  • Byoung-Seon Yang
    • Biomedical Science Letters
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    • v.9 no.4
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    • pp.183-187
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    • 2003
  • Pseudomonas aerugionsa is a commonly isolated nosocomial pathogen. DNA fingerprinting of P. aerugionsa is examined by randomly amplified polymorphic DNA (RAPD). In this study, P. aeruginosa were isolated from environmental and clinical specimens and the molecular typing of the microorganisms was investigated by RAPD. Thirty strains of P. aeruginosa were selected from the strains isolated formerly and submitted for type identification to the University Hospital. 15 strains of P. aeruginosa were received from Chungnam University Hospital and 14 strains from Gyeongsang University Hospital. DNA of P. aeruginosa was extracted by Qiagen genomic DNA kit. PCR mixtures were set up and incubated, Reactions mixtures were made to be optimal for P. aeruginosa. RAPD typing analysis was carried out by the multivariate statistical program (MVSP) V3.0. RAPD type I was the most common pattern and included 23 strains. Most of strains from Gyeongsang University Hospital belonged to RAPD type lb and 15 strains from Chungnam University Hospital to RAPD type I or II. RAPD typing of P. aeruginosa isolated from the environmental and clinical specimens was very simple and reproducible.

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Random Amplified Polymorphic DNA (RAPD) Variation in Porhyra yezoensis and P. tenera

  • Beom-Kyu Kim;Gyu-Hwa Chung;Yuji Fujita
    • Journal of Aquaculture
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    • v.10 no.3
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    • pp.321-326
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    • 1997
  • The random amplified polymorphic DNA (RAPD) technique was used to anlayze six isolates of two species of Porphyra, P. yezoensis and P. tenera. Four 21-mer prrmers were combined randomly into six groups of double primers and screened for DNA amplification using nuclear and chloroplast tempate DNA. The RAPD patterns resulting from RnRc and CnCc primers provided evidence for both genetically homo-and heterogeneous populations of P. yezoensis and P. tenera. Similarity values obtained by RnRc primer analysis of nuclear DNA varied from 0.364 to 0.714 and those of chloroplast DNA were high, ranging from 0.727 to 1.000, except for P. yezoensis (Enoura).

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DNA Polymorphism and Genetic Similarity in Cultured Catfish by Polymerase Chain Reaction-Random Amplified Polymorphic DNAs

  • Yoon, Jong-Man;Park, Kwan-Ha;Park, Sung-Woo
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2001.05a
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    • pp.530-531
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    • 2001
  • Out of 20 primers, 6 generated 1349 highly reproducible RAPD markers, producing approximately 5.2 polymorphic bands per primer in catfish(Sizurus asotus) population from Kunsan. The electrophoretic analysis of polymerase chain reaction-random amplified polymorphic DNAs (PCR-RAPD) products showed the middle levels of similarity between different individuals in population from Kunsan. That is to say, the degree of similarity varied from 0.40 to 0.54, with the average of 0.46, as calculated by bandsharing analysis. (omitted)

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Use of RAPD-PCR(Random Amplified Polymorphic DNA-Polymerase Chain Reaction) Method for a Detection of Pathogenic Listeria monocytogenes (RAPD-PCR(Random Amplified Polymorphic DNA - Polymerase Chain Reaction) 방법을 이용한 Listeria monocytogenes의 검색)

  • Park Bum-Joon;Sihn Eon-Hwan
    • The Korean Journal of Food And Nutrition
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    • v.17 no.3
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    • pp.254-259
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    • 2004
  • Rapid detection of foodbome pathogens is becoming increasingly important. The requirement for faster, more reliable tests has lead to the development of a wide range of rapid methods. Among these methods, the use of systems based on nucleic acid based detection has been increasing since they offer advantages of reduction in test time and more reliable detection or identification. Random Amplification Polymorphic DNA(RAPD) method has been used to fingerprint foodbome microorganisms; Listeria monocytogenes. In this study, 10-mer primer OPG-13(5'-CTCTCCGCCA-3') was used to generate RAPD-PCR for detection of pathogenic L. monocytogenes of Listeria spp. Among 20 primers tested, OPG-13 showed on acceptable result for the differentiation of a pathogenic Listeria from non-pathogenic microorganisms. Pathogenic Listeria, L. monocytogenes(ATCC 15313, 19111, 19112, 19113) showed two bands for 700 bp and 1,500 bp while non-pathogenic bacteria, L. ivanovii, L. grayi, L. murrayi, L. innocua, L. welshimeri, and L. seeligeri had only one band sizing from 2,000 to 2,300 bp. This RAPD method proved to be a valuable to gain important information on sources of pathogenic bacteria in food industry.

Genotyping of Agaricus bisporus Strains by PCR Fingerprints

  • Min, KyongJin;Oh, YounLee;Kang, HeeWan
    • 한국균학회소식:학술대회논문집
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    • 2014.10a
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    • pp.41-41
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    • 2014
  • Agaricus bisporus, commonly known as the button mushroom, is the most widely cultivated species of edible fungi. Low frequency of recombination ratio and homokaryotic or monokaryotic spore on meiotic basidia form obstacles for breeding programs. Since the first hybrid varieties for white button mushrooms were released in Europe, new varieties released afterwards were either identical of very similar to these first hybrids on morphologies. Therefore, different DNA markers have been used to define unique varieties of A. bisporus strains. Aim of this study is to assess the genetic diversity of different A. bisporus strains in Korea. Twelve UFP (Universal fungal primer, JK BioTech. Ltd), 12 simple sequence repeat (ISSR) and 30 SSR primers were used to assess genetic diversity of monokaryotic and dikaryotic Agaricus bisporus strains including other 19 Agaricus spp. Of them, four UFP, four SSR primers, $(GA)_8T$, $(AG)_8YC$, $(GA)_8C$ and $(CTC)_6$ and seven SSR markers produced PCR polymorphic bands between the Agaricus species or within A. bisporus strains. PCR polymorphic bands were inputted for UPGMA cluster analysis. Forty five strains of A. bisporus are genetically clustered into 6 groups, showing coefficient similarity from 0.75 to 0.9 among them. In addition, genetic variations of monokaryotic and dikaryotic Agaricus bisporus strains were partially detected by PCR technologies of this study. The varieties, Saea, saedo, Saejeong and Saeyeon that have recently been developed in Korea were involved in the same group with closely genetic relationship of coefficient similarity over 0.96, whereas, other strains were genetically related to A. bisporus strains that were introduced from USA, Eroupe and Chinese.

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Genetic Variability Based on Randomly Amplified Polymorphic DNA in Mistletoe Fig (Ficus deltoidea Jack) Collected from Peninsular Malaysia

  • Bhore, Subhash Janardhan;Arneida H., Nurul;Shah, Farida Habib
    • Journal of Forest and Environmental Science
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    • v.25 no.1
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    • pp.57-65
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    • 2009
  • Ficus deltoidea Jack is an important and popular medicinal plant species found in the Malaysia. Plants are being collected and used based on morphology and authentication to prevent adulteration is not in practice. In this study, twenty-six accessions of F. deltoidea Jack were collected from Kelantan and Terengganu states of Peninsular Malaysia to examine their genetic similarities and differences using randomly amplified polymorphic DNA (RAPD) technique. Out of 20 arbitrary primers, two primers (D-10 and D-11) were selected which produced reliable DNA polymorphism. D-10 and D-11 primers generated 138 RAPD bands ranging from 250 bp to 3000 bp. Ninety-nine of them were polymorphic loci (72%) and thirty-nine were nonpolymorphic loci (28%). A total of 56 bands with polymorphic loci were amplified with primer D-10 and analyzed by cluster analysis and UPGMA to present a dendrogram depicting the degree of genetic relationship among 26 accessions. Eight RAPD markers were sequenced to determine their identity. RAPD analysis showed the genetic diversity among 26 accessions of F. deltoidea Jack. The RAPD profile and RAPD marker sequences reported in this paper could be used in plant and/or plant material authentication. This study also suggested that RAPD can be a useful technique to study DNA polymorphism in F. deltoidea Jack.

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Genetic Relationship among Garlic Cultivars Based on RAPD Analysis (RAPD에 의한 마늘의 유연관계 분석)

  • 권순태;오세명
    • Journal of Life Science
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    • v.9 no.6
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    • pp.671-676
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    • 1999
  • RAPD analysis using random primers were tried to evaluate the genetic variation and diversity of the nine garlic cultivars including two foreign varieties. Thirty-two primers out of 70 primers screened were used to amplify genomic DNA of garlic cultivars using polymerase chain reaction(PCR). Among a total of 151 bands amplified by 32 primers, 125 polymorphic bands were subjected to analysis for genetic relationship of garlic cultivars. The estimated size of amplified PCR products were in the range of 932 to 4,060 base pairs. Nine garlic cultivars were classified into two groups, such as group I corresponded to Changnyung and Hungary cultivars, and group II, Namdo, Sandong from China, Yecheon, Euiseong, Youngweol, Danyang, Jeongsun cultivars, with the genetic distance value of 0.271. The major ecological types of garlics, so called southern and northern types, was grouped in the genetic distance value of 0.200. The results presented in this study suggest that RAPD analysis are likely to be useful for identification of cultivars and evaluation of genetic origin in garlics.

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Association between PCR-RFLP of Melatonin Receptor 1a Gene and High Prolificacy in Small Tail Han Sheep

  • Chu, M.X.;Ji, C.L.;Chen, G.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.12
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    • pp.1701-1704
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    • 2003
  • Melatonin regulates circadian rhythms and reproduction changes in seasonally reproductive mammals through binding to high-affinity, G-protein-coupled receptors. Small Tail Han sheep that has significant characteristics of high prolificacy and nonseasonal ovulatory activity is an excellent local sheep breed in P. R. China. The exon 2 of the ovine melatonin receptor 1a (MTNR1A) gene was amplified and a uniform fragment of 824 bp was obtained in 150 ewes of Small Tail Han sheep. The 824 bp PCR product was digested with restriction endonucleases Mnl I and Rsa I, and genetic polymorphism was detected by PCR-RFLP. Polymorphic Mnl I site was detected at base position 605 of the exon 2 of the MTNR1A gene. There were two kinds of genotypes in Small Tail Han sheep, AB (303 bp, 236 bp/67 bp) and BB (236 bp/67 bp, 236 bp/67 bp). The results indicated that genotype AA (303 bp, 303 bp) at Mnl I-RFLP site did not exist in non-seasonal estrous Small Tail Han sheep, which suggested that there was an association between genotype AA (303 bp, 303 bp) and reproductive seasonality in sheep. Polymorphic Rsa I site was detected at base position 604 of the exon 2 of the MTNR1A gene. Three kinds of genotypes were found in Small Tail Han sheep, AA (290 bp, 290 bp), AB (290 bp, 267 bp/23 bp) and BB (267 bp/23 bp, 267 bp/23 bp). Least squares means of litter size in the first parity and the second parity for genotype AA (290 bp, 290 bp) at Rsa I-RFLP site were 0.43 and 1.06 more than those for genotype AB (290 bp, 267 bp/23 bp) in Small Tail Han sheep.