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Golembiewski, Robert Craig. "Identification and characterization of creeping bentgrass using randomly amplified polymorphic DNA (RAPD) markers /." The Ohio State University, 1997. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487942739809001.
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Woodburn, Mary Alice. "Random amplified polymorphic DNA (RAPD) analysis of Bacillus sphaericus." Thesis, This resource online, 1994. http://scholar.lib.vt.edu/theses/available/etd-07102009-040429/.
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Robert, Florence. "Typage de "Candida albicans" par Random Amplified Polymorphic DNA (RAPD)." Paris 5, 1992. http://www.theses.fr/1992PA05P198.
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Wei, Ling. "Identification of DNA Markers in Triticum aestivum-Aegilops caudata Additions Lines by Randomly Amplified Polymorphic DNA (RAPD) Technology." DigitalCommons@USU, 1995. https://digitalcommons.usu.edu/etd/4543.
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The objective of this study was to identify DNA markers for each of six added C-genome chromosomes in Triticum aestivum L. cv. 'Alceso'-Aegilops caudata L. addition lines using the randomly amplified polymorphic DNA (RAPD) technique. DNA from Ae. caudata, T. aestivum, amphiploid of T. aestivum X Ae. caudata, and six disomic addition lines of wheat having a pair of Ae. caudata chromosomes was used as the template for the amplification of RAPD markers with a total of 58 random 10-mer oligonucleotide primers. Two primers, OPC-08 and OPJ-16, produced one intense band each from the amphiploid of T. aestivum X Ae. caudata and Ae. caudata, which was absent in all six addition lines. Each of these two primers produced a chromosome marker that could be tentatively located to the chromosome CA of Ae. caudata. OPJ-02, OPD-12, OPD-02, OPJ-12, OPD-20, and OPJ-14 produced a marker each for CB, CC, CD, CE, CF, and CG, respectively. OPJ-09 produced C-genome chromosome-specific RAPD markers. Also, OPC-05 and OPJ-19 produced RAPDs from both wheat and Ae. caudata genomes.
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Papadopoulos, Sarantos. "Untersuchungen genomischer Veränderungen von Mammakarzinomzellen mittels Random Amplified Polymorphic DNA." Doctoral thesis, Humboldt-Universität zu Berlin, Medizinische Fakultät - Universitätsklinikum Charité, 2001. http://dx.doi.org/10.18452/14631.
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Im ersten Teil der Arbeit wurde die random amplified polymorphic DNA (RAPD) Methode für die Anwendung humaner DNA optimiert, indem die Konzentrationen der einzelnen Agentien und das thermische Profil der PCR-Reaktion verändert wurden. Des weiteren wurde der Einfluss von Polymerasen und PCR-Maschinen auf die RAPD, insbesondere auf die Erweiterung des Spektrums der Amplimere und die Reproduzierbarkeit der Reaktionen untersucht. Unsere Untersuchungen haben gezeigt, dass die RAPD eine sehr robuste und reproduzierbare Methode ist. Im zweiten Teil wurden qualitative und quantitative Unterschiede zwischen DNA von Brustkrebszellen und DNA von Leukozyten detektiert. Die dazu benutzten Primer basieren auf Sequenzen die in den Mechanismen der Tumorgenese involviert sind. Unsere Studie hat gezeigt, dass random priming in der Abschätzung von genomischen Schäden im Brustkrebs sehr nutzbar sein kann.In the first part of this work, we have optimized random amplified polymorphic DNA (RAPD) for the use of human DNA in altering the concentration of the reaction components and the steps of the thermal profile in the polymerase chain reaction, by testing a large number of polymerases and multiple combinations with respect to their ability to increase the spectrum of amplimers and by examining the performance of various thermocyclers. We conclude that RAPD is a robust and reproducible method that could prove very useful for scientists and physicians. In the second part we used primers that were designed by choosing sequences involved in the development of DNA mutations, to successfully detect qualitative and quantitative differences between breast cancer DNA/normal DNA pairs. Our study showed that random priming proves very useful for assessing genomic damage in breast cancer.
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Campos, Lázara Pereira. "Genome relationships among Lotus species based on random amplified polymorphic DNA (RAPD)." Thesis, McGill University, 1992. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=56888.
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The usefulness of RAPDs (Random Amplified Polymorphic DNA) to distinguish among different taxa of Lotus was evaluated. The following species were included: L. corniculatus, L. tenuis, L. alpinus, L. japonicus, and L. uliginosus. Several accessions for each species were studied. Following DNA extraction, amplification reactions were performed in a Hybaid DNA Thermal Cycler, and the product visualized according to a standard procedure. Twenty primers were used for each species/accession. Clear bands and several polymorphisms were obtained for all primers. A phenogram was drawn based on the genetic distance among the species. L. alpinus appears as the most distant species from L. corniculatus, followed by L. uliginosus, L. tenuis, and L. japonicus. With the exception of L. alpinus, these findings are in agreement with previous experimental studies in the L. corniculatus group. The use of a greater number of primers and increased number of species may provide a greater resolution of the systematics of these taxa.
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Rinaldi, Catherine. "Authentication of the Panax genus plants used in Traditional Chinese Medicine (TCM) using Randomly Amplified Polymorphic DNA (RAPD) analysis." University of Western Australia. Centre for Forensic Science, 2007. http://theses.library.uwa.edu.au/adt-WU2008.0054.
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[Truncated abstract] Traditional medicines are used by millions of people throughout the world as their primary source of medical care. A range of materials are in used traditional medicines including plant and animal parts. Even though the traditional medicine trade is estimated to be worth sixty billion dollars annually the trade remains largely unregulated. Unscrupulous practices by vendors to increase their profit margins such as substituting and adulterating expensive material with cheaper varieties go unchecked. This can be dangerous to consumers because some substitutions involve poisonous material. Also, animal parts from endangered species can find their way into traditional medicines, therefore there needs to be a way to identify them in traditional medicines to prosecute poachers. The traditional techniques used for the identification of material used in Traditional Chinese Medicine (TCM) include, morphological, histological, chemical and immunological analysis. However, these techniques have their limitations. This makes applying multiple techniques essential to provide thorough authentication of the material. DNA profiling provides a technique well suited to analysing material used in TCM. DNA profiling is advantageous over other techniques used to authenticate material used in TCM because it requires only a small sample amount, can determine the cultivator, be used on all forms of TCM and potentially distinguish the components of mixtures. ... Therefore, profiles of different species/individual are different and species? can be distinguished. Commercially sold traditional medicines are processed which is likely to degrade the DNA of the sample making extraction and amplification difficult. Here an organic Phenol:Chloroform extraction technique extracted DNA from commercial dried root samples. The extracted DNA was amplifiable using RAPD primers. The RAPD primers used here produced enough polymorphic bands to distinguish different plant species. They were used to distinguish commercial samples that were sold as three different species within the Panax genus, Panax ginseng, Panax quinquefolium and Panax notoginseng and genetically unrelated plant material; Potato and Eleutherococcus senticosus. Liquid samples and mixtures were also profiled with the RAPD primers to determine whether the RAPD primers provide enough distinguishing ability to analyse these forms of TCM. DNA was extracted from the liquid samples, one a ginseng drink and the other an ginseng extractum. However, there was no reliability in the production of PCR products. The analysis of the mixture samples found that not enough polymorphic bands were produced by the RAPD primers used here to identify Panax species within mixtures of two Panax species. While when P. ginseng was mixed with a genetically unrelated sample there was enough polymorphism to differentiate the two samples in the mixture. The results of this research show that RAPD analysis provides a simple and inexpensive technique to begin analysis of materials used in TCM. Using RAPD analysis it is possible to distinguish Panax plant species from each other. However, the RAPD primers used here did not provide enough reproducibility or polymorphism to analyse liquid and mixtures of Panax species plants.
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Patel, Sushma M. "Ribosomal RNA genes and RAPD for Cryptosporidium species and subspecies discrimination." Thesis, Coventry University, 1997. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.389769.
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Hughes, Greta. "Population structure and genetics of the European lobster Homarus gammarus." Thesis, Bangor University, 2000. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.341219.
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Góis, Plácia Barreto Prata. "Aplicação do RAPD (Randomly Amplified Polymorphic dna) para tipagem molecular de amostras de Salmonella isoladas de diversas fontes da cidade de Aracaju-SE." Universidade Federal de Sergipe, 2006. https://ri.ufs.br/handle/riufs/3673.
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Coordenação de Aperfeiçoamento de Pessoal de Nível SuperiorThe Salmonella infection is a relevant problem in the public health, being one of the most important causes of the world´s enteric pathologies, with 1,3 million ill people, resulting in 600 deaths/year. The Salmonella spp. transmission occurs especially through water consumption and contaminated food. The diagnosis is realized using biochemical, serologic, and molecular tests. The RAPD (Randomly Amplified Polymorphic DNA) test is able to detect the genetic variation present in the isolated Salmonella spp. samples, allowing a molecular typing. This research aimed at achieving molecular typing from Salmonella spp. samples isolated from various resources (oyster, chicken, potable water, blood, and human feces) utilizing the RAPD technique. 33 Salmonella spp. samples, that came from bacteria located at the Laboratório de Virologia Comparada-DMO/CCBS/UFS, and two standard samples were utilized. Six randomized primers were used from the Ready-To-Go System RAPD; the products amplified were submitted to an electrophoretic run in a 5% polyacrilamid gel, and silver dyed. The band standard observed was analyzed by the NTSYS program. After a computational analysis it was possible to discriminate the 35 Salmonella spp. samples, resulting in 35 RAPD individual and distinct patterns, showing that the samples are genetically diversed and that there is an ample genetic biodiversity in the circulating samples in Aracaju-SE. To the grouping the Salmonella spp. samples was observed the epidemiological relationship between human samples and chicken.
A infecção por Salmonella é um relevante problema de saúde pública, sendo uma das mais importantes causas de patologias entéricas do mundo, com 1,3 milhões de doentes, resultando em aproximadamente 16000 hospitalizações e 600 mortes/ano. A transmissão da Salmonella spp. ocorre principalmente através do consumo de água e alimentos contaminados. O diagnóstico é realizado através de testes bioquímicos, sorológicos e moleculares. A técnica de RAPD (Randomly Amplified Polymorfhic DNA) é capaz de detectar as variações genéticas presente nos isolados, permitindo a tipagem molecular. Este estudo teve como objetivo realizar a tipagem molecular das amostras de Salmonella spp. isoladas de diversas fontes (ostra, frango, água residual, sangue e fezes humanas) utilizando a técnica de RAPD. Foram utilizadas 33 amostras de Salmonella spp. da bacterioteca do Laboratório de Virologia Comparada DMO/CCBS/UFS e duas amostras padrões. Foram utilizados seis primers randômicos do Sistema Read-To-Go RAPD, os produtos amplificados foram submetidos à corrida eletroforética em gel de poliacrilamida 5% e corado pela prata. O padrão de bandas observadas foi analisado pelo programa NTSYS. Após análise computacional foi possível discriminar as 35 amostras de Salmonella spp., resultando em 35 padrões de RAPD individuais e distintos, mostrando que as amostras são geneticamente diversas e que existe uma ampla biodiversidade genética nas amostras circulantes em Aracaju-SE. Ao grupar as amostras de Salmonella spp. observou-se o relacionamento epidemiológico entre as amostras humanas e de frango.
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Allnutt, Theodore Richard. "The study of genetic variation in trees using the random amplified polymorphic DNA (RAPD) technique." Thesis, Liverpool John Moores University, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.319848.
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Atienzar, Franck Andre. "Development of the random amplified polymorphic DNA (RAPD) technique to measure the effects of genotoxinsin aquatic organisms." Thesis, University of Plymouth, 2000. http://hdl.handle.net/10026.1/1757.
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Studies were undertaken to evaluate the potential of random amplified polymorphic DNA (RAPD) to detect DNA effects (including DNA damage and mutations) in aquatic invertebrates, following their exposure to a variety of environmental contaminants under laboratory conditions. After rigorous optimisation of the RAPD method, the protocol, which used a high annealing temperature (50"C for 10-mer primers), was found to generate good-quality DNA profiles from groups of organisms belonging to the bacterial, plant and animal kingdoms. The RAPD method was initially used to detect benzo(a)pyrene [B(a)P] and copper-induced DNA effects in the water flea Daphnia magna and ultraviolet-mediated DNA effects in the marine alga Palmaria palmata. The results clearly showed that changes occurred in RAPD profiles obtained from the exposed populations when compared to controls. In these studies, the effect of the genotoxins at higher levels of biological organisation (e.g. Darwinian parameters and/or fitness parameters) were also investigated and were compared with genomic DNA template stability (GTS), a qualitative index representing clear changes in panems compared to control RAPD profiles. The results from these experiments revealed that GTS could be more sensitive than growth parameters and showed at least equal or even greater sensitivity than other measures of fitness. Changes in RAPD profiles were believed to be the result of DNA effects, namely adduct formation, DNA breakage, oxidative damage and mutations and possibly other effects (e.g. variation in gene expression). Nevertheless, the nature and amount of DNA effects could only be speculated because diverse events may induce the same category of changes (i.e. variation in band intensity, appearance of bands, and disappearance of amplicons) in RAPD patterns. Further studies confirmed that RAPD had the potential to qualitatively detect oestrogen and xeno-oestrogen -induced DNA effects in barnacles. Additional experiments emphasised that oxygen radicals and variation in gene expression may induce significant changes in RAPD profiles. To further understand the effects of DNA lesions and mutations on RAPD patterns, individual types of DNA damage were created in vitro. The results clearly indicated that BaP DNA adducts, DNA photoproducts. and DNA breakages had significant effects on RAPD profiles but that diverse types of DNA damage may induce the same category of changes in RAPD patterns which render the interpretation of the results difficult. It was also concluded that mutations could be detected provided they do not arise in a random fashion. Finally, an attempt was made to determine the kinetics of DNA damage and DNA repair and whether changes in patterns obtained from B(a)P exposed Daphnia magna could be transmitted to successive generations. This strategy was developed to distinguish between mutations and DNA damage. The results showed that some bands obtained from the exposed populations were transmitted to the first and/or second generation but not to the third. It was concluded that the transmission of modified genetic material to the offspring was more likely to be the result of large genomic rearrangements and/or base methylation (epigenetic processes) rather than point mutations. In conclusion, the results presented in this research project show the potential of the RAPD assay as a useful method for the qualitative assessment of DNA effects including genotoxicity and changes in gene expression. The main advantage of this technique is that it can be applied to any species without requiring any information about the nucleotide sequence. In the field of ecogenotoxicology, its main advantage lies in its sensitivity and speed to detect a wide range of DNA damage including DNA breakage, DNA adducts, oxidative damage as well as mutations (including point mutations and large rearrangements). On the other hand, RAPD only allows a qualitative assessment of the DNA effects and the nature of the changes occurring in profiles can only be speculated. Finally, a great deal of further experimentation and validation are required in order to assess the applicability of the technique to a variety of other species and pollutants, particularly under field conditions.
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Dias, Paula Menna Barreto. "Caracterização de espécies brasileiras de Adesmia DC. por RAPD." reponame:Biblioteca Digital de Teses e Dissertações da UFRGS, 2003. http://hdl.handle.net/10183/4379.
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Dentro do gênero Adesmia, as técnicas moleculares ainda não foram empregadas na caracterização de germoplasma e na análise da diversidade genética das espécies brasileiras que compôem o gênero. Portanto os objetivos deste trabalho foram: caracterizar, com a utilização de marcador molecular do tipo RAPD, as espécies brasileiras do gênero Adesmia DC; com base nestas informações estabelecer relações de diversidade genética entre as espécies e os acessos analisados; relacionar dados de diversidade com dados morfológicos e de reprodução.
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Kongkiatngam, Prasert. "Genetic studies of red clover (Trifolium pratense L.) using morphological, isozyme and random amplified polymorphic DNA (RAPD) markers." Thesis, McGill University, 1995. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=29066.
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Genetic variation within and between two cultivars of red clover (Trifolium pratense L.), Essi from Europe and Ottawa from Canada was estimated using morphological, isozyme and random amplified polymorphic DNA (RAPD) markers. A total of 21 enzyme-coding loci with 43 alleles was detected using twelve enzyme systems. The mean number of alleles per locus was 1.81 in Essi and 1.67 in Ottawa. Nine 10-mer primers were used to assay 20 individuals from each cultivar for RAPD markers. Each primer gave from 7 to 20 amplified bands with an average of 14.8 bands per primer. High within-cultivar variation was observed in both cultivars using both isozyme and RAPD markers. The mode of inheritance of seven isozyme loci: Aat-2, Amy-1, Est-4, Est-7, Pgd-1, Pgd-2 and Skd-1, in red clover was verified. The genetic basis of banding patterns for 16 other isozyme loci: Aat-3, Adh-1, Dia-1, Dia-2, Dia-3, Est-1, Est-2, Gpi-2, Idh-1, Mdh-1, Mdh-2, Mdh-3, Mdh-4, Me-1, Me-2 and Pgm-2, was also postulated, based on the segregation patterns observed within cultivars. Two pairs of linked enzyme-coding loci, Est-4/Est-7 and Pgd-2/Skd-1, were found with joint segregation analysis. Estimates of genetic variability of 15 red clover cultivars from three different origins indicated that within-cultivar variation was much higher than between-cultivar variation. Allele frequencies of these isozymes could discriminate the five North American cultivars assayed, but they could not differentiate cultivars from Europe and Japan. The use of RAPD markers obtained from bulked samples was investigated for cultivar identification in red clover. Pooled samples were examined in order to minimize variation within cultivars. Twenty was found to be an appropriate number of red clover individuals per bulk for homogenizing genetic variation within cultivars. Fourteen 10-mer primers were used to amplify genomic DNA from combined leaf samples of 15 red clover cultivars from European, Japanese and North American origin
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Bardakci, F. "Applications of the random amplified polymorphic DNA (RAPD) technique in tilapia (Pisces cichlidae) : species, subspecies and sex identification." Thesis, Swansea University, 1995. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.636032.
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The RAPD reaction was optimised and factors affecting RAPD results were examined to obtain reproducible and reliable DNA markers for the tilapia genome. Homogeneity between RAPD samples during the preparation, amplification and electrophoresis is essential for obtaining reproducible RAPD fingerprint patterns. RAPD markers were used to discriminate between several commercially important species and subspecies of tilapia. The technique was efficient in revealing genetic variability both within and between populations. Phylogenetic relationships between these species and subspecies showed broad agreement with results from other studies. The technique was used to try to develop sex-linked RAPD markers in O. niloticus for individual sex identification. In an attempt to increase the efficiency of the technique, Bulk Segregant Analysis was initially used for three genotypic sexes (YY, XY and XX) of O. niloticus. As this approach did not produce reproducible sex-linked RAPD markers, individual DNA samples were also used. Discriminant function analysis based on the results for several primers allowed discrimination of the three genotypic sexes of O. niloticus. Three non-random short primers of repeat sequences were also tested in an attempt to search for sex-specific markers. One of these produced a RAPD-like fingerprint pattern but there were not polymorphisms between sexes. Although none of the individual primers tested produced reproducible and reliable RAPD markers, some primers gave polymorphism in some amplifications between genotypic sexes of O. niloticus. DNA from some of these marker bands was cloned and sequenced to make highly specific PCR primers. This experiment was carried out to detect genetic variation between sexes using direct sequencing, restriction fragment analysis and Single Strand Conformational Polymorphism analysis of these specific RAPD bands. None of these methods showed any difference between sexes for the RAPD bands examined. Amplification of these markers from different species of tilapia showed the potential and benefits, for diagnostic purposes, of amplification of single RAPD markers using these specific primers.
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Alamri, Sarah. "COMPARATIVE ANALYSIS OF SOYBEAN (GLYCINE MAX) ACCESSIONS USING INTER SIMPLE SEQUENCE REPEAT (ISSR) AND RANDOM AMPLIFIED POLYMORPHIC DNA (RAPD) MARKERS." Thesis, Laurentian University of Sudbury, 2014. https://zone.biblio.laurentian.ca/dspace/handle/10219/2201.
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Soybean (Glycine max) is an important crop in the world in terms of total production and usage. It is also among the least diverse species. The main objectives of the present study were 1) to determine differences between ISSR and RAPD marker systems in detecting genetic variation in soybeans and 2) to identify and characterize accession- diagnostic molecular markers in G. max accessions. Genomic DNAs from 108 G. max accessions from 11 different gene pools were analyzed using several ISSR and RAPD primers. The levels of polymorphic loci detected with the two marker systems were in general moderate and similar.. Overall, 82% of genetic distance values were above 0.40 based on ISSR analysis. However, RAPD data revealed that the accessions from different countries are closely related with 64% genetic distance values below 0.40. The dendrograms constructed with ISSR data revealed that the South Korean accessions formed an out-group while the RAPD analysis showed that accessions from Sweden were separate from the other 10 gene pools. One variety-diagnostic marker generated with ISSR 5 primer was identified in the accession Kao Chien Tao from China. This marker was cloned, and sequenced. Although RAPD and ISSR marker systems detected similar levels of genetic variability, they target different regions of the soybean genome, resulting in different clustering of the 11 gene pools indicating different genetic relatedness among them. This finding demonstrates the usefulness of both marker systems in assessing diversity and relatedness among Glycine max gene pools.
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Monteiro, Alexandre Amorim. "Padronização de metodologia para caracterização molecular por RAPD ? Random Amplified Polymorphic DNA, de bactérias ácido láticas isoladas de leite cru." Universidade Estadual de Londrina. Centro de Ciências Agrárias. Programa de Pós-Graduação em Ciência Animal, 2008. http://www.bibliotecadigital.uel.br/document/?code=vtls000129296.
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No Brasil, de modo geral, o leite é obtido sob condições higiênico-sanitárias deficientes e, em conseqüência, apresenta baixa qualidade microbiológica, constituindo um risco à saúde da população quando consumido sem tratamento térmico. Desta forma, vários grupos de microrganismos indesejáveis podem ser detectados no leite cru, gerando alterações nas mais variadas condições. Em resposta a esta situação, diferentes estratégias estão em estudo para melhoraria da qualidade. Uma dessas estratégias é a bioconservação, que vem sendo cada vez mais estudada, devido seu enorme potencial de aplicação nos mais variados tipos de alimentos. As Bactérias Ácido Láticas (BAL) são microrganismos adequados para o uso como bioconservadores devido a sua reconhecida importância como microrganismos com capacidade antagonista a diversos patógenos presentes no leite. Contudo, no Brasil, os dados sobre a presença de BAL no leite cru são limitados, sendo assim necessário o estudo de metodologias que facilitem a identificação e caracterização destes microrganismos. Entretanto, entre as diferentes espécies de bactérias lácticas ocorre uma grande similaridade em relação às necessidades nutricionais e condições de crescimento, que dificulta o uso de métodos microbiológicos clássicos para identificação de gêneros e espécies. Pesquisas têm focado a aplicação de técnicas de biologia molecular para a rápida detecção e diferenciação deste grupo de bactérias. As BAL têm sua classificação baseada em propriedades fenotípicas, incluindo parâmetros fisiológicos e padrões de fermentação de açúcares. Contudo, a correta classificação e identificação das BAL são deficientes sem o suporte das técnicas genéticas. O objetivo deste trabalho foi a avaliação de um protocolo alternativo para caracterização molecular de BAL com marcadores de RAPD (Random Amplified Polymorphic DNA), utilizando-se 265 cepas de Bactérias Ácido Láticas, com capacidade antagonista a patógenos, isoladas de 27 amostras de leite cru, de propriedades dos municípios de São Bento do Una e Saloá, do Agreste Pernambucano. A técnica foi padronizada com sucesso e apresentou reprodutibilidade, entretanto os resultados obtidos com os dois primers utilizados não foram suficientes para uma boa caracterização molecular, sendo necessária a realização de novos RAPDs com diferentes marcadores, para melhor caracterização dos isolados.The milk production in Brazil is usually conducted under poor hygienic conditions, resulting in low microbiological quality, therefore constituting a risk for the population when this milk is consumed without thermal treatment. In this form, some groups of microorganisms can be detected in raw milk, and in general, the microbiota that contaminates the milk has high capacity of multiplication, generating alterations in the most varied conditions. In reply to this situation, different strategies are in study for improve of the quality. One of these strategies is the bioconservation, that comes more being each studied at time, had its enormous potential of application in the most varied types of foods. The Lactic Acid Bacteria (LAB) are microorganisms adjusted for the use as bioconservadores due its recognized importance as microorganisms with antagonistic capacity the diverse pathogens present in milk. However in Brazil, the data of presence of BAL in raw milk are limited, being thus necessary the study of methodologies that facilitate to the identification and characterization of these microorganisms. Between the different species of lactic bacteria occur a great similarity in relation to the nutritional needs and conditions of growth, which it makes the use of classic microbiological methods limited for the identification when it comes to the genus level. Scientific works have focused the application of molecular biology techniques for the fast detection and differentiation of the LAB group. The LAB has its classification on phenotypic standards, including physiological parameters and standards of sugar fermentation. However, the correct classification and identification of the LAB group are deficient without the support of the genetic techniques. However, the correct classification and identification of the LAB group are deficient without the support of the genetic techniques. The aim of this work was the standardization of alternative methodology for molecular characterization by RAPD - Random Amplified Polymorphic DNA , 265 isolates of lactic acid bacteria from 27 samples of raw milk, from properties of the municipalities of São Bento do Una and Saloá from the Agreste of Pernambuco. The standardized technique presented to successfully reproduce, however the results obtained with the two primers used were not enough for a good molecular characterization, therefore it will be necessary to do further RAPDs with different primers, for better characterization of isolates.
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CORREIA, Janaina. "Clonagem e sequenciamento de um fragmento de DNA específico de um isolado virulento de Paracoccidioides brasiliensis." Universidade Federal de Pernambuco, 2004. https://repositorio.ufpe.br/handle/123456789/706.
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Previous issue date: 2004Paracoccidioides brasiliensis é um fungo dimórfico e agente etiológico da paracoccidioidomicose, uma micose sistêmica de evolução aguda ou crônica que se não diagnosticada e tratada a tempo pode ser fatal. Um método molecular para caracterização e detecção de P. brasiliensis foi desenvolvido a partir da clonagem e do sequenciamento de um fragmento de DNA de ~750 pb, obtido por RAPD (Random Amplified Polymorphic DNA), presente em isolados virulentos e ausente em isolados avirulentos deste fungo. Uma região interna do fragmento de DNA seqüenciado foi usada para desenhar primers que posteriormente foram utilizados em uma reação de hemi-nested PCR em tubo único. A reação de PCR específica foi capaz de amplificar DNA de três isolados de P. brasiliensis reconhecidamente virulentos e três isolados recentemente obtidos de pacientes com paracoccidioidomicose. A especificidade desta PCR foi confirmada pela ausência de produtos amplificados com DNA genômico de isolados de Histoplasma capsulatum, Blastomyces dermatitidis, Coccidioides immitis, Sporothrix schenckii, Cryptococcus neoformans, Candida albicans, Aspergillus fumigatus, Mycobacterium tuberculosis, Leishmania braziliensis, Trypanosoma cruzi, Schistosoma mansoni, DNA genômico humano (leucócitos) e de isolados de P. brasiliensis reconhecidamente avirulentas. A amplificação de cDNA de um isolado virulento sugere tratar-se de um gene expresso. A detecção específica de isolados virulentos de P. brasiliensis sugere ser este um candidato a marcador de virulência para este fungo. O potencial diagnóstico da PCR específica foi verificado com DNA extraído de aspirado de linfonodo de um paciente com paracoccidioidomicose
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Ivgin, Tunca Rahsan. "Determination And Comparison Of Genetic Variation In Honey Bee (apis Mellifera L.)populations Of Turkey By Random Amplified Polymorphic Dna And Microsatellite Analyses." Phd thesis, METU, 2009. http://etd.lib.metu.edu.tr/upload/3/12610443/index.pdf.
Full textAbstract:
We analyzed a total of 760 worker bees, two samples per colony, 390 colonies in 26
provinces in Turkey to determine and compare the genetic variation of Turkish
honey bee (Apis mellifera L.) populations using 10 primers for RAPD and 6
microsatellite loci. Mean gene diversity levels ranged from 0.035 (Sanliurfa) to 0.175
(Antalya) for RAPD and 0.449 (Mugla) to 0.739 (Artvin) for microsatellite markers.
Private band patterns and alleles, pairwise FST values support that the Anatolian
honey bees belong to C lineage except for Hatay and Sanliurfa populations illustrated
from previous findings of mitochondrial DNA studies. Genetic differentiation (GST)
from RAPD data ranged from 0.060 (Bilecik and Mugla) to 0.395 (Gökç
eada and Sanliurfa). The genetic diversity (FST) for microsatellites ranged from -0.068 (Gö
kç
eada and &
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zmir) to 0.347 (Konya and Mugla). The results of the present research are in agreement to that of previous study in Turkish honey bee populations which used different microsatellite loci. That is the genetic variation was the highest in African, the lowest in European and intermediate in the Mediterranean honey bee populations. The data presented here indicate that in spite of extensive migratory beekeeping, there is still a large genetic differentiation among honey bee populations. These results should be considered in establishment of conservation plans particularly in moving of colonies between regions. The most importantly introduction of bees with foreign origin and distribution queen bees from one center to all over the country which will homogenize the gene pool of the populations should be prevented
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Sovic, Michael G. "An Evaluation of Fluorescent Randomly Amplified Polymorphic DNA (FRAPD) as a Tool for Identifying Species Hybrids, and the Application of these Markers to Questions of Hybridization in Two Groups of Ohio River Basin Fishes." The Ohio State University, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=osu1308056342.
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Sarre, Caroline. "Utilisation de la méthode Random Amplified Polymorphic DNA (RAPD) pour le typage moléculaire du genre Scedosporium : application à des isolats collectés chez des patients atteints de mucoviscidose." Paris 5, 1999. http://www.theses.fr/1999PA05P039.
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Barcelos, Ivanildes Solange da Costa. "Polimorfismo por Random Amplified Polymorphic DNA (RAPD) em metacestódeos de Taenia solium provenientes de diferentes áreas geográficas do Brasil e a reatividade de anticorpos IgG séricos de pacientes com neurocisticercose frente aos isolados obtidos." Universidade Federal de Uberlândia, 2006. https://repositorio.ufu.br/handle/123456789/16559.
Full textAbstract:
Neurocysticercosis (NC) is a polymorphic disease and the immune response in
human carrier is heterogenic. In this study, 35 primers were used for amplifications by
Random Amplified Polymorphic DNA (RAPD) of Taenia solium metacestodes, from five
different geographic areas in Brazil: 1) Distrito Federal (DF), Center West; 2) Barreiras
(BA), Northeast and Southeast; 3) Hydro Basin of the Mosquito River (North of Minas
Gerais, RM-MG), 4) São Paulo (SP) and 5) Uberaba (Minas Gerais, UB-MG).
Metacestodes saline crude extracts of four populations (DF, BA, RM-MG e SP) were used
for the detection of specific IgG antibodies by ELISA and Western Blotting (WB). A total
of 157 serum samples of three groups, (G1): 49 NC patients; (G2): 68 patients with other
helminthiasis: hydatidosis (10), taeniasis (20), strongyloidiasis (20), schistosomiasis (10)
and hymenolepiasis (8); and (G3): 40 healthy individuals; were analyzed by ELISA. From
these, the 98 serum samples were assayed by WB; G1 (49), G2 (39) and G3 (10). The
genetic distances, in disagreement percentage, between the metacestode populations were
calculated from of 15 RAPD markers and showed 49.5% (DF), 48% (BA), 38.5% (UBMG)
and 28% (RM-MG and SP) of genetic distances. Six primers identified polymorphic
fragments and the primers 26 (GGGTTTGGCA) and 29 (TCGCCAGCCA) allowed a
better differentiation of populations. The fragments of 1000, 500 and 326 pb (pairs of
bases) in the UB-MG and of 600 and 244 pb in RM-MG were amplified by primer 26. The
fragments generated by primer 29 were 500, 800 and 1191 pb, 300 and 1377 pb, 1000 pb
and 244 and 434 pb in SP, UB-MG, DF and BA populations, respectively. In G1, the
positivity by ELISA was: 90% (DF), 69% (BA), 71% (MG) and 67% (SP). The DF extract
was more antigenic than others (p=0.02). In WB, the 64-68 kDa antigens were recognized
in all extracts, exclusively, in serum samples from active NC patients (p=0.001). Variation
in banding pattern was detected between the extracts (p<0.05). In G2, the serum samples
of hydatidosis patients presented from 70 to 90% positivity by ELISA in antigenic
extracts (p<0.05); however, the bands recognition pattern in WB was different from that
presented in G1 samples. The 77 kDa band was significantly identified in hydatidosis
samples (p=0.0001). In conclusion, the T. solium populations analyzed showed genetic
variability and antigenic differences.A neurocisticercose (NC) constitui doença polimórfica, apresentando heterogeneidade da resposta imune no hospedeiro humano. Nesse estudo, o teste Random Amplified Polymorphic DNA (RAPD) foi utilizado com 35 primers na detecção de polimorfismo em metacestódeos de Taenia solium provenientes de cinco áreas geográficas distintas do Brasil: 1) Distrito Federal (DF), região Centro-Oeste; 2) Barreiras (BA), região nordeste e da região sudeste: 3) Bacia Hidrográfica do Rio Mosquito (norte de Minas Gerais, RM-MG), 4) São Paulo (SP) e 5) Uberaba (Minas Gerais, UB-MG). Os extratos salinos totais de metacestódeos de quatro populações (DF, BA, RM-MG e SP) foram utilizados na detecção de anticorpos IgG específicos pelo ELISA e Western Blotting (WB). As 157 amostras de soro de três grupos (G) de indivíduos: G1: 49 pacientes com NC; G2: 68 pacientes com outras helmintíases, sendo hidatidose (10), teníase (20), estrongiloidíase (20), esquistossomose (10) e himenolepíase (8) e G3: 40 indivíduos saudáveis (controles); foram analisadas pelo ELISA. Foram ensaiadas 98 amostras de soro: G1 (49), G2 (39) e G3 (10) pelo WB. As distâncias genéticas, por porcentagem de desacordo, foram de 49,5% (DF), 48% (BA), 38,5% (UB-MG) e 28% (RM-MG e SP) nas populações de metacestódeos, calculadas a partir de 15 marcadores de RAPD. Seis primers geraram fragmentos polimórficos nos isolados analisados, sendo que os primers 26 (GGGTTTGGCA) e 29 (TCGCCAGCCA) permitiram melhor diferenciação entre as populações, o primer 26 gerou os fragmentos de 1000, 500 e 326 pb (pares de bases) na amostra de UB-MG, e de 600 e 244 pb em RM-MG. O 29 gerou fragmentos em quatro das populações analisadas, sendo 500, 800 e 1191 pb em SP, 300 e 1377 pb em UBMG, 1000 pb no DF e 244 e 434 pb na BA. No G1, as freqüências de positividade no ELISA, foram: 90% (DF), 69% (BA), 71% (MG) e 67% (SP), sendo o extrato do DF mais antigênico que os demais (p = 0,02). No WB, o peptídeo de 64-68 kDa foi reconhecido em todos os extratos, exclusivamente, em amostras de pacientes com NC ativa (p=0,001). Detectou-se variação no padrão de reconhecimento de bandas entre os extratos (p<0,05). No G2, as amostras de soro de pacientes com hidatidose apresentaram de 70 a 90% de positividade no ELISA frente aos extratos analisados (p<0,05); porém, o padrão de reconhecimento de bandas no WB diferiu do apresentado pelas amostras do G1, sendo que a banda de 77 kDa foi significativamente identificada pelas amostras de pacientes com hidatidose (p=0,0001). Concluiu-se que as populações de T. solium analisadas nesse estudo, apresentaram variabilidade genética e diferenças de antigenicidade.
Doutor em Imunologia e Parasitologia Aplicadas
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Beeson, Keri Elizabeth. "Differentiation of plasmids in marine diazotroph assemblages determined by randomly amplified polymorphic DNA analysis." Thesis, Georgia Institute of Technology, 2001. http://hdl.handle.net/1853/25366.
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Corniquel, Béatrice. "Construction de banques d'ADNc de palmier dattier (Phoenix dactylifera L. ). Différenciation de cultivars par RFLP à l'aide des ADNc : isolement et caractérisation d'un ADNc polymorphe. Isolement et caractérisation d'un ADNc codant pour la glutamine synthetase cytosolique." Angers, 1994. http://www.theses.fr/1994ANGE0014.
Full textAbstract:
La caractérisation des cultivars de palmier dattier a été réalisée par des analyses RFLP et RAPD à partir des feuilles internes des rejets. Des sondes ont été isolées d'une banque d’ADNc construite à partir d'ARNM extraits de cals organogènes du cultivar bou sthammi noire. La sonde ADNc 1 a permis d'obtenir neuf profils d'hybridation distincts pour chacun des neuf cultivars étudiés. Des profils d'amplification distincts pour trois cultivars ont été obtenus par RAPD avec des amorces oligonucléotidiques. Des analyses RFLP ont été aussi réalisées sur du matériel maintenu in vitro ou issu de culture tissulaire afin de déterminer si d'éventuelles variations induites par la culture in vitro affectaient le génome du palmier dattier. L'analyse moléculaire de la sonde ADNc 1 a été abordée. Le transcrit qui comprend 2400 nucléotides est exprimé dans les feuilles et les cals organogènes. Sa séquence partielle a été établie, l'absence d'homologie entre cette séquence de 1456 nucléotides et les séquences enregistrées dans les banques de données suggère que nous avons isole un ADNc codant pour une nouvelle protéine. Un ADNc codant pour la glutamine synthétase a été isolé. L'absence au niveau de la séquence des aminoacides de cet ADNc de l'extension c-terminale qui caractérise la structure primaire des séquences des sous-unités chloroplastiques montre que l’ADNc isole code pour la sous-unité cytosolique de la glutamine synthétase.
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MARTINS, Walter Fabrício Silva. "Diversidade genética de populações naturais de Anthonomus grandis Boheman (Coleoptera : Curculionidae)." Universidade Federal de Pernambuco, 2006. https://repositorio.ufpe.br/handle/123456789/716.
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Previous issue date: 2006O bicudo do algodoeiro, Anthonomus grandis Boheman, é considerado a maior praga da cotonicultura mundial, ocasionando danos que repercutem principalmente na produtividade, qualidade do algodão colhido e gasto com medidas de controle. Neste estudo foi realizada pela primeira vez, uma análise da diversidade e estrutura genética das populações naturais de A. grandis do Brasil. Doze populações coletadas em seis estados brasileiros (Paraíba, Ceará, Bahia, Pará, Mato Grosso e Goiás) em áreas onde são praticadas a agricultura em escala empresarial e agricultura familiar, foram avaliadas pelas técnicas de Polimorfismo do DNA Amplificado Randomicamente (RAPD), Isoenzimas e Microssatélite. Os resultados obtidos em seis populações pela técnica de RAPD baseados em 25 loci, revelaram uma heterozigosidade média de 0,262, com polimorfismo (P) variando de 52 a 84%. A diferenciação genética entre as populações foi extremamente elevada e significativa (GST = 0,258; p < 0,05), refletindo a existência de baixo fluxo gênico entre as mesmas (Nm = 0,72). A análise de cinco populações com 6 loci alozímicos mostrou uma heterozigosidade média de 0,212 e polimorfismo (P) variando entre 25 e 100%. O índice de diferenciação genética FST obtido por este marcador entre as populações correspondeu a 0,544 (p < 0,05), sugerindo a ocorrência de baixo fluxo gênico (Nm = 0,210) entre as populações. A heterozigosidade e o polimorfismo (P) observados em onze populações pela análise de 8 loci de microssatélites variaram entre 0,038 e 0,224 e de 37,5 a 75%, respectivamente. O FST entre as populações correspondeu a 0,220, produzindo um Nm de 0,8. Os três marcadores moleculares utilizados revelaram que as populações de A. grandis dos estados brasileiros avaliados apresentam baixa diversidade genética, em comparação às populações dos Estados Unidos, México e demais países da América do Sul, sugerindo que a colonização deste inseto ocorreu em uma ou poucas áreas. Os resultados obtidos relativos à diversidade genética também permitiram distinguir populações oriundas de regiões onde é praticada a agricultura em escala empresarial das áreas de agricultura familiar, assim como mostrou que as populações do nordeste podem estar servindo de fonte para colonização de novas áreas e de áreas já tratadas
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Wang, Jau-Yueh, and 王昭月. "Identifications of Capsicum spp. by Isozymes and Randomly Amplified Polymorphic DNA (RAPD) Markers." Thesis, 1995. http://ndltd.ncl.edu.tw/handle/94172284351782431844.
Full textAbstract:
碩士國立中興大學
園藝學系
83
Morphological traits,isozymes and randomly amplified polymor- phic DNA (RAPD) markers were used to study the identification of Capsicum species.The purposes of this study were to establish molecular markers for classification of the Capsicum spp., to establish the genetic linkage relationships and a quick, technique for the identification of seed purity of Capsicum spp. The results of isozyme analysis indicated that there were significantly different in the zymograms of EST, GOT, PGI, PGM, PRX, SkDH and SOD between C. annuum L. and C. chinense Jacquin. Distinctly polymorphic patterns were exhibited among the C. annuum L., but only three lines can be distinguished. Besides, the isozyme patterns might vary with different extracting buffers and sampling positions. For example, the expressions of PRX, SOD and PGI were clearer in lower leaf, GOT, PGM and SkDH in middle leaf and EST in upper leaf. A total of 10 primers was selected from 300 random primers that showed the polymorphic patterns in Capsicum spp.. Among , the RAPD patterns of six primers,i.e."UBC 313","UBC 327" , "UBC 346", "UBC 457","UBC 483" and "UBC 484", could clearly eight tested Capsicum lines and might be used as an effective for the identification of F1 hybrids and cultivated varieties of Capsicum spp.. The results of linkage cluster analysis revealed that the genetic background of 12 cultivars was very similar, and might originate from P852. Among the five inbred lines of sweet pepper, P1717 had a closer relation with P852, next by P1657 and P1740, but P 859 was farthest.The three hot pepper lines belonged to different population, when compared with these five inbred lines of sweet pepper. The results suggested that the RAPD markers is preciser and more sensitive than morphological and isozyme identifications and can be used as an effective tool to distinguish plant germplasms.
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Wang, Zhao-Yue, and 王昭月. "Identifications of capsicum spp. by isozymes and randomly amplified polymorphic DNA (RAPD) markers." Thesis, 1995. http://ndltd.ncl.edu.tw/handle/25557591573302930114.
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Wei, Jun-hung, and 魏俊宏. "Cloning Sex Relatine Genes of Carica papaya L. by Randomly Amplified Polymorphic DNA (RAPD)." Thesis, 1998. http://ndltd.ncl.edu.tw/handle/84072335020026873496.
Full textAbstract:
碩士東吳大學
微生物學系
86
Carica papaya L. is a trimonoecious (polygamous) species that individualplants can be classified to three basic sex types:hermaphrodite, pistillate(female) and staminate (male).The results of cross- and self-pollinations in papaya have showed that the determination of sex type was caused by inherited factors.In this work, we try to clone the different DNA fragments between staminate and pistillate by randomly amplified polymorphic DNA (RAPD).In order to isolate the sex determination genes of papaya, we have screenedwith 200 primers and have cloned two specific DNA fragmentsamplified by respective two primers.A genomic DNA blot analysis by using these two fragments as probes indicates no restriction fragments length polymorphism(RFLP) between pistillate and staminate genomes.In order to isolate thesex determined gene of papaya, it is necessary to use a large number of10-meroligo primer to perform RAPD experiments.
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Liu, Yi-Chin, and 劉怡君. "Identification of Cynodon dactylon (L.) Pers. by Morphological Traits and Randomly Amplified Polymorphic DNA (RAPD) Markers." Thesis, 1996. http://ndltd.ncl.edu.tw/handle/67411824389673206103.
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碩士國立中興大學
園藝學系
84
Summary The clusis analysis with both the morphological traits and the RAPD marker data were used to identify 16 bermudagrass cultivars. It was the first time RAPD method been used on bermudagrass for cultivar identification. The goals for this experiment are: 1)To build up a database for bermudagrass morphological characters, and use the data for cluster analysis to construct relationship data between cultivars. 2)Testing the reliability of RAPD test compare to the morphological traits. 3)Develop an accurate and rapid techniquefor cultivar identification. The morphological traits were measured based on the categories for Gramineaegrasses. Both qualitative and quantitative characters including: ligual, hairs on the edges of collar, pubescence on leaf blade and on leaf back, stem color,plant height, texture of the leaf and the canopy of the grass were used to describe the characteristics of 16 bermudagrass collected. We quantitized those qualitative characters then added up with the quantitative charactersfor similarity cluster analysis. With the cluster analysis, 16 bermudagrass cultivars were separated into three groups. Selected from 100 primers tested, four were chosen to be useful forcultivar identification using RAPD marker. Four primers were able to produce polymorphic DNA fragments to clearly identify N1, 0035 and 0036 varieties.With cluster analysis, 16 bermudagrass cultivars were separated into threegroups. Menbers of the three groups are corresponding to three types of bermudagrass according to their plant heitht and genetic constituent. The use of RAPD marker data for bermudagrass identification were proved to be useful by comparing the results with morphological traits data. Except for Tifway,15 cultivars were easily categorized into three types (common, dwarf, or semi-dwarf) by either RAPD or morphological traits method. The primerbank may need to be expanded for the selection of right primer used to separate every bermudagrass cultivars. From our experiments, use of the RAPD marker for cultivar identification is a quick and reliable method. A large scale selection of primers suitable for RAPD on bermudagrass has to be held before the method can be efficiently used.
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Blue, Gillian Margaret. "Assessment of genetic diversity and DNA fingerprinting of the Cape parrot (Poicephalus robustus) using randomly amplified polymorphic DNA (RAPD)." Thesis, 2004. http://hdl.handle.net/10413/10126.
Full textAbstract:
The Cape parrot (Poicephalus robustus) is South Africa's only endemic parrot. It has
become increasingly rare in recent years, with fewer than 500 birds left in the wild,
and is now regarded as endangered. Possible factors contributing to this rapid decline
in numbers include habitat loss, food shortage, disease and illegal trafficking and
trading in the species. Habitat loss and food shortage have been brought about by the
rapid destruction of the yellowwood trees in the afromontane forests in South Africa
and have played a role in reducing the population numbers. The Psittacine beak and
feather disease virus (PBFDV) has also contributed to the loss of some individuals,
however it is the illegal trafficking of this rare and valuable species that has become
of great concern. As the Cape parrot is becoming increasingly rare and therefore
highly sought after, its commercial value has multiplied to the extent that illegal black
market trapping is on the rise.
The industry involved in breeding and conservation of endangered bird species, has a
need for the proper establishment of studbooks, containing all available information
on captive as well as tagged birds. Most of the information found in studbooks is
based on morphological attributes of individual birds. Although this is useful, there is a
need to add molecular information in order for complete identification of individuals,
particularly in a species threatened by illegal trading and theft. A preliminary analysis
of the amount of variation present in the population of interest is therefore required so
that appropriate methods and techniques can be developed to identify individual
birds. A RAPD analysis was conducted to assess the amount of variation in the Cape
parrot and lay the foundations for the establishment of individual identification in the
species.
Blood samples from 30 parrots, consisting of both related and unrelated individuals,
were obtained from three separate locations: Amazona in Assagay, Rehoboth Farm
in Dargle, as well as from the Eastern Cape. 15 random primers were selected and used to conduct a randomly amplified polymorphic DNA (RAPD) analysis. RAPDs are
extremely useful in situations where relatively inexpensive first approximations of the
genetic variation are needed, such as in rare and endangered species. After
successful optimisation of the technique in the species, the 15 primers were screened
for all 30 individuals and the individual DNA fingerprints, analysed.
Clear, distinctive and reliable DNA fingerprints were obtained for all individuals
however, it was interesting to note despite the analysis of 85 loci using the 15 primers
almost identical DNA fingerprints were produced between the individual birds. A
population analysis into the amount of variation present between and within the three
populations, as well as for the representative population as a whole, was conducted.
Using various statistical programmes such as POPGENE and ARLEQUIN,
heterozygosities, genetic distance measures, diversity indices, Wright's fixation index
and AMOVAs were estimated.
The amount of polymorphism detected in this investigation was 33 % and the
heterozygosity, 0.37, which is a relatively high value for the uniformity displayed in the
DNA profiles. The high GC content of the primers however, could be a possible
explanation thereof. Relationship and kinship determination, sex determination as
well as population assignment was possible despite not being able to identify each
individual based on unique DNA fingerprints.
The AMOVA analysis indicated significant variation on both the between (5.59 %) and
within (94.41 %) levels of analysis. Little variation or differentiation was observed
between the three subpopulations, which was confirmed with an FST value of 0.056.
The variation experienced within each subpopulation was analysed using Shannon's
index of phenotypic diversity. The Amazona population displayed the most variation
with a value of 0.286 and the Rehoboth population, the least with 0.195. This was
expected, with the individuals from the latter population comprising one extended
family. Nei's measures of genetic identity revealed that the individuals from Amazona were more similar to the Eastern Cape population, which was again expected with
regular exchanging of chicks between the two breeders.
RAPD technology was successful in laying the foundations for individual identification
in the Cape parrot. It was also successful in producing reproducible DNA fingerprints
in the species that were able to determine relatedness to some extent, determine the
sex of individuals and identify individuals from a particular subpopulation.
Furthermore RAPD analysis gave a good indication of the variation found in the Cape
parrot population, which is important for conservation purposes. In order to maximize
conservation efforts and strategies in an endangered species, determining the level of
genetic diversity and variation found in the remaining individuals of the population is
of great importance. This information could provide powerful insight for conservation
purposes and depending on the level of diversity detected, appropriate breeding
programmes could be set up in order to increase the genetic variation and thereby
reduce the chance of extinction of the species.
The following important findings emerged from this investigation:
• RAPD technology, once optimised for the species of interest, is successful in
producing clear and reliable DNA fingerprints, provided the same protocol is
followed carefully throughout the investigation.
• An optimised protocol for fingerprinting the Cape parrot using RAPDs was
established.
• Possible sex identification, population assignment and a degree of kinship
determination was determined using RAPDs.
• Little variation was found within the representative Cape parrot population as a
whole due to small population size and possible inbreeding.
• As expected for an avian species, little genetic sub-division or differentiation
was observed between the three populations analysed.Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2004.
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Nagaraja, G. M. "Genetic analysis of Bombyx mori using Random amplified Polymorphic DNA (RAPD) markers." Thesis, 1999. http://hdl.handle.net/2009/1354.
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Liu, Chi-An, and 劉啟安. "Application of Random Amplified Polymorphic DNA (RAPD) for Species-Specific Identification in Parrots." Thesis, 2016. http://ndltd.ncl.edu.tw/handle/42374419271822770641.
Full textAbstract:
碩士國立中興大學
動物科學系所
104
In taxonomy, parrots belong to gegnus Psittaciformes and distribute mainly between north and sough latitudes. Until now, it is known that there are about 374 kinds of parrots around the world. Although Taiwan is not the original habitat of parrots, parrots are one of the most popular companion animals in Taiwan due to their beautiful plumage and high intelligence. However, all of the parrot species, except for Agapornis roseicollis, Melopsittacus undulates, Nymphicus holandicus and Psittacula krameri, are endangered and protected by CITES (Convention on International Trade in Endangered Species of Wild Fauna and Flora) under different degrees of protection by listed on Appendix I or Appendix II. Therefore, it is necessary to develop molecular diagnosis methods with smuggled eggs or trace of material evidence to identify species, and the polymerase chain reaction (PCR)-based methods would be the first reasonable choice. Among various methods, the PCR-based random amplified polymorphic DNA (RAPD) analysis is conducted by short random primer sequences at low annealing temperature to produce multiple-band PCR products without knowing the sample information. Hence, RAPD was applied for identification of parrot species in this study. A total of 120 random primers from 6 series were used to identify the species-specific fragment in genomic DNA from 8-15 parrot species, and 52 possible bands were obtained after RAPD. After further analysis, two pairs of sequence characterized amplified region (SCAR) primers, called GCC-F/R and Monk-F/R, were designed according to sequencing results of the 437 bp RAPD product obtained from Pyrrhura molinae by random primer OPD15 and 529 bp RAPD product obtained from Myiopsitta monachus by random primer OPD7, respectively. GCC-F/R primer sequences were identical to the first 26 bp and the last 27 bp from the 437 bp RAPD product of Pyrrhura molinae, and Monk-F/R primer sequences were from the 529 bp RAPD product of Myiopsitta monachus. The GCC-F/-R and Monk-F/-R were applied to evaluate their species specificity. The electrophoresis result revealed that only genomic DNA from green-cheeked conure could be amplified with GCC-F/-R primers, and only monk parakeets genomic DNA could be amplified with Monk-F/-R primers after validation among 16/ 12 intra- and 15 inter species, indicating that these two pairs of SCAR primers could be applied for identification of monk parakeets and green-cheeked conure.
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Edwards, Nicola Rachel. "Optimisation of the randomly amplified polymorphic DNA (RAPD) technique for the characterisation of selected South African maize (Zea mays L.) breeding material." Thesis, 2000. http://hdl.handle.net/10413/9812.
Full textAbstract:
Maize (Zea mays L.) is an important agronomic crop with the maize industry forming an
important component of the South African economy. Considerable effort has been directed
towards the genetic improvement of maize through both conventional breeding and
biotechnology. Genotype identification by DNA fingerprinting is becoming an important activity
in plant breeding. A widely used molecular based and relatively inexpensive method for DNA
fingerprinting is the randomly amplified polymorphic DNA (RAPD) technique. The RAPD
technique was tested in this study for its potential use in maize breeding programmes. Initial
results using the technique showed a low degree of reproducibility, therefore both the DNA
isolation and RAPD protocols were extensively optimised. DNA quality and quantity, and choice
of Taq polymerase buffer were three of the variables found to be influential in ensuring
reproducibility. The ability of the RAPD technique to characterise seven maize genotypes was
evaluated. Sixty random oligonucleotide primers were screened. Forty two primers scored a
total of 233 fragments (an average of 5.5 per primer), but not all primers gave reproducible
profiles. Eighteen primers scored a total of 110 loci for the presence (1) and absence (0) of DNA
fragments. RAPD markers were able to distinguish between all seven genotypes with five primers
producing specific fragments for four genotypes. Genetic similarity matrices were calculated
using two software programmes i.e. Genstat 5™ release 4.1 (1993) and PAUP (Phylogenetic
Analysis Using Parsimony) 4.0 beta version (Swafford, 1998). Cluster analysis was used to
generate dendrograms to visualise the genetic relationships of the seven maize genotypes (only
minor differences were observed between the Genstat or PAUP method of analysis). Genetic
diversity ranged from 0.62 to 0.96. The estimation of genetic relationship was in accordance with
the presumed pedigree of the genotypes showing that the RAPD technique demonstrates potential
for genome analysis of maize. The applicability of the technique for marker assisted selection was
also evaluated. Near-isogenic lines (NILs) for leaf blight (Helminthosporium spp.) were screened
for polymorphisms using a total of 120 primers. Ten primers identified polymorphisms between
the NILs. Four primers produced five polymorphic fragments present in the resistant inbred
K0315Y and absent in the susceptible inbred D0940Y. A small F2 population of 14 individuals
was produced by selfing the F1 of a cross between K0315Y and D0940Y. To speed up the generation time, the F1 and F2 plants were cultured by embryo rescue from 18d old harvested
seed. One fragment of 627 base pairs produced by primer OPB-01 (5' GTTTCGCTCC 3')
showed a 3: 1 segregation in the small F2 population and was considered putatively linked to the
HtN gene for leaf blight resistance. This study shows that the RAPD technique does have
application in maize breeding programmes.Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2000.
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Mailer, Rodney J. "Cultivar identification of oilseed rape, Brassica napus, by high performance liquid chromatography (HPLC) and randomly amplified polymorphic DNA (RAPDs)." 1993. http://hdl.handle.net/1993/17765.
Full text
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Wu, Hui-Ju, and 吳慧如. "Availability of Using RAPD(Random Amplified Polymorphic (DNA) Segments as Hybridization Probes in Sorghum RFLP analysis." Thesis, 1993. http://ndltd.ncl.edu.tw/handle/96177597261715815246.
Full textAbstract:
碩士國立臺灣大學
農藝學系
81
The molecular markers had been developed as a powerful tool for the studies in plant genetics and breeding. There are two types of molecular markers : one is restriction fragment length polymorphism (RFLP) marker, and the other is random amplified polymorphic DNA (RAPD) marker. In order to obtain the RFLP markers, it should take a long time and spend much resources to construct and screen a genetic library. However, since the polymerase chain reaction (PCR) technique was developed, it has revolutionized the RAPD marker production. RAPD markers are polymorphic DNA segments separated by gel electrophoresis after PCR amplification using random primers. Genetic analysis with RAPD markers is fast and involves no radioactivity and hybridization. For our current sorghum RFLP map construction, we had screened the informative probes from a PstI and a HpaII genomic libraries. In this study, we exploited the availability of using polymorphic DNA segments appeared in RAPD analysis as probes in RFLPs. Two sorghum parents, V-3 and V-160, were used as sources of DNA. We had screened out 192 amplified segments in 221 arbitrary 10-mer primers. And 142 of them had been used in RFLP analysis. Most of them were with a fragment size larger than 1.0 kb and shown to be multiple copies. There were 54 segments shown to be suitable as RFLP probes in these 142 segments. To explore the features of RFLP in sorghum, the relationship between the probability that a given enzyme detected polymorphism with a given probe in this study and the number of other enzymes detecting polymorphism with the same probe was plotted for regression. The result of regression suggested that insertion/ deletion was the major mechanism for generating RFLPs in the sorghum genome, however, the base replacement was not excluded.
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Naguran, Riann. "Fingerprinting of full and half-sib black wattle (Acacia mearnsii) progenies using Random Amplified Polymorphic DNA (RAPD)." Thesis, 2005. http://hdl.handle.net/10413/4745.
Full textAbstract:
Black wattle (Acacia mearnsii), which belongs to the genus Acacia, is one of the many species of trees or hardwoods grown commercially in South Africa. Black wattle is a species indigenous to Australia and was introduced into South Africa by the van der Plank brothers in 1864. These trees are grown in South Africa because of its tannin-rich bark, the extract of which is used by the leather tanning industry. Black wattle is also grown for its timber, timber products and pulp. The introduction and cultivation history of
black wattle suggests that the South African plantations contain limited genetic variation with relatedness amongst groups estimated to be high, thus implying a narrow genetic base in the South African black wattle population. In this investigation, Random Amplified Polymorphic DNA (RAPD) was used to estimate the genetic variation between seven different black wattle groups. A total number of 34 individuals obtained from different areas in South Africa were examined; Piet Retief (group 47 and 50: half-sibs), Kumbula (group 85: unrelated individuals), Howick (group 400: unrelated individuals) and an unknown area (groups 88, 89, 91: full-sibs). As this investigation was the first of its kind, a DNA isolation method as well as a PCR-RAPD protocol had to be modified. Total genomic DNA was successfully extracted using the CTAB DNA extraction method. This method removed large amounts of tannin present in the cells of the black wattle
leaves and extracted high quality DNA to conduct between 50-100 RAPD reactions. The DNA purities ranged from 0.1 to 1.8, with an average of 1.46. A total of fourteen 10-mer RAPD primer sequences were randomly selected from the Operon Technologies primer list A, and tested in this investigation. Of the 14 primers used, only nine primers produced clear, single and repeatable bands. Therefore nine primers were selected for
subsequent analyses. Ninety one loci that generated bands ranging from 300-3050 base pairs were produced. Seven to 13 loci per primer were generated. A total of 95.6 % of the loci were polymorphic. The overall expected mean heterozygosity (H = 0.3) obtained in this study was high in comparison to other studies conducted on acacias. The high levels of genetic variation were attributed to mating systems, dissortative mating and geographic distribution. The statistical packages POPGENE and ARLEQUIN were used to analyse the RAPD fingerprints. The genetic measures, Nei's diversity and Shannon's Information Index, showed that there was greater diversity exhibited (Nei's gene diversity = 32.09 % and Shannon's = 48.31 %), in the whole population than in each of the groups (with average of Nei's gene diversity = 20.33 % and Shannon's = 34.64 %).
With regards to individual group analyses, low levels of genetic variation was obtained in group 400 (unrelated), from the Howick region, and group 85 (unrelated), from the Kumbula region, (mean 0.14 and 0.17 respectively). The low genetic values were attributed to limited gene exchange occurring in these two areas, bottlenecks and selection pressures. Groups 88, 89 and 91, from the unknown region (full-sib groups), were the most variable in comparison to the other groups, with means of (0.27,0.24 and 0.18 respectively). These high genetic variation values could be due to the fact that gene migration could have occurred between these groups and others in the area. It is thought that most acacias are insect-pollinated and this could have lead to gene migration between groups or populations, thereby explaining the high mean values. The gene flow obtained for the seven groups (FST = 0.174) indicated that great genetic differentiation existed in this population of black wattle studied. This value is higher in comparison to other woody species; however it is similar to other acacia species. UPGMA cluster analysis using Nei's unbiased genetic distance, revealed four distinct clusters of groups corresponding to the distribution areas represented in this study. The
Howick (group 400: unrelated) and Kumbula (group 85: unrelated) were more closely related to each other than to the other groups, since both these groups are from Natal. The Piet Retief groups (groups 47 and 50: half-sibs), branched-off together, indicating that they are distinct from the other groups. The pairwise analysis of identity showed that
the relationship between the group from Howick (group 400: unrelated) and all the other groups from the other regions was the lowest, ranging from 64 % to 79 %. The relationship between all the groups beside the group from Howick (group 400: unrelated) was reasonably high, ranging from 78 % to 90 %. This distance displayed by group 400 (unrelated) from Howick in relation to the groups, is attributed to the fact that it is frost
resistant and the other groups not. Genetic variation was also detected and partitioned, between and within groups, by Analysis of Molecular Variance (AMQVA). Majority of the variation existed within groups (82.65 %) but significant differentiation was recorded between groups (17.44 %). This high level of within group differentiation may be explained by many aspects, such as the species breeding system, genetic drift or genetic isolation of groups or populations. The application of RAPD fingerprinting in
black wattle has provided a more in depth understanding of the genetic variation residing in the South African population. The results achieved implementing this technique has shown that significant genetic variation exists within the black wattle population in South Africa. The results obtained in this study are also important since it is contrary to the expectation that the black wattle population in South Africa has low
genetic variation. This knowledge is of great value to genetically discriminate between individuals or groups, to improve the selection of superior genotypes and allowing improved quality control in breeding programmes and seed orchard management.Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2005.
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Lee, Shao-Pei, and 李紹珮. "Analysis of random amplified polymorphic DNA (RAPD) markers associated with yield traits of tomato under heat stress." Thesis, 2004. http://ndltd.ncl.edu.tw/handle/82955925450323541644.
Full textAbstract:
碩士中國文化大學
生物科技研究所
92
Tomato contains many kinds of chemical constituents, such as Vitamin C and lycopene, which are considered as free radicals scavenger and thus protects people from some chronic disorders or reduces the rate of some cancers. Most tomato varieties are adaptive to warm and dry weather, and don’t show heat tolerance. The optimum temperature for growing tomato is 15-20/20-26 °C night/ day. During summer time in Taiwan, the production of tomato is decreased due to high temperature and high rainfall. Heat tolerance of tomato is a quantitatively inherited trait and easily affected by environmental conditions. A total of 43 F7 recombinant inbred lines derived from a heat tolerant CL 5915 crossed with a heat sensitive L4422 (L. Pimpinellifolium) were grown in the screenhouse at PCCU in the summer of 2002. Five heat tolerance-related traits were scored: flower number, fruit number, fruit set, fruit weight and fruit yield. RAPD analysis was used to identify any primer associated with the flower number, fruit number and fruit weight by bulking DNAs of skewed lines (bulked segregant analysis). A total of 200 random primers were screened. Only 14 RAPD-PCR products generated by OPERON primer with DNA template from L4422, CL5915 and F7 RILs (Recombinant inbred line) were found with polymorphism. Among them, 88 bands were amplified, in which 23 bands showed informative and were used for χ2 test. An average of 1.6 bands out of 6.3 bands was generated by a primer and thought to be informative. Combination of the mentioned 5 traits and the informative markers, we were able to identify and locate the RAPD markers linked to the QTL of traits using Mapmaker/EXP 3.0b and Mapmaker/QTL 1.1b softwares. Two linkage groups were constructed from Mapmaker: C09 and S13; K06, K14, and P08. Nevertheless, all these 5 primers didn’t show any linkage to the five heat tolerance-related traits.
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Hou, ya-wen, and 侯雅文. "Construction of a linkage map based on RAPD (Random Amplified Polymorphic DNA)markers in Cucumis melo L." Thesis, 1996. http://ndltd.ncl.edu.tw/handle/71442632451340748353.
Full textAbstract:
碩士輔仁大學
生物學系
84
This research project is intended to construct a RAPD (random amplified polymorphic DNAs) linkage map of Cucumis melo L. Conditions such as the amount of DNA template,the concentration of Mg+2 ions available,the annealing temperature of the primer used and its duration were optimized to generate reproducible RAPDpatterns.Furthermore,a F2 population was established from a F1 plant obtained from a cross of a Eastern melon ♀ parent (cv. SLK-V-052) and a musk melon ♂ parent (cv. PI-414723) .Fourty five prescreened from 300 random 10-mer oligonucleotides primers (with GC% between 50% and 90%) were able to produce DNA polymorphisms among parental lines and F1 in a preliminary survey. Their segregation patterns in RAPDs were subsequently examined on a F2 mapping population comprising of 98 individuals. So far, 20 polymorphic RAPD markers were analyzed. The mapping distances of 19 RAPD markers were computed using the software "Linkage1" and their plausible map locations were determined.In summary, 7 linkage groups spanning 422.61 cM was recognized.
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Lee, Huey Jen, and 李慧真. "Analysis of Genetic Variation of Pseudocercospora fuligena and P. atromarginalis by Random Amplified polymorphic DNA (RAPD) technique." Thesis, 1996. http://ndltd.ncl.edu.tw/handle/31131496415158966156.
Full textAbstract:
碩士國立中興大學
植物病理學系
84
The fungi tested in this study included each 33 isolates of Pseudocercosporafuligena (on Lycopersicon esculentum ) and P. atromarginalis (on Solanumnigrum), and 20 other isolates of Cercospora spp. and Pseudocercospora spp. wereused to perform analysis of their genetic variation by RAPD. Based on thepreliminary tests of searching suitable RAPD conditions, the results revealedthat optimal reaction mixtures contained DNA ( 50 ng ), 1 x buffer ( 1.5 mM Mg+2), primer ( 0.5 μ M), dNTPs ( 0.2 mM each ), DynaZyme I DNA polymerase ( 0.6 U/ μ l), and BSA( 0.5 mg / ml ) and the suitable amplification reaction is 2cycles at 94 ℃ for 60 s, 42 ℃ for 7 s, and 72 ℃ for 70 s, then 35 cycles at94 ℃ for 3 s, 42 ℃ for 7 s, and 72 ℃ for 70 s, and one cycle at 72 ℃ for 4min. Preliminary 100 different 10 mer primers (Operon Technologies Inc.,Alameda, CA, U. S. A. ) were tested for each 3 isolates of P. fuligena ( pf-10,pf-12, and pf-18 ) and P. atromarginalis ( pa-22, pa-24, and pa-29 ) among them,13 primers gave no banding pattern or poor amplification, the other 87 primersconsistantly produced common banding patterns. Therefore, 30 reproducibleprimers were selected for assessing genetic variation among isolates of P.fuligena, P. atromarginalis and other Cercospora spp. and Pseudocercospora spp.The banding patterns of RAPDs obtained from application of these primers wereused to calculate the genetic similarity of those fungi, and genetic cluster ofthose fungi were analyzed with the average linkage method of hierarchialclustering method in SAS (Statistic Analysis System Inc. ). The results of RAPDpatterns, genetic similarity and dendrogram indicated that isolates of P.fuligena formed a homogenous group, so did P. atromarginalis, except the 4isolates of P. atromarginalis ( pa-15, pa-30, pa-35 and pa-39 ); intraspecificand interspecific variation were not detected between isolates of P. fuligenaand P. atromarginalis from different areas; the 3 isolates (P. solani-melogenicola, P. piricola and P. eriobotryae ) showed the similar RAPD patternsas P. fuligena and P. atromarginalis did; isolates of other Cercospora spp. andPseudocercospora spp. were the host specific fungi. In conclusion, the twospecies, P. fuligena and P. atromarginalis were homogeneous based on RAPDanalysis could be a synonym. The RAPD is a very potential technique in using theunique RAPD bands as genetic markers for identification of interspecies andintergenus and assessing the genetic diversity and relationships among isolatesof Cercospora spp. and Pseudocercospora spp. This model should be asupplementary tool when the traditional taxonomy method can not solve theproblems caused by host specificity or morphology as the criteria in the fungusclassification.
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Lin, Cheng Chieh, and 林政潔. "Application of Morphological Traits and Random Amplified Polymorphic DNA (RAPD) Markers on Cultivar Identification of Capsicum annuum L." Thesis, 1998. http://ndltd.ncl.edu.tw/handle/75770954719698825603.
Full textAbstract:
碩士國立中興大學
園藝學系
86
Morphological traits and randomly amplified polymorphic DNA (RAPD) markers are used to identify their relationship of 49 pepper(Capsicum annuum L.) cultivars from Know You and Evergrow Seed Companies. Twenty five horticultural traits based on the classification system of National Plant Genetic Resource Center showed that the morphological traits of seed, leaf, flower, and fruit can be used to distinguish 49 Capsicum annuum L. cultivars. By utilization of the differences of 14 fruit characteristics to test these 49 cultivars that they could not be fully distinguished by the morphological traits along.A total of 10 DNA primers are selected from 110 random primers which has shown the polymorphic patterns among 7 pepper cultivars from Know You Seed Company. Ten of the 110 primers, i. e. UBC-327, UBC-484, UBC-002, UBC-005, UBC-019, UBC-020, UBC-042, UBC-043, UBC-066, UBC-084, showed polymorphism and could distinguish all 49 Capsicum annuum L. cultivars. The least similarity of cluster analysis is about 81% among cultivars by 10 primers.Pepper cultivars with UBC-019m(1330bp) or UBC-020j(1800bp) band belong to group of hot pepper cultivars with UBC-043e(1300bp) band had strong pungency, narrower fruit shape, and smooth fruit surface. Hot pepper only showed cultivars with UBC-020j(1800bp) band but without UBC-043e(1300bp) band are slight pungency , wider fruit shape, and groove fruit surface, such as ‵Peace Star′ and ‵ Group Stars′ cultivars of Know You Seed Company. The dendrograms displayed sweet pepper and hot pepper can be distinguished through the cluster analysis of using RAPD markers. Cultivar ‵Uranus′ showed proximate genetic relationship with ‵Big Star′ of Know You Seed Company. And Know You cultivars of ‵Peace Star′ and ‵Group Stars′ were clustered in the same group with Evergrow Seed Company cultivars of ‵Lady Square′ and ‵Summer Bell′. The same as ‵Vega′ and ‵Beauty Bell′ cultivars of Know You Seed Company and ‵Square Lamuyo′, ‵Rich Square′, and ‵Blocky Red′ cultivars of Evergrow Seed Company. The result of genetic relationship among 49 Capsicum annuum L. cultivars based on RAPD markers are similarity with cluster analysis of morphological traits.
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Shing, Jong，Ming, and 鍾明勳. "Use Random Amplified Polymorphic DNA (RAPD) Analysis on the Genetic Relationship among Phyllostachys Species and among Bambusa Species." Thesis, 1996. http://ndltd.ncl.edu.tw/handle/62824528169573568914.
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Tsai, Tzung-Yu, and 蔡宗育. "Genetic Diversity in butterhead and crisphead lettuce (Lactuca sativa L.) by using Morphological Traits, Random Amplified Polymorphic DNA (RAPD) and Sequence Characterized Amplified Regions (SCAR) Markers." Thesis, 2008. http://ndltd.ncl.edu.tw/handle/59226251002783909910.
Full textAbstract:
碩士國立中興大學
園藝學系所
96
The genetic diversity of 31 butterhead (BUT) and 26 crisphead (CRP) accessions were evaluated by morphological traits, RAPD (Random Amplified Polymorphic DNA) and SCAR (Sequence Characterized Amplified Regions). Data for 19 morphological traits were subjected to a genetic diversity analysis, after which a UPGMA cluster analysis was performed. The 57 accessions were clustered into 2 groups according to type with genetic similarity of 0.38. The first group included 15 accessions form Tainan and 3 accessions form the National Germplasm of the USA, which were clustered into 5 subgroups with the similarity beyond 0.61. The second group included 10 crisphead and 26 butterhead accessions, which were clustered into 6 subgroups with the similarity beyond 0.70. A total of 271 RAPD bands, with a mean band of 9.3 for each primer, were generated using 29 out of 130 primers in the RAPD analysis. The polymorphism was 53.9%. All accessions were clustered into 3 groups in the RAPD analysis. The first group included 13 crisphead accessions form the Taiwan agricultural research institute(TARI) with the similarity 0.86. The second group including 15 accessions form the Tainan district agricultural research and extention station(TDARES)、3 accessions form USA and 3 butterhead accessions with the similarity 0.88. The first, second and fourth subgroups included crisphead accessions. The third and fifth subgroups included 3 butterhead accessions. The third group was composed of 23 butterhead accessions with a the similarity of 0.82. In analyzing results from SCAR of butterhead and crisphead accessions, a total of 100 bands were generated using 21 out of 51 primer pairs. The means of bands generated by a primer pairs was 4.8 and the polymorphism was 73%. The 57 accessions were clustered into 3 groups. The first group included 13 crisphead accessions form the TARI with the similarity 0.76. The second group including 15 accessions form TDARES with a similarity of 0.88. The third group was composed of 26 butterhead and 3 accessions the USA with a similarity of 0.88. The analysis of RAPD and SCAR result in the main group of the dendrogram showed a similarity of 0.78~0.86 based on RAPD markers and which the SCAR markers displayed a similarity 0.68~0.78. The analysis of RAPD and SCAR result in the subgroup of the dendrogram showed a similarity of 0.84~0.96 based on RAPD markers and which the SCAR markers displayed a similarity 0.88~0.97. Lettuce germplasm could be identified by morphological characteristics such as leaf color, leaf texture and head type. The molecular markers could be used to cluster accessions into groups by SCAR and subgroups by RAPD successfully.
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Min, Tee See, and 鄭思敏. "Genetic Diversity in Colocasia and Xanthosoma Based on Morphological Traits, Random Amplified Polymorphic DNA (RAPD) and Simple Sequence Repeats (SSR) Markers." Thesis, 2005. http://ndltd.ncl.edu.tw/handle/28240061297483500453.
Full textAbstract:
碩士國立中興大學
園藝學系
93
Abstract The genetic diversity of 30 Colocasia (KCC) and 14 Xanthosoma (KCX) accessions collected from Taiwan and other places were evaluated by morphological traits, RAPD (randomly amplified polymorphic DNA) and SSR (simple sequence repeats). Data for 28 characters of shoot, 9 characters of corm and 2 indexes were subjected to a genetic diversity analysis and the UPGMA cluster analysis was performed. The 44 accessions were clustered into 2 groups according to genus with genetic similarity of 0.32. The Colocasia accessions were clustered into 3 subgroups. KCC175 was independence from other accessions with 0.57 similarities. The second subgroup was composed of 14 accessions with the genetic similarity of 0.52 which included accessions with whitish or green leaf vein, corm flesh with slightly fibrous and white in color. The others comprised the third subgroup with the genetic similarity beyond 0.66. Most of the accessions with purple leaf vein, corm flesh with very fibrous and purple in color were included in this group. A total of 221 RAPD bands, which a means of 10.4 band by each primer, were generated using 15 out of 88 primers in the RAPD analysis. The polymorphism was 98.2%. All accessions were clustered into 2 groups in the RAPD analysis. The similarity within KCX accessions was 0.85 compared to 0.71 within KCC accessions. All KCX accessions except KCX002 were indistinguishable. All KCC accessions were clustered into 8 subgroups. KCC131 and 132 were in the first subgroup with genetic similarity 0.71. KCC002, 004, 062 and 064 were the only accession in subgroup with 0.77, 0.81, 0.78 and 0.84 similarities respectively. The 4th subgroup consisted of KCC029 and 050 which similarity was 0.85. The 6th subgroup included KCC145, 169 and CHC01 with similarity 0.84. The largest subgroup included other 19 accessions with similarity 0.85. In analyzing KCC accessions from SSR results, a total of 73 bands were generated using 13 out of 49 primer pairs. The means of bands generated by a primer pairs was 5.2 and the polymorphism was 93.2%. Thirty accessions were clustered into 5 groups with similarity 0.72. The similarity of the first subgroup included KCC029, 050 and 062 was 0.74. KCC131 and 132 comprised the second group with similarity 0.96. The third group consisted of 6 accessions that had similarity 0.78 and the forth group consisted of KCC002, 004 and 064 with similarity 0.74. The similarity of the fifth group that composed of other 16 accessions had 0.72 similarity. Taro germplasm could be identified by morphology such as leaf blade margin color, leaf vein color, color of corm flesh, flesh fibre and number of cormels. The molecular markers could be used to analyze the accessions within groups. The genetic diversity of the 44 taro accessions could be identified by at least 2 RAPD primers or 4 SSR primer pairs.
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Shen, Zi Ru, and 沈姿如. "Stduy on the taxonomic identity of schima superba var. superba and S. superba var. kankaoensis by random amplified polymorphic DNA (RAPD)." Thesis, 1996. http://ndltd.ncl.edu.tw/handle/27494675779896588354.
Full textAbstract:
碩士國立中興大學
森林學研究所
84
Schima superba var. superba is an important native and plantation species in Taiwan and S. superba var. kankaoensis is a variety of S. superba var. superba. Since there is no reli-able morphological characteristic to distinguish the two of them, we tried to use a new technique, random amplified polymorphic DNA(RAPD), to study the taxonomic identity of the two species. Twenty arbitrary sequenced primers(10 nucleotides in length) were selected to proceed RAPD-PCR. One hundred and forty seven bands were recorded and 90(%) of them were polymorphic bands. DNA markers generated by primers U2, T1, V5 and X11 have specific bands that can differentiate S. superba var. superba from S. superba var. kankaoensis. These results indicate that the two species do have some levels of genetic variation.In Nei''s similarity matrix, S. superba var. superba has higher variation within population(1－F＝0.150∼0.314) than that in S. superba var. kankaoensis(1－F＝0.110∼0.289). The cause of this variation may due to the former species spreading widely in Taiwan that may generate higher genetic variation than the latter species that is restricted in Heng-Chun peninsula. Based on the dendrogram generated by the RAPDistance program, when the genetic distance is equal to 0.400, the two species can be separated intotwo groups. However when the genetic distance reduced to 0.213, the S. superba var. superba population can be further divided into three subgroups. This may due to the samples originally collected from plots in Hue-Sun Experimental Forest Station were geographically separated long enough to diverge. On the other hand, samples collected from the S. superba var. kankaoensis population in Heng-Chun peninsula also geo-graphically separate enough to diverge into three subgroups.
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Sewpersad, Yaksha. "Estimation of genetic variation and marker identification in black wattle (Acacia mearnsii De Wild) with RAPD fingerprinting." Thesis, 2004. http://hdl.handle.net/10413/10016.
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Chiou, Chin-Chun, and 邱金春. "Identification of Cymbidium spp. by randomly amplified polymorphic DNA markers." Thesis, 1997. http://ndltd.ncl.edu.tw/handle/00190219285461565041.
Full textAbstract:
碩士國立中興大學
園藝學系
85
Morphological traits and randomly amplified polymorphic DNA (RAPD) markerswere used to study the identification and discrimination of Cymbidium species.The purposes of this study were to establish the molecular markers for discrimination of the Cymbidium spp. and cultivars and to explore the relationships of genetic linkage among the Cymbidium spp. and cultivars. The data of phenotype characters and growth habit showed that the tenCymbidium spp. can be discriminated followed the order of serrulated marginof leaf, leaf shape, leaf size, and growth habit. However, fifteen cultivars ofCymbidium can only be parly discriminated with leaf stripes and leaf shape. Twenty DNA primers, which generated clear and discriminative polymorphicDNA fragments, were selected from 300 random primers to discriminate ten Cymbidium species. Of 79 polymorphic DNA fragments, 46 fragment were species-specific and 33 fragments were presented between two Cymbidium spp. Therewere averaged 12.5 amplified DNA fragments a-nd 3.9 DNA polymorphic markersgenerated by each primer. The results of linkagof cluster analysis revealedthat the ten Cymbidium spp. can be classified into four groups, the first group includes terrestrial plants of C. rubrigemmum, C. ensifolium var. misericors, C. karan, C. sinense and C.tortisepalum, the second group includes C. lancifolium form aspidistrifolium and C. formosanum, the third group includes epiphytic plants of C. Sensation and C. pumilumm, and the fourth group includes C. dayanum var. austro-japonicum with the lowest similar-ity with other Cymbidium spp. Eleven DNA primers were selected from 300 random primers to discriminate fifteen cultivars of Cymbidium. Of 21 polymorphic DNA fragments which discriminated eleven cultivars of Cymbidiu sinense, 11 fragments were cultivar-specific and 10 were shared among two to four cultivars. There were averaged 11 amplified DNA fragments and 1.9 DNA polymorphic markers gener-ated by each primer. The results of linkage cluster analysis revealed that the eleven cultivars of Cymbidium sinense can be classified into four groups, the first group includes C. sinense and `taur-ji', the second group includes `rueih-baau', `yarng-ming-jiin', `jin- shan', `shyueh-bair-jaau' and`shyuh-huarng', thethird group includes ` shyr-mern', and the fourth groupincludes `jin-huah- shan' and `shih-ba-hsiieh-shih'.
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Lin, Yin-Shan, and 林吟珊. "Identification of Peanut Genetic diversity by Randomly Amplified Polymorphic DNA markers." Thesis, 2001. http://ndltd.ncl.edu.tw/handle/77913625353644316682.
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碩士國立高雄師範大學
生物科學研究所
89
Abstract The 115 peanut varieties as materials in this research include important released breeding varieties and common used progenitors of breeding in the past in Taiwan, introduced varieties with rust and leafspot resistance, and the breeding varieties recently released from Tainan District Agricultural Improvement Station. Observing the phenotypes, characters of pods and seeds of these 115 varieties, varieties in each group have above 90% relativity in four traits, including procumbent, decumbent or erect growth habits, early or late flowering, leaf size and seed dormancy. Therefore, we may classify them by these four characters first. And with other assistant traits, we may further classify them into four botanical types as Spanish, Valencia, Virginia Bunch and Virginia Runner type. Using SDS-PAGE analysis of the 115 varieties, we planned to separate them into six groups (the four botanical types, leafspot resistant group and high oil content group) by their protein profiles. Although no obvious differences among the six groups were found, we still can distinguish individual varieties by their own specific protein patterns, except 12 varieties. Furthermore, the differences of peanut genomes were directly detected by RAPD (Randomly amplified polymorphic DNA) in DNA level. We selected 5 suitable random primers with their polymorphism and well reappearance from 44 random primers. We combined RAPD analysis of OPA-09, OPA-12, OPB-03, and OPB-08 primers whose genetic similarity coefficients are between 44~63 to calculate their combined genetic similarity coefficient of the 115 varieties, and to get dendrograms by UPGMA cluster analysis. By the dendrogram, we classified 115 peanuts varieties into I, II,Ⅲand Ⅳ groups. There are 17 varieties in group I, and most of them are Virginia Bunch or Virginia Runner type (only Tainan 14 is Spanish type ) and have disease resistance. Group II are complicated. Group Ⅲ are all Valencia type. Group IV are Spanish type and have high oil content, except Pongfu 3. In conclusion, this study shows that traditional investigation of phenotypes, characters of pods and seeds is a possible way to classify them into four botanical types. The protein profile can be used to distinguish individual variety of peanuts. In addition, with proper combination of OPA-09, OPA-12, OPB-03, and OPB-08 random primers, RAPD analysis can be used to explore the genetic diversity among peanut varieties and group them initially. Above results would be helpful to peanut breeders.
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Lee, Shough-Peng, and 李碩朋. "Identification of native Dendrobium species by Randomly Amplified Polymorphic DNA Markers." Thesis, 2002. http://ndltd.ncl.edu.tw/handle/35571663803232979822.
Full textAbstract:
碩士國立中興大學
園藝學系
90
Genetic diversity and relatedness were assessed among 19 Dendrobium species such as D. chameleon, D. erumenatum, D. candidum, etc., using 10 randomly amplified polymorphic DNA (RAPD) primers. Of these Dendrobium studied, 14 are native in Taiwan, 2 from the Philippinges and 3 from mainland China. The phalaenopsis aphrodite seedlings derived from tissue culture were included for comparison. The plant morphology and biological traits were also investigated to study ecological effects on genetic composition. The plant growth was influenced by its epiphytic site and intercepted light intensity. Grown on rocks, the plant has smaller pseudobulb and less leaves than that grown in woods. The latter may be two times of the size of the former. Different Dendrobium species could be distinguished by the color,size and shape of flower. Other morphological traits could also be used to separate different species, such as angle of leaf growth, color of pseudobulb, internode length and growth habit. The brilliant orange color of D. clavatum is the most beautiful, follower by pink series of D. falconeri, D. linawianum and D. miyakei. The bright white color of D. crumenatum. and yellow series of D. tosaense are also valuable. The flowering period of individual flower was longest in D. tosaense (15 days) , followed by D. linawinanum (11 d) and D. leptocladum and D. moniliforme (10 d), The shortest flowering period was observed in D. crumenatum and Flickingeria comata with life span less than one day. Self-pollination was found successful in D. clavatum , D. tosaense and F. comate. About 50% of fruit set from self- pollination was obtained in D. equitans, D. leptocladum, D. linawianum and Epigeneium nakaharaei. Cross pollination enhanced fruit set in D. crumenatum, D. falconeri, D. linawinanum and D. moniliforme. Twenty Operon primers (OPE-01~20) were used to screen for polymorphism. The results showed interspecific divergency. The Dendrobium species surveyed were classified into six groups. D. crumenatum from both Green Island and the Philippines as a group, D. miyakei(from both the Philippines and Orchid Island), D. chameleon and Epigeneium sanseiense in group Ⅱ, D. moniliforme, D. candidum, D. tosaense, and D. falconeri (from Ku-kuan, Sun-Link —Sea & Yun-Nan) in group Ⅲ, D. clavatum and D. linawianum in group Ⅳ, D. somai and D. leptocladum in groupⅤ, Flickingeria comata, F. tairukountia and F. finbriata in group Ⅵ. Low similarity value detected from D. miyakei suggested it as another genus. Species in group Ⅱ and Ⅴ may also be treated as independent genera. Genetic diversity as well as morphological difference existed between accessions of the same species from different regions. As in D. falconeri, accessions from Ku-kuan and Sun-Link —Sea are genetically more similar in between than to that from Yun-Nan, China. Geographical distance might affect intra specific similarity.
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Zhang, Cai-Yuan, and 張財源. "Using RAPO(random amplified polymorphic DNA) fragment analysis in brassica." Thesis, 1994. http://ndltd.ncl.edu.tw/handle/99021615354849068420.
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Lopez-Buesa, Otilia. "Randomly amplified polymorphic DNA markers as tags for the major virus-resistance genes in cucumber." 1994. http://catalog.hathitrust.org/api/volumes/oclc/33036230.html.
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Thesis (M.S.)--University of Wisconsin--Madison, 1994.Typescript. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 10-12).