The aim of this study was to obtain the molecular marker of Taiwania cryptomerioides Hayata based on DNA sequence data of PCR- sequencing and inter-simple sequence repeat (ISSR), and to evaluate the genetic diversity of populations of Taiwania cryptomerioides Hayata and molecular phylogeny of T. cryptomerioides and Taiwania flousiana Gaussen. The sequence data based on the internal transcribed spacer (ITS) of a total of 108 samples of T. cryptomerioides were determined. Eight different populations of T. cryptomerioides and 12 samples of T. flousiana from Yunnan, China were analyzed. The finding of the study showed that heterogeneity of ITS region within individuals of T. cryptomerioides was high by showing high nucleotide diversity among ITS sequences both in T. cryptomerioides ( £k = 0.18153) and T. flousiana ( £k = 0.19751). The findings fit in Tajima¡¦s D test of neutrality based on DNA sequence variation in the ITS region of T. cryptomerioides and T. flousiana. It is not obvious to incorporate into different population through clustering analysis based on data of the ITS region of T. cryptomerioides and T. flousiana. However, slightly genetic differentiation between T. cryptomerioides and T. flousiana was found, which figured of Fst (Fst = 0.0441~ 0.0856, an average value = 0.0611). On the other hand, the samples were studied by using ISSR markers. Of the 100 primers screened, 4 produced highly reproducible ISSR bands, and 24 discernible DNA fragments were generated with 17 being polymorphic. Based on cluster analysis of molecular data, the cluster is not clear among populations of T. cryptomerioides and T. flousiana. The analysis of AMOVA revealed that the variance component between species of T. cryptomerioides and T. flousiana was 38.54¢H (P < 0.001); however, the variance component within species is 61.46 (P < 0.001). The variation within population of T. cryptomerioides was 84.74¢H (P < 0.001) and the variance between populations is 15.26¢H (P < 0.001), indicating that the genetic diversity of individuals within population was high. The aforementioned data suggest that gene flow among different populations of T. cryptomerioides was high, indicating that the genetic diversity was high among individuals of T. cryptomerioides but was low between populations. Furthermore, it is concluded both species are genetically closer and could be grouped into the same species.

The objective of this study is to elucidate the efficiency of enation-structure (at node) recognition method at pre-flowing stage and to understand the population genetic variation of the Mikania weeds in Taiwan. The plant materials collected by recognizing enation-structure symptom method from North, Central, South and East Taiwan and off-shore islands. Using PCR¡V sequencing marker techniques, the sequencing revealed that nrDNA ITS region could identify three Mikania weeds and the 97% similarity of phylogenetic relationship between M.cordata and M.micrantha are more closer than that between M.cordata and M.scandens, whose ITS sequence is obtained from GeneBank. However ,the sequences of chloroplast DNA of M.cordata and M.micrantha at trnL intron¡]436bp¡^or trnL-trnF IGS¡]345 bp¡^are almost the same and could not be used as molecular markers. The recognizing techniques of enation-structure was supported by the nrDNA ITS region and ISSR results at the end, thus the finding can be recommended to the Council of Agriculture in order to eliminate the weed and to reduce the impact on M.cordata, which is native in Taiwan. Moreover, the findings of ISSR analysis in the aspect of population genetic variation indicated that high genetic differentiation ¡]Gst>0.5¡^was found among the M. cordata and M.micrantha populations. Based on the Mantle test, there was no relationship between genetic distance and geographic distance in M.micrantha¡]r=0.0053,p=0.47¡^.This phenomenon revealed that populations of M.micrantha had complex population variability within the short-term invaded into Taiwan that might be resulted from the random dispersion of human activities. The population of M.micrantha was established by few individuals (founders) and grown rapidly in Taiwan, resulting in population differentiation via genetic drift. In contrast to M.micrantha ,there was relationship between genetic distance and geographic distance in M.cordata ¡]r=0.44,p=0.025¡¯¡^.It revealed that populations of M.cordata agreed to the concept of isolation by distance model, which might be evolved from the result of natural dispersion. In conclusion, the population structure of M.micrantha in Taiwan is stable , suggesting that control of Mikania population should be based on different populations where have large differentiation among them .

Several oomycete species of the genus Saprolegnia are recognized as devastating fish pathogens and are responsible for the loss of millions of fish annually for the aquaculture industry. Until recently, these pathogens were kept in check using malachite green; however, due to its toxicity, this chemical has now been banned from use. Saprolegnia parasitica is recognized as the major pathogen of aquaculture fish species. The industry is struggling to predict and control S. parasitica outbreaks in fish hatcheries and there is a need for new knowledge regarding the population genetic structure of this pathogen. Random amplified microsatellites were used to compare isolates of S. parasitica collected from a variety of hatchery locations during the period of November 2009 - August 2011, in order to determine the level of genetic variability and determine changes in genetic diversity over time. Allele frequencies of scored characters were graphically compared. Population genetic diversity was measured using Nei’s genetic distance, Shannon’s Information Index, number of polymorphic loci and phylogenetic trees. Due to the presence of Saprolegnia parasitica in the facilities tested, it appears to be ubiquitous in aquaculture facilities and treatment and prevention will be an ongoing concern in aquaculture management. Overall, genetic diversity of S. parasitica isolates was determined to be low with at least some sexual recombination occurring over time. There was a diversity of genotypes collected from the same hatchery on a single day, indicating there was not a single genotype present at a given time point. Genetic profiling, such as used here, could provide facility managers with a new approach to develop a series of best practices to control sporadic outbreaks of disease. Use of these genetic markers and close monitoring of S. parasitica genotypes will permit early detection and sanitation protocols. / Graduate / 2015-04-24 / 0476 / 0792 / 0369 / cren06@uvic.ca

Resumo: A variabilidade entre progênies é criada pela segregação cromossômica independente dos genes e pela recombinação genética intracromossomal durante a meiose. O objetivo deste estudo foi analisar a variabilidade derivada de crossing-overs em cruzamentos biparentais (G2 e J2), quádruplos (G4 e J4) e óctuplos (G8 e J8), avaliados em populações segregantes derivadas de parentais contrastantes para resistência ao nematóide de cisto da soja (raça 3) - NCS - e ao oídio - O. A análise foi realizada em populações F2, através de marcadores SSR (single sequence repeat) concentrados em uma região de 55 cM ao redor do gene rmd (resistência ao oídio) e rhg1 (resistência ao NCS). Após o teste dos marcadores, quanto ao polimorfismo, apenas marcadores polimórficos foram utilizados para detectar crossing-over. Todos os marcadores analisados foram não significativos pelo teste de qui-quadrado (P > 0,05), indicando que os valores observados se ajustam à proporção genotípica esperada em F2 (1:2:1). As maiores médias de crossing-over por genótipo foram obtidas para G4 (4,00), no grupo G, e J8 (2,91), no grupo J. Por outro lado, as maiores médias de crossing-over considerando o número de gerações para formar cada população, foram para G2 (2,02) e J8 (0,97). A recombinação entre alelos ocorreu em algumas populações, entretanto para G4 e J8 em 1,89% dos genótipos não ocorreram. Em geral, nos cruzamentos com maior número de parentais envolvidos a ocorrência de crossingover foi maior, sendo satisfatórios na criação de variabilidade. O progresso no melhoramento de soja tem sido alcançado em partes pela criação de novas combinações alélicas dentro dos cromossomos. / Abstract: The variability among the progenies is created by chromosome segregation, independent assortment of genes, and intra-chromosomal genetic recombination during meiosis. The objective of this study was to analyze the variability derived from crossovers in soybean biparental (G2 and J2), quadruple (G4 and J4) and octuple (G8 and J8) crosses, measured in segregant population derived from contrasting parental regarding their resistance to cist nematode (race 3) - SCN and powdery mildew - PM. The analyses were made in F2 population through SSR (single sequence repeat) markers located in a 55CM region around Rmd (powdery mildew) and Rhg1 (cist nematode) resistance genes. After screening makers for their polymorphism, only polymorphic markers were used to detect crossovers. All markers were not significant by chi-square test (P > 0.05), showing that observed values corroborates to genotypic inheritance ratio expected in F2 population (1:2:1). Thus, the higher average of crossovers for some populations were observed for G4 (4.00), at linkage group G and J8 (2.91), at linkage group J. On the other hand, the higher average of crossovers considering the generation number to form each population, was found for G2 (2.02) and J8 (0.97). The recombination between alleles occurred in some populations, however, to G4 and J8, in 1.89% of the genotypes not showing crossover. In general, the crosses with larger numbers of parents showed higher number of crossovers, being very satisfactory for the creation of genetic variability. Soybean breeding progress has been accomplished in part by creating on new within_chromossome allele combinations. / Orientador: Antonio Orlando Di Mauro / Coorientador: Todd Pfeiffer / Banca: Natal Antonio Vello / Banca: Osvaldo Toshiyuki Hamawak / Banca: José Roberto Môro / Banca: Janete Apparecida Desidério Sena / Doutor

Submitted by Amanda Silva (amanda.osilva2@ufpe.br) on 2015-04-08T12:53:20Z No. of bitstreams: 2 Dissertação Nathália Leite.pdf: 1163803 bytes, checksum: ed3fcb6e6b34f0c3e3452518a29c3263 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) / Made available in DSpace on 2015-04-08T12:53:20Z (GMT). No. of bitstreams: 2 Dissertação Nathália Leite.pdf: 1163803 bytes, checksum: ed3fcb6e6b34f0c3e3452518a29c3263 (MD5) license_rdf: 1232 bytes, checksum: 66e71c371cc565284e70f40736c94386 (MD5) Previous issue date: 2012-02-29 / CNPq / O presente estudo foi realizado com os objetivos de identificar acessos de feijão-caupi tolerantes ao caruncho (C. maculatus) e caracterizar esses acessos a nível molecular, em conjunto a acessos de V. unguiculata ssp. cylindrica e V. radiata, por meio de marcadores DAF (DNA Amplification Fingerprinting) e ISSR (Inter Simple Sequence Repeat), visando identificar parentais promissores para futuros trabalhos de melhoramento, bem como para mapeamento, pela associação com o nível de resposta ao caruncho. Para identificação da tolerância ao caruncho em feijão-caupi, 27 acessos foram avaliados em ensaios de laboratório, utilizando-se o Delineamento Experimental Inteiramente Casualizado. Para cada acesso, uma amostra com 30 grãos de feijão-caupi foi infestada com cinco casais de caruncho com até 24 h de idade, sendo os resultados obtidos submetidos à análise estatística e suas médias comparadas pelo teste de Tukey a 5% de probabilidade. Para as duas variáveis estudadas, o acesso INHUMA comportou-se como suscetível, enquanto os acessos PATATIVA e MNC99-537F-4 apresentaram-se tolerantes para ambas as variáveis. Os demais acessos testados apresentaram comportamento intermediário, o que não os diferencia em relação à INHUMA, PATATIVA e MNC99-537F-4. Na caracterização molecular dos acessos, 25 primers geraram um total de 239 amplicons, dos quais 163 foram polimórficos. Os fragmentos amplificados foram incluídos em uma matriz de dados para análise pelo método de Neighbor-Joining. No fenograma obtido, V. radiata e V. unguiculata ssp. cylindrica assumiram uma posição basal isolada dos subgrupos formados, enquanto que os acessos de V. unguiculata subdividiram-se em dois grupos, nos quais os acessos contrastantes quanto à tolerância ao caruncho se distribuíram em subgrupos distintos, o que evidencia a existência de diferenças genéticas significativas entre alguns candidatos. Nesse sentido, os resultados obtidos demonstraram que os ensaios de tolerância ao caruncho associados à aplicação de marcadores moleculares foram capazes de identificar genótipos contrastantes de feijão-caupi promissores para aplicação em trabalhos de melhoramento vegetal, bem como para a geração de populações segregantes adequadas ao mapeamento genético.

Phytophthora sojae, the causal organism of stem and root rot, and <i>Soybean mosaic virus</i> (SMV) cause two of the most highly destructive diseases of soybean (<i>Glycine max</i> L. Merr). <i>P. sojae</i> can be managed either through deployment of race-specific resistance or through quantitative resistance termed partial resistance. In the current study, partial resistance to <i>P. sojae</i> was mapped in an interspecific recombinant inbred line (RIL) population of <i>Glycine max</i> by <i>Glycine soja</i>. One major quantitative trait loci (QTL) on molecular linkage group (MLG)-J (chromosome 16) and two minor QTL on MLG-I (chromosome 20) and -G (chromosome 18) were mapped using conventional molecular markers. Additionally, partial resistance to <i>P. sojae</i> was mapped in the same RIL population using single feature polymorphism (SFP) markers that further fine mapped the <i>P. sojae</i> QTL and identified potential candidate genes contributing to resistance. In a separate study, race-specific resistance was characterized in PI96983 discovering a potentially new allele of <i>Rps4</i> on MLG-G. Finally, using the newly available whole-genome shotgun sequence of soybean, <i>Rsv4</i> conferring resistance to strains of SMV known in the US, was localized to an approximately 100 kb region of sequence on chromosome 2 (MLG-D1B). Newly designed PCR-based markers permit for efficient selection of <i>Rsv4</i> by breeding programs. Identified candidate genes for <i>Rsv4</i> are discussed. Genomic resources developed in all of these studies provide breeders the tools necessary for developing durable resistance to both SMV and <i>P. sojae</i>. / Ph. D.

No description available.

Previously seed coat color in flax has been used as a phenotypic marker for specialty quality traits and currently there is an increasing demand to use seed coat color in flax to market flax for human and animal nutrition uses. Seed coat color was studied to 1) understand the inheritance of seed coat color conditioned by the b1 locus, to 2) understand the relationship of other important flax traits with seed coat color as well as to 3) identify markers that are linked to seed coat color for future marker assisted selection of seed coat color. Spearman’s rank correlation and an allelism test was used to show the inheritance of the alleles at the b1 locus. Bulked segregant analysis (BSA) was used to identify putatively linked markers with the b1 locus, these were then screened on the CDC Bethune x M96006 recombinant inbred line population. Furthermore, the CDC Bethune x M96006 and CDC Bethune x USDA-ARS Crystal recombinant inbred line populations were used to identify any important flax traits that had a significant relationship with seed coat color. It was shown that seed coat color conditioned by the b1 locus was stably inherited and that b1vg and b1 are allelic to one another. The results of the BSA showed that there were 17 candidates for linkage but when these markers were screened on the population only the Lu456 from linkage group (LG) six was identified to have linkage (χ²=3.90; P<0.05) with the b1 locus. Additionally, it was shown that the b1 seed coat color allele of the b1 locus had a pleiotropic effect on flower color and flower shape and that seed coat color was associated with linolenic fatty acid content. None of the traits examined were found to be associated with the b1vg allele of this locus. These results show that the b1 locus is likely present on linkage group six, more marker coverage on linkage group six of markers that are polymorphic between the two seed coat color parents would increase the accuracy of detection. Lastly, this study showed that plant breeders should consider using the b1vg allele that conditions the variegated seed coat color to mark unique lines with important combinations of traits because it sorted independently for seed quality traits. Whereas, the yellow seed coat color conditioned by the b1 allele was found to be associated with higher linolenic fatty acid content and the semi-lethality of this allele would make it not suitable for use in parental lines.

Made available in DSpace on 2014-06-11T19:32:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-09-28Bitstream added on 2014-06-13T18:43:25Z : No. of bitstreams: 1 muniz_frs_dr_jabo.pdf: 1182648 bytes, checksum: 94ba816afe15c07503cde37414bc5a99 (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A variabilidade entre progênies é criada pela segregação cromossômica independente dos genes e pela recombinação genética intracromossomal durante a meiose. O objetivo deste estudo foi analisar a variabilidade derivada de crossing-overs em cruzamentos biparentais (G2 e J2), quádruplos (G4 e J4) e óctuplos (G8 e J8), avaliados em populações segregantes derivadas de parentais contrastantes para resistência ao nematóide de cisto da soja (raça 3) – NCS – e ao oídio - O. A análise foi realizada em populações F2, através de marcadores SSR (single sequence repeat) concentrados em uma região de 55 cM ao redor do gene rmd (resistência ao oídio) e rhg1 (resistência ao NCS). Após o teste dos marcadores, quanto ao polimorfismo, apenas marcadores polimórficos foram utilizados para detectar crossing-over. Todos os marcadores analisados foram não significativos pelo teste de qui-quadrado (P > 0,05), indicando que os valores observados se ajustam à proporção genotípica esperada em F2 (1:2:1). As maiores médias de crossing-over por genótipo foram obtidas para G4 (4,00), no grupo G, e J8 (2,91), no grupo J. Por outro lado, as maiores médias de crossing-over considerando o número de gerações para formar cada população, foram para G2 (2,02) e J8 (0,97). A recombinação entre alelos ocorreu em algumas populações, entretanto para G4 e J8 em 1,89% dos genótipos não ocorreram. Em geral, nos cruzamentos com maior número de parentais envolvidos a ocorrência de crossingover foi maior, sendo satisfatórios na criação de variabilidade. O progresso no melhoramento de soja tem sido alcançado em partes pela criação de novas combinações alélicas dentro dos cromossomos. / The variability among the progenies is created by chromosome segregation, independent assortment of genes, and intra-chromosomal genetic recombination during meiosis. The objective of this study was to analyze the variability derived from crossovers in soybean biparental (G2 and J2), quadruple (G4 and J4) and octuple (G8 and J8) crosses, measured in segregant population derived from contrasting parental regarding their resistance to cist nematode (race 3) – SCN and powdery mildew – PM. The analyses were made in F2 population through SSR (single sequence repeat) markers located in a 55CM region around Rmd (powdery mildew) and Rhg1 (cist nematode) resistance genes. After screening makers for their polymorphism, only polymorphic markers were used to detect crossovers. All markers were not significant by chi-square test (P > 0.05), showing that observed values corroborates to genotypic inheritance ratio expected in F2 population (1:2:1). Thus, the higher average of crossovers for some populations were observed for G4 (4.00), at linkage group G and J8 (2.91), at linkage group J. On the other hand, the higher average of crossovers considering the generation number to form each population, was found for G2 (2.02) and J8 (0.97). The recombination between alleles occurred in some populations, however, to G4 and J8, in 1.89% of the genotypes not showing crossover. In general, the crosses with larger numbers of parents showed higher number of crossovers, being very satisfactory for the creation of genetic variability. Soybean breeding progress has been accomplished in part by creating on new within_chromossome allele combinations.

Made available in DSpace on 2014-08-20T14:06:15Z (GMT). No. of bitstreams: 1 Dissertacao_Luciano_Carlos_da_Maia.pdf: 2299013 bytes, checksum: 1ea0e6e151a64a565fee2caf64a2b9d7 (MD5) Previous issue date: 2007-04-05 / The classic plant breeding methods are responsible for the major advances of modern agriculture. However, molecular biology and genomic techniques have provided insights into how further advance in genetic gains. Molecular markers have been successfully applied in genetic mapping and marker assisted selection in many plant species. Rice after the complete sequence of its genome, has been more and more used as a model for cereal improvement. Currently, strategies have been relying on the transference of information between the model genome (rice) and other grasses, making that the information generated in rice and for other major crops such as maize, wheat and rice can be also used to improve orphan grass crops. Microsatellites (SSRs) have been described as the preferred type of marker to be used in these studies. Given these features of rice and SSRs and aiming to evaluate the abundance of these markers on the rice genome and their availability for public use, a computational tool was developed. This tool search’s and characterizes these loci, applies primer design and searchs for anchoring sites for the primers in genomic databases of any species. It also, evaluates the affectivity of transposition of these markers by simulating a PCR. All SSRs found in the rice genome were analyzed and primers were designed for all loci in chromosome 1 and simulated against the other rice chromosomes, giving an idea of potentially duplicated regions across the genome. / O uso de várias classes de marcadores moleculares tem sido implementado em mapeamento genético e na seleção assistida de várias espécies vegetais. O arroz, após o sequenciamento completo do seu genoma, tem sido proposto como um modelo genético entre as várias espécies gramíneas de importância agronômica. Modernamente, uma estratégia tem sido adotada com base na transferência de informações da genômica estrutural dessas gramíneas, sendo que, desta forma, o conhecimento obtido em espécies com maiores investimentos técnicos como o milho, trigo e o arroz, possam ser utilizados também nas gramíneas com menores níveis tecnológicos de pesquisa. A classe dos marcadores moleculares conhecida como microssatélites é atualmente descrito como preferencial nestes estudos. Dadas essas características do arroz e dos microssatélites, com objetivo de se conhecer a riqueza desses marcadores no genoma do arroz e a disponibilização desses para a utilização pública, foi desenvolvido uma ferramenta computacional para busca e caracterização desses locos, desenho de primers e busca por sítios de ancoramento para os primers em bancos de dados genômicos de qualquer espécie, avaliando dessa forma, a transposição de marcadores moleculares entre espécies através da simulação da PCR. Foram analisadas e descritas todas as possíveis ocorrências de microssatélites no genoma do arroz e desenhados primers para os locos encontrados no cromossomo 1, que posteriormente foram usados para a simulação da PCR contra os demais cromossomos do arroz, resultando o número de amplificações possíveis para cada conjunto de primers e suas respectivas regiões genômicas.