Crown gall disease is an agricultural problem caused by the soil-borne bacterium, Agrobacterium tumefaciens. A. tumefaciens oncogenes cause transformed plant cells to overproduce the hormones, auxin and cytokinin. High hormone levels cause unorganized plant cell growth resulting in a gall. Control of crown gall disease is difficult because after plant cell transformation has occurred, the bacterium is no longer required for the disease to progress. Apple trees engineered to express double-stranded RNA of two A. tumefaciens oncogenes, ipt and iaaM, silenced the expression of the wild-type oncogenes and prevented crown gall disease. Only the iaaM oncogene was targeted for posttranscriptional gene silencing (PTGS) as measured by biological assays and by quantitative reverse-transcription polymerase chain reaction (q-RTPCR) on transgenic tissue. However, if the translation initiation sequence of the iaaM construction was eliminated, gall formation was not prevented, indicating that translatable RNA initiates silencing whereas untranslatable RNA does not. Other data indicate that the Arabidopsis thaliana micro-RNA pathway gene is involved in A. tumefaciens-mediated tumorigenesis. A. thaliana plants with a mutation in HEN1, a gene required for micro-RNA maturation, demonstrated a tenfold reduction in tumorigenesis upon A. tumefaciens infection compared to wild-type. The same plant line showed no difference in T-DNA transfer and nuclear uptake. / Graduation date: 2006

This thesis concerns the phylogenetic relationships and the evolution of the gall-inducing wasps belonging to the family Cynipidae. Several previous studies have used morphological data to reconstruct the evolution of the family. DNA sequences from several mitochondrial and nuclear genes where obtained and the first molecular, and combined molecular and morphological, analyses of higher-level relationships in the Cynipidae is presented. A Bayesian approach to data analysis is adopted, and models allowing combined analysis of heterogeneous data, such as multiple DNA data sets and morphology, are developed. The performance of these models is evaluated using methods that allow the estimation of posterior model probabilities, thus allowing selection of most probable models for the use in phylogenetics. The use of Bayesian model averaging in phylogenetics, as opposed to model selection, is also discussed. It is shown that Bayesian MCMC analysis deals efficiently with complex models and that morphology can influence combined-data analyses, despite being outnumbered by DNA data. This emphasizes the utility and potential importance of using morphological data in statistical analyses of phylogeny. The DNA-based and combined-data analyses of cynipid relationships differ from previous studies in two important respects. First, it was previously believed that there was a monophyletic clade of woody rosid gallers but the new results place the non-oak gallers in this assemblage (tribes Pediaspidini, Diplolepidini, and Eschatocerini) outside the rest of the Cynipidae. Second, earlier studies have lent strong support to the monophyly of the inquilines (tribe Synergini), gall wasps that develop inside the galls of other species. The new analyses suggest that the inquilines either originated several times independently, or that some inquilines secondarily regained the ability to induce galls. Possible reasons for the incongruence between morphological and DNA data is discussed in terms of heterogeneity in evolutionary rates among lineages, and convergent evolution of morphological characters.

La galle du collet, caractérisée par la formation d'une tumeur, représente la bactériose la plus répandue chez la vigne et entraine d'importantes pertes économiques à travers le monde. L'agent responsable de cette maladie est le plasmide Ti (pTi) qui confère à ces bactéries hôtes, telles que des Allorhizobium vitis, leur pouvoir pathogène. En plus du pTi, les A. vitis sont aussi les hôtes d'autres plasmides dont les fonctions et les modes de dissémination sont peu documentés. Afin d'étudier l'adaptation particulière des souches d'A. vitis à la vigne, nous nous sommes intéressés à la diversité de ces pathogènes ainsi qu'au rôle de ces plasmides et à la régulation de leur dissémination. Tout d'abord, nous avons analysé la diversité des isolats provenant de vignobles du Maroc atteints par la galle du collet. Nous avons pu mettre en évidence que l'ensemble des isolats pathogènes (e.g. porteur d'un pTi) forme 4 groupes génétiques se distinguant par le type d'opine produit. Nous nous sommes par la suite intéressés à la souche A. vitis S4 qui héberge 5 plasmides dont 3 possèdent un mécanisme de transfert pouvant être régulé par un système de communication bactérienne nommé quorum sensing (QS). Nous avons montré que le système QS du plasmide p130 (renommé pApi) contrôle non seulement le transfert conjugatif du pApi mais aussi d'autres fonctions plasmidiques non caractérisées. Ce système QS nécessite la synthèse d'une molécule signal de type N-acyl-homosérine lactone qui d'après nos travaux de caractérisation possède une structure atypique. De plus, des analyses génomiques couplées à des tests de résistance au cuivre nous ont permis d'identifier, sur le plasmide p79 d'A. vitis S4, un îlot de gènes potentiellement impliqués dans la résistance au cuivre de cette souche. Via des analyses de diversité ainsi que l'étude d'une souche modèle, nos travaux fournissent des connaissances sur l'adaptation d'Allorhizobium vitis à son unique plante hôte, la vigne / Crown gall disease, characterized by the formation of a tumor, represents the most widespread bacteriosis in the vineyard and causes significant economic losses throughout the world. The causing agent is the Ti plasmid (pTi) which confers pathogenicity to their bacterial hosts, such as Allorhizobium vitis. In addition to pTi, A. vitis harbours other plasmids whose functions and ways of dissemination are poorly documented. In order to study the peculiar adaptation of A. vitis strains to grapevine, we studied the diversity of these pathogens as well as the role of plasmids and the regulation of their dissemination.First, we analyzed the diversity of isolates from vineyards of Morocco affected by crown gall. We highlighted that all the pathogenic isolates (e.g. carrying a pTi) form 4 genetic groups that are distinguished by the type of opine produced and the ability to induce a hypersensitive response. We then focused on strain A. vitis S4, which contains 5 plasmids, including 3 possessing a transfer mechanism that can be regulated by a bacterial communication system called quorum sensing (QS). We demonstrated that the QS system of plasmid p130 (renamed pApi) controls not only the conjugative transfer of the pApi but also other uncharacterized plasmid-encoded functions. This QS system requires the synthesis of a N-acyl-homoserine lactone signal molecule with an atypical structure. In addition, genomic analyzes combined with copper resistance assays allowed us to identify, on plasmid p79 of A. vitis S4, a genomic island that is likely involved in copper resistance of this strain.Through analyzes of diversity and study of a model strain, our work provides insight into adaptation of Allorhizobium vitis to its unique host plant, grapevine

Este trabalho teve como objetivos estudar aspectos da morfologia, biologia e ecologia de Epichrysocharis burwelli, uma micro-vespa recentemente introduzida no Brasil e que induz galhas em folhas de Corymbia citriodora. Foram verificados neste trabalho, porcentagem média de infestação de galhas em folhas, horário de emergência e longevidade de adultos, razão sexual e caracterização do macho, duração do período de desenvolvimento, sazonalidade de adultos, nível de infestação no campo e influência da pilosidade de folhas sobre a ocorrência de galhas e adultos. Além disso, a influência da formação de galhas sobre o rendimento de óleo essencial de C. citriodora foi verificada. 65% das folhas avaliadas apresentaram algum grau de infestação de galhas de E. burwelli, com uma densidade média de sete galhas/cm2, variando entre 0,04 e 44,10 galhas/cm2. Foram encontradas diferenças significativas entre as árvores avaliadas quanto ao nível de infestação de galhas nas folhas. O período da manhã foi o preferencial para a emergência de adultos, com um pico às 9:00 horas. A longevidade da espécie foi significativamente maior quando o mel foi incorporado à dieta, tanto com água como com folhas jovens de C. citriodora. A razão sexual encontrada foi de 165 fêmeas para cada macho e não foram encontradas diferenças entre os sexos além dos caracteres sexuais da genitália e uma pequena estrutura na antena do macho, visível apenas sob microscópio. A duração total do ciclo, de ovo a adulto é de aproximadamente 75 dias em condições de campo. As galhas passam por cinco estágios de crescimento, que provavelmente são acompanhados por mudanças no estágio de vida do inseto. Os adultos ocorrem durante todo o ano e sua ocorrência está relacionada com a liberação de ramos novos pela planta. A espécie é multivoltina e ocorre sobreposição de gerações. A pilosidade das folhas quando a planta se encontra no estágio juvenil funciona como uma barreira mecânica para a oviposição do adulto e teve influência significativamente negativa sobre a ocorrência desses e a formação de galhas. Altos níveis de galhas nas folhas causam efeito significativamente negativo sobre o rendimento de óleo essencial das folhas de C. citriodora, que ainda é potencializado devido à queda prematura de folhas, quando estas estão sob altas infestações. A pesquisa permitiu ter um maior conhecimento sobre aspectos da biologia e ecologia da espécie que podem auxiliar no manejo desta em condições de campo, além de quantificar a redução de óleo causada pela presença das galhas. / This research deals with aspects of morphology, biology and ecology of Epichrysocaris burwelli a micro-wasp inducing galls on leaves of Corymbia citriodora, and recently introduced in Brazil. The following parameters were studied: percentage of galls on the leaf, time of adult emergence, adult longevity, sex ratio and characterization of the male, duration of the developmental period, seasonal occurrence of adults, level of infestation in the field and the influence of leaf hairs on the occurrence of galls and adults. The influence of galls on the yield of C.citriodora oil was studied as well. From the total of leaves evaluated, 65% presented galls ranging between 0.04 and 44.10 galls/cm2, with a mean density of 7 galls per square centimeter. One also observed significant differences among the evaluated trees as to the number of galls on the leaves. Adult emergence occurred during the morning hours, with a peak at 9 am and adult longevity was longer when honey solution was available. The sex ratio was 165 females: 1 male. The external morphology of the genitalia and a microscopic structure in the male antenna were the only differences between the sexes. The life cycle, from egg to adult, lasted 75 days in field conditions. The galls presented five growing stages which probably are related to the changes in the immature insect periods. The adults are seen all year round and such occurrence is related to the continuous flushing of sprouts by the plant. This insect species is multivoltine and thus present overlapping generations. Young plants present hairy leaves acting as a mechanical barrier to oviposition, preventing the occurrence of adults and galls. The yield of C.corymbia oil leaves is negatively affected not only by the high levels of gall infestation but also by the following premature defoliation.

Thesis (PhD)--Stellenbosch University, 2008. / ENGLISH ABSTRACT: Introduced species displace native species and alter ecological communities, affect agriculture as well as human health and are economically costly to eradicate. Long term monitoring of introduced species including the documentation of levels of genetic variation is therefore of the utmost importance. This study investigated the distribution of genetic variation in two introduced species distributed across South Africa the bud gall-forming wasp Trichilogaster acaciaelongifoliae and the Argentine ant Linepithema humile. The bud gall-forming wasp was introduced into South Africa as a biological control agent to curb the spread of the invasive long-leaved wattle Acacia longifolia. In addition to the intended (target) host, the bud gall-forming wasp has also colonised A. floribunda, a noninvasive ornamental plant. Limited genetic variation was found across South Africa based on the mitochondrial DNA cytochrome oxidase subunit I (COI) gene. Only 3 haplotypes characterized 53 individuals collected from 23 localities (nucleotide diversity £k = 0.002 ¡Ó 0.001, haplotype diversity h = 0.482 ¡Ó 0.045). No significant partitioning of genetic variation was found across South Africa including between host plants (target host = A. longifolia, non target host = A. floribunda) or between the core (sites of introduction) and edge (naturally dispersed) sites (ƒ¶ST = 0.094, P = 0.288). The limited genetic variation and the absence of significant genetic structure are congruent with patterns described for many other introduced species and may suggest that propagule pressure plays only a minor role in species establishment and spread of the gall-forming wasp across South Africa. Mitochondrial and nuclear markers were used to describe the distribution of genetic variation within Argentine ants across their introduced range in South Africa. For the mitochondrial DNA, low genetic diversity was found for the COI gene with only five haplotypes, separated by single mutational changes, characterizing 101 specimens from 35 localities (nucleotide diversity π = 0.001 ± 0.001, haplotype diversity h = 0.151 ± 0.048). Notwithstanding the low levels of genetic diversity, mitochondrial variation was significantly structured (ST = 0.54, P < 0.001) across the landscape. In contrast, microsatellite analyses of 230 ants from 23 localities, employing six polymorphic microsatellite markers, revealed a relatively high amount of genetic diversity (HE = 0.51 ± 0.22). Significant population structure was similarly evident (RST = 0.14, P < 0.001) with the localities of Elim2, Porterville2 and Bloemfontein2 clustering as a distinct population from the remainder of the localities. Importantly, individuals from these localities also had a unique mitochondrial haplotype and, when taken with the nuclear results, may indicate the occurrence of more than one introduction event (and possibly more than one colony) in South Africa. This is further underscored by the presence of unique microsatellite alleles in these three populations. In an attempt to establish the source populations for the introduction of Argentine ants into South Africa, mitochondrial cytochrome b sequences were generated for a subset of ants representing the two major genetic clades across South Africa. A comparison with the published data from across the world including the native range of the Argentine ant in South America grouped Argentine ants from South Africa with three potential source populations namely Ocampo and Rosario in Argentina and Passo do Lontra in Brazil. The results of this study underscore the role of human-mediated dispersal in shaping the levels of genetic variation in both species. Human-mediated dispersal can lead to genetic homogenization across populations. / AFRIKAANSE OPSOMMING: Indringer spesies verplaas of verander ekologiese gemeenskappe, beinvloed landbou asook menslike gesondheid en is ekonomies duur om te verwyder. Langtermyn monitering van indringer spesies asook die dokumentasie van genetiese variasie is dus baie belangrik. Hierdie studie bestudeer die verspreiding van genetiese variasie in twee indringer spesies wat regoor Suid-Afrika voorkom, naamlik die kroongal-vormende wespe Trichilogaster acaciaelongifoliae en die Argentynse mier Linepithema humile. Die kroongal-vormende wesp is na Suid Afrika gebring as biologiese beheeragent om die verspreiding van indringer lang-blarige wattle Acacia longifolia te beveg. Die kroongalvormende wespe het sowel die teiken spesies, asook A. floribunda, ‘n nie-indringer ornamentele plant gekoloniseer. Beperkte genetiese variasie is gevind regoor Suid Afrika gebasseer op die mitochondriale DNA sitokroom-oksidasie subeenheid I (COI) geen. Slegs 3 haplotipes karakteriseer 53 indiviue van 23 bevolkings (nukleotied diversiteit π = 0.002 ± 0.001, haplotiep diversiteit h = 0.482 ± 0.045). Geen beduinde groepering van genetiese variase is gevind regoor Suid Afrika nie (ST = 0.094, P = 0.288). Hierdie bevinding geld onafhanklik van die gasheer plant (teiken gasheer = A. longifolia, nie-teiken gasheer = A. floribunda). Ook is geen beduidende genetiese groepering gevind tussen die sentrale (plek van inisiele blootstelling) en perifêre (natuurlik verspreide) lokaliteite nie. Die kleinskaalse genetiese variase en die afwesigheid van beduidende genetiese struktuur wat hier gevind is, verskil van die patrone wat voorheen vir baie ander indringer-spesies beskryf is. Dit mag daarop dui dat ‘propagule’ druk slegs ’n klein rol speel in spesies-vestiging en verspreiding van die galvormende wespe regoor Suid-Afrika. Mitochondriale asook kern merkers is gebruik om die verspreiding van genetiese variasie in Argentynse miere in Suid Afrika te beskryf. Vir die mitochondriale DNA is lae genetiese variase gevind vir die COI geen, met slegs 5 haplotipes, gedifferensieer deur enkele mutasie veranderinge wat 101 monsters van 35 lokaliteite karakteriseer (nukleotied diversiteit π = 0.001 ± 0.001, haplotiep diversiteit h = 0.151 ± 0.048). Desondanks die lae genetiese variasie, is gevind dat mitochondriale variasie beduidend gestruktureerd is (ST = 0.54, P < 0.001) oor die landskap. Hierteenoor het mikrosatelliet analises van 230 miere van 23 lokaliteite, deur gebruik te maak van ses polimorfiese mikrosatelliet merkers, ’n relatiewe hoë hoeveelheid genetiese diversiteit aangedui. Beduidende bevolkingstruktuur was ook gevind (RST = 0.14, P < 0.001) in die areas Elim2, Porterville2 en Bloemfontein2 wat saam groepeer as eiesoortige bevolkings vergeleke met die res van die areas. Ook van belang is dat individue van die areas ’n unieke mitochondriale haplotipe besit, en in kombinasie met die kern resultate, mag dit die voorkoms van meer as een blootstellingsgeleentheid (en moontlik meer as een kolonie) in Suid Afrika aandui. Hierdie bevinding word verder beklemtoon deur die teenwoordigheid van unieke mikrosatelliet allele in die drie bevolkings. In ‘n poging om die oorsprong van die oorsprong-bevolking vir die blootstelling van Argentynse miere in Suid Afrika vas te stel, is mitochondriale sitokroom b volgordes gegenereer vir ’n substel miere wat die twee hoof genetiese klades in Suid Afrika voorstel. Vergelyking met gepubliseerde data van regoor die wêreld, insluitende die endemiese gebied van die Argentynse mier in Suid Amerika, het die Argentynse mier van Suid Afrika met drie potensiele oorsprong-bevolking verbind, naamlik Ocampo en Rosario in Argentina en Passo do Lontra in Brazil.

La notion de « bile », exprimée par le substantif χολή, représente l’un des plus importants et des plus célèbres concepts de la médecine hippocratique, inséparable dans la pensée moderne de la fameuse théorie humorale. Au premier abord, les choses semblent donc claires. Cependant, lorsqu’on se plonge dans la lecture des écrits hippocratiques, la notion de cholè s’avère moins simple et évidente. Notre analyse des textes hippocratiques montre, en effet, que la cholè diffère d’un traité à l’autre et que chaque auteur hippocratique élabore sa propre notion de cholè. Nous nous sommes posé alors la question de l’origine de ce concept médical, ainsi que de l’origine de son cadre, qui est la théorie humorale. Notre quête des origines nous a amenée jusqu’aux idées homérique de chólos et aristophanique de cholè, qui se présentent toutes les deux comme fondamentalement différentes de l’idée médicale de cholè, unissant en elles-mêmes les notions de substance et d’état d’esprit. C’est justement cet écart entre les concepts non-médicaux et les concepts médicaux qui nous a intéressée au plus haut point, car il permet de voir comment les médecins hippocratiques élaborent leurs idées et leur discours. L’essentiel de notre travail consiste, donc, en un examen approfondi des procédés par lesquels les hippocratiques s’approprient des idées non-médicales : ce qu’ils retranchent, ce qu’ils rajoutent et ce qu’ils remanient. Nous espérons ainsi mettre en évidence les chemins par lesquels passe la pensée médicale ancienne, dans son processus d’émancipation de la culture traditionnelle, mais aussi des autres « sciences » de l’époque, telle que la philosophie. / The notion of « gall », expressed by the noun χολή, is one of the most important as well as the most celebrated concepts of the hippocratic medicine, inseparable for the modern mind from the humoral theory. At first sight then, the idea of « gall » seems fairly obvious. However, reading hippocratic treatises in detail, one realizes that the notion of cholè turns out to be far more complex and intricate than expected. Our analysis of the most relevant hippocratic texts shows indeed that the concept of cholè varies according to the texts involved, as every author tends to develop his own concept of cholè. We tried to find out whether the complex nature of the medical concept known as cholè could be elucidated by a survey of its origins, and a survey of the origins of the humoral system as a whole. Our search for the origins of cholè has led us to the Homeric concept of chólos and to the Aristophanic concept of cholè. The prerequisites of both notions conspicuously differ from the medical concept of cholè, because they unite the substance with a state of mind. This discrepancy between medical and non-medical concepts was of utmost importance for us, since it helped to understand how hippocratic authors developed their ideas and their discourse. The main asset of our work consists, therefore, in an in-depth analysis of the ways in which hippocratic authors take over some non-medical ideas to frame concepts of their own : what are the components they cut out, add or modify. Our goal is to show how ancient medical thought proceeds, in its endeavour to emancipate itself from the tradition as well as from the other contemporary “sciences”, as philosophy.

Foi realizado o estudo prospectivo de uma amostra da população portadora de Síndrome de Down que procurou o Departamento de Genética do Hospital Infantil Darcy Vargas,no período de 1959 a 1996. Dos 2816 portadores de Síndrome de Down que compareceram a este Serviço, neste período , a amostra estudada consistiu de 518 pacientes que mantiveram períodos de retomo ambulatorial conforme previsto pelo Protocolo de segmento clínico adotado. O objetivo do estudo foi caracterizar a incidência de colecistopatia litiásica em pacientes portadores de Síndrome de Down . Para desenvolver este estudo, após o diagnóstico citogenético e clínico da Síndrome de Down, procurou-se correlacionar faixa etária dos genitores, uso de anticoncepcionais e principalmente promover a utilização de exames subsidiários específicos que permitiram melhores condições diagnósticas e definição de mecanismos fisiopatológicos eventuais relacionados à colilitiase. Verificou-se a presença de cálculos de vesícula biliar em 27 pacientes ( 3,28%) comparando-se à incidência descrita na população como um todo (0,07%). Estes dados caracterizam a prevalência de cálculos de vesícula biliar em portadores de Síndrome de Down ( a distribuição das probabilidades da frequência de cálculo de vesícula biliar em Sindrome de Down está dentro do intervalo de 95%, entre 3,6% e 7,7%). As correlações realizadas vieram a excluir,na amostra estudada, algumas etiologias comumente descritas para colelitíase e provavelmente vincula o fenômeno da colelitíase a mecanismos fisioembriopatológicos ligados à hipotonia e estase do conteúdo da vesícula biliar( e/ou redução na velocidade do seu esvaziamento). Em consequência torna-se importante propor uma investigação rotineira para esta patologia, com ênfase nos mecanismos fisiopatológicos, prováveis responsáveis pela incidência aumentada na população com Síndrome de Down. / The purpose of this study has been the prospective analysis of the Down syndrome population received by the Genetic Department of \"Hospital Infantil Darcy Vargas\", in the period from 1959 to 1996. The Department has received 2816 Down Syndrome patients\' in this period, and this study has selected 518 patients, clinical and cytogenetically diagnosed, which were studied following the specific clinical protocol, in periodical evaluations. The incidence of cholelithiasis was verified and it has been analysed the possible relationships among this disease and parentaI age, contraceptive use and clinical features leading to the discussion of eventual aetiologic mechanisms causing gallstones. It was verified 27 patients affected by Down Syndrome and presenting cholelithiasis in the survey of 518 patients ( 3.28 %) compared to the frequency of 7/10000 in the non Down population ( 0.07% ). The statistical analysis has shown that the distribution of the frequency probabilities have been between 3.6% and 7.7%, These studies have verified that the aetiology of the gallstones in the Down syndrome patients could be related to the hypotonic characteristic of the patients, leading to cholestasis, It is very important the suggestion of routine evaluation for this pathology, related to the eventual aetiologic features.

The African Rice Gall Midge (AfRGM), Orseolia oryzivora Harris and Gagné (Diptera: Cecidomyiidae), is an endemic rice pest found throughout Africa. The failure of most other control methods imposes the need to use crop resistance. This study was initiated: (1) to develop an accurate method for assessing damage caused by AfRGM; (2) to determine AfRGM resistance genes’ modes of action, the heritability estimates of their resistance to AfRGM and the behavioural pattern of progenies with resistance to AfRGM attack; (3) to reveal convergent evolution of same or similar resistance gene(s) in geographically distinct landraces, or divergent evolution of genotypes carrying the same gene, by analysing the genetic diversity among five AfRGM parental lines; (4) to build a core sample of progenies to be used as a reduced mapping population, largely reflecting the entire genome of the whole population, after an estimate of the heritability of 15 agro-morphological descriptors and; (5) determine Simple Sequence Repeat (SSR) markers flanking genes or quantitative trait loci (QTLs) linked to resistance to AfRGM. A method of accurately assessing damage caused by AfRGM was determined by comparing four methods of assessment including the International Rice Research Institute’s (IRRI) Standard Evaluation System (SES) for rice and three methods based on resistance index (RI) assessments differing in the computing of the percentage of tillers with galls on a resistant check variety. The RI-based assessment (RI-BA) methods consistently provided a better evaluation of AfRGM damage than the SES, regardless of the trial size. Within RI-BA methods, RI-BA2 was always more accurate than RI-BA1 and RI-BA3 when the plot was large. RI-BA2 and RI-BA3 were equally accurate when the plot size was small, and they provided better estimates than RI-BA1. When the plot was of medium size, RI-BA2 was more accurate than RI-BA3; RI-BA3 also surpassed RI-BA1. Overall, the best method of assessing AfRGM damage was RI-BA2, regardless of the plot size. Five rice populations including F1, F2 and F3 generations involving ITA306, a susceptible variety of Oryza sativa subsp. indica, and four varieties having different reactions against AfRGM were used to determine the genetic basis of resistance and estimate the heritability of resistance to AfRGM. All the F1s were susceptible, suggesting recessive gene inheritance. The F2 generations’ segregation pattern of 1R:15S in both ITA306-TOS14519 and ITA306-TOG7106 crosses as well as the segregation of 1R:8Seg:7S in ITA306-TOS7106 F3 families indicated that the AfRGM resistance expression being studied is governed by two genes. The deviation of the segregation patterns of crosses involving ITA306 and the tolerant parental lines from Mendelian segregation ratios suggests that the tolerance to AfRGM shown by BW348-1 and Cisadane is under complex mechanisms of control rather than under simple genetic control. The narrow-sense heritability estimates of resistance to AfRGM were low in populations involving tolerant varieties and were high in populations involving resistant varieties. They ranged from 0.086 in the ITA306-Cisadane population, to 0.4 in the ITA306-TOG7106 population. Conversely, the broad-sense heritability estimates ranged from 0.23 (ITA306-Cisadane) to 0.63 (ITA306-TOS14519). The behavioural patterns of progenies against AfRGM attack were evaluated for 532, 413 and 479 F2 progenies from ITA306-BW348-1, ITA306-Cisadane and ITA306-TOS14519 crosses, respectively, in addition to 90 BC1F2 progenies from the ITA306 and TOG7106 cross. One F3 generation of 649 families from a cross between ITA306 and TOS14519 was also tested. Four types of behavioural pattern categories were observed: (1) progenies were more resistant than the resistant check entry at 45 DAT and 70 DAT; (2) progenies were more resistant at 45 DAT and became susceptible at 70 DAT; (3) progenies were susceptible at both 45 DAT and 70 DAT; (4) progenies were susceptible at 45 DAT but reverted to resistant at 70 DAT. The first three categories were the most frequently observed and occurred in all cross combinations. The last category was observed only for a few progenies from the ITA306-TOS14519 F2 and F3 generations and, surprisingly, many from the ITA306 and BW348-1 cross. Heritability estimates were calculated for 15 major traits in an F3 population in order to predict the genetic gain associated with each trait, together with the resistance to AfRGM and to estimate the influence of the environment on phenotypic values. Broad-sense heritability (H2) estimates were high for the penultimate leaf length (PLL) - 0.99, penultimate leaf width (PLW) – 1.0, flag leaf length (FLL) - 0.99, flag leaf width (FLW) – 1.0, ligule length (LigL) - 0.99, tillering ability (Til) - 0.99, number of days to booting (DB) - 0.95, number of days to first heading (DFH) - 0.96, number of days to heading (DH) - 0.89, number of days to maturity (DM) - 0.98, culm length (CL) - 0.99, plant height (PH) - 0.99, panicle length (PanL) - 0.95, secondary branching (SB) - 0.95 and the thousand grains weight (TGW) - 0.71. Conversely, narrow-sense heritability estimates were very low (nearly 0) in PLL, FLL, Lig, DB, DFH, DM and SB or low (at most 0.267) in PLW, FLW, DH and PH, with a high value of 0.727 for TGW. Inheritance of the traits studied was therefore under non-additive gene effects rather than additive genetic effects and can therefore be improved using pedigree breeding schemes along with breeding for AfRGM resistance. Fine genetic evaluation of five AfRGM parental lines was studied in terms of polymorphisms using 303 SSR primers covering the rice genome. Of the 178 polymorphic primers identified, 60 were highly polymorphic and informative. The number of alleles amplified by these primers ranged from one to five for a total of 1,041 alleles. The polymorphism rate was globally high, ranging from 45.2% to 66.8%. The mean of the polymorphism information content (PIC) was 0.553. Factorial analysis, based on the allelic diversity, demarcated the parental lines into Oryza glaberrima Steud, Oryza sativa subsp. japonica and O. sativa subsp. indica groups, while a cluster analysis distinguished them into four groups: AfRGM resistant, susceptible, moderately resistant and tolerant. BW348-1 and Cisadane showed the least diversity, despite their distant geographical origins. TOS14519 and TOG7106 showed more divergence to ITA306 despite their common West African origin. This variability amongst the genotypes tested is the result of farmer-based selection for AfRGM resistance rather than direct breeding efforts through breeder intervention. A method of selecting individuals for a mapping population, based on a core sample, was developed in order to speed up the mapping procedure. A diversity study amongst F2 and F3 generations involving 15 quantitative and 26 qualitative agro-morphological characters was carried out and led to the dropping of seven non-discriminant descriptors. The diversity index (H) was calculated for each remaining character and the discriminant descriptors were selected based on a diversity index threshold value above 0.4. Four descriptors of H values less than 0.35 were therefore dropped. The sizing of the core collection of 64 individuals and the selection of these individuals were done using MSTRAT version 4.1 package in redundancy mode, a construction run of 100 times with an iteration number of 500. The core sample was similar to the whole population for clustering pattern, minimum and maximum quantitative values and diversity index, while mean values and coefficient of variation distinguished them. The core sample, which represents 10% of the whole population, also revealed the same phenotypic variation and the same genotypic segregation according to two SSR markers. It can therefore efficiently reflect the whole population as a mapping population. Finally, a study was undertaken to identify flanking markers to the gene/QTL involved in the resistance against AfRGM using bulked segregant analysis (BSA). A polymorphism study between ITA306 and TOS14519 displayed 145 polymorphic SSR markers, which were used to screen the bulks that originated from the two tails, and depicted only two SSRs as candidate markers linked to gall midge resistance. These markers included RM317 and RM17303 which displayed strong significance after an analysis of variance using an F test, meaning that they were segregating with the resistant alleles. / Thesis (Ph.D.)-University of KwaZulu-Natal, Pietermaritzburg, 2012.

Foi realizado o estudo prospectivo de uma amostra da população portadora de Síndrome de Down que procurou o Departamento de Genética do Hospital Infantil Darcy Vargas,no período de 1959 a 1996. Dos 2816 portadores de Síndrome de Down que compareceram a este Serviço, neste período , a amostra estudada consistiu de 518 pacientes que mantiveram períodos de retomo ambulatorial conforme previsto pelo Protocolo de segmento clínico adotado. O objetivo do estudo foi caracterizar a incidência de colecistopatia litiásica em pacientes portadores de Síndrome de Down . Para desenvolver este estudo, após o diagnóstico citogenético e clínico da Síndrome de Down, procurou-se correlacionar faixa etária dos genitores, uso de anticoncepcionais e principalmente promover a utilização de exames subsidiários específicos que permitiram melhores condições diagnósticas e definição de mecanismos fisiopatológicos eventuais relacionados à colilitiase. Verificou-se a presença de cálculos de vesícula biliar em 27 pacientes ( 3,28%) comparando-se à incidência descrita na população como um todo (0,07%). Estes dados caracterizam a prevalência de cálculos de vesícula biliar em portadores de Síndrome de Down ( a distribuição das probabilidades da frequência de cálculo de vesícula biliar em Sindrome de Down está dentro do intervalo de 95%, entre 3,6% e 7,7%). As correlações realizadas vieram a excluir,na amostra estudada, algumas etiologias comumente descritas para colelitíase e provavelmente vincula o fenômeno da colelitíase a mecanismos fisioembriopatológicos ligados à hipotonia e estase do conteúdo da vesícula biliar( e/ou redução na velocidade do seu esvaziamento). Em consequência torna-se importante propor uma investigação rotineira para esta patologia, com ênfase nos mecanismos fisiopatológicos, prováveis responsáveis pela incidência aumentada na população com Síndrome de Down. / The purpose of this study has been the prospective analysis of the Down syndrome population received by the Genetic Department of \"Hospital Infantil Darcy Vargas\", in the period from 1959 to 1996. The Department has received 2816 Down Syndrome patients\' in this period, and this study has selected 518 patients, clinical and cytogenetically diagnosed, which were studied following the specific clinical protocol, in periodical evaluations. The incidence of cholelithiasis was verified and it has been analysed the possible relationships among this disease and parentaI age, contraceptive use and clinical features leading to the discussion of eventual aetiologic mechanisms causing gallstones. It was verified 27 patients affected by Down Syndrome and presenting cholelithiasis in the survey of 518 patients ( 3.28 %) compared to the frequency of 7/10000 in the non Down population ( 0.07% ). The statistical analysis has shown that the distribution of the frequency probabilities have been between 3.6% and 7.7%, These studies have verified that the aetiology of the gallstones in the Down syndrome patients could be related to the hypotonic characteristic of the patients, leading to cholestasis, It is very important the suggestion of routine evaluation for this pathology, related to the eventual aetiologic features.

Submitted by Edineia Teixeira (edineia.teixeira@unioeste.br) on 2018-03-02T14:50:51Z No. of bitstreams: 2 Adriana_Walerius2017.pdf: 932754 bytes, checksum: cc0c99130f89646094f3b4fee12e13b8 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-03-02T14:50:51Z (GMT). No. of bitstreams: 2 Adriana_Walerius2017.pdf: 932754 bytes, checksum: cc0c99130f89646094f3b4fee12e13b8 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-03-10 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Gyropsylla spegazziniana (Hemiptera: Psyllidae) is considered an important pest of the yerba mate (Ilex paraguariensis), causing considerable economic losses. Most insects use chemical, visual, or acoustic signals in interspecific communication. Sex pheromones are the main signals involved in mating behavior. In this sense, this work aimed to describe the mating behavior of G. spegazziniana, to verify the attractiveness between males and females in Y olfactometer bioassays, and to identify the sex pheromone of this species. To describe the mating behavior virgin couples were placed in cages containing a yerba mate seedling. The insect behavior during the photophase and scotophase was observed. For behavioral tests in Y-olfactometer, virgin males or virgin females and a branch of yerba mate were used as odor source. To verify the presence of a sexual pheromone, body extracts of virgin males or virgin females were obtained and were analyzed by using gas chromatograph coupled to mass spectrometer (GC-MS). Mating occurred only during the photophase, in the scotophase the couples remained immobile in the abaxial surface of the leaves. The mating sequence showed that the female walks on the new leaves and the male goes to meet her, standing face to face with her. After a brief antennal touch between the couple, the male turns the abdomen and attaches to the female. As a result of Y olfactometer bioassays, males and females were attracted to females in the olfactometer but were not attracted to males, suggesting a presence of volatiles released by females for sexual attraction. There was a presence of a single female-specific compound, identified as 2-nonadecanone, that may act as sex pheromone.The results suggest that G. spegazziniana females release sex pheromone to attract males for mating. The compounds released by the females can interact with the volatiles emitted by the yerba mate, resulting in the attractiveness of G. spegazziniana males. / Gyropsylla spegazziniana (Hemiptera: Psyllidae) é um psílideo considerado uma das principais pragas da cultura da erva-mate (Ilex paraguariensis), causando grandes prejuízos econômicos. A maioria dos insetos utiliza sinais químicos, visuais ou acústicos na comunicação interespecífica. Os feromônios sexuais são os principais sinais envolvidos no acasalamento. Neste sentido, este trabalho teve como objetivo descrever o comportamento de acasalamento de G. spegazziniana, verificar a atratividade entre machos e fêmeas em bioensaios com olfatômetro em Y e identificar o feromônio sexual da espécie. Para a descrição do acasalamento, casais virgens foram colocados em gaiolas contendo uma muda de erva-mate. Foi observado o comportamento dos casais durante a fotofase e escotofase. Para os ensaios de atratividade em olfatômetro em Y, foram utilizados insetos vivos (machos e fêmeas virgens) e um ramo de erva-mate como fonte de odor. Para analisar a presença do feromônio sexual foram feitos extratos do corpo de machos e fêmeas virgens, que foram analisados em cromatógrafo gasoso acoplado à espectrômetro de massas (CG-MS). Foi verificado que a cópula ocorreu somente durante a fotofase, sendo que na escotofase os casais permaneciam imóveis na parte abaxial da folhas. A sequência de acasalamento demonstrou que a fêmea caminha principalmente pela brotação e o macho vai ao seu encontro, ficando frente a frente com a mesma; após um breve toque de antenas entre o casal, o macho vira o abdome e rapidamente ocorre a cópula. Nos bioensaios de atratividade, machos e fêmeas foram atraídos para fêmeas vivas no olfatômetro, porém não foram atraídos para machos vivos, o que sugere a presença de voláteis liberados pelas fêmeas para atração sexual. Foi constatada a presença de um composto fêmea-específico, identificado como 2-nonadecanona, com o possível papel de feromônio sexual. De acordo com os resultados, sugere-se que as fêmeas de G. spegazziniana liberam feromônios sexuais para atrair machos para o acasalamento. Os compostos liberados pelas fêmeas podem interagir com os voláteis emitidos pela erva-mate, resultando na atratividade dos machos de G. spegazziniana.