Made available in DSpace on 2015-04-20T12:31:49Z (GMT). No. of bitstreams: 1 Carlos Gustavo Nunes da Silva.pdf: 7132119 bytes, checksum: 87b96758db3807b14b707d1380997db6 (MD5) Previous issue date: 2008-03-24 / Conselho Nacional de Desenvolvimento Científico e Tecnológico / The presence of castes among females and a sytem of seasonal production of males in order to fund new daughter colonies or replace the Queen, is essencial to life in society for the high eusocial bees. The haplodiploidy has unique consequences to the populational ecology and social oveolutinos for the social bees. Studies have shown, in depleted popultions, that consecutive mating between indviduals with equal allele composition for the sex determination gene can cause its extinction. The sex determination genes csd (complementary sex determiner) and fem (feminizer) seems to be at the top of a regulatory cascade in which will originate the different phenotypes, which is indeed a essential way to mainten the eusociability in bees. The sex determination system as well as the several distincts caste determination systems existing among the high eusocial bess are yet poorly undertood in their genetic basis and seems to be fairly complex as diverse. The goal of this thesis was to sequence genes envolved in the sex or caste determination in the Melipona bees. Using the RACE (Rapid Amplification of cDNA Ends) technique, the gene fem (feminizer) was cloned and sequenced in M. compressipes and analysed with help of bioinformatcis tools. The analysis show in Melipona compressipes that the predict gene product have 64% of identity and 75% of similarity with its equivalent (Fem) in Apis mellifera, posesing the same protein domains: Arginime-Serine domain (RS-domain) and proline domain (proline rich dormain). The gene fem in M. compressipes as in Apis, do not have hyper variable region which characterizes its paralog gene csd Thus, as this work proposes would indicate that the fem product would have function in the activation of genes in Melipona switching on other secondary feminizer genes in contrast with the masculining genes, which are constitutive.We propose a model that that put together literature information with the new gene fem findings in Melipona, aiming a discussion about the relulatory cascade which surpass from the sex determination / A presença de castas entre as fêmeas e um sistema de produção sazonal de machos para renovação da rainha ou fundação de colônias filhas é fundamental para a vida em sociedade nas abelhas eusociais. Um sistema peculiar de determinação sexual é encontrado nesses insetos considerados organismos haplodiplóides, no quais, machos viáveis reprodutivamete são haplóides nascidos por partenogêne arrenótica (ovos não fecundados dando origem a machos), enquanto fêmeas são originadas de ovos fecundados. A haplodiploidia tem conseqüências fundamentais para a ecologia populacional e evolução social. A determinação das castas, por sua vez, em abelhas do gênero Melipona é sustentada pela hipótese do sistema genético-alimentar proposta por (KERR,1974), sendo dependente da dupla heterozigose, dos genes xa e xb (cada um com dois alelos). O objetivo desse trabalho foi o de seqüenciar genes envolvidos na determinação de sexo e ou castas em abelhas do gênero Melipona. Por meio de técnicas de isolamento gênico, utilizando o método de amplificação de extremidades para a obtenção de cDNAs completos (Rapid Amplification of Cdna Ends RACE). O gene fem (feminizer) foi clonado e seqüenciado em M. compressipes e analizado com o auxílio de ferramentas de bioinformática. Os resultados da análise mostram, que em Melipona compressipes, o produto predito do gene tem 64% de identidade e 75% de similaridade com o equivalente (fem) em Apis mellifera. Possuindo os mesmos domínios ricos em prolina (proline rich domain). Além disso, o gene fem é expresso em todos os instares imaturos e inclusive em adultos e igualmente encontrado em cDNAs de machos, operárias e rainhas, mostrando, da mesma forma que em A. mellifera, que esse é um gene expresso em todas as fases do desenvolvimento e em ambos sexos e castas, além de ser muito conservado interespecificamente. O gene fem, não possui a região hipervariada. Isso, segundo proposta desse trabalho, indicaria outros genes além de fem teria funções na ativação dos genes de castas em Melipona e dispararia outros genes feminizantes em contraposição aos genes masculinizantes, esses, por sua vez, constitutivos. É proposto um modelo, que reúne as informações da literatura com a descoberta do gene fem em Melipona

Altruism in its extreme form is seen in social insects where most individuals give up their own reproduction and work to rear the offspring of their queen. The origin and evolution of such sterile worker castes remains a major unsolved problem in evolutionary biology. Primitively eusocial polistine wasps are an attractive model system for investigating this phenomenon. Ropalidia marginata (Lep.) (Hymenoptera: Vespidae) is one such tropical primitively eusocial wasp, in which new nests are initiated either by a single foundress or by a group of female wasps. Worker behaviour in Ropalidia marginata cannot be satisfactorily explained by the haplodiploidy hypothesis due to the existence of polyandry and serial polygyny which reduce intra-colony genetic relatedness to levels lower than the value expected between a solitary foundress and her offspring. Besides, wasps appear to move frequently between newly initiated nests, perhaps further reducing intracolony genetic relatedness. To study social organization and examine the possibility of kin recognition and task specialization under conditions of low intra-colony relatedness, genetically mixed colonies were created by introducing alien one-day old wasps onto recipient nests. As a first step I have tried to determine the factors that influence the acceptance of foreign wasps onto established colonies. I have introduced wasps between 1 to 20 days of age from donor colonies located at least 10 km away onto 12 different recipient colonies, observed these wasps for a period of 10 hours and later dissected them to examine their ovarian condition. Observations were carried out in the blind i.e. the observer was unaware of the identity of the wasps. Wasps upto 6 days of age were accepted by the alien nests. Older wasps may have been rejected because their relatively better ovarian condition may have been perceived as a reproductive threat to the recipient nest. Alternatively, younger wasps may have been accepted because they may be more easily moulded to the desired roles or due to some other correlate of age per se independent of ovarian condition. Although ovarian condition appeared to influence the probability of acceptance, it was not statistically significant in the presence of age in multiple regression models, making a favourable case for the 'ease of moulding hypothesis' or 'age per se hypothesis' over the 'reproductive threat hypothesis'. In any case these findings gave me a method to create genetically mixed colonies. On 12 different nests Ropalidia marginata, I similarly introduced one-day old wasps and thus created genetically mixed colonies. Such an introduction simulates the eclosion of distantly related individuals which is quite common on nests of R. marginata due to the presence of serial polygyny. About 7 such wasps were introduced per colony and the introductions were so arranged as to matched with natural eclosions on the recipient nest. After 7 days following the last introduction, colonies were observed for 20 hours each. Alien wasps became well integrated and performed most of the behaviours and tasks shown by the natal wasps. There was no evidence of kin recognition or task specialization between natal and introduced wasps. The introduced wasps also sometimes became replacement queens. In an attempt to test the costs in terms of brood rearing efficiency, of living in such genetically variable groups, I created kin and non-kin pairs of wasps in plastic containers. They were provided with ad libitum food, water and building material. The nests initiated were monitored till an adult offspring eclosed. There were no detectable differences in either the productivities or the developmental periods of immature stages in the kin and nonkin pairs suggesting that there is no apparent cost of living with unrelated or distantly related individuals. To compare the extent of cooperation between the two wasps in kin and non-kin pairs, I conducted behavioural observations on 12 pairs each of kin and nonkin wasps. I found no difference in the rates at which the non-egg layers brought food and pulp, fed larvae and built the nest in the kin and nonkin pairs suggesting that cooperative nest building and brood rearing was common to the kin as well as non-kin pairs. The results reported here strengthen the idea that factors other than genetic relatedness must play a prominent role in the maintenance of worker behaviour in Ropalidia marginata.

<p>  </p> <p>Mutations drive evolution, generating variation that selection can act upon. Germline mutations are heritable genetic alterations that occur in the gametes prior to fertilization and until embryogenesis. The study of germline mutations is vital to understanding the genetic basis of heritable diseases and evolution. Somatic mutations are genetic alterations across the body that arise post-fertilization in non-gametic cells. Although somatic mutations in most animals cannot be inherited, they can still significantly affect an organism's reproductive success. In humans, for example, cancers can be the result of somatic mutations. Somatic mutations originate from both exogenous mutagens (e.g. UV radiation) and endogenous processes (e.g. DNA replication, aging). Beyond their origins, we know little about the distribution and frequency of somatic mutations across Animalia. Honey bees provide a unique model for the study of somatic mutations as they are haplodiploid: males come from unfertilized eggs and are haploid, while females come from fertilized eggs and are diploid. It is therefore possible to sequence and robustly identify somatic mutations in haploid males. I have developed a unique exploratory study to elucidate the distribution of somatic mutation accumulation in honey bee drones. With this, I aim to investigate processes generating somatic mutations. Our findings demonstrate that variance in somatic mutational load is better captured across individuals rather than within individuals (across tissues). I provide a comprehensive tissue atlas of somatic mutagenesis in haploids. Our findings drive us to enhance our view of mutagenesis from the tissue level down to the cellular level. </p>

Genomische Selektion ist ein Routine-Verfahren bei verschiedenen Nutztierarten, aber noch nicht bei der Honigbiene wegen der Besonderheiten dieser Spezies. Für die Zuchtwertschätzung bei der Honigbiene ist eine spezielle genetische Verwandtschaftsmatrix erforderlich, da die Paarungsbiologie dieser Spezies ungesicherte Vaterschaft, diploide Königinnen und haploide Drohnen umfasst. Die Arbeit präsentiert einen neu-entwickelten Algorithmus zur effizienten Berechnung der Inversen der genetischen Verwandtschaftsmatrix und der Inzuchtkoeffizienten auf großen Datensätzen. Die Methode wurde zur Voraussage von genomischen und Stammbaum-basierten Zuchtwerten in einer Simulationsstudie genutzt. Die Genauigkeit und die Verzerrung der geschätzten Zuchtwerte wurden ausgewertet unter Berücksichtigung verschiedener Größen der Referenzpopulation. Außerdem wurde der Zuchtfortschritt im ersten Durchlauf von Zuchtprogrammen ausgewertet, die Zuchtschemata mit genomischer oder Stammbaum-basierter Selektion nutzten. Ein erheblich größerer Zuchtfortschritt als bei Stammbaum-basierter Selektion wurde mit genomischer Vorselektion erzielt, für die junge Königinnen genotypisiert wurden, und nur die Kandidaten mit den höchsten genomischen Zuchtwerten zur Anpaarung oder Leistungsprüfung zugelassen wurden. Für einen realen Datensatz von ungefähr 3000 genotypisierten Königinnen wurden Stammbaum-basierte und genomische Zuchtwerte für sechs wirtschaftlich bedeutende Merkmale vorhergesagt. Drei Merkmale zeigten eine signifikant höhere Vorhersagegenauigkeit bei genomischer Zuchtwertschätzung gegenüber Stammbaum-basierten Verfahren und die Unterschiede zwischen allen sechs Merkmalen konnten im Wesentlichen aus den genetischen Parametern der Merkmale und der begrenzten Größe der Referenzpopulation erklärt werden. Damit zeigt die Arbeit, dass die genomische Selektion bei der Honigbiene genutzt werden kann, den Zuchtfortschritt zu erhöhen. / Genomic selection is a routine practice for several important livestock species but not yet in honey bees, due to the peculiarities of this species. For honey bees, a specialized genetic relationship matrix is required for the prediction of breeding values, since their mating biology involves uncertain paternity, diploid queens, and haploid drones. The thesis presents a novel algorithm for the efficient computation of the inverse of the numerator relationship matrix and the coefficients of inbreeding on large data sets. The method was used to estimate genomic and pedigree-based breeding values in a simulation study. The accuracy and bias of the estimated breeding values were evaluated and various sizes of the reference population were considered. Subsequently, the genetic gain in the initial cycle of breeding programs was evaluated for several breeding schemes employing genomic or pedigree-based selection. A considerably higher genetic gain than with pedigree-based selection was achieved with genomic preselection, for which queens were genotyped early in life, and only the candidates of high genomic breeding value were admitted for mating or phenotyping. On a real data set of about 3000 genotyped queens, pedigree-based and genomic breeding values were predicted for six economically relevant traits. Three traits showed significantly higher prediction accuracy with genomic compared to pedigree-based methods, and the differences between all the six traits could be explained mainly from their genetic parameters and the limited size of the reference population. The results show that genomic selection can be applied in honey bees, and the thesis provides appropriate breeding schemes and mathematical methods for its implementation.