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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
31

The Genetic Basis for Seed Coat Polymorphisms In Lupinus Perennis

Wilson, Rachel Erin 26 November 2019 (has links)
No description available.
32

Characterization of the physiological and behavioral effects of dehydration on mosquito vectorial capacity

Holmes, Christopher 23 August 2022 (has links)
No description available.
33

Zic3 and the embryonic mouse node: Defining early processes involved in left-right patterning and heart development

Sutherland, Mardi J. January 2013 (has links)
No description available.
34

The genomic architecture of sex-biased gene expression in Xenopus borealis

Song, Xue-Ying January 2019 (has links)
Most vertebrates have separate sexes, and sex-specific traits that are regulated by genes with sex-biased expression patterns. In many species with genetic sex determination system, genetic recombination is suppressed in genomic regions linked to the master regulator of sex determination – the gene or set of linked loci that orchestrate sexual differentiation. Natural selection may favour alleles with sex-specific effects - including those with sexually antagonistic (SA) fitness effects (e.g., beneficial to females but harmful to males) – to become fixed in or be translocated to these non-recombining regions of sex chromosomes, because sex-specific or sex-biased modes of inheritance can resolve genomic conflict associated with SA. Sexually antagonism may also be resolve by sex-biased gene expression, and in theory these two mechanism (sex-linkage and sex-biased gene expression) could operate synergistically. However, there are relatively few empirical studies that test whether genes with sex biased expression patterns are indeed more abundant on sex chromosomes – and especially on newly evolved sex chromosomes. We explored this question with an African frog species Xenopus borealis, whose sex chromosome evolved within the last 25 million years (my) and have a large (~50Mbp) region of suppressed recombination, making it a young sex chromosome system compared to many other intensively studied systems, such as the sex chromosomes of mammals. We tested the possibility that a higher proportion of genes with sex-biased expression would be located on the sex-linked region of the sex chromosome of this species. By examining gene expression in adult liver and gonad and also tadpole gonad/mesenephros at two developmental stages, we found that the sex-linked region of these sex chromosome do have a higher proportion of sex biased genes compared to the non-sex-linked region of the same sex chromosomes, compared to (i) a homeologous genomic region in the tetraploid genome of X. borealis, and also (ii) the autosomes of this species. We did not observe the same pattern in a closely related frog species, Xenopus laevis, which has sex chromosome that are not homologous to those of X. borealis and, unlike X. borealis, lacks a large region of suppressed recombination on its sex chromosome. Using Brownian Motion model, we found as well that expression divergence evolution of genes in the sex-linked region of X. borealis is faster compared to its non-sex-linked homeologs (within X. borealis), and also compared to orthologous regions that are also non-sex-linked. One possible explanation for these observations is that natural selection favoured an expansion of recombination suppression (via unknown mechanisms) on chromosome such that polymorphic regulatory regions became linked (or unlinked) to the sex determining locus in such a way to resolve SA. Alternatively, it is possible that these sex-biased expression pattern evolved rapidly after recombination suppression. / Thesis / Master of Science (MSc) / Sexual selection favours the evolution of distinctive traits in each sex in order to optimize the reproductive success of each one. However, because most of the genome is shared between the sexes, sexual selection may result in genomic conflict when mutations are beneficial to one sex but harmful to the other; this conflict is known as sexual antagonism. Genomic conflict associated with alleles with sexually antagonistic (SA) fitness effects can be resolved via the origin of sex-biased expression patterns and this may be catalyzed by genetic linkage to a sex-determining locus on a sex chromosome. Consequently, one might predict there to be an enrichment of genes with sex-biased expression patterns on the sex chromosome as compared to the autosomes. We tested this expectation in an African frog species Xenopus borealis, which has a relatively young sex chromosomes and a large region of recombination suppression on the female-specific W-chromosome. We found enrichment of sex-biased genes on the nonrecombining region of the sex chromosomes of this species in adult liver and gonad tissue and also tadpole mesenephros/gonads at two developmental stages. Additionally, we found that expression divergence of genes in the non-recombining region have a faster rate of evolution as compared to the rate of expression divergence of genes in other genomic regions. One possible explanation for these observations is that natural selection favours an expansion of recombination suppression (via unknown mechanisms) on sex chromosome such that polymorphic regulatory region become linked (or unlinked) to the sex determining locus in such a way as to resolve SA.
35

苔類ゼニゴケにおけるデータベース整備及びそれを活用した植物特異的なメチル基転移酵素ファミリー遺伝子の解析

川村, 昇吾 25 September 2023 (has links)
京都大学 / 新制・課程博士 / 博士(生命科学) / 甲第24945号 / 生博第507号 / 新制||生||67(附属図書館) / 京都大学大学院生命科学研究科統合生命科学専攻 / (主査)教授 河内 孝之, 教授 荒木 崇, 教授 中野 雄司 / 学位規則第4条第1項該当 / Doctor of Philosophy in Life Sciences / Kyoto University / DFAM
36

Immunité innée et multi-infections chez le moustique (Diptera, Culicidae) : étude fonctionnelle des interactions Wolbachia-arbovirus-Aedes albopictus / Innate immunity and multiple infection in mosquito (Diptera, Culicidae) : functional study of Wolbachia-arbovirus-Aedes albopictus interaction

Raquin, Vincent 18 December 2012 (has links)
On assiste actuellement à l'émergence et la ré-émergence mondiale d'arboviroses comme le chikungunya, la dengue ou la fièvre de la vallée du Rift. Ces maladies, responsables d'environ 30 000 décès par an, sont dues à des virus principalement transmis à l'homme par des moustiques vecteurs. En l'absence de vaccins efficaces et face aux limites de l'utilisation d'insecticides, nocifs pour les écosystèmes et entrainant des résistances chez les vecteurs, il est nécessaire de développer des moyens alternatifs de lutte. La découverte récente du potentiel antiviral de certaines bactéries symbiotiques du moustique, comme le genre Wolbachia, représente un outil de lutte biologique prometteur face aux arboviroses. Ce projet de thèse porte sur la relation tripartite moustique-bactéries endosymbiotiques-arbovirus, en prenant pour modèle le moustique-tigre, Aedes albopictus. Cette espèce originaire d'Asie envahit progressivement l'Europe. Elle transmet notamment le virus de la dengue et du chikungunya, et est naturellement infectée par Wolbachia. Les résultats obtenus ont permis d'observer un phénotype antiviral chez les moustiques infectés par Wolbachia, contrairement aux moustiques aposymbiotiques. L'utilisation d'une méthode de transcriptomique haut-débit (RNAseq) a permis de déterminer certains mécanismes cellulaires et moléculaires majeurs du moustique spécifiquement impliqués dans l'interaction avec les arbovirus, Wolbachia, et les deux partenaires simultanément. Le développement d'une lignée cellulaire d'Ae. albopictus stablement infectée par Wolbachia a permis de mettre en évidence le rôle central de l'autophagie dans l'interaction Wolbachia-arbovirus chez Ae. Albopictus / Arthropod-borne virus (arbovirus) are important cause of human diseases worldwide, leading to nearly 30.000 deaths every year. Many arboviruses like dengue virus (DENV), chikungunya virus (CHIKV) or Rift valley Fever virus (RVFV) are transmitted by mosquitoes, and global changes like climate warming or international trade increase vectors geographic range, thus facilitating the emergence of arbovirosis. Very few vaccines are currently available, and the use of insecticides remains the only way to prevent arbovirosis but cause adverse effects on ecosystems, and lead to resistance phenotypes in vector populations. Recent work showed that mosquito bacterial flora, especially bacteria from the genus Wolbachia, can modulate viral infection, a phenotype called microbial interference. These results provide a promising tool to limit transmission of arboviruses, but little is known about mosquito-Wolbachia-arbovirus interaction especially at the cellular level. We characterized for the first time this multipartite interaction in Aedes albopictus, an important mosquito vector of DENV and CHIKV, which is naturally infected by Wolbachia. Results showed an antiviral phenotype in Wolbachia-infected mosquitoes, compared to aposymbiotic insects. We used RNAseq to decipher the major mosquito pathways implemented during mono-infection by virus, bacteria or during bi-infection. Moreover, we developed an Ae. albopictus cell line stably infected by Wolbachia to go further in mechanical aspects, and showed that autophagy is a major pathway involved in Wolbachia-arbovirus interaction in Ae. albopictus
37

Flower evolution in species of Croton L. (Euphorbiaceae): ontogeny and global profile of gene expression / Evolução floral em espécies de Croton L. (Euphorbiaceae): ontogênese e perfil global da expressão gênica

Gagliardi, Karina Bertechine 27 July 2018 (has links)
The Euphorbiaceae are notable for floral and inflorescence diversity and evolutionary complexity. Croton, is the second largest genus in the family and exhibits particular diversity in its flowers, especially regarding perianth and number of stamens, besides the inflorescences, which are also very diverse. Considering Croton\'s great variability in the reproductive structures, the aim of this thesis was to study flowers and inflorescences with an evolutionary approach, including morphology, ontogeny, vasculature, auxin regulation and genetic expression. Flowers in several stages of development were analyzed using light microscopy and scanning electron microscopy. Inflorescences were analyzed in stereomicroscope and the traits were plotted on the most recent phylogeny of the genus. The genetic expression was tested using RNAseq. In the first chapter the flowers showed similarity in the initiation of sepals and the presence of filamentous, petaloid structures in Croton lundianus (Didr.) Müll. Arg., interpreted here as staminodes. In Croton sphaerogynus Baill., staminodes were described for the first time. The staminodes reported here could be interpreted as transitional structures that we considered as evolutionary reductions. In the second chapter, the staminate flowers showed polystemonous androecium and the delay in petals\' initiation and the antesepalous nectaries development interfered in the development of the stamens, characterizing obdiplostemony. Vasculature corroborated obdiplostemony and revealed a central stamen in C. fuscescens with carpelar features, interpreted here as a homeosis case. Glandular staminodes were registered and interpreted as a heterotopy case. The obdiplostemony may be related to modulation of the free IAA concentrations during floral developmental steps and Croton flowers can be used as good models for obdiplostemony, homeosis and heterotopy. In the third and fourth chapter we studied Croton inflorescences, which showed 17 patterns with differences on the organization and distribution of pistillate flowers. The inflorescence traits analyzed were very homoplastic, most likely determined by convergent evolution in distantly related lineages distributed in similar habitats. The genetic expression of C. fuscescens was particularly analyzed and the transcriptome showed that the different zones have their development guided through the same transcripts set. Each zone has different expression level and these variations and gradient could be interpreted as the boundary between each inflorescence zone. The floral developmental novelties and evolutionary links identified here raise the importance of future floral studies with the genus, what would bring a better understanding on how the reproductive structures evolved in the history of the group / Euphorbiaceae é uma família que recebe destaque quanto à diversidade de flores e inflorescências, além de sua complexidade evolutiva. Croton L. é o segundo maior gênero da família e exibe particular diversidade floral, em especial quanto a o perianto e número de estames, além das inflorescências, que também se apresentam muito diversas. Considerando a grande variação nas estruturas reprodutivas de Croton, o objetivo desta tese foi estudar as flores e inflorescências com abordagem evolutiva, incluindo morfologia, ontogênese, vascularização, regulação hormonal e expressão gênica. Flores em diversos estágios de desenvolvimento foram analisadas em microscopia der luz e varredura. Inflorescências foram estudadas em estereomicroscópio e os caracteres observados foram analisados nas filogenias mais recentes do grupo. A expressão gênica foi analisada com a técnica RNAseq. No primeiro capítulo as flores apresentaram semelhanças na iniciação das sépalas e presença de filamentos, estruturas petaloides em Croton lundianus (Didr.) Müll. Arg., interpretadas como estaminódios. Em Croton sphaerogynus Baill., estaminódios foram descritos pela primeira vez. Estas estruturas podem ser interpretadas como estruturas de transição evolutiva e reduções florais. No segundo capítulo as flores estaminadas apresentaram androceu polistêmone e o retardo na iniciação das pétalas e o desenvolvimento antessépalo dos nectários foram considerados como fatores chave para o desenvolvimento do androceu como obdiplostêmone. A vascularização corroborou a obdiplostemonia e revelou um estame central com características carpelares em C. fuscescens, interpretado aqui como um caso de homeose. Nectários glandulares foram registrados e interpretados como uma mudança heterotópica. A obdiplostemonia pode estar relacionada com as diferentes concentrações de auxina ao longo das etapas de desenvolvimento e as flores de Croton podem ser consideradas como bons modelos de obdiplostemonia, homeose e heterotopia. No terceiro e quarto capítulo nós investigamos as inflorescências de Croton, que apresentaram 17 padrões com diferenças na organização e distribuição das flores pistiladas especialmente. Os caracteres das inflorescências se mostraram homoplásticos e provavelmente determinados por evolução convergente em linhagens distantes distribuídas em habitats semelhantes. A expressão gênica de C. fuscescens foi particularmente analisada e o transcriptoma demonstrou que o desenvolvimento das diferentes zonas é regulado pelo mesmo conjunto gênico. Cada zona, pistilada ou estaminada, apresenta níveis distintos de expressão diferencial e o gradiente na expressão pode ser o delimitador entre as zonas. Os novos relatos quanto ao desenvolvimento floral em Croton e os links evolutivos identificados nesta tese levanta a importância de estudos para uma melhor compreensão sobre a evolução das estruturas reprodutivas neste grupo tão importante
38

Mecanismos de interação do carrapato Rhipicephalus microplus com o fungo acaropatogênico Metarhizium anisopliae

Araujo, Anelise Webster de Moura Vieira January 2017 (has links)
O carrapato Rhipicephalus microplus é o principal ectoparasita de bovinos. O controle de R. microplus baseia-se principalmente no uso de acaricidas químicos, o que contribuiu para o problema emergente da seleção de populações de carrapatos resistentes. Portanto, há a necessidade do desenvolvimento de métodos mais eficientes/sustentáveis de controle, como o controle biológico utilizando fungos acaropatogênicos. A eficácia do fungo Metarhizium anisopliae de forma isolada ou em associação com acaricida químico para controle do carrapato bovino já foi evidenciada em condições de campo utilizando uma cepa de carrapatos resistente à acaricidas. No entanto, pouco se sabe sobre os mecanismos moleculares de R. microplus envolvidos na sua interação com M. anisopliae e com acaricidas. Assim, o objetivo do presente estudo foi avaliar a resposta de larvas de R. microplus expostas ao fungo, ao acaricida e à associação de ambos. Primeiramente foi realizado um estudo para determinar a metodologia mais indicada para avaliar o efeito in vitro do fungo M. anisopliae sobre larvas de R. microplus. Para isso, comparamos o Teste de Pacote de Larvas modificado (TPL) e o Teste de Imersão de Larvas (TIL). Os valores de tempo letal mediano (TL50) obtido na maior concentração de M. anisopliae (108conídios/mL) foram 24,8 e 9,2 dias para o TPL e TIL, respectivamente. A mortalidade após 21 dias foi de 38% e 98% para o TPL e TIL respectivamente, na mesma concentração. O TIL demonstrou ser o teste mais indicado a ser utilizado, sendo, portanto, escolhido para realização dos experimentos futuros. Em seguida foi realizado um estudo para comparar a suscetibilidade de diferentes isolados de R. microplus ao fungo M. anisopliae. Foram avaliados 67 isolados de campo. Para tanto, as larvas de R. microplus foram imersas em uma suspensão de M. anisopliae (108conídios/mL) durante 5 min. Os tempos letais medianos (TL50) variaram de 2,6 a 24,9 dias A mortalidade observada no 15º dia após o tratamento variou de 26,3 a 100% nas amostras testadas. Esses resultados demonstraram que as populações de campo de R. microplus apresentam uma alta variação em sua suscetibilidade a M. anisopliae. Por fim, foi realizada uma análise transcricional (RNAseq) de larvas de R. microplus expostas a M. anisopliae, cipermetrina, associação de ambos e do controle (não-tratado). A análise dos transcritos dos quatro grupos gerou um total de 507.792 sequências com um tamanho total de 303.160.891 pb. Foram encontrados 31 genes diferencialmente expressos no grupo controle quando comparado com M. anisopliae, 39 com cipermetrina e 73 com M. anisopliae + cipermetrina. M. anisopliae e o grupo da associação apresentaram 81 genes diferencialmente expressos e M. anisopliae e cipermetrina, 46. Houve 177 genes diferencialmente expressos ao comparar M. anisopliae + cipermetrina com larvas expostas à cipermetrina Os resultados deste estudo demonstraram que a associação M. anisopliae + cipermetrina aumentou a expressão de genes relacionados a resposta à agressão, provavelmente pelo fato da associação provocar uma agressão maior ao carrapato e este responder de forma mais rápida, levando à superexpressão de genes de defesa. As descobertas deste estudo fornecem informações sobre as vias e mecanismos moleculares de R. microplus ativados em resposta à interação de M. anisopliae e a acaricida. Além disso, esses estudos estabelecem as bases para pesquisas futuras sobre genes-chave que controlam a suscetibilidade de R. microplus a M. anisopliae. O sinergismo evidenciado em nossos estudos comprova a ideia de que o controle integrado, utilizando o controle biológico com o controle químico é uma opção tanto para o controle de cepas de carrapatos resistentes à acaricidas, quanto para um controle mais rápido de cepas de carrapatos que não apresentam resistência ou que apresentem resistência intermediária. / The tick Rhipicephalus microplus is the major cattle ectoparasite. The control of R. microplus is based mainly on the use of chemical acaricides, which contributed to the emerging problem of the selection of tick populations resistant to acaricides. Therefore, there is a need for the development of more efficient/sustainable methods of control, such as biological control using acari pathogenic fungi. The effectiveness of the fungus Metarhizium anisopliae alone or in association with chemical acaricide to control the bovine tick has already been evidenced under field conditions using an acaricide resistant tick strain. However, to date, little is known about the molecular pathways of R. microplus involved with its interaction with M. anisopliae and with acaricides. Thus, the aim of the present study is to evaluate the response of R. microplus larvae exposed to the fungus, the acaricide and the association of both. Firstly, a study was carried out to determine the most appropriate methodology to evaluate the in vitro effect of the fungus M. anisopliae on larvae of R. microplus tick. For this, we compared the Modified Larval Packet Test (LPT) and the Larval Immersion Test (LIT). The values of lethal time (LT50) obtained in the highest concentration of M. anisopliae (108conidia / mL) were 24.8 and 9.2 days for LPT and LIT, respectively. Mortality after 21 days was 38% and 98% for LPT and LIT, respectively, at the same concentration. The LIT proved to be the most indicated test to be used and was therefore chosen to carry out the following experiments Then, a study was performed to compare the susceptibility of different R. microplus isolates to M. anisopliae fungus. Sixty-seven field isolates were evaluated. For this, R. microplus larvae were immersed in a suspension of M. anisopliae (108conidia/mL) for 5 min. Lethal times (LT50) ranged from 2.6 to 24.9 days. Mortality observed on the 15th day after treatment ranged from 26.3 to 100% in the tested samples. These results demonstrated that R. microplus field populations showed a high variation in their susceptibility to M. anisopliae. Finally, a RNAseq analysis of R. microplus larvae exposed to M. anisopliae, cypermethrin, association of both or untreated (control) was performed. Analysis of the transcripts of the four groups generated a total of 507,792 sequences with a total length of 303,160,891 bp. We found 31 differentially expressed genes in the control group when compared to M. anisopliae, 39 with cypermethrin and 73 with M. anisopliae + cypermethrin. M. anisopliae and the association group had 81 differentially expressed genes and M. anisopliae and cypermethrin, 46 There were 177 differentially expressed genes when comparing M. anisopliae + cypermethrin with cypermethrin exposed larvae. The results of this study demonstrated that the association of M. anisopliae + cypermethrin increased the expression of genes related to response to aggression, possibly because the M. anisopliae + cypermethrin association causes a greater aggression to the tick and, then it could respond quickly, leading to overexpression of defense genes. The findings of this study provide information on the pathways and molecular mechanisms of R. microplus in response to the interaction of M. anisopliae and acaricides. In addition, these studies establish the basis for future research on key genes that control the susceptibility of R. microplus to M. anisopliae. The synergism evidenced in our studies confirms the idea that integrated control using biological control and chemical control is an option both for the control of acaricide resistant tick strains and for a faster control of tick strains that do not show resistance or exhibit intermediate resistance.
39

Mecanismos de interação do carrapato Rhipicephalus microplus com o fungo acaropatogênico Metarhizium anisopliae

Araujo, Anelise Webster de Moura Vieira January 2017 (has links)
O carrapato Rhipicephalus microplus é o principal ectoparasita de bovinos. O controle de R. microplus baseia-se principalmente no uso de acaricidas químicos, o que contribuiu para o problema emergente da seleção de populações de carrapatos resistentes. Portanto, há a necessidade do desenvolvimento de métodos mais eficientes/sustentáveis de controle, como o controle biológico utilizando fungos acaropatogênicos. A eficácia do fungo Metarhizium anisopliae de forma isolada ou em associação com acaricida químico para controle do carrapato bovino já foi evidenciada em condições de campo utilizando uma cepa de carrapatos resistente à acaricidas. No entanto, pouco se sabe sobre os mecanismos moleculares de R. microplus envolvidos na sua interação com M. anisopliae e com acaricidas. Assim, o objetivo do presente estudo foi avaliar a resposta de larvas de R. microplus expostas ao fungo, ao acaricida e à associação de ambos. Primeiramente foi realizado um estudo para determinar a metodologia mais indicada para avaliar o efeito in vitro do fungo M. anisopliae sobre larvas de R. microplus. Para isso, comparamos o Teste de Pacote de Larvas modificado (TPL) e o Teste de Imersão de Larvas (TIL). Os valores de tempo letal mediano (TL50) obtido na maior concentração de M. anisopliae (108conídios/mL) foram 24,8 e 9,2 dias para o TPL e TIL, respectivamente. A mortalidade após 21 dias foi de 38% e 98% para o TPL e TIL respectivamente, na mesma concentração. O TIL demonstrou ser o teste mais indicado a ser utilizado, sendo, portanto, escolhido para realização dos experimentos futuros. Em seguida foi realizado um estudo para comparar a suscetibilidade de diferentes isolados de R. microplus ao fungo M. anisopliae. Foram avaliados 67 isolados de campo. Para tanto, as larvas de R. microplus foram imersas em uma suspensão de M. anisopliae (108conídios/mL) durante 5 min. Os tempos letais medianos (TL50) variaram de 2,6 a 24,9 dias A mortalidade observada no 15º dia após o tratamento variou de 26,3 a 100% nas amostras testadas. Esses resultados demonstraram que as populações de campo de R. microplus apresentam uma alta variação em sua suscetibilidade a M. anisopliae. Por fim, foi realizada uma análise transcricional (RNAseq) de larvas de R. microplus expostas a M. anisopliae, cipermetrina, associação de ambos e do controle (não-tratado). A análise dos transcritos dos quatro grupos gerou um total de 507.792 sequências com um tamanho total de 303.160.891 pb. Foram encontrados 31 genes diferencialmente expressos no grupo controle quando comparado com M. anisopliae, 39 com cipermetrina e 73 com M. anisopliae + cipermetrina. M. anisopliae e o grupo da associação apresentaram 81 genes diferencialmente expressos e M. anisopliae e cipermetrina, 46. Houve 177 genes diferencialmente expressos ao comparar M. anisopliae + cipermetrina com larvas expostas à cipermetrina Os resultados deste estudo demonstraram que a associação M. anisopliae + cipermetrina aumentou a expressão de genes relacionados a resposta à agressão, provavelmente pelo fato da associação provocar uma agressão maior ao carrapato e este responder de forma mais rápida, levando à superexpressão de genes de defesa. As descobertas deste estudo fornecem informações sobre as vias e mecanismos moleculares de R. microplus ativados em resposta à interação de M. anisopliae e a acaricida. Além disso, esses estudos estabelecem as bases para pesquisas futuras sobre genes-chave que controlam a suscetibilidade de R. microplus a M. anisopliae. O sinergismo evidenciado em nossos estudos comprova a ideia de que o controle integrado, utilizando o controle biológico com o controle químico é uma opção tanto para o controle de cepas de carrapatos resistentes à acaricidas, quanto para um controle mais rápido de cepas de carrapatos que não apresentam resistência ou que apresentem resistência intermediária. / The tick Rhipicephalus microplus is the major cattle ectoparasite. The control of R. microplus is based mainly on the use of chemical acaricides, which contributed to the emerging problem of the selection of tick populations resistant to acaricides. Therefore, there is a need for the development of more efficient/sustainable methods of control, such as biological control using acari pathogenic fungi. The effectiveness of the fungus Metarhizium anisopliae alone or in association with chemical acaricide to control the bovine tick has already been evidenced under field conditions using an acaricide resistant tick strain. However, to date, little is known about the molecular pathways of R. microplus involved with its interaction with M. anisopliae and with acaricides. Thus, the aim of the present study is to evaluate the response of R. microplus larvae exposed to the fungus, the acaricide and the association of both. Firstly, a study was carried out to determine the most appropriate methodology to evaluate the in vitro effect of the fungus M. anisopliae on larvae of R. microplus tick. For this, we compared the Modified Larval Packet Test (LPT) and the Larval Immersion Test (LIT). The values of lethal time (LT50) obtained in the highest concentration of M. anisopliae (108conidia / mL) were 24.8 and 9.2 days for LPT and LIT, respectively. Mortality after 21 days was 38% and 98% for LPT and LIT, respectively, at the same concentration. The LIT proved to be the most indicated test to be used and was therefore chosen to carry out the following experiments Then, a study was performed to compare the susceptibility of different R. microplus isolates to M. anisopliae fungus. Sixty-seven field isolates were evaluated. For this, R. microplus larvae were immersed in a suspension of M. anisopliae (108conidia/mL) for 5 min. Lethal times (LT50) ranged from 2.6 to 24.9 days. Mortality observed on the 15th day after treatment ranged from 26.3 to 100% in the tested samples. These results demonstrated that R. microplus field populations showed a high variation in their susceptibility to M. anisopliae. Finally, a RNAseq analysis of R. microplus larvae exposed to M. anisopliae, cypermethrin, association of both or untreated (control) was performed. Analysis of the transcripts of the four groups generated a total of 507,792 sequences with a total length of 303,160,891 bp. We found 31 differentially expressed genes in the control group when compared to M. anisopliae, 39 with cypermethrin and 73 with M. anisopliae + cypermethrin. M. anisopliae and the association group had 81 differentially expressed genes and M. anisopliae and cypermethrin, 46 There were 177 differentially expressed genes when comparing M. anisopliae + cypermethrin with cypermethrin exposed larvae. The results of this study demonstrated that the association of M. anisopliae + cypermethrin increased the expression of genes related to response to aggression, possibly because the M. anisopliae + cypermethrin association causes a greater aggression to the tick and, then it could respond quickly, leading to overexpression of defense genes. The findings of this study provide information on the pathways and molecular mechanisms of R. microplus in response to the interaction of M. anisopliae and acaricides. In addition, these studies establish the basis for future research on key genes that control the susceptibility of R. microplus to M. anisopliae. The synergism evidenced in our studies confirms the idea that integrated control using biological control and chemical control is an option both for the control of acaricide resistant tick strains and for a faster control of tick strains that do not show resistance or exhibit intermediate resistance.
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Adaptation à la pression hydrostatique chez les bactéries sulfato-réductrices piézophiles du genre Desulfovibrio / Adaptation to hydrostatic pressure in two sulfate reducing bacteria of the genus Desulfovibrio

Amrani, Amira 20 October 2015 (has links)
Les bactéries sulfato-réductrices sont largement présentes dans les environnements marins profonds. Ces écosystèmes sont caractérisés par un paramètre physique très important, la pression hydrostatique. Afin de mieux comprendre l'adaptation des bactéries sulfatoréductrices à la pression hydrostatique, la technique RNAseq ainsi que des analyses métaboliques ont été utilisées pour étudier la réponse à différentes pressions de croissance chez deux bactéries piézophiles du genre Desulfovibrio, D. hydrothermalis et D. piezophilus. L’analyse transcriptomique révèle que le métabolisme et le transport des acides aminés, le métabolisme énergétique et la transduction du signal jouent un rôle important dans l'adaptation à la pression hydrostatique chez les souches étudiées. Le dosage du glutamate intracellulaire a montré son accumulation à haute pression hydrostatique, suggérant son rôle de piézolyte chez les espèces du genre Desulfovibrio. Les analyses métaboliques ont montré que la production d'énergie chez ces bactéries est optimisée pour la vie en profondeur et qu’elle est plus efficace à la pression optimale de croissance de chacune des souches. De plus, ces analyses nous ont fourni des informations sur les mécanismes adoptés par la souche D. piezophilus pour modifier son métabolisme énergétique afin de s’adapter à des conditions de haute pression hydrostatique. L’analyse de gènes co-régulés à haute pression chez la souche D. piezophilus nous a permis d'identifier un motif de régulation en amont de gènes essentiellement impliqués dans la production d’énergie et qui serait impliquée dans l'adaptation à des pressions hydrostatiques élevées chez D. piezophilus. / Sulfate-reducing bacteria are widely distributed in deep marine environments These biotopes are characterized by a very important physical parameter, the hydrostatic pressure, which increases by 1 MPa each 100 m depth. To better understand adaptation of sulfate-reducing bacteria to high pressure, RNAseq and metabolic analyses were used to study the response of the piezophilic strains of the genus Desulfovibrio, D. hydrothermalis and D. piezophilus to various hydrostatic pressure growth conditions. The transcriptomic datasets obtained revealed that amino acids metabolisms and transport, energy metabolism and signal transduction are important for adaptation to hydrostatic pressure. Glutamate quantification showed its accumulation at high hydrostatic pressure in both strains, suggesting its role as piezolyte in Desulfovibrio species. Metabolic analyses showed that energy production of those bacteria is optimized for deep-sea life conditions and more efficient at the optimum pressure growth of each strain. Moreover, these analyses provide new insights into mechanism of metabolism pathway modification in response to hight hydrostatic pressure in the strain D. piezophilus. Analysis of hight pressure co-regulated genes in the strain D. piezophilus, allowed us to identify a regulatory motif in the upstream of genes essentially involved in energy production. This motif could be a binding site for a regulatory protein involved in the adaptation to high hydrostatic pressure in D. piezophilus that need to be identified.

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