Plusieurs espèces fongiques dites « technologiques » jouent un rôle primordial, via leurs activités métaboliques, dans le développement des caractéristiques organoleptiques et la typicité des fromages. Cependant, malgré une utilisation et une consommation ancestrale, un manque de connaissances persiste que ce soit aux niveaux génétique ou fonctionnel concernant certaines de ces espèces, notamment Penicillium roqueforti. Principalement utilisé dans la production de fromages à pâte persillée, c’est lors de l’affinage que P. roqueforti participe au développement des qualités organoleptiques typiques de ces fromages via, entre autres, ses activités lipolytiques et protéolytiques. Cette espèce est également caractérisée par une diversité morphologique importante, largement révélée au cours de ces travaux, et ayant donné lieu à certaines dénominations technologiques distinctes par le passé conduisant à une interrogation sur la possible existence de plusieurs espèces au sein de l’entité P. roqueforti. Dans ce contexte, une étude phylogénétique basée sur le principe de concordance généalogique (GC-PSR) a été menée sur une vaste collection mondiale de P. roqueforti en utilisant 8 loci. Cette étude a montré l’existence d’une espèce unique. La diversité intraspécifique des isolats de P. roqueforti a ensuite été évaluée en utilisant 4 marqueurs microsatellites qui ont permis de révéler l’existence de 28 haplotypes répartis en 3 populations génétiquement différenciées et corrélées à différents types de fromages à pâte persillée. Sur la base de ces résultats, 55 isolats représentatifs ont par la suite été criblés afin d’évaluer la possible existence d’un caractère haplotype-dépendant concernant les activités protéolytiques, lipolytiques et la production de métabolites secondaires volatils et non-volatils. Cette étude a permis de mettre en évidence une diversité fonctionnelle relativement importante avec, entre autres, l’identification de 52 molécules volatiles, les méthyl-cétones étant majoritaires dans le volatilome des cultures de P. roqueforti, et des profils de production de mycotoxines parfois contrastés. Cependant, une bonne corrélation a été observée entre les groupes d’isolats issus de fromages AOP ou IGP et les métabolites produits. Suite à l’observation de niveaux de productions contrastées d’acide mycophénolique retrouvées chez P. roqueforti, le cluster de gènes impliqué dans sa biosynthèse a été identifié in silico chez P. roqueforti avant que des expériences d'ARN interférence ne soient réalisées sur le gène mpaC codant une polycétide synthase. La fonctionnalité de l’ensemble du cluster a pu être confirmée par différentes approches et des comparaisons de séquences entre différentes souches ayant des productions d’acide mycophénolique contrastées ont permis de mettre en évidence une délétion de 174 pb au niveau du gène mpaC limitant cette production à des niveaux non détectables chez certaines souches. Ce dernier résultat a ainsi apporté un premier élément de réponse quant à la nature a priori souche-dépendante de la production de ce métabolite. / Several « technological » fungal species play an important role in the development of organoleptic properties and typicity of cheeses via their metabolic activities. However, despite an ancestral use and consumption of cheeses, there is still a lack of knowledge about both genetic and functional traits of certain mold species, in particular Penicillium roqueforti. This species, mainly used in blue-veined cheese production, largely contributes to the specific organoleptic properties in the final product via its intense lipolytic and proteolytic activities during ripening. P. roqueforti is characterized by high morphological diversity, largely revealed during this study, and has been associated with different technological names in the past raising the question about several species within P. roqueforti. Consequently, a phylogenetic study based on genealogical concordance principle (GC-PSR) was performed using eight loci on a large worldwide P. roqueforti collection and confirmed the presence of one single species. Taking into account this result, P. roqueforti intraspecific diversity was then assessed using 4 microsatellite markers and 28 haplotypes were identified and distributed in three genetically differentiated populations linked to blue-veined cheese types. Based on these results, 55 representative isolates were further screened to determine potential haplotype-dependent traits related to proteolytic and lipolytic activities and volatile and non-volatile secondary metabolite production. This study highlighted the outstanding functional diversity among strains with, among others, 52 volatile molecules identified, mainly methyl-ketones, and contrasted mycotoxin production profiles. Noteworthy, a clear relationship between PDO/PGI group isolates and produced metabolites was observed. Following the contrasted production of mycophenolic acid among isolates, the mycophenolic acid biosynthetic gene cluster was identified in silico in P. roqueforti and RNA interference experiments targeting the mpaC gene, encoding a polyketide synthase, were performed. The cluster was shown to be fully functional in this species. Moreover, the entire mycophenolic acid cluster was compared between three strains characterized by contrasted mycophenolic acid production and a 174 bp deletion was detected in the mpaC gene of non-producing strains. This last result provided first insight into the a priori strain-dependent character for production.

Introdução - A malária é uma das principais doenças humanas do mundo e afeta principalmente as populações pobres em regiões tropicais e subtropicais onde as condições ambientais são favoráveis tanto para a proliferação dos agentes etiológicos como dos mosquitos vetores. No Brasil, An. darlingi é considerado vetor primário de plasmódios humanos. Devido à importância médica, esse inseto tem sido objeto de campanhas de controle populacional. No entanto, não foi considerada a ocorrência de microevolução em An. darlingi e consequentemente, a possibilidade de populações diferentes apresentarem características biológicas distintas. Objetivo Verificar a presença de variabilidade genética e morfológica em populações de An. darlingi no Brasil. Metodologia - Foram analisados e comparados 26 atributos da morfologia externa de ovos de nove populações, bem como a morfometria geométrica da forma alar de espécimes de 10 localidades diferentes de Anopheles darlingi. Além disso, foram empregadas sequências do gene mitocondrial citocromo oxidase subunidade I (COI) para analisar a estrutura populacional de An. darlingi. Resultados e Conclusão Apesar dos atributos dos ovos apresentarem variação, somente as amostras de Tocantins e Pará foram diferenciadas das demais populações. As variações nas estruturas externas dos ovos são provavelmente adaptativas, com influência de fatores ambientais, como temperatura, umidade e disponibilidade de alimento para as fêmeas. A comparação da morfometria geométrica da asa demonstrou que existe maior similaridade entre as populações da costa (estados do Espírito Santo , do cerrado (estados de Goiás e Tocantins), do norte do rio Amazonas (estados do Amazonas e Amapá), e do sul do rio Amazonas (estados de Mato Grosso e Pará). As análises de COI demonstraram diferenciação das populações da costa da Mata Atlântica em relação às demais populações do Brasil central, Amazônia e Mata Atlântica do Interior. O padrão da diversidade de haplótipos de COI, os testes de neutralidade e a distribuição de diferenças pareadas indicaram recente expansão demográfica. A distância geográfica, as ecorregiões e fatores ambietais podem provavelmente limitar a dispersão da espécie, tendo alguma influência sobre a estrutura populacional de An. darlingi / Introduction Malaria is a major human disease that primarily affects poor persons in tropical and subtropical regions where environmental conditions are favorable to the proliferation of the etiological agent as well as vector mosquitoes. In Brazil, Anopheles darlingi is considered a primary vector of human plasmodia and has been the target of control campaigns because of its medical importance. However, the occurrence of microevolution in An. darlingi was not considered and, consequently, the possibility of different populations exhibit distinct biological characteristics. Objective Verify the presence of morphological and genetic variability in Brazilian Anopheles darlingi populations. Methods Twenty six attributes of external egg morphology were analyzed and compared in nine populations. Similarly, the geometric morphometry of wing shape was evaluated in ten An. darlingi localities. In addition, the mitochondrial gene cytochrome oxidase subunit I (COI) was employed to analyze the population structure of An. darlingi. Results and Conclusion - Although, egg attributes showed variation, only samples from Tocantins and Pará were differentiated from other populations. Variation in external egg structure is arguably adaptive in light of environmental factors such as temperature, humidity and food availability for females. Comparisons of wing geometric morphometry indicated that there is greater similarity between Coastal (Espírito Santo and Rio de Janeiro states) and Inner (São Paulo and Paraná states) populations of the Atlantic Forest, and differentiation of those from (Goiás and Tocantins states) and regions both north (Amazonas and Amapá states) and south (Pará and Mato Grosso states) of the Amazon Region. Cytochrome oxidase I analyses indicated differentiation between coastal Atlantic Forest populations (Espírito Santo and Rio de Janeiro states) and those from the cerrado, the Amazon and the interior Atlantic Forest. The pattern of COI haplotype diversity, neutrality tests and distribution of pairwise differences suggest a recent demographic expansion. Geographical distance, ecoregions and potentially environmental constraints may limit the species´ dispersal and influence the population structure of An. darlingi

Introdução - A malária é uma das principais doenças humanas do mundo e afeta principalmente as populações pobres em regiões tropicais e subtropicais onde as condições ambientais são favoráveis tanto para a proliferação dos agentes etiológicos como dos mosquitos vetores. No Brasil, An. darlingi é considerado vetor primário de plasmódios humanos. Devido à importância médica, esse inseto tem sido objeto de campanhas de controle populacional. No entanto, não foi considerada a ocorrência de microevolução em An. darlingi e consequentemente, a possibilidade de populações diferentes apresentarem características biológicas distintas. Objetivo Verificar a presença de variabilidade genética e morfológica em populações de An. darlingi no Brasil. Metodologia - Foram analisados e comparados 26 atributos da morfologia externa de ovos de nove populações, bem como a morfometria geométrica da forma alar de espécimes de 10 localidades diferentes de Anopheles darlingi. Além disso, foram empregadas sequências do gene mitocondrial citocromo oxidase subunidade I (COI) para analisar a estrutura populacional de An. darlingi. Resultados e Conclusão Apesar dos atributos dos ovos apresentarem variação, somente as amostras de Tocantins e Pará foram diferenciadas das demais populações. As variações nas estruturas externas dos ovos são provavelmente adaptativas, com influência de fatores ambientais, como temperatura, umidade e disponibilidade de alimento para as fêmeas. A comparação da morfometria geométrica da asa demonstrou que existe maior similaridade entre as populações da costa (estados do Espírito Santo , do cerrado (estados de Goiás e Tocantins), do norte do rio Amazonas (estados do Amazonas e Amapá), e do sul do rio Amazonas (estados de Mato Grosso e Pará). As análises de COI demonstraram diferenciação das populações da costa da Mata Atlântica em relação às demais populações do Brasil central, Amazônia e Mata Atlântica do Interior. O padrão da diversidade de haplótipos de COI, os testes de neutralidade e a distribuição de diferenças pareadas indicaram recente expansão demográfica. A distância geográfica, as ecorregiões e fatores ambietais podem provavelmente limitar a dispersão da espécie, tendo alguma influência sobre a estrutura populacional de An. darlingi / Introduction Malaria is a major human disease that primarily affects poor persons in tropical and subtropical regions where environmental conditions are favorable to the proliferation of the etiological agent as well as vector mosquitoes. In Brazil, Anopheles darlingi is considered a primary vector of human plasmodia and has been the target of control campaigns because of its medical importance. However, the occurrence of microevolution in An. darlingi was not considered and, consequently, the possibility of different populations exhibit distinct biological characteristics. Objective Verify the presence of morphological and genetic variability in Brazilian Anopheles darlingi populations. Methods Twenty six attributes of external egg morphology were analyzed and compared in nine populations. Similarly, the geometric morphometry of wing shape was evaluated in ten An. darlingi localities. In addition, the mitochondrial gene cytochrome oxidase subunit I (COI) was employed to analyze the population structure of An. darlingi. Results and Conclusion - Although, egg attributes showed variation, only samples from Tocantins and Pará were differentiated from other populations. Variation in external egg structure is arguably adaptive in light of environmental factors such as temperature, humidity and food availability for females. Comparisons of wing geometric morphometry indicated that there is greater similarity between Coastal (Espírito Santo and Rio de Janeiro states) and Inner (São Paulo and Paraná states) populations of the Atlantic Forest, and differentiation of those from (Goiás and Tocantins states) and regions both north (Amazonas and Amapá states) and south (Pará and Mato Grosso states) of the Amazon Region. Cytochrome oxidase I analyses indicated differentiation between coastal Atlantic Forest populations (Espírito Santo and Rio de Janeiro states) and those from the cerrado, the Amazon and the interior Atlantic Forest. The pattern of COI haplotype diversity, neutrality tests and distribution of pairwise differences suggest a recent demographic expansion. Geographical distance, ecoregions and potentially environmental constraints may limit the species´ dispersal and influence the population structure of An. darlingi

Le charançon de la carotte (Listronotus oregonensis) s’avère un ravageur important des cultures d’apiacées en Amérique du Nord. Une recrudescence des dommages a été observée ces dernières années, et ce, malgré toutes les mesures de contrôle mises en place. Cette étude visait à déterminer la structure génétique des populations du charançon de la carotte en Amérique du Nord et d’évaluer le rôle de la distance géographique et de la plante hôte sur leur niveau de différenciation génétique. La préférence olfactive envers la plante hôte sur laquelle les charançons d’une même population s’y sont développés a aussi été analysée. La caractérisation de la structure génétique de L. oregonensis s’appuyait sur la discrimination des haplotypes (ADNmt COI) et des nucléotides (SNPs) par génotypage-par-séquençage (GBS). Dix-huit populations incluant 220 individus ont été échantillonnées au Québec, en Ontario, en Nouvelle-Écosse (Canada) et en Ohio (États-Unis). L’olfactométrie examinait la réponse olfactive de trois populations du Québec (195 femelles) en fonction de quatre plantes hôtes (carotte, céleri, céleri-rave et persil). Nos résultats ont montré que la distance géographique s’avère un facteur important de différenciation génétique entre les régions échantillonnées. Les analyses de GBS identifient la Nouvelle-Écosse comme étant la région la plus différenciée de toutes les populations analysées contrairement aux analyses de l’ADNmt COI qui suggèrent une différenciation récente. La plante hôte n’entraîne pas de signature génétique distincte chez le ravageur au Québec, et les trois populations testées n’ont pas présenté de préférence marquée pour une plante hôte, hormis la population issue du champ de carotte. Ces résultats suggèrent une capacité de dispersion limitée du charançon de la carotte à travers l’Amérique du Nord. / The carrot weevil (Listronotus oregonensis) is a major pest of apiaceae crops in North America. An upsurge in damage has been observed in recent years, despite all the control measures deployed. This study aimed to determine the genetic structure of carrot weevil populations in North America and to assess the role of geographic distance and host plants on their level of genetic differentiation. The olfactory preference of carrot weevils for the host plant on which the populations were collected was also analyzed. The characterization of the genetic structure of L. oregonensis was based on the discrimination of haplotypes (COI mtDNA) and nucleotides (SNPs) by genotyping-by-sequencing (GBS). Eighteen populations including 220 individuals were sampled in Quebec, Ontario, Nova Scotia (Canada) and Ohio (United States). Olfactometry examined the olfactory response of three populations in Quebec (195 females) according to four host plants (carrot, celery, celeriac and parsley). Our results showed that geographic distance was an important factor in genetic differentiation between the regions sampled. GBS analyzes identify Nova Scotia as the most differentiated region of all populations, unlike COI mtDNA analyzes, which suggest recent differentiation. The host plant did not demonstrate a distinct genetic signature in Quebec, and the three populations tested did not show a marked preference for a host plant, apart from the carrot field population. Finally, these results suggest limited dispersal of the carrot weevil across North America.