Els resultats d'aquesta Tesi posen de manifest el paper del cervell en el control del desenvolupament larvari de Sesamia nonagrioides. Així, S. nonagrioides és el primer lepidòpter on s'ha trobat que es pot desenvolupar de larva a adult sense cervell. Les nostres dades mostren que les mudes depenen de l'alliberament d'ecdisteroides per les glàndules protoràciques (GPs) però poden tenir lloc sense la hormona protoracicotròpica (PTTH) del cervell.Mentre que les larves decapitades (larves sense cervell ni corpora allata, (CA)) pupen, les larves de les que s'ha privat del cervell (però s'ha mantingut el CA) sofreixen sovint varies mudes successives; la primera muda pot ser de larva o pupa segons l'edat a la que la larva ha estat privada del cervell i el fotoperíode al qual la larva s'ha desenvolupat. Moltes de les larves L6 privades del cervell un dia després de la muda van mudar a larva independentment de les condicions de fotoperíode durant el desenvolupament, de dia llarg (DL) o curt (DC), mentre que 5 dies després totes les larves desenvolupades en DL van pupar i les de la mateixa edat però desenvolupades en DC van mudar a larva. Els implants de cervell acceleren les mudes a pupa però no alteren les mudes a larva. S. nonagrioides no sembla tenir cap font alternativa als CA de hormona juvenil (HJ). Les larves decapitades no van mostrar quantitats significades de HJ mentre que les larves privades de cervell van mostrar concentracions que es poden detectar fins i tot 10 dies després de la manipulació quirúrgica, però els implants de cervell no activen els CA essent aparentment neural l'activació d'aquests factors. El cervell podria ser el responsable del manteniment de l'estat larvari per inhibició neural de la pupació; quan les larves d'edat variada van ser privades del cervell, la majoria van pupar. De manera similar, el cervell també podria ser el responsable del manteniment de la diapausa per inhibició de la pupació, conseqüentment, quan les larves van ser privades del cervell (tot mantenint o no els CA) les diferencies entre larves dipausants o no diapausants van desaparèixer.El nivell d'ecdisteroides en les larves decapitades augmentà 10 dies després de la manipulació, aproximadament el temps necessari per a pupar en aquelles larves en absència de HJ, encara que l'extracció de les GPs va impedir la muda, i això probà que la presencia de GPs és essencial per al procés de muda. Les GPs en les larves de S. nonagrioides poden funcionar sense estimulació del cervell i la PTTH pot ser alliberada per una font de fora del cap; a S. nonagrioides hem identificat la PTTH mRNA i una font alternativa en l'intestí. La qPCR confirmà que el gen de la PTTH a S. nonagrioides s'expressa molt al cervell de l'instar 6è amb un màxim al dia 5è i un mínim en la prepupa, però el nivell d'expressió de la PTTH es va detectar també en l'intestí de les larves intactes i encara més en el de las decapitades amb una expressió màxima en la prepupa.La més gran part de les larves decapitades muden a pupa sense cap senyal de desenvolupament d'adult mentre que la majoria de pupes provades de cervell sofreixen metamorfosi a adult, lo que suggereix que la transformació de pupa a adult depèn d'un factor desconegut present en les larves privades de cervell però no en les decapitades. En S. nonagrioides la HJ aplicada tòpicament no solament no va inhibir la metamorfosi de pupa a adult però la pogué haver afavorit mentre que l'aplicació d'un agonista d'ecdisteroides a les pupes no va tenir cap efecte en el desenvolupament d'adult.La ingestió per les larves de S. nonagrioides de quantitats subletals de la proteïna Cry1Ab continguda en les fulles de panís o en la dieta va produir un perllongament del seu desenvolupament acompanyat d'un augment en el nombre de mudes abans de pupar però només en les larves criades en condicions de DL, no en les criades en condicions DC. Aquests resultats són deguts a un augment del nivell de HJ en la hemolimfa de les larves no diapausants (DL) alimentades amb fulles de panís Bt o amb la proteïna Bt afegida a la dieta; pel contrari, no es va detectar el possible lleuger increment de HJ causat per la ingestió de la proteïna Bt en les larves diapausants (DC). A més, l'efecte de la proteïna Bt en la concentració d'ecdisteroides en les larves no diapausants va ser suprimir l'augment de la hormona que era necessari per la pupació i per tant va retardar la pupació en les larves tractades. Aquestes respostes poden ser considerades com un mecanisme de defensa que permet que algunes larves puguin mudar i sobreviure a la ingestió de la toxina. / Los resultados de esta Tesis ponen de manifiesto el papel del cerebro en el control del desarrollo larvario de Sesamia nonagrioides. Así, S. nonagrioides es el primer lepidóptero en el que se demuestra que el desarrollo de larva a adulto puede producirse sin la presencia del cerebro. Nuestros datos demuestran que aunque las mudas dependen de la liberación de ecdisteroides por las glándulas protorácicas (GPs), estas pueden ser activadas sin hormona protoracicotrópica (PTTH) del cerebro.Mientras que las larvas decapitadas (larvas sin cerebro ni corpora allata, (CA)) pupan, las larvas descerebradas, de las que se ha extraído el cerebro pero mantienen el CA, sufren a menudo varias mudas sucesivas, la primera a otra larva o pupa según la edad a la que la larva ha sido desprovista del cerebro y según el fotoperiodo bajo el cual la larva se había desarrollado. Muchas larvas desprovistas del cerebro un día después de la muda a sexto estadio mudan a larva independientemente de las condiciones de fotoperiodo, día largo (DL) o corto (DC), recibidas durante su desarrollo, mientras que 5 días después todas las larvas desarrolladas en día largo puparán y las de la misma edad pero desarrolladas en día corto mudaran a larva. Los implantes de cerebro aceleran las mudas a pupa pero no alteran las mudas a larva. S. nonagrioides no parece tener ninguna fuente de hormona juvenil (HJ) alternativa a los CA. Las larvas decapitadas no mostraron cantidades significativas de HJ mientras que las desprovistas de cerebro mostraron cantidades de HJ detectables incluso 10 días después de la manipulación quirúrgica indicando así que los CA siguieron liberando HJ en ausencia del cerebro. Los implantes de cerebro no activaron los CA por lo que su activación es aparentemente neural. El cerebro debe ser el responsable del mantenimiento del estado larvario por inhibición neural de la pupación ya que cuando se extrajo el cerebro a larvas de diferentes edades la mayoría puparon. De manera similar, el cerebro también debe ser el responsable del mantenimiento de la diapausa ya que cuando las larvas fueron desprovistas del cerebro (manteniendo o no los CA) las diferencias entre larvas dipausantes o no diapausantes desaparecieron.El nivel de ecdisteroides en las larvas decapitadas aumentó 10 días después de la extracción del cerebro, este tiempo es aproximadamente el tiempo que las larvas de último estadio larvario necesitan para pupar. La extracción de las GPs impidió la muda, lo que demuestra que su presencia es esencial para que tenga lugar el proceso de la muda. Ante la evidencia de que las GPs de las larvas de S. nonagrioides podían activarse sin la estimulación por hormona PTTH del cerebro se buscaron fuentes alternativas de la hormona y se identificó PTTH mRNA en el intestino de la larva. La qPCR confirmó que el gen de la PTTH en S. nonagrioides se expresa de forma elevada en el cerebro de la larva de 6º estadio con un máximo el 5º día del mismo y un mínimo en la prepupa, pero la expresión de la PTTH se detectó también en el intestino de las larvas intactas y mucho más en las decapitadas con máxima expresión en el periodo de prepupa.De forma general se asume que la metamorfosis pupa-adulto en los insectos se produce en ausencia de HJ y en el caso de S. nonagrioides la mayoría de las larvas decapitadas mudan a pupa sin mostrar posteriormente ningún indicio de desarrollo a adulto mientras que la mayoría de pupas desprovistas de cerebro pero que mantiene su CA sufren metamorfosis a adulto. Este hecho sugirió que la transformación de pupa a adulto en esta especie depende de algún factor presente en las larvas descerebradas pero no en las decapitadas. En S. nonagrioides la HJ aplicada tópicamente no solamente no inhibió la metamorfosis de pupa a adulto sino que la favoreció mientras que la aplicación de un agonista de ecdisteroides a las pupas no tuvo efecto sobre el desarrollo a adulto.La ingestión por las larvas de S. nonagrioides de cantidades subletales de la proteína Cry1Ab contenida en hoja de maíz o en dieta produjo un prolongamiento de su desarrollo acompañado de un aumento en el número de mudas larvarias antes de pupar, pero sólo en las larvas desarrolladas en condiciones de DL, no en las desarrolladas en condiciones de DC. Estos resultados son consecuencia del aumento de HJ en la hemolinfa de las larvas no diapausantes alimentadas con hoja de maíz Bt o con la proteína Bt añadida. Sin embargo, no se detectó el posible ligero aumento de HJ causado por la ingestión de la proteína Bt en las larvas diapausantes (desarrolladas en DC). Otro efecto de la ingestión de la proteína Bt en las larvas no-diapausantes fue suprimir el aumento en la concentración de ecdisteroides necesario para la pupación que por tanto se retrasó en las larvas tratadas. Estas respuestas pueden ser consideradas como un mecanismo de defensa que permite que algunas larvas puedan mudar y así sobrevivir a la ingestión de la toxina. / The results of this Thesis highlight the role of brain in the control of larval development in Sesamia nonagrioides. Thus, S. nonagrioides is the first lepidopteran found to develop from larvae to adult without brain. Our data show that molts depend on the release of ecdysteroids by prothoracic glands (PGs) but they can occur without prothoracicotropic hormone (PTTH) from the brain.While all decapitated larvae (larvae without brain nor corpora allata, (CA)) pupate, the debrained larvae (larvae with no brain but with CA) often undergo several successive molts; first molt could be to larva or pupa, depending on the age at which the larva has been debrained and on the photoperiod under which the larvae have developed: many of the L6 larvae debrained 1 day after molting, molted to larvae independently of the photoperiod conditions of development, long (LD) or short (SD) day, but 5 days later all larvae developed under LD conditions pupated whereas larvae of the same age developd under SD conditions molted to larvae. Brain implants slightly accelerate pupal molts but do not alter the timing of larval molts. S. nonagrioides does not seem to have any alternative source of juvenile hormone (JH) to the CA. Decapitated larvae did not show noticeable amounts of JH while debrained larvae showed detectable concentrations of JH still 10 days after the surgical manipulation but the brain implants do not activate CA, apparently being neural the activation of these factors. The brain might be responsible for larval stage maintenance by neural inhibition of pupation; when the larvae of any age were deprived of their brain, the majority pupated. In the same way, the brain might be also responsible of diapause maintenance by neural inhibition of pupation; consequently, when the larvae were deprived of their brain (maintaining or not their CA) differences between diapausing and non-diapausing larvae disappeared.The level of ecdysteroids in the decapitated larvae increased ten days after manipulation, approximately the time needed to pupate in these larvae in absence of JH, but the removal of PGs prevented molting, proving that the presence of PGs is essential for the molting process. The PGs of S. nonagrioides larvae can function without brain stimulation and PTTH could be released by a source outside the head; in S. nonagrioides we have identified the PTTH mRNA and an alternative PTTH source in the gut. The qPCR confirmed that the PTTH gene of S. nonagrioides is strongly expressed in the brain of the 6th instar with a maximum on day 5 and a minimum in prepupa, but the level of PTTH expression was also detected in the gut of intact and even more in decapitated larvae with a maximum expression in prepupa.Most decapitated larvae molt to pupa with no sign of adult development while the majority of debrained pupae suffer metamorphosis to adult thus suggesting that pupal-adult transformation depends on an unknown factor present in the debrained but not in the decapitated larvae. In S. nonagrioides JH applied topically not only did not inhibit the pupal-adult metamorphosis but could have favored it while the application of an ecdysteroids agonist to the pupae had no effect on the adult development.Ingestion by S. nonagrioides larvae of sub-lethal amounts of Cry1Ab protein contained in maize leaves or the diet produced a prolonged development accompanied by an increase in the number of molts before pupating only in the larvae reared under LD conditions but not in the larvae reared under SD conditions. These results are due to an increase of the level of JH in the hemolymph in the non-diapausing larvae fed with Bt maize leaves or with Bt protein in the diet; on the contrary, in diapausing (SD) larvae the possible low increase of JH due to the Bt toxin ingested was not detected. In addition, the effect of Bt toxin on the ecdysteroids titer in non diapausing larvae was to suppress the increase of the hormone necessary for the pupation of and thus delaying pupation in the treated larvae. These responses may be considered as a defense mechanism allowing some larvae to molt and to survive to the toxin ingestion.

No description available.

Un objectif important de la biologie du développement est de comprendre la base cellulaire de la morphogenèse, notamment le destin des différentes cellules souches dans l'embryon en développement. En décrivant la morphogenèse des espèces représentatives des différents groupes ou embranchements, on fournit une base solide pour comparer les processus similaires dans les différents organismes, et pour tirer des conclusions sur l'évolution des plans d'organisation des animaux. À cette fin, les scientifiques développent des techniques chez des espèces sélectionnées - celles-ci comprennent la manipulation de la fonction des gènes, le marquage et le suivi des populations distinctes de cellules, et l'imagerie in vivo.Dans cette thèse, je présente mes efforts pour améliorer la boîte à outils génomique du coléoptère Tribolium castaneum. J'utilise une technique d'édition du génome récemment découverte, CRISPR/Cas, pour introduire des modifications précises dans le génome de Tribolium, y compris l'introduction de grands fragments par recombinaison homologue. Je montre que l'expression de tous les composants de CRISPR/Cas est induite de manière efficace par des promoteurs endogènes de Tribolium. En me basant sur ces résultats, je développe VALCYRIE, une approche transgénique pourmarquer des clones de cellules uniques dans l'embryon de Tribolium.Ce travail me permet d'enquêter sur le devenir des cellules dans la région terminale postérieure du blastoderme de Tribolium. En utilisant une approche de marquage clonal, je montre que ces cellules donnent naissance à des cellules germinales primordiales et aumésoderme postérieur. Avec la même méthode, je montre que l'intestin postérieur de Tribolium se développe à partir d'une population distincte de cellules au début de la bandelette germinale. En utilisant une technique de microscopie timelapse haute résolution, je décris le sort de cellules individuelles dans le blastoderme de Triboliumet je fais la lumière sur le plan de développement des segments gnathal et thoracique de l'embryon à ce stade. En outre, je montre que l'amnios de Tribolium augmente considérablement au cours du développement précoce. En me basant sur des données d'imagerie, je passe en revue la cartographie du devenir de la bandelette germinale en ce qui concerne l'amnios et l'ectoderme embryonnaire / An important objective of developmental biology is to understand the cellular basis of morphogenesis, including fates of distinct progenitor cells in the developing embryo. Describing morphogenesis in representative species of different groups or phyla provides a solid basis for comparing similar processes in different organisms, and for drawing conclusions about the evolution of animal body plans. To this end, scientists develop techniques in selected species - these include manipulation of gene function, marking and tracking of distinct populations of cells, and in vivo imaging.In this thesis, I present my efforts to enhance the genomic toolkit of the beetle Tribolium castaneum. I use a recently discovered genome editing technique, CRISPR/Cas, to introduce precise alterations in the Tribolium genome, including the introduction of large fragments by homologous recombination. I show that all CRISPR/Cas components are driven efficiently by endogenous Tribolium promoters. Based on these results, I develop VALCYRIE, a transgenic approach to mark single cell clones in developing Tribolium embryos.This work allows me to investigate the fates of the cells in the posterior terminal region of the Tribolium blastoderm. Using a clonal labeling approach, I demonstrate that these cells give rise to primordial germ cells and posterior mesoderm. With the same technique, I demonstrate that the hindgut of Tribolium develops from a distinct cell population in the early germband. Using high-resolution time lapse microscopy, I describe the fates of single cells in the Triboliumblastodermand shed new light on the fatemap of gnathal and thoracic segments of the embryo at this stage. Furthermore, Ishow that the amnion of Tribolium expands greatly during early development. Based on imaging data, I review the fate map of the early germ band with regard to the amnion and the embryonic ectoderm

京都大学 / 0048 / 新制・課程博士 / 博士(農学) / 甲第21140号 / 農博第2266号 / 新制||農||1057(附属図書館) / 学位論文||H30||N5114(農学部図書室) / 京都大学大学院農学研究科森林科学専攻 / (主査)教授 藤井 義久, 教授 吉村 剛, 教授 松浦 健二 / 学位規則第4条第1項該当 / Doctor of Agricultural Science / Kyoto University / DFAM

This thesis comprises three published papers and one manuscript, all focused on the role of juvenile hormone (JH), the JH receptor Methoprene-tolerant (Met) and its target gene Krüppel-homolog 1 (Kr-h1) in insect development and reproduction. The JH-Met-Kr-h1 pathway is critical for metamorphic transition in hemimetabolan Pyrrhocoris apterus (Hemiptera) and holometabolan Bombyx mori (Lepidoptera) but seems to be dispensable during early larval postembryonic development. The results also show that Met and its heterodimeric partner Taiman (Tai) but not Kr-h1 are critical for ovarian development and vitellogenesis in Pyrrhocoris females. In vitro, in vivo and cell-based techniques in Drosophila melanogaster have demonstrated that Met and its paralog Gce are a bona fide receptor for JH. Only Gce capable of binding JH rescued Drosophila deficient for Met and Gce proteins, and the capacity of Gce to bind JH was necessary for JH-dependent transcriptional activation by Gce and Tai.

O presente trabalho foi subdividido em quatro capítulos com os seguintes objetivos: (i) apresentar revisão bibliográfica abordando as principais referências presentes na literatura sobre a cultura da batata, Solanum tuberosum L. (Solanaceae), os aspectos bioecológicos da traça-da- -batatinha, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae), os hormônios no desenvolvimento dos lepidópteros e a atuação de alguns reguladores de crescimento de insetos (RCIs); (ii) avaliar a ingestão dos carboidratos por adultos de P. operculella e sua influência na reprodução da espécie; (iii) testar parâmetros reprodutivos devido à ingestão de alguns ingredientes ativos presentes em RCIs; (iv) avaliar a ingestão de carboidratos aplicados sobre plantas de batata de maneira a ser possível a aplicação de RCIs como método de redução populacional da praga. Inicialmente, para comprovação da ingestão dos carboidratos, foi utilizada a técnica de marcadores coloridos, em que se ofereceu, por capilaridade via rolete dental, solução de mel a 10% contendo corante líquido artificial de coloração azul anis. Os insetos foram confinados em um recipiente cilíndrico construído com tubo de PVC rígido. Após três dias de oferecimento da alimentação, analisou-se a presença do corante no sistema digestivo dos insetos. Para verificação da ingestão de carboidrato em condições mais próximas ao ambiente das culturas agrícolas, foi realizada nova avaliação em recipientes com diferentes dimensões: 0,0018; 0,054 e 9,6 m3. Posteriormente, simulando um borrifamento na cultura, foi realizada uma terceira avaliação no ambiente de 9,6 m3, na qual a solução de mel foi oferecida aos insetos em gotas grandes no limbo foliar de plantas de batata. Para análise da influência dos carboidratos na reprodução da espécie, foram verificados os seguintes parâmetros reprodutivos: ritmo de postura, fecundidade, viabilidade dos ovos e índice de ovigenia. Para análise dos efeitos dos RCIs, foram utilizados quatro compostos com diferentes ingredientes ativos: azadiractina, lufenurom, metoxifenozida e piriproxifem. Estes compostos foram oferecidos aos insetos em solução de mel a 10%, nas concentrações de 5, 10 e 20 ppm. Durante 10 dias após o acasalamento, foram analisados os parâmetros reprodutivos. O corante colorido mostrou-se adequado para os estudos visuais do sistema digestivo de P. operculella. A ingestão dos carboidratos pelos insetos ocorreu somente quando estes foram confinados em ambientes menores, 0,0018 e 0,054 m3. No ambiente maior, de 9,6 m3, a ingestão ocorreu somente quando a solução foi oferecida via gotas em plantas de batata. Quanto aos efeitos dos RCIs, verificou-se diminuição significativa na fecundidade das fêmeas de P. operculella que ingeriram solução de mel contendo azadiractina e lufenurom à concentração de 5 ppm e piriproxifem à concentração de 20 ppm. Quanto aos parâmetros viabilidade dos ovos e período embrionário não foram verificadas diferenças significativas entre os tratamentos. Este fato sugere que a fertilidade não foi alterada, demonstrando que os RCIs não afetam os machos. A comprovação da ingestão dos carboidratos oferecido via gotas no limbo foliar de plantas de batata e os resultados obtidos na redução da fecundidade de P. operculella indicaram que esta técnica pode ser utilizada em programas de manejo integrado da traça-da-batatinha. / The present work has been subdivided into four chapters, with the following objectives: (i) to present a bibliographic review of the main aspects related to the potato crop, Solanum tuberosum L. (Solanaceae), the bioecological aspects of potato tuber moth, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae), the hormones related to lepidopterans development and the role of some insect growth regulators (IGRs); (ii) to evaluate the carbohydrate intake by P. operculella adults and its influence on the species reproduction; (iii) to test the effects of active ingredients of IGRs on reproductive parameters; (iv) to evaluate the ingestion of carbohydrates applied on potato plants, in order to allow IGRs application as a population reduction strategy for this pest. Initially, confirmation of carbohydrate consumption was by a means colored marker technique, which offers a 10% honey solution containing an artificial liquid food dye, blue anise, by capillarity via a dental roll wick. The insects were caged in a cylindrical container made of a rigid PVC tube. After three days of food offering, dye presence was analyzed within the insects digestive system. For confirmation of carbohydrate intake under simulated field conditions, an additional evaluation was conducted in containers with different sizes: 0.0018, 0.054 and 9.6 m3. Later, a third evaluation was carried out in the 9.6 m3 container, in which the honey solution was offered to insects as big drops applied on the leaf blade of potato plants, simulating a spray application. To analyze carbohydrate influence on species reproduction, the following reproductive parameters were assessed: oviposition rate, fertility, egg viability and ovigeny index. Four compounds with different active principles were utilized to analyze IGR effects: azadirachtin, lufenuron, methoxyfenozide and pyriproxyfen. These compounds were offered to the insects in a 10% honey solution at concentrations of 5, 10 and 20 ppm. During 10 days after mating the reproductive parameters were evaluated. The colored dye method proved to be efficient for visual studies of P. operculellas digestive system. Carbohydrate ingestion by the insects occurred only when they were caged in small environments, 0.0018 and 0.054 m3. In larger ones 9.6 m3, feeding occurred when the solution was offered by dripping over potato plants. Results showed a significant reduction of P. operculella fertility among females that ingested the honey solution containing azadirachtin and lufenuron, at 5 and 20 ppm, respectively. Regarding egg viability and embrionary period, no significant differences were found between the treatments. This fact suggests that fertility was not affected, thus demonstrating that IGRs did not affect males. Confirmation of carbohydrate ingestion offered by dripping over potato leaf blades and the results of fertility reduction in P. operculella, indicated that this technique may be utilized in integrated management programs against the potato tuber moth.

O presente trabalho foi subdividido em quatro capítulos com os seguintes objetivos: (i) apresentar revisão bibliográfica abordando as principais referências presentes na literatura sobre a cultura da batata, Solanum tuberosum L. (Solanaceae), os aspectos bioecológicos da traça-da- -batatinha, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae), os hormônios no desenvolvimento dos lepidópteros e a atuação de alguns reguladores de crescimento de insetos (RCIs); (ii) avaliar a ingestão dos carboidratos por adultos de P. operculella e sua influência na reprodução da espécie; (iii) testar parâmetros reprodutivos devido à ingestão de alguns ingredientes ativos presentes em RCIs; (iv) avaliar a ingestão de carboidratos aplicados sobre plantas de batata de maneira a ser possível a aplicação de RCIs como método de redução populacional da praga. Inicialmente, para comprovação da ingestão dos carboidratos, foi utilizada a técnica de marcadores coloridos, em que se ofereceu, por capilaridade via rolete dental, solução de mel a 10% contendo corante líquido artificial de coloração azul anis. Os insetos foram confinados em um recipiente cilíndrico construído com tubo de PVC rígido. Após três dias de oferecimento da alimentação, analisou-se a presença do corante no sistema digestivo dos insetos. Para verificação da ingestão de carboidrato em condições mais próximas ao ambiente das culturas agrícolas, foi realizada nova avaliação em recipientes com diferentes dimensões: 0,0018; 0,054 e 9,6 m3. Posteriormente, simulando um borrifamento na cultura, foi realizada uma terceira avaliação no ambiente de 9,6 m3, na qual a solução de mel foi oferecida aos insetos em gotas grandes no limbo foliar de plantas de batata. Para análise da influência dos carboidratos na reprodução da espécie, foram verificados os seguintes parâmetros reprodutivos: ritmo de postura, fecundidade, viabilidade dos ovos e índice de ovigenia. Para análise dos efeitos dos RCIs, foram utilizados quatro compostos com diferentes ingredientes ativos: azadiractina, lufenurom, metoxifenozida e piriproxifem. Estes compostos foram oferecidos aos insetos em solução de mel a 10%, nas concentrações de 5, 10 e 20 ppm. Durante 10 dias após o acasalamento, foram analisados os parâmetros reprodutivos. O corante colorido mostrou-se adequado para os estudos visuais do sistema digestivo de P. operculella. A ingestão dos carboidratos pelos insetos ocorreu somente quando estes foram confinados em ambientes menores, 0,0018 e 0,054 m3. No ambiente maior, de 9,6 m3, a ingestão ocorreu somente quando a solução foi oferecida via gotas em plantas de batata. Quanto aos efeitos dos RCIs, verificou-se diminuição significativa na fecundidade das fêmeas de P. operculella que ingeriram solução de mel contendo azadiractina e lufenurom à concentração de 5 ppm e piriproxifem à concentração de 20 ppm. Quanto aos parâmetros viabilidade dos ovos e período embrionário não foram verificadas diferenças significativas entre os tratamentos. Este fato sugere que a fertilidade não foi alterada, demonstrando que os RCIs não afetam os machos. A comprovação da ingestão dos carboidratos oferecido via gotas no limbo foliar de plantas de batata e os resultados obtidos na redução da fecundidade de P. operculella indicaram que esta técnica pode ser utilizada em programas de manejo integrado da traça-da-batatinha. / The present work has been subdivided into four chapters, with the following objectives: (i) to present a bibliographic review of the main aspects related to the potato crop, Solanum tuberosum L. (Solanaceae), the bioecological aspects of potato tuber moth, Phthorimaea operculella (Zeller) (Lepidoptera: Gelechiidae), the hormones related to lepidopterans development and the role of some insect growth regulators (IGRs); (ii) to evaluate the carbohydrate intake by P. operculella adults and its influence on the species reproduction; (iii) to test the effects of active ingredients of IGRs on reproductive parameters; (iv) to evaluate the ingestion of carbohydrates applied on potato plants, in order to allow IGRs application as a population reduction strategy for this pest. Initially, confirmation of carbohydrate consumption was by a means colored marker technique, which offers a 10% honey solution containing an artificial liquid food dye, blue anise, by capillarity via a dental roll wick. The insects were caged in a cylindrical container made of a rigid PVC tube. After three days of food offering, dye presence was analyzed within the insects digestive system. For confirmation of carbohydrate intake under simulated field conditions, an additional evaluation was conducted in containers with different sizes: 0.0018, 0.054 and 9.6 m3. Later, a third evaluation was carried out in the 9.6 m3 container, in which the honey solution was offered to insects as big drops applied on the leaf blade of potato plants, simulating a spray application. To analyze carbohydrate influence on species reproduction, the following reproductive parameters were assessed: oviposition rate, fertility, egg viability and ovigeny index. Four compounds with different active principles were utilized to analyze IGR effects: azadirachtin, lufenuron, methoxyfenozide and pyriproxyfen. These compounds were offered to the insects in a 10% honey solution at concentrations of 5, 10 and 20 ppm. During 10 days after mating the reproductive parameters were evaluated. The colored dye method proved to be efficient for visual studies of P. operculellas digestive system. Carbohydrate ingestion by the insects occurred only when they were caged in small environments, 0.0018 and 0.054 m3. In larger ones 9.6 m3, feeding occurred when the solution was offered by dripping over potato plants. Results showed a significant reduction of P. operculella fertility among females that ingested the honey solution containing azadirachtin and lufenuron, at 5 and 20 ppm, respectively. Regarding egg viability and embrionary period, no significant differences were found between the treatments. This fact suggests that fertility was not affected, thus demonstrating that IGRs did not affect males. Confirmation of carbohydrate ingestion offered by dripping over potato leaf blades and the results of fertility reduction in P. operculella, indicated that this technique may be utilized in integrated management programs against the potato tuber moth.

<div>Forensic entomology is a subdiscipline of entomology that involves the use of insect behavior and developmental data to aid in criminal investigations. Genetic data has become increasingly important to the field as there has been a push for DNA-based species identification methods of forensically relevant insects. Genetic data can also elucidate population structure and relatedness of these insects, and such knowledge can contribute to the development of more specific datasets for insects in different regions. The first study presented here investigated the phylogenetics of sister species <i>Lucilia cuprina</i> and <i>Lucilia sericata</i> to identify possible subspecies divisions and issues with DNA-based identifications in the United States. The initial aim of this study was to identify genetic differences between specimens of <i>L. cuprina</i> that preferred live versus carrion flesh. Flies collected from Indiana, USA and South Africa were sequenced and analyzed. Upon sequencing of the genes <i>COI, Period,</i> and <i>28s,</i> our results indicated that <i>L. cuprina</i> from Indiana possess a unique combination of nuclear and mitochondrial haplotypes that suggest a unique lineage, possibly indicating modern hybridization with <i>L. sericata. </i>The inability of both nuclear and mitochondrial genes to distinguish between <i>L. cuprina</i> and <i>L. sericata</i> raises questions about the capabilities of DNA-based species identifications within this genus. Additionally, the inability of these genes to distinguish between specimens that preferred live versus carrion flesh highlights a need for continued research of these behavioral differences. The second study presented here investigated the population structure and relatedness of house flies in the American southwest in relation to a civil lawsuit where neighbors of a poultry farm alleged that flies were emanating from the farm to their homes. <i>Musca domestica</i> (house fly) specimens were collected from the chicken farm and from locations in varying directions and distances from the farm. Amplified fragment length polymorphism (AFLP) analysis was performed and the data were used in a number of analyses. Population reallocation simulations generally indicated that samples from different locations were not genetically different enough from other locations to allocate to their true origin population over others. Kinship analysis showed differences in samples collected in a later season that indicate a genetic bottleneck over time. Population structure analysis indicated the presence of two intermixing genetic populations in the dataset. AMOVA revealed that the majority of genetic variation laid within, rather than among, populations. A Mantel test revealed no significant correlation between genetic and geographic distances. These results indicate that the <i>M. domestica</i> population in this region of southwestern America is large and intermixing, with no clear genetic distinctions between specimens collected at the poultry farm versus the surrounding locations. In regard to the civil lawsuit, it was not possible to conclude that the flies did not emanate from the poultry farm. In a broader perspective, these data can be utilized to develop pest management strategies in this region. Overall, the data from both studies presented here will be useful to forensic investigations, development of more specific and detailed data and identification techniques, and pest control measures.</div>

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