Los cohetes de la verbena : novela madrileña /

Mode of access: Internet.

Verbena Camellia Stephanotis, etc.

Verbena Camellia Stephanotis--The doubts of Dives--The demoniac--The doll's house, and after.

Combinación de diferentes estrategias de aplicación de hongos entomopatógenos para el control de Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) y Bemisia tabaci (Gennadius) (Hemiptera:Aleyrodidae)

Los ascomicetos mitospóricos entomopatógenos, que actúan por vía tegumentaria, han sido utilizados para el control microbiano de plagas mediante distintas estrategias que permiten el contacto de sus conidios con los insectos diana. Sin embargo, se conocen nuevos aspectos sobre su función ecológica tales como su capacidad secretora de compuestos con actividad insecticida, así como su sorprendente comportamiento como endófitos, que complementan su empleo clásico y que pueden pantag ermitir el desarrollo de nuevas estrategias de control de plagas. El empleo conjunto de estos hongos y sus extractos para el control de un insecto polífago de importancia mundial Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae) es abordado en el capítulo II, donde se pone de manifiesto la existencia de variación intra- e interespecífica en la virulencia y estrategias patogénicas de las cepas de Beauveria bassiana y Metarhizium brunneum evaluadas frente a larvas del noctuido. La aplicación conjunta de cepas con distintas estrategias patogénicas junto con sus extractos a larvas del lepidóptero tuvo efecto aditivo, y alguno antagónico, de lo que depende su empleo conjunto. Los capítulos III y IV revelan la existencia de colonización endofítica transitoria tras la aplicación foliar de suspensiones de conidios de cepas de las especies mencionadas en alfalfa, tomate y melón, así como el impacto de esta colonización sobre dos fitófagos con diferentes hábitos de alimentación, uno masticador S. littoralis y otro picador-suctor Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). El capítulo III muestra la contribución aditiva de la mortalidad causada por la alimentación de larvas de S. littoralis a expensas de material vegetal colonizado endofíticamente por cepas de B. bassiana y M. anisopliae con la debida al tratamiento tópico de las mismas con las suspensiones fúngicas. El origen de la mortalidad iniciada a través de la vía digestiva en larvas de este lepidóptero permanece incierto para las cepas de B. bassiana, pero podría estar asociado a la presencia en la planta de metabolitos fúngicos en las cepas de M. brunneum como denota la presencia de destruxina (dtx) A en el 11,0% de los cadáveres. El capítulo IV revela que cepas de ambas especies fúngicas pueden iniciar ciclos de infección en insectos picadores-suctores cuando se alimentan a expensas de sustrato vegetal colonizado endofíticamente, si bien, ambas especies presentan estrategias diferentes. Así, B. bassiana muestra una gran capacidad para colonizar el melón, incluso con efecto traslaminar, que causa la infección de ninfas de B. tabaci por contacto con el tegumento. Sin embargo, la mortalidad de las mismas causada por M. brunneum, de crecimiento mucho más localizado en la hoja, está relacionada una vez mas con la presencia de dtx A en el 43,0% de los cadáveres. Estos resultados deben ser considerados para la evaluación del impacto real de los tratamientos con hongos entomopatógenos, y abren nuevas vías en el control de plagas.

Insect eggs suppress plant defence against chewing herbivores.

Plants activate direct and indirect defences in response to insect egg deposition. However, whether eggs can manipulate plant defence is unknown. In Arabidopsis thaliana, oviposition by the butterfly Pieris brassicae triggers cellular and molecular changes that are similar to the changes caused by biotrophic pathogens. In the present study, we found that the plant defence signal salicylic acid (SA) accumulates at the site of oviposition. This is unexpected, as the SA pathway controls defence against fungal and bacterial pathogens and negatively interacts with the jasmonic acid (JA) pathway, which is crucial for the defence against herbivores. Application of P. brassicae or Spodoptera littoralis egg extract onto leaves reduced the induction of insect-responsive genes after challenge with caterpillars, suggesting that egg-derived elicitors suppress plant defence. Consequently, larval growth of the generalist herbivore S. littoralis, but not of the specialist P. brassicae, was significantly higher on plants treated with egg extract than on control plants. In contrast, suppression of gene induction and enhanced S. littoralis performance were not seen in the SA-deficient mutant sid2-1, indicating that it is SA that mediates this phenomenon. These data reveal an intriguing facet of the cross-talk between SA and JA signalling pathways, and suggest that insects have evolved a way to suppress the induction of defence genes by laying eggs that release elicitors. We show here that egg-induced SA accumulation negatively interferes with the JA pathway, and provides an advantage for generalist herbivores.

Diagnosi de l’estat ecològic de les poblacions de nàiades als recs i rieres de l’Estany de Banyoles: propostes de gestió per la seva conservació

Estudi sobre la situació actual de les nàiades dels recs de l’estany de Banyoles, concretament de les espècies unio elongatulus penchinatianus, potomida littoralis subreniformis i anodonta cygnea. Amb les dades recollides es fa una proposta de pla de gestió per a la conservació, protecció i repoblació d’aquestes espècies a la zona

Characterisation of "fou2", a constitutive wound-activated mutant and analysis of the proteome and leaf physiology in the region proximal to wounds in Arabidopsis

Résumé : Les jasmonates (JA), une famille d'hor1none végétale, jouent un rôle central dans la réponse à la blessure, et aux attaques d'insectes et de pathogènes. Les JA sont principalement dérivés d'un acide gras, l'acide linolénique. L'addition par une lipoxygénase d'une molécule d'oxygène à l'acide linolénique initie la synthèse de JA. Cependant les mécanismes régulant l'activation de la biosynthèse de JA ne sont pas encore connus. C'est pour cette raison que dans ce travail, nous avons caractérisé chez Arabidopsis thaliana (l'Arabette des Dames) un mutant fou2 dont l'activité lipoxygénase est plus élevée que celle d'une plante sauvage. Les niveaux de JA sont constitutivement plus élevés et l'activation de la synthèse de JA après blessure est fortement plus induite chez fou2 que chez le type sauvage. En outre, fou2 est plus résistant au pathogène Botrytis cinerea et à la chenille Spodoptera littoralis. Afin de comprendre quel mécanisme chez fou2 génére ce phénotype, nous avons cloné le gène responsable du phénotype de fou2. Le mutant fou2 porte une mutation dans le gène d'un canal à deux pores transportant probablement du potassium, du lumen de la vacuole végétale vers le compartiment cytosolique. L'analyse du protéome de fou2 a permis d'identifier une expression plus élevée de sept protéines régulées par les JA ou le stress. La découverte de l'implication d'un canal dans le phénotype de fou2 renforce l'hypothèse que les flux de cations pourraient être impliqués dans les étapes précoces de la synthèse des JA. Nous avons également étudié le protéome et la physiologie d'une feuille blessée, Pour évaluer les changements d'expression protéique en réponse à la blessure et contrôlés par les JA, nous avons quantifié l'expression de 5937 protéines chez une plante d'Arabidopsis sauvage et chez un mutant incapable de synthétiser des JA. Parmi ces 5937 protéines, nous avons identifié 99 protéines régulées par la blessure chez le type sauvage. Nous avons observé pour 65% des protéines dont l'expression protéique changeait après blessure une bonne corrélation entre la quantité de transcrits et de protéines. Plusieurs enzymes de la voie des chorismates impliquées dans la biosynthèse des acides aminés phénoliques étaient induites par les JA après blessure. Une quantification des acides aminés a montré que les niveaux d'acides aminés phénoliques augmentaient significativement après blessure. La blessure induisait aussi des changements dans l'expression de protéines impliquées dans la réponse au stress et particulièrement au stress oxydatif. Nous avons quantifié l'état réduit et oxydé du glutathion, un tripeptide qui, sous sa forme réduite, est l'antioxydant majeur des cellules. Nous avons trouvé une quantité significativement plus élevée de glutathion oxydé chez le type sauvage blessé que chez la plante aus blessée. Ce résultat suggère que la génération d'un stress oxydatif et la proportion relative de glutathions réduits et oxydés sont contrôlés par les JA après blessure. Abstract : Plants possess a family of potent fatty acid-derived wound-response and developmental regulators: the jasmonates. These compounds are derived from the tri?unsaturated fatty acid a-linolenic-acid (18:3). Addition of an oxygen molecule to 18:3 by 13-lipoxygenases (13-LOX) initiates JA biosynthesis. Actually components regulating the activation of JA biosynthesis are poorly defined. Therefore we characterized in Arabidopsis thaliana the fatty acid Qxygenation upregulated 2 (fou2) mutant, which was previously isolated in a screen for mutants with an enhanced 13-LOX activity. As a consequence of this increased 13-LOX activity, JA levels in fou2 are higher than in wild type (WT) and wounding strongly increased JA biosynthesis compared to WT. fou2 was more resistant to the fungus Botrytis cinerea and the generalist caterpillar Spodaptera littomlis, The fou2 mutant carries a missense mutation in the Two Pore Channel 1 gene (TPCJ), which encodes a vacuolar cation channel transporting probably K* into the cytosol. Patchclamp analysis of fou2 vacuolar membranes showed faster time-dependent conductivity and activation of the mutated channel at lower membrane potentials than wild-type. Proteomic analysis of fou2 leaves identified increased levels of seven biotic stress- and JA- inducible proteins. The discovery of the implication of a channel in the fou2 phenotype strenghtens the hypothesis that cation fluxes might be implicated in early steps of JA synthesis. We further concentrated on the proteome and leaf physiology in the region proximal to wounds in Arabidopsis using the WT and the aos JA-biosynthesis deficient mutant in order to find JA- induced proteins changes. We used two successive proteomic methods to assess protein changes in response to wounding Arabidopsis leaves, two dimensional electrophoresis (2DE) and linear trap quadrupole ion-trap mass spectrometry. In total 5937 proteins were quantified. We identified 99 wound-regulated proteins in the WT. Most these proteins were also wound-regulated at the transcript level showing a good correlation between transcript and protein abundance. We identified several wound-regulated enzymes involved in amino acid biosynthesis and confirmed this result by amino acid quantification. Proteins involved in stress reponses were upregulated, particularly in redox species regulation. We found a significantly higher quantity of oxidized glutathione in wounded WT relative to wounded aos leaves. This result suggests that levels of reduced glutathione are controlled by JA after wounding.

Meliponini neotropicais: o gênero Partamona Schwarz, 1939 (Hymenoptera, Apidae, Apinae) - bionomia e biogeografia

This work, dedicated to the study of nesting habits of the species of the Neotropical genus Partamona Schwarz, is a sequence to the taxonomic revision recently published elsewhere. A total of 214 nests and nest aggregations of 18 species [Partamona epiphytophila Pedro & Camargo, 2003; P. testacea (Klug, 1807); P. mourei Camargo, 1980; P. vicina Camargo, 1980; P. auripennis Pedro & Camargo, 2003; P. combinata Pedro & Camargo, 2003; P. chapadicola Pedro & Camargo, 2003; P. nhambiquara Pedro & Camargo, 2003; P. ferreirai Pedro & Camargo, 2003; P. pearsoni (Schwarz, 1938); P. gregaria Pedro & Camargo, 2003; P. batesi Pedro & Camargo, 2003; P. ailyae Camargo, 1980; P. cupira (Smith, 1863); P. mulata Moure in Camargo, 1980; P. seridoensis Pedro & Camargo, 2003; P. criptica Pedro & Camargo, 2003; P. helleri (Friese, 1900)] were studied , including data about habitat, substrate, structural characteristics, construction materials and behavior. The descriptions of the nests are illustrated with 48 drawings. Partial data of the nests of P. bilineata (Say, 1837), P. xanthogastra Pedro & Camargo, 1997, P. orizabaensis (Strand, 1919), P. peckolti (Friese, 1901), P. aequatoriana Camargo, 1980, P. musarum (Cockerell, 1917) and P. rustica Pedro & Camargo, 2003 are also presented. Nests of P. grandipennis (Schwarz, 1951), P. yungarum Pedro & Camargo, 2003, P. subtilis Pedro & Camargo, 2003, P. vitae Pedro & Camargo, 2003, P. nigrior (Cockerell, 1925), P. sooretamae Pedro & Camargo, 2003 and P. littoralis Pedro & Camargo, 2003 are unknown. The species of Partamona build notable nest entrance structures, with special surfaces for incoming / exiting bees; some of them are extremely well-elaborated and ornamented, serving as flight orientation targets. All species endemic to western Ecuador to Mexico with known nesting habits (P. orizabaensis, P. peckolti, P. xanthogastra, P. bilineata, P. aequatoriana and P. musarum) build their nests in several substrates, non-associated with termitaria, such as cavities and crevices in walls, among roots of epiphytes and in bases of palm leaves, in abandoned bird nests, under bridges, and in other protected places, except P. peckolti that occasionally occupies termite nests. In South America, on the eastern side of the Andes, only P. epiphytophila and P. helleri nest among roots of epiphytes and other substrates, non-associated with termitaria. All other species studied (P. batesi, P. gregaria, P. pearsoni, P. ferreirai, P. chapadicola, P. nhambiquara, P. vicina, P. mourei, P. auripennis, P. combinata, P. cupira, P. mulata, P. ailyae, P. seridoensis, P. criptica and P. rustica) nest inside active termite nests, whether epigeous or arboreous. The only species that builds obligate subterranean nests, associated or not with termite or ant nests (Atta spp.) is P. testacea. Nests of Partamona have one vestibular chamber (autapomorphic for the genus) closely adjacent to the entrance, filled with a labyrinth of anastomosing pillars and connectives, made of earth and resins. One principal chamber exists for food and brood, but in some species one or more additional chambers are filled with food storage pots. In nests of P. vicina, there is one atrium or "false nest", between the vestibule and the brood chamber, which contains involucral sheaths, cells and empty pots. All structures of the nest are supported by permanent pillars made of earth and resins (another autapomorphy of the genus). The characters concerning nesting habits were coded and combined with morphological and biogeographic data, in order to hypothesize the evolutive scenario of the genus using cladistic methodology. The phylogenetic hypothesis presented is the following: (((((P. bilineata (P. grandipennis, P. xanthogastra)) (P. orizabaensis, P. peckolti)) (P. aequatoriana, P. musarum)) P. epiphytophila, P. yungarum, P. subtilis, P. vitae) (((((P. testacea (P. mourei, P. vicina)) (P. nigrior (P. auripennis, P. combinata))) (P. ferreirai (P. pearsoni (P. gregaria (P. batesi (P. chapadicola, P. nhambiquara)))))) ((((P. ailyae, P. sooretamae) P. cupira, P. mulata) P. seridoensis) P. criptica, P. rustica, P. littoralis)) P. helleri))). One area cladogram is presented. Dates of some vicariance / cladogenesis events are suggested. For bilineata / epiphytophila group, which inhabits the Southwestern Amazonia and the Chocó-Mexican biogeographical components, the origin of ancestral species is attributed to the Middle Miocene, when the transgressions of the Maracaibo and Paranense seas isolated the tropical northwestern South America from the eastern continental land mass. The next cladogenic event in the history of the bilineata / epiphytophila group is attributed to the Plio-Pleistocene, when the Ecuadorian Andes reached more than 3000 m, and the ancestral species was fragmented in two populations, one occupying the western Andes (ancestral species of the bilineata subgroup) and other the southwestern Amazon (ancestral species of the epiphytophila subgroup). Other aspects of the history of Partamona are also discussed.

Meliponini neotropicais: o gênero Partamona Schwarz, 1939 (Hymenoptera, Apidae)

Neotropical Meliponini: the genus Partamona Schwarz, 1939 (Hymenoptera, Apidae). The systematics and biogeography of Partamona Schwarz, a Neotropical genus of stingless bees (Meliponini, Apinae, Apidae), are revised. Seventeen new species are described: P. epiphytophila sp. nov., P. subtilis sp. nov., P. nhambiquara sp. nov., P. batesi sp. nov., P. yungarum sp. nov., P. vitae sp. nov., P. ferreirai sp. nov., P. gregaria sp. nov., P. auripennis sp. nov., P. nigrilabris sp. nov., P. combinata sp. nov., P. chapadicola sp. nov., P. seridoensis sp. nov., P. littoralis sp. nov., P. criptica sp. nov., P. rustica sp. nov. and P. sooretamae sp. nov. Partamona pseudomusarum Camargo, 1980, is considered as junior synonym of P. vicina Camargo, 1980. Types of P. grandipennis (Schwarz, 1951), P. xanthogastra Pedro & Camargo, 1996-1997, P. pearsoni (Schwarz, 1938), P. ailyae Camargo, 1980, P. pseudomusarum, P. vicina, P. mulata Moure in Camargo, 1980, P. aequatoriana Camargo, 1980, P. mourei Camargo, 1980, P. peckolti, (Friese, 1901), P. testacea (Klug, 1807), P. helleri (Friese, 1900) and P. musarum (Cockerell, 1917) were examined. Lectotypes of P. orizabaensis (Strand, 1919), and P. cupira (Smith, 1863) are designated. An identification key for the species and drawings of morphological characters are presented. A phylogenetic hypothesis, based mainly on morphological characters is proposed. Four groups are defined, considering the shape of mandible of workers and sternum VII of males: bilineata / epiphytophila group (western Amazon to México), including P. bilineata (Say), P. grandipennis, P. xanthogastra P. orizabaensis P. peckolti P. epiphytophila sp. nov., P. subtilis sp. nov., P. nhambiquara sp. nov., P. batesi sp. nov., P. yungarum sp. nov. and P. vitae sp. nov.; musarum group (Central Brazil, north of South America to Central America), including P. musarum, P. aequatoriana, P. vicina, P. mourei, P. pearsoni, P. ferreirai sp. nov., P. gregaria sp. nov. and P. testacea; nigrior group (Central Brazil to northeast of South America) including P. nigrior (Cockerell, 1925), P. auripennis sp. nov., P. nigrilabris sp. nov., P. combinata sp. nov., P. chapadicola sp. nov., P. seridoensis sp. nov. and P. littoralis sp. nov., and cupira group (southeastern and Central Brazil), including P. cupira, P. mulata, P. ailyae, P. sooretamae sp. nov., P. criptica sp. nov., P. rustica sp. nov. and P. helleri. Some geographic distribution patterns, congruent with that of other Meliponini bees, are commented.

Four 13-lipoxygenases contribute to rapid jasmonate synthesis in wounded Arabidopsis thaliana leaves: a role for lipoxygenase 6 in responses to long-distance wound signals.

Damage-inducible defenses in plants are controlled in part by jasmonates, fatty acid-derived regulators that start to accumulate within 30 s of wounding a leaf. Using liquid chromatography-tandem mass spectrometry, we sought to identify the 13-lipoxygenases (13-LOXs) that initiate wound-induced jasmonate synthesis within a 190-s timeframe in Arabidopsis thaliana in 19 single, double, triple and quadruple mutant combinations derived from the four 13-LOX genes in this plant. All four 13-LOXs were found to contribute to jasmonate synthesis in wounded leaves: among them LOX6 showed a unique behavior. The relative contribution of LOX6 to jasmonate synthesis increased with distance from a leaf tip wound, and LOX6 was the only 13-LOX necessary for the initiation of early jasmonate synthesis in leaves distal to the wounded leaf. Herbivory assays that compared Spodoptera littoralis feeding on the lox2-1 lox3B lox4A lox6A quadruple mutant and the lox2-1 lox3B lox4A triple mutant revealed a role for LOX6 in defense of the shoot apical meristem. Consistent with this, we found that LOX6 promoter activity was strong in the apical region of rosettes. The LOX6 promoter was active in and near developing xylem cells and in expression domains we term subtrichomal mounds.

TRANSCRIPTION FACTORS INVOLVED IN PLANT DEFENCE AGAINST INSECT HERBIVORES IN ARABIDOPSIS THALIANA

Afin de pouvoir se défendre contre les insectes nuisibles, les plantes ont développé plusieurs stratégies leur permettant de maximiser leurs chances de survie et de reproduction. Parmi elles, les plantes sont souvent pourvues de barrières physiques telles que les poils urticants, les épines et la cuticule. En plus, les plantes sont capables de produire des protéines anti-digestives et des métabolites secondaires insecticides tels que la nicotine, les tannins ou les glucosinolates (GS). La mise en place de ces barrières physiques et chimiques comporte un coût énergétique au détriment de la croissance et de la reproduction. Par conséquent, en absence d'insectes, la plante investit la majeure partie de son énergie dans le développement et la croissance. A l'inverse, une blessure causée par un insecte provoquera une croissance ralentie, une augmentation de la densité de poils urticants ainsi que la synthèse de défenses chimiques. Au niveau moléculaire, cette défense inductible est régulée par l'hormone végétale acide jamsonique (AJ). En réponse à l'attaque d'un insecte, la plante produit cette hormone en grande quantité, ce qui se traduira par une forte expression de gènes de défense. Pendant ma thèse, j'ai essayé de découvrir quels étaient les facteurs de transcription (FT) responsables de l'expression des gènes de défense dans Arabidopsis thaliana. J'ai ainsi pu démontrer que des plantes mutées dans les FTs comme MYC2, MYC3, MYC4, ZAT10, ZAT12, AZF2, WRKY18, WRKY40, WRKY6, ANAC019, ANAC55, ERF13 et RRTF1 deviennent plus sensibles aux insects de l'espèce Spodoptera littoralis. Par la suite, j'ai également pu montrer que MYC2, MYC3 et MYC4 sont probablement la cible principale de la voie de signalisation du AJ et qu'ils sont nécessaires pour l'expression de la majorité des gènes de défense dont la plupart sont essentiels à la biosynthèse des GS. Une plante mutée simultanément dans ces trois protéines est par conséquent incapable de synthétiser des GS et devient hypersensible aux insectes. J'ai également pu démontrer que les GS sont uniquement efficaces contre les insectes généralistes tels S. littoralis et Heliothis virescens alors que les insectes spécialisés sur les Brassicaceae comme Pieris brassicae et Plutella xylostella se sont adaptés en développant des mécanismes de détoxification. - In response to herbivore insects, plants have evolved several defence strategies to maximize their survival and reproduction. For example, plants are often endowed with trichomes, spines and a thick cuticule. In addition, plants can produce anti-digestive proteins and toxic secondary metabolites like nicotine, tannins and glucosinolates (GS). These physical and chemical barriers have an energetic cost to the detriment of growth and reproduction. As a consequence, in absence of insects, plants allocate their energy to development and growth. On the contrary, an attack by herbivore insects will affect plant growth, increase trichome density and induce the production of anti-digestive proteins and secondary metabolites. At the molecular level, this inducible defence is regulated by the phytohormone jasmonic acid (JA). Thus, an attack by herbivores will be followed by a burst of JA that will induce the expression of defence genes. The aim of my thesis was to characterize which transcription factors (TF) regulate the expression of these defence genes in Arabidopsis thaliana. I could show that plants mutated in various TFs like MYC2, MYC3, MYC4, ZAT10, ZAT12, AZF2, WRKY18, WRKY40, WRKY6, ANAC019, ANAC55, ERF 13 and RRTFl were more susceptible to the herbivore Spodoptera littoralis. Furthermore, I could demonstrate that MYC2, MYC3 and MYC4 are probably the main target of the JA-signalling pathway and that they are necessary for the insect-mediated induction of most defence genes including genes involved in the biosynthesis of GS. A triple mutant myc2myc3myc4 is depleted of GS and consequently hypersensitive to insects. Moreover, I showed that GS are only efficient against generalist herbivores like S. littoralis and Heliothis virescens whereas specialized insects like Pieris brassicae and Plutella xylostella have evolved detoxification mechanisms against GS.

" Arabidopsis Thaliana " response to insect feeding : new components controlling defense gene expression and plant resistance

Abstract: Plants cannot run away to escape attacking herbivores, but they defend themselves by producing anti-digestive proteins and toxic compounds (for example glucosinolates). The first goal of this thesis was to study changes in gene expression after insect attack using microarrays. The responses of Arabidopsis thaliana to feeding by the specialist Pieris rapae and the generalist Spodoptera liffora is were compared. We found that the transcript profiles after feeding by the two chewing insects were remarkably similar, although the generalist induced a slightly stronger response. The second goal was to evaluate the implication of the four signals jasmonic acid (JA), salicylic acid (SA), ethylene (ET), and abscisic acid (ABA) in the control of insect-regulated gene expression. Using signaling mutants, we observed that JA was the predominant signal and that ABA modulated defense gene expression. In contrast, SA and ET appeared to control slightly gene expression, but only after feeding by S. litforalis. The third goal was to establish whether plant responses are really effective against insects. In accordance with the transcript profile, both insects were affected by the JA-dependent defenses, as they performed better on the JA-insensitive mutant. S. littoralis also performed better on ABA-deficient mutants, providing evidence for the role of ABA in defense against insects. When testing indole or aliphatic glucosinolate deficient mutants, we found that they were also more susceptible to insect feeding, providing some of the first genetic evidence for the defensive role of glucosinolates in planta. Finally, a glutathione-deficient mutant, pad2-1, was also more susceptible to insect feeding and we could attribute this phenotype to a lowered accumulation of the major indole glucosinolate. In this thesis, we provide a comprehensive list of insect-regulated genes, including many transcription factors that constitute interesting candidate genes for the further study of insect-induced expression changes. Understanding how the plant responses to insects are regulated will provide tools for a better management of insect pest in the field. Résumé: Les plantes ne peuvent s'échapper pour fuir les insectes qui les attaquent, mais elles se défendent en produisant des protéines anti-digestives et des composés toxiques (par exemple des glucosinolates). Le premier but de cette thèse était d'étudier les changements de l'expression génétique lors d'attaque par des insectes en utilisant des puces à ADN. Nous avons comparé la réponse d'Arabidopsis thaliana à deux espèces d'insectes avec des habitudes alimentaires différentes : le spécialiste Pieris rapae et le généraliste Spodoptera littoralis. Nous avons trouvé que les profils de transcription après l'attaque par les deux insectes sont remarquablement similaires, bien que le généraliste induise une réponse légèrement plus forte. Le deuxième but était de déterminer l'implication de quatre signaux dans le contrôle de la réponse :l'acide jasmonique (JA), l'acide salicylique (SA), l'éthylène (ET), et l'acide abscissique (ABA). En utilisant de mutants de signalisation, nous avons montré que l'acide jasmonique était le signal prédominant et que l'acide abscissique modulait l'expression génétique. D'autre part, l'acide salicylique et l'éthylène contrôlent à un degré moindre l'expression génétique, mais seulement après l'attaque par S. littoralís. Le troisième but était d'établir si les réponses des plantes sont efficaces contre les insectes. En accord avec le profil de transcription, les deux espèces d'insectes se sont mieux développées sur un mutant insensible au JA, indiquant que les défenses contrôlées par ce signal sont cruciales pour la plante. De plus, les larves de S. littorales se sont mieux développées sur des mutants déficients en ABA, ce qui fournit une preuve du rôle de l'acide abscissique dans la défense contre les insectes. En testant des mutants déficients en glucosinolates de type indole ou aliphatique, nous avons trouvé qu'ils étaient plus sensibles aux insectes, démontrant ainsi le rôle défensif des glucosinolates in planta. Finalement, le mutant déficient en glutathion pad2-1 était aussi plus sensible à l'attaque des insectes, et nous avons pu attribuer ce phénotype à une plus faible augmentation d'un indole glucosinolate dans ce mutant. Dans cette thèse, nous avons mis en évidence un nombre important de gènes contrôlés par les insectes, comprenant de nombreux facteurs de transcription qui constituent des candidats intéressants pour`étudier plus en détail les changements d'expression génétique induits par les insectes. Une meilleure compréhension de la réponse des plantes contre l'attaque des insectes devrait nous permettre de développer de nouvelles stratégies pour mieux gérer les ravageurs des cultures.

A conserved transcript pattern in response to a specialist and a generalist herbivore.

Transcript patterns elicited in response to attack reveal, at the molecular level, how plants respond to aggressors. These patterns are fashioned both by inflicted physical damage as well as by biological components displayed or released by the attacker. Different types of attacking organisms might therefore be expected to elicit different transcription programs in the host. Using a large-scale DNA microarray, we characterized gene expression in damaged as well as in distal Arabidopsis thaliana leaves in response to the specialist insect, Pieris rapae. More than 100 insect-responsive genes potentially involved in defense were identified, including genes involved in pathogenesis, indole glucosinolate metabolism, detoxification and cell survival, and signal transduction. Of these 114 genes, 111 were induced in Pieris feeding, and only three were repressed. Expression patterns in distal leaves were markedly similar to those of local leaves. Analysis of wild-type and jasmonate mutant plants, coupled with jasmonate treatment, showed that between 67 and 84% of Pieris-regulated gene expression was controlled, totally or in part, by the jasmonate pathway. This was correlated with increased larval performance on the coronatine insensitive1 glabrous1 (coi1-1 gl1) mutant. Independent mutations in COI1 and GL1 led to a faster larval weight gain, but the gl1 mutation had relatively little effect on the expression of the insect-responsive genes examined. Finally, we compared transcript patterns in Arabidopis in response to larvae of the specialist P. rapae and to a generalist insect, Spodoptera littoralis. Surprisingly, given the complex nature of insect salivary components and reported differences between species, almost identical transcript profiles were observed. This study also provides a robustly characterized gene set for the further investigation of plant-insect interaction.

Identity and combinations of arbuscular mycorrhizal fungal isolates influence plant resistance and insect preference

1. Accumulating evidence indicates that plant resistance against above-ground herbivores can be affected by the presence of arbuscular mycorrhizal fungi (AMF) in association with the host plant. Little is known, however, about how AMF composition can influence herbivore choice to feed on a particular plant. 2. Unravelling the preference-performance hypothesis in a multitrophic context is needed to expand our knowledge of complex multitrophic interactions in natural systems. If given mycorrhizal fungal genotypes increase attractiveness for a herbivore (reduced plant resistance), then the benefits of increased unpalatability provided by the mycorrhizal fungi (increased plant resistance) might be outweighed by the increased herbivore recruitment. 3. This was addressed by designing three experiments to test the effects of different AMF genotypes, inoculated either alone or in combination, to measure intraspecific AMF effects on plant resistance and insect herbivore preference. Using strawberry (Fragaria vesca L.) plants that were colonised by eight different combinations of Rhizophagus irregularis isolates, we measured effects on plant growth, insect growth and survival, as well as feeding preferences of a generalist herbivore caterpillar (Spodoptera littoralis Boisduval). 4. Overall, it was found that: (i) AMF influenced plant resistance in an AMF genotype-specific manner; (ii) some AMF inoculations decreased insect performance; (iii) insects preferentially chose to feed more on leaves originating from non-mycorrhizal plants; but also that (iv) in a whole plant bioassay, insects preferentially chose the biggest plant, regardless of their mycorrhizal status. 5. Therefore, AMF-mediated trade-offs between growth and resistance against herbivores have been shown. Such trade-offs, particularly driven by plant attractiveness to herbivores, buffer the positive effects of the mycorrhizal symbiosis on enhanced plant growth.

Signaling pathways controlling induced resistance to insect herbivores in Arabidopsis.

Insect attack triggers changes in transcript level in plants that are mediated predominantly by jasmonic acid (JA). The implication of ethylene (ET), salicylic acid (SA), and other signals in this response is less understood and was monitored with a microarray containing insect- and defense-regulated genes. Arabidopsis thaliana mutants coi1-1, ein2-1, and sid2-1 impaired in JA, ET, and SA signaling pathways were challenged with the specialist small cabbage white (Pieris rapae) and the generalist Egyptian cotton worm (Spodoptera littoralis). JA was shown to be a major signal controlling the upregulation of defense genes in response to either insect but was found to suppress changes in transcript level only in response to P. rapae. Larval growth was affected by the JA-dependent defenses, but S. littoralis gained much more weight on coi1-1 than P. rapae. ET and SA mutants had an altered transcript profile after S. littoralis herbivory but not after P. rapae herbivory. In contrast, both insects yielded similar transcript signatures in the abscisic acid (ABA)-biosynthetic mutants aba2-1 and aba3-1, and ABA controlled transcript levels both negatively and positively in insect-attacked plants. In accordance with the transcript signature, S. littoralis larvae performed better on aba2-1 mutants. This study reveals a new role for ABA in defense against insects in Arabidopsis and identifies some components important for plant resistance to herbivory.