Дисертації з теми "Defense stimulator"

La protection intégrée, qui vise à réduire l’usage des pesticides, est un défi majeur pour l’agriculture du XXIème siècle. Le développement de nouvelles approches agronomiques qui concilient environnement et agriculture est une condition indispensable pour l’agriculture de demain. Dans ce contexte, l’utilisation d’éliciteurs capables de mimer une attaque pathogène et de promouvoir un état de résistance chez les plantes face à des maladies représente une alternative naturelle à la lutte chimique. Ces éliciteurs sont nommés les stimulateurs de défense des plantes (SDP). Ils peuvent provenir de différentes sources et être extraits à partir de macroalgues comme c’est le cas des SDP à base de polysaccharides d’algues tels que la laminarine utilisée pour stimuler l’immunité de plantes agronomiques. Toutefois, l’exploitation de ces ressources dans leur milieu naturel et les difficultés de production liées à leur cycle de développement constituent des freins à leur utilisation. La valorisation des microalgues comme source de SDP pourrait permettre de s’affranchir de ces contraintes. Cependant la recherche et de molécules SDP chez les microalgues est encore peu abordée. Au cours de ce travail, le potentiel d’une culture de microalgue, Phaeodactylum tricornutum, à induire des réactions de défense chez les plantes a été évalué. Un broyat cellulaire a été appliqué sur des plantules d’Arabidopsis thaliana. Le caractère éliciteur de ce broyat a été testé et caractérisé par des approches microscopiques, physiologiques et moléculaires. Les résultats ont montré que les plantes traitées présentaient des niveaux d’expression des gènes PR-1, PAD3, ACS6 et WRKY40 et un niveau de protection contre la bactérie Pseudomonas syringae DC3000 (Pst) plus élevés que les plantes non traitées. De plus, un effet bactéricide in vitro sur la bactérie Pst a été observé. Ces résultats offrent de nouvelles perspectives pour le développement de produits SDP d’origine naturelle capables de protéger les cultures
Integrated plant protection, which aims to reduce the use of pesticide, is a major challenge for the agriculture of the 21st century. The development and application of new agronomic approaches is a prerequisite for crop protection in a sustainable agriculture system. In this context, the use of elicitors capable of mimicking a pathogenic attack and promoting a plant resistance state against diseases is a natural alternative to the use of agro-chemicals. These elicitors are also called plant defense stimulators (PDS). These can be obtained from different sources including macroalgae as it the case for the polysaccharide-based PDS laminarin that is currently used for the protection of a number of crops. However, the exploitation of these natural resources and the difficulties of their production due to their development cycle do hamper their use at a large scale. One of the possibilities to overcome these difficulties is the use of microalgae as a source of PDS. But this possibility and the potential of microalgaederived PDS for crop protection are currently under investigated. In the present work, we have used a cell extract from the microalgae Phaeodactylum tricornutum and assessed its defense response-eliciting activities on Arabidopsis thaliana seedlings by using microscopic, physiological and molecular approaches. The results show that treated plants exhibit higher levels of expression of the PR-1, PAD3, ACS6 and WRKY40 genes and a higher level of protection against the pathogenic bacterium Pseudomonas syringae DC3000 (Pst) than nontreated plants. An In vitro antibacterial activity on the Pst bacteria was also observed. Our findings suggest that P. tricornutum cell extracts are able to activate plant immune responses and offer new perspectives for the development of novel plant defense stimulators

Les co-produits industriels de la première étape d’extraction d’agar (de l’algue rouge Gelidium sesquipedale) constituent au sein de l’entreprise SETEXAM des volumes importants potentiellement valorisables. L’objectif de ce travail est de démontrer qu’ils peuvent être utilisés en tant qu‘éliciteur, ou stimulateur des défenses naturelles (SDN). D’abord, la composition élémentaire du co-produit alcalin a révélé une teneur en minéraux de 44 % (Na, K éléments majeurs) avec des traces de métaux lourds, mais en dessous de la limite autorisée. Les principaux composés organiques détectés sont des glucides (12,5 %) et une fraction riche en floridoside a été obtenue. Ce composé a été identifié par une méthode originale, la CPG-MS. Le co-produit alcalin, recyclé au cours du procédé industriel, voit sa teneur en glucides diminuer avec l’augmentation du nombre de cycles. Ces co-produits, appliqués sur des plants de tomates cultivées en serre sous stress biotique, sont capables de stimuler des réponses de défense (activités enzymatiques, expression des gènes). La dose optimale (50 mg.L-1) a été déterminée ainsi que les conditions d’applications et la durée d’activation des réponses. En plein champ, les co-produits ont été testés sur vigne et sur tomate, pour leur capacité à réduire les symptômes du mildiou, ou de la cladosporiose. Des résultats prometteurs ont été obtenus comparés à des éliciteurs déjà commercialisés, en conditions de pression modérée. Le co-produit issu de la première étape d’extraction industrielle d’agar a montré une activité comme SDN et peut constituer une solution de protection des cultures permettant une réduction des pesticides pour une agriculture durable et plus respectueuse de l’environnement. Ce travail constitue une base pour un dossier d’homologation du produit qui transformerait ces co-produits de coût en ressource pour l’entreprise
The industrial by-products of the first step of agar extraction (from red alga Gelidium sesquipedale) constitute large volumes at SETEXAM company that might be valorized. The objective of this work is to prove that they can be used as elicitor, or plant defense stimulator (PDS). Firstly, the elemental composition of the alkaline by-product revealed a mineral content of 44% (Na, K major components) with heavy metal traces, but under authorized limits. The principal organic components detected are carbohydrates (12.5%) and a fraction rich in floridoside was obtained. This molecule was identified with an original method, through GC-MS. The alkaline by-product, recycled through the industrial process, has its carbohydrate content diminished as the number of cycles increases. These by-products, applied on tomato plants grown in greenhouse under biotic stress, are capable of stimulating defense responses (enzymatic activities, gene expression). The optimal dose (50 mg.L-1) was determined together with the application conditions and time span of responses. In field, the by-products were tested on grapevine and on tomato, for their capacity to reduce downy mildew, or leaf mold symptoms. Promising results were obtained compared to already commercialized elicitors, for moderate disease pressure. The by-product obtained from the first step of industrial agar extraction showed a PDS activity and can be a solution for culture defense allowing a reduction of pesticides for a durable and environmentally friendly agriculture. This work is a base for a homologation file of the product that would transform these by-products from cost to resource for the company

Le lin, principale plante à fibres cultivée en France possède un intérêt industriel pour la qualité de ses fibres. Les cultures de lin sont régulièrement attaquées par un champignon tellurique, Fusarium oxysporum f. sp. lini (Fol), responsable des plus grandes pertes dans les cultures de lin. Les PGPR (Plant Growth Promoting Rhizobacteria) sont des bactéries réputées pour leurs capacités à améliorer la croissance et le développement des plantes, mais également pour leur pouvoir compétiteur au sein de la rhizosphère et leur aptitude à induire une réponse immunitaire chez les plantes. Parallèlement, l’utilisation de SDP (Stimulateur des Défenses des Plantes), molécules capables d’éliciter les mécanismes de défenses des plantes est une autre alternative pour limiter l’utilisation des pesticides. Dans le cadre de ce projet mené sur deux variétés de lin, Aramis et Mélina, il a pu être montré que Fol induisait un remodelage de la paroi au niveau des racines et des tiges, impliquant les hémicelluloses et les pectines, seulement deux jours après inoculation avec le champignon. L’utilisation de la souche ATCC 6633 de Bacillus subtilis comme agent de biocontrôle a permis de réduire de façon significative l’apparition des symptômes de la fusariose. Il a également été montré qu’en plus d’avoir un effet fongicide sur Fol, cette bactérie est capable d’induire l’expression de deux gènes de défense (Pathogenesis-Related) codant pour une β-(1,3)-glucanase (PR-2) et codant pour une chitinase-like (CTL-10), de gènes impliqués dans la voie des phénylpropanoïdes (PHENYLALANINE AMONIA LYASES, PAL-3 et PAL-4) et dans le remodelage pariétal (PECTIN METHYLESTERASE-3, PME-3) au niveau racinaire. Des analyses biochimiques ont également permis de montrer que B. subtilis provoque des modifications se traduisant par un renforcement pariétal au niveau des tiges chez les deux variétés. Enfin, l’association de la PGPR avec une molécule élicitrice (pregnénolone sulfate) a eu un effet synergique sur l’expression de gènes de défense
In France, flax (Linum usitatissumum) is a principal fibers crop. Fusarium oxysporum f sp lini (Fol), a soil-borne fungus, is responsible for the major losses in crop yield. PGPR (Plant Growth Promoting Rhizobacteria) are known for their abilities to promote plant growth and health. These bacteria are also good competitors in the rhizosphere and can induce a plant defense response. The use of compounds able to elicit plant defense mechanisms is also an alternative to limit the use of pesticides. In this project, it has been shown that F. oxysporum f. sp. lini induces only two days after inoculation cell wall remodeling in the root and the stem involving hemicelluloses and pectins on two flax varieties, Aramis and Mélina, . The use of the Bacillus subtilis strain ATCC 6633 as biocontrol agent significantly reduced fusarium wilt appearance. In addition to its antifungal effect against Fol, this bacteria is able to induce the expression of two Pathogenesis-Related genes coding for a β-(1,3)-glucanase (PR-2) and a chitinase-like (CTL-10), genes involved in the phenylpropanoid pathway (PHENYLALANINE AMONIA LYASES, PAL-3 and PAL-4) and also in cell wall remodeling (PECTIN METHYLESTERASE-3, PME-3) in the root. Biochemical analyses show that B. subtilis causes modifications resulting in cell wall reinforcement in the stem in both varieties. Finally, the association of B. subtilis with an elicitor (pregnenolone sulfate) had a synergistic effect on the expression of defense-related genes

O congelamento a um contexto previamente associado ao choque nas patas é atenuado pela lesão da matéria cinzenta periaquedutal ventrolateral (MCPvl). Por outro lado, a estimulação elétrica ou microinjeção de compostos que diminuem a neurotransmissão GABAérgica na matéria cinzenta dorsolateral (MCPdl) provocam congelamento e fuga. O presente estudo examinou a possibilidade deste congelamento provocado pela estimulação da MCPdl ser resultado da ativação indireta da MCPvl. Ratos com lesão da MCPvl ou falsooperados foram eletricamente estimulados na MCPdl para a aferição dos limiares de resposta para o congelamento e a fuga. Os mesmos animais foram também submetidos ao condicionamento aversivo ao contexto através de choque nas patas, visando a validação de nossas condições experimentais. Um segundo grupo de ratos lesados e falso-operados receberam microinjeções de semicarbazida, um bloqueador da síntese do GABA, na MCPdl. Os resultados mostram que a lesão da MCPvl atenua o congelamento condicionado ao contexto, mas é ineficaz em alterar os efeitos da estimulação elétrica ou química da MCPdl sobre o congelamento e a fuga. A MCPvl é o alvo preferencial do núcleo central da amígdala na MCP, enquanto que a MCPdl recebe aferências principalmente dos núcleos do hipotálamo relacionados à defesa. Estas evidências são coerentes com a participação da MCPvl nas respostas de defesa ao perigo potencial, e da MCPdl ao perigo imediato.
Freezing to a context previously associated to footshock is attenuated by ventrolateral periaqueductal gray (vlPAG) lesion. Moreover, electrical stimulation or microinfusion of compounds that interfere with GABA neurotransmission in the dorsolateral periaqueductal gray (dlPAG) provoke freezing and escape. The present study examined the possibility of this freezing being the result of an indirect activation of vlPAG through dlPAG stimulation. Rats bearing vlPAG or sham lesions were electrically stimulated at dlPAG sites to have their freezing and escape threshold currents measured. The same animals were also submitted to a contextual fear-conditioning paradigm through footshock to validate our experimental setting. A second group of vlPAG- and sham-lesion rats received infusions of semicarbazide, a GABA-synthesis blocker, in the MCPdl. The results obtained show that vlPAG lesions do attenuate conditioned freezing, but are ineffective against dlPAG-stimulation freezing and escape. The vlPAG is the main PAG target to central nucleus of amygdala projections, while the dlPAG receives afferents primarily from hypothalamic nuclei related to defense. This evidence is coherent with vlPAG mediating responses to potential danger, while dlPAG would be more related to immediate danger.

Mestrado em Biomedicina Molecular
The present paper presents a review and compilation of all the scientifically relevant bibliography to date, regarding the antiviral signalling pathways implicated in the cellular innate immune system in humans. Emphasizing the mitochondrial antiviral signalling adaptor (MAVS), this paper explores the special features of the signal transduction pathways and their components in two specific organelles: mitochondria and peroxisomes. These pathways, ultimately, result in the expression of interferon-stimulated genes (ISGs), which are primarily responsible for fighting against viral replication, viral particle assembly and virion release within the cell. In this paper, several proposals for further investigation are also presented, since there is still a lot to learn about the role of peroxisomes in the antiviral innate immune responses.
O presente trabalho propõe-se a rever e compilar toda a bibliografia cientificamente relevante até à data, no que respeita as vias de sinalização antivirais implicadas na imunidade celular inata em células humanas. Com ênfase na proteína adaptadora MAVS, este trabalho explora as particularidades das vias de transdução de sinal e respetivos intervenientes em dois organelos celulares específicos: mitocôndrias e peroxissomas. Estas vias, em última instância, resultam na expressão de genes estimulados por interferões (ISGs), principais responsáveis pelo combate celular eficaz contra a replicação viral, montagem de partículas virais e libertação de viriões na célula infetada. Neste trabalho são ainda apresentadas propostas para investigações futuras, uma vez que ainda muito pouco se sabe sobre o papel dos peroxissomas nas respostas imunitárias inatas contra infeções virais.

La pourriture grise causée par Botrytis cinerea est l'une des principales maladies de la vigne. La principale solution pour faire face à cette maladie est l'utilisation de produits chimiques, mais la lutte chimique cause des dommages environnementaux et conduit au développement de souches de B. cinerea résistantes. Une stratégie alternative consiste à stimuler les mécanismes de défense naturelle de la vigne. Cependant, B. cinerea est connu pour contourner les défenses de la plante (El Oirdi et Bouarab, 2011) et à ce jour aucun éliciteur n'a montré d'effet protecteur probant contre B. cinerea. De plus, les résultats prometteurs observés au laboratoire se révèlent souvent décevants au vignoble et à ce jour aucun produit stimulateur de défense ne confère à la vigne une protection acceptable contre la pourriture grise. Pour développer des éliciteurs efficaces contre la pourriture grise, il s'avère donc essentiel de caractériser des marqueurs de protection qui permettraient de distinguer stimulation des défenses et protection efficace de la vigne contre B. cinerea. Pour ce faire des analyses comparatives ont été réalisées par 2D-PAGE afin de comparer au niveau protéique l'effet d'éliciteurs induisant une protection et l'effet d'éliciteur n'induisant pas une protection. Les recherches se sont concentrées sur l'apoplaste, qui est un réseau continu dans les plantes créant une interface avec l'environnement. Après la cuticule, l'apoplaste est ainsi la première barrière contre les attaques d'agents pathogènes (Agrawal et al., 2010). Afin d'obtenir un aperçu du protéome constitutif de l'apoplaste (secrétome), une méthode d'infiltration-centrifugation a été optimisée afin de recueillir le fluide apoplastique à partir de feuilles de vigne. Les profils protéiques apoplastiques ont ensuite été comparés aux profils protéiques de feuilles entières par 2D-PAGE et les protéines identifiées par spectrométrie de masse. Cette approche nous a permis d'établir une carte protéique des feuilles entières et du secrétome de la vigne. A notre connaissance, cette étude fournit la première carte protéique du secrétome de la vigne. Cette étude nous donne ainsi un aperçu des protéines les plus abondantes de l'apoplaste de feuilles de vigne. Une prédiction de la fonction des protéines nous a permis de conclure que l'apoplaste de vigne contient principalement (i) des protéines liées aux stress, (ii) des protéines impliquées dans le métabolisme de la paroi cellulaire et (iii) des protéases. Pour confirmer la qualité des extractions, l'analyse prédictionnelle de la sécrétion des protéines a révélé une forte proportion de protéines secrétées de manière classique et non classique (Leaderless Secreted Proteins, LSP). Cette approche offre un grand nombre de protéines candidates impliquées dans les diverses fonctions physiologiques de l'apoplaste. (…)Parallèlement cette étude présente les travaux préliminaires entrepris, également par 2D-PAGE, pour (i) mettre en évidence des marqueurs de potentialisation chez la vigne, (ii) mettre en évidence les bouleversements, au niveau protéique, causés par une infection de B. cinerea sur la vigne et (iii) comprendre comment un traitement avec un éliciteur protecteur pourrait limiter ces effets néfastes. La validation de ces marqueurs par différentes approches permettra d'améliorer la compréhension des mécanismes de défense des plantes et le développement d'outils pour le criblage de nouveaux éliciteurs candidats pour leurs effets protecteurs contre la pourriture grise et pour le suivi des défenses de la vigne directement au vignoble
Grey mould caused by Botrytis cinerea infection is one of the main diseases affecting grapevine. The main solution to cope with this disease is the use of chemicals, but chemical control cause environmental damages and lead to the development of B. cinerea resistant strains. An alternative strategy to prevent diseases consists in stimulating plant defense mechanisms. Nevertheless B. cinerea is known to manipulate plant defenses (El Oirdi and Bouarab, 2011) and to date no elicitors have shown expected protective effect against B. cinerea even if they stimulate defense markers. Moreover despite that numerous studies showed an excellent efficacy of elicitors in laboratory conditions, in vineyard the obtained results are often disappointing.Thus it is necessary to distinguished elicitors inducing protection (protective elicitor) from elicitors inducing defense but no protection (non protective elicitors). For that it appears crucial to characterize markers which would enable to discriminate grapevine defense stimulation from effective protection against B. cinerea. To reach this goal, comparative analyses were performed by 2D-PAGE in order to compare the effect of protective elicitors from non protective elicitors against B. cinerea on grapevine apoplastic fluids. Those biomarkers could be defined as protection biomarkers.Researches were focused on the apoplast, which is a continuous network in plants and creates an interface with the environment. After the cuticle, apoplast is the first barrier against pathogen attacks (Agrawal et al., 2010). In order to obtain an overview of the constitutive apoplastic proteome (secretome), a vacuum-infiltration-centrifugation method was optimized to collect the apoplastic fluid from grapevine leaves. Apoplastic protein profiles were compared to whole-leaf protein profiles by 2D-PAGE and protein identifications were performed by tandem mass spectrometry. This approach allowed us to establish a well-defined proteomic map of whole-leaf and apoplastic leaf. To our knowledge, it is the first time that the apoplastic fluid is recovered from grapevine to characterize its protein content. This study provides a comprehensive overview of the most abundant proteins present in grapevine apoplast. Protein function prediction allowed us to conclude that the grapevine apoplast mainly contains a high proportion of (i) stress-related proteins, (ii) proteins involved in cell wall metabolism, (iii) proteases. To confirm quality of extractions, proteins secretion prediction tools revealed a high proportion of classical and non-classical secreted proteins namely Leaderless Secreted Proteins (LSP). This approach provides thus a large number of candidate proteins involved in physiological functions of the apoplast under various stresses.Differential analyses let us to highlight 7 putative markers, namely an aspartyl protease, a β-1,3 glucanase, an isoflavone reductase for induced markers and an another aspartyl protease, an another β-1,3 glucanase, a germin-like protein and a serine pyruvate amino transferase for repressed markers. This proteins could act in a concert manner to (i) regulate reactive oxygen species homeostasy and dercease programmed cell death, (ii) counteract B. cinerea virulence factors, (iii) increase plant cell wall stiffening, (iv) cause fungal membrane leakage and (v) participate in the induction of systemic defence responses.In addition, this study provides preliminary research performed to highlight (i) priming biomarkers in grapevine, (ii) damages caused by B. cinerea infection on grapevine proteome and (iii) how a protective elicitor treatment could limit those damages.Further functional analyses of these proteins will improve the understanding of plant defense mechanisms and tools development for large scale screening of new elicitors regarding their protective effects against grey mould disease and for following grapevine defenses in vineyard

PONTIFÍCIA UNIVERSIDADE CATÓLICA DO RIO DE JANEIRO
COORDENAÇÃO DE APERFEIÇOAMENTO DO PESSOAL DE ENSINO SUPERIOR
PROGRAMA DE SUPORTE À PÓS-GRADUAÇÃO DE INSTS. DE ENSINO
Este estudo investiga o envolvimento do córtex pré-frontal medial ventral nos comportamentos de defesa inatos e aprendidos em paradigmas de condicionamento de medo e estimulação elétrica intracraniana em ratos. A lesão cortical aumentou significativamente o comportamento defensivo condicionado. No comportamento defensivo incondicionado, a lesão cortical diminuiu significativamente o congelamento pós-fuga dos animais. Os resultados replicam os dados da literatura científica a respeito do papel do córtex infralímbico como uma estrutura inibitória do estímulo condicionado em um circuito amidaloide de medo condicionado, e indicam uma participação do córtex pré-frontal na modulação dos comportamentos de defesa originários da estimulação da MCPd, em especial a sustentação do congelamento motor pós fuga.
This study investigates the role of the prefrontal cortex in the innate and conditioned defensive behaviors in rats during classical conditioning and intracranial electrical stimulation procedurals. It was found that the cortical lesion augmented the conditioned freezing behavior to contextual fear cues. On the other hand, the lesions impaired the motor freezing presented after the escaping provoked by dPAG stimulation. These results replicate the findings from the literature about a prefrontal cortex role as an inhibitory structure in the aversive classic conditioning circuitry, as well as presenting a role for it in modulating freezing behavior in a panic circuitry involving the dPAG, especially regarding its function as a possible short term memory device for innate fear expression.

Nos travaux de recherches de thèse portent sur une étude comparative entre le déclanchement des mécanismes de défense in planta de la plante de melon après élicitation biotique, et un traitement avec un produit ayant des propriétés de Stimulateur des Défenses Naturelles (SDN). Le pathosystème étudié est Cucumis melo - Fusarium Oxysporum melonis. L'éliciteur abiotique SDN utilisé codé FEN560 est un extrait de graines de fenugrec (Trigonella foenum-graecum). Les principaux travaux sont : 1) L'analyse des activités enzymatiques relatives au stress et caractérisation d'enzymes PR. 2) L'étude des métabolites du métabolisme secondaire, notamment la production l'accumulation des composés organiques volatiles (COV). 3) Une étude de l'expression de gènes candidats impliqués dans la production de ces PR-protéines et/ou dans les voie de signalisation est aussi entreprise ; Les principaux résultats se traduisent par (i) une précocité et intensité de l'induction des activités enzymatiques étudiées pour les deux variétés et pour les organes d'un même plant (ii) Le priming est généralement observé au niveau du site de la seconde élicitation (infection FOM) avec une induction remarquable des PR-protéines. (iii) L'élicitation est Systémique, car l'induction se transmet mais à moindre intensité sur les intervalles de temps étudiés. (iv) L'augmentation de l'émission des COV coïncide avec l'induction de la LOX. Il est aussi noté l'émission de nouvelles molécules connus par leurs propriétés antibiotique ou répulsive des insectes herbivores ou attractive des prédateurs d'insectes herbivores. Les résultats sur les expressions des gènes candidats sont corrélés avec les activités enzymatiques et la production des composés organiques. Il en est ressorti que l'induction de la résistance par le traitement par pulvérisation de FEN560 est similaire au phénomène de « primig » dans la résistance systémique induite
This work is a comparative study between defense mechanisms in melon plant induced by biotic elicitor and treatment by plant extract having simulative proprieties of plant defense mechanisms. The studied pathosystème is Cucumis melo - Fusarium oxysporum fsp melonis. The used plant extract elicitor is named FEN560, it is from fenugreek seeds (Trigonella foenum-graecum). The main works were: 1) Analysis of the enzymatic activities relating to the stress and characterization of PR-proteins sch as peroxidases and chitinases. The lipoxygenase (LOX), the key enzyme of oxylipins biosynthesis and phenylalanine ammonia-lyase, the key enzyme of the phenylpropanoids pathway, were also studyed. 2) The study of the metabolites of the secondary metabolism, in particular accumulation of volatile organic compounds (VOC). 3) A study of expression candidate genes implied in the production of Pr-proteins and the induced resistance pathways is also undertaken. The main results: (i) precocity and intensity of enzymatic activities induction for the two varieties after inoculation of pretreated plants (ii) the priming is generally observed in the site of the second elicitation with a remarkable induction of PR-proteins. (iii) The elicitation induced by FEN560 is systemic; because induction is transmitted between roots and shoots (iv) The modification of VOC emissions coincides with the induction of the LOX activity. It is also noticed the emission of new molecules known for their antibiotic properties or repulsive of the insects herbivorous or attractive of predatory of herbivorous insects. The results on the genes candidates' expression are in general correlated with the enzymatic activities and the production of the volatile organic compounds through LOX activity. In brief, the induction of resistance by the treatment by FEN560 is similar to the phenomenon of "primig" in induced systemic resistance

PONTIFÍCIA UNIVERSIDADE CATÓLICA DO RIO DE JANEIRO
CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO
A matéria cinzenta periaquedutal dorsal (MCPD) é associada com comportamento defensivo e ataques de pânico em humanos. Estimulações elétricas da MCPD induzem a um repertório de defesa: o congelamento pré fuga, reação de fuga e congelamento pós fuga. Evidências já apresentadas suportam a hipótese que enquanto o congelamento pré fuga seria um modelo de ataque de pânico o congelamento pós fuga seria um modelo de transtorno de pânico. Os resultados no experimento 1 indicaram que as duas linhagens de animais selecionadas para alto (Carioca alto congelamento) e baixo (Carioca baixo congelamento) congelamento em resposta ao contexto associado com choque nas patas, demonstraram que embora a ansiedade antecipatória possa exercer um efeito inibitório na expressão do congelamento pré fuga e na reação de fuga, pode por sua vez facilitar o congelamento pós fuga. No experimento 2, o efeito de lesões eletrolíticas no Córtex Cingulado Anterior Rostral não alteraram o congelamento pré fuga e a reação de fuga, porém as mesmas exercem um efeito inibitório no congelamento pós fuga demonstrando que os tipos de congelamento no medo incondicionado são mediados por circuitarias de defesa independentes. No experimento 3, o efeito de lesões eletrolíticas no Córtex Cingulado Anterior Rostral (CCAr) aumentou a sensibilidade a dor (comportamento recuperativo) de animais submetidos ao teste de formalina no paradigma de analgesia condicionada demonstrando a importância do CCAr na modulação da circuitaria de dor.
The dorsal portion of the periaqueductal Gray (DPAG) is notably associated with defensive behavior and panic attacks in humans. Electrical stimulation of the DPAG induces a repertoire of defense: dPAG-evoked freezing, escape reaction and dPAG post-stimulation freezing. Past evidence support the hypothesis that whereas dPAG-evoked freezing would serve as model of panic attack, the dPAG post-stimulation freezing appears to be a model of panic disorder. The experiment 1 used two lines of animals selectively bred for high (Carioca High-Freezing) and low (Carioca Low-Freezing) freezing in response to contextual cues associated with footshock. The results suggest that although anticipatory anxiety might exert an inhibitory effect on the dPAG-evoked freezing and escape reaction, it might also facilitate the dPAG post-stimulation freezing. The experiment 2 studied the effect of electrolytic lesions on rostral anterior cingulated cortex (rACC). The results of electrolytic lesions on rACC suggest that although rACC lesions did not change the dPAG-evoked freezing and escape threshold, it might exert an inhibitory effect on the dPAG post-stimulation freezing, reinforcing the hypothesis that dPAG-evoked freezing and dPAG post- stimulation freezing are modulated by two independent circuitry of defense. The experiment 3 studied pain sensibility of rACC lesioned animals submitted to formalin test on conditioned analgesia paradigm. The results suggest that rACC lesions might exert an inhibitory effect on conditioned analgesia and consequently exacerbates recuperative behavior. The results also support the hypothesis of the rule of rACC on pain modulation.

Artikelserien "Hudhunger" som Dagens Nyheter publicerat under våren 2004 har varit en inspiration för oss när vi har skrivit vår uppsats. Serien har belyst begreppet beröring och dess betydelse för oss människor. I litteraturgenomgången presenterar vi först begreppet beröring kopplat till barnkonventionen och förskolans läroplan, varför beröring är så viktigt för oss leveande varelser genom hela livet. Vi tar även upp vad beröringsbrist kan leda till och dess effekter. Vidare presenterar vi hur beröringskänslighet, eller taktilt försvar kan yttra sig och hur man kan arbeta med dessa barn som har dessa svårigheter. Det kan också finnas misstänktsamhet vid beröring av barn. I det avsnittet presenterar vi varför beröring är viktigt för att motverka dessa misstänksamheter då barnen bygger upp mer förståelse för bra och dålig beröring. Slutligen tar vi upp om det finns någon skillnade mellan pojkar och flickor, hur de upplever beröing, enligt forskare.

I vår undersökning har vi gjort en enkät genom kvalitativ metod där vi undersökte hur pedagoger arbetar med beröring i förskolan. Undersökningen är gjord på fyra olika förskolor i två mellanstora städer i Sverige. Resultatet visar att de pedagoger som besvarat vår enkät har kunskap om beröring och beröringskänslighet. Samt att de har en positiv syn då de arbetar aktivt med beröring i form av exempelvis massage och taktila lekar. Vi anser att beröring ska vara en naturlig del i förskolans verksamhet och att vår uppsats uppmärksammar vikten av beröring och ett fortsatt arbete med beröring.

Dans le cadre de la promotion d'une viticulture durable, le développement d'alternatives écologiques à l’utilisation de pesticides de synthèse pour le contrôle des maladies de la vigne (Vitis vinifera) gagne en importance. Une des méthodes de bio-contrôle proposée est l'induction de l'immunité des plantes par l’application de substances biodégradables, non toxiques pour la santé et l'environnement, appelées éliciteurs ou stimulateurs de défense des plantes (SDP). La résistance conférée contre divers agents pathogènes peut être obtenue grâce à l’emploi de molécules utilisant le plus fréquemment les voies de signalisation de l'acide jasmonique (JA), de l'acide salicylique (SA) et/ou de l'éthylène (ET). De telles voies peuvent déclencher l'induction de gènes liés à la défense tels ceux codant des enzymes responsables de la biosynthèse des stilbènes, des métabolites phénoliques antimicrobiens parmi les plus importants des Vitacées. Au vignoble, pour contrôler les maladies, les éliciteurs peuvent être appliqués en complément des pesticides de synthèse et non en remplacement total car leur efficacité est souvent variable selon les agents pathogènes et les conditions environnementales. Afin de développer leur utilisation, des études supplémentaires qui pourraient notamment élucider leur mécanisme d'action sont nécessaires. L’objectif de cette thèse était d'étudier les réponses de la vigne à des éliciteurs de différents modes d'action, comme le jasmonate de méthyle (MeJA), impliqué dans la voie de signalisation du JA, l’acide S-méthyl ester 2,1,3-benzothiadiazole-7-carbothioïque (BTH), un analogue synthétique du SA, des phosphonates (PHOS), molécules à double action stimulateur-fongicide. Le profil des stéroïdes et des triterpénoïdes pentacycliques a été caractérisé par des analyses en chromatographie en phase gazeuse couplée à la spectrométrie de masse (GC-MS). Dans un premier temps, l'effet éliciteur du MeJA a été évalué dans des suspensions cellulaires (in vitro) de plusieurs cultivars de V. vinifera. Une surproduction de triterpénoïdes pentacycliques a été observée avec des différences selon la variété considérée : induction de l’accumulation de la bétuline et de l'acide oléanolique ou des phytostérols pour le Petit Verdot, le Gamay Teinturier) et le Cabernet Sauvignon, respectivement. Puis les élicitations ont été effectuées au niveau des feuilles de boutures de serre de V. vinifera cv. Cabernet Sauvignon. Un effet stimulant sur les triterpénoïdes pentacycliques liés à la défense a été démontré au détriment de la biosynthèse des stérols, composants structurels essentiels des membranes cellulaires. Par l’utilisation de puces NeoVigen et la chromatographie liquide à ultra haute performance couplée à la spectrométrie de masse (UHPLC-MS), l’induction de l’expression de gènes liés à la défense et l'accumulation de polyphénols (stilbènes, flavanols et flavonols) ont été notées suite aux trois traitements éliciteurs. La protection de la vigne conférée par les éliciteurs a été confirmée par des biotests sur disques foliaires inoculés par l’oomycète biotrophe Plasmopara viticola, l’agent responsable du mildiou. Par ailleurs, il est important d’avoir connaissance de l’impact des éliciteurs sur le métabolisme général afin d'obtenir l’effet optimal entre croissance, rendement et défense. Ainsi, une approche métabolomique utilisant la spectroscopie de résonance magnétique nucléaire du proton (RMN 1H) a été menée. Une reprogrammation similaire et/ou spécifique selon l'éliciteur considéré a été notée en particulier au niveau des glucides, des acides aminés et de certains intermédiaires du cycle de Krebs. Les recherches présentées dans cette thèse, démontrent que la compréhension approfondie de l'interaction entre l'éliciteur, les réponses moléculaires et métaboliques de la plante et le pathogène, est cruciale pour le développement de stratégies de protection efficaces basées sur l'utilisation des SDP pour contrôler les maladies de la vigne
In the frame of promoting sustainable vitiviniculture, the development of eco-friendly alternatives to synthetic chemical products for phytosanitary treatments against grapevine (Vitis vinifera) pests is gaining importance. One of the bio-control methods that can be proposed is the induction of plant immunity by using elicitors, also called plant defense stimulators (PDS), as these substances are biodegradable and, non-toxic to health and environment. A conferred resistance against various pathogens can be obtained with natural molecules acting most frequently through jasmonic acid (JA), salicylic acid (SA), and/or ethylene (ET) signaling pathways. These pathways are involved in the induction of defense-related genes such as those encoding enzymes responsible for the biosynthesis of stilbenes, which are the most important polyphenolic antimicrobial metabolites (phytoalexins) in Vitaceae. For vineyard protection, PDS can be applied as a complement for pesticides and not as a full replacement since their effectiveness is often variable according to pathogens and environmental conditions. In order to develop the strategies based on PDS use, more studies which could elucidate their mechanism of action are needed. The aim of this thesis was to examine the responses of grapevine to elicitors of different mode of action, as methyl jasmonate (MeJA), implicated in JA signaling pathway, 2,1,3-benzothiadiazole-7-carbothioic acid S-methyl ester (BTH), a synthetic analogue of SA, and phosphonates (PHOS), molecules of a double stimulator-fungicide action. Due to scarce information about steroids and pentacyclic triterpenoids in grapevine, their profile after PDS treatment were characterized in different grapevine experimental models using gas chromatography-mass spectrometry (GC-MS) analyses. Firstly, the effect of elicitation with MeJA was evaluated in cell suspension cultures (in vitro) of V. vinifera. An overproduction of bioactive pentacyclic triterpenoids occurred with differences according to the cultivar studied, i.e., acumulation of betulin and oleanolic acid or phytosterols was noted in respectively Petit Verdot, Gamay Teinturier and Cabernet Sauvignon cell suspension cultures. Then, elicitations were effectuated on the leaves of V. vinifera cv. Cabernet Sauvignon greenhouse cuttings. A stimulatory effect on the potentially defense-related pentacyclic triterpenoids at the expense of the biosynthesis of sterols, which are essential structural components of cell membranes, was shown. By the use of NeoVigen microarrays, and ultra-performance liquid chromatography-mass spectrometry (UHPLC-MS), the accumulation of defense-related transcripts and polyphenols (stilbenes, flavanols and flavonols) were noted after the three elicitors treatments. Grapevine protection conferred by these elicitors was confirmed on foliar discs against the biotrophic oomycete Plasmopara viticola, the causal agent of downy mildew. Furthermore, the impact of PDS on primary metabolism should be evaluated in order to ensure, in the longer term, the best trade-off between growth, yield and defense. Thus, a thorough metabolomic approach using proton nuclear magnetic resonance spectroscopy (1H-NMR) was performed. A reprogramming similar and/or specific to the elicitor applied was noted, particularly within carbohydrates, amino acids, and some of the Krebs cycle intermediates. The research presented in the current dissertation revealed that the thorough comprehension of the interaction between elicitor, plant molecular and metabolic responses and pathogen, is crucial for the development of effective protection strategies based on the use of PDS for grapevine diseases control

O medo e a ansiedade são emoções que apresentam claro valor adaptativo, e que tem suas origens nas reações de defesa que os animais exibem em resposta a situações de ameaça que podem comprometer sua integridade física ou sobrevivência. Recentes estudos têm indicado que a matéria cinzenta periaquedutal dorsal (MCPD) está envolvida na organização e expressão de comportamentos intempestivos do tipo fuga e luta, os quais são relacionados ao estado de medo, e também participa, juntamente com estruturas prosencefálicas (ex. córtex pré-frontal, sistema septo-hipocampal e amígdala), do controle de comportamentos defensivos mais elaborados e orientados relacionados à ansiedade. O presente estudo investigou a participação da MCPD na modulação de diferentes comportamentos defensivos (p. ex. fuga, esquiva e avaliação de risco) induzidos por métodos artificial (estimulação química) e naturalístico (exposição ao predador) em camundongos. Na primeira etapa, investigamos o padrão de resposta comportamental induzida pela infusão do ácido D,L-homocistéico (DLH, estímulo aversivo químico) na MCPD em diferentes situações ou ambientes, com e sem grande disponibilidade de espaço - o Mouse Defense Test Battery (MDTB) e a Arena (Experimento 1), respectivamente. Além disso, o presente estudo avaliou a habilidade dos animais de reagirem a estímulos aversivos (predador) durante o período inicial (nos 60 s iniciais) do efeito do ácido DLH (fuga explosiva) (Experimento 3), e imediatamente após esse período, no qual o animal apresente comportamento de congelamento ou imobilidade (Experimento 2). Nossos resultados indicaram que a fuga desencadeada pela estimulação química é a resposta predominante de camundongos e que sua exibição depende da disponibilidade de espaço, uma vez que a maioria dos saltos observados na arena está intimamente relacionada ao contato tátil do animal com as paredes do aparato. Esse perfil de respostas de fuga explosiva e saltos parece não representar o padrão comportamental defensivo natural, tal como acontece diante de uma ameaça proximal (ex. um predador), uma vez que durante a estimulação química os camundongos apresentaram um déficit na estratégia antipredador. A segunda etapa do estudo avaliou os efeitos da injeção intra-MCPD do agonista glutamatérgico ácido N-metil-D-aspártico (NMDA), do inibidor da enzima de síntese do óxido nítrico neuronial (NOSn), N?-propil-L-arginina (NPLA) (Experimento 4), e do agonista não seletivo de receptores do fator de liberação de corticotrofina (CRF), CRF ovino (oCRF) (Experimento 5), no comportamento defensivo de camundongos submetidos ao MDTB e ao teste de exposição ao rato (Rat Exposure Test; RET). Os resultados da segunda etapa demonstraram que a ativação de receptores NMDA na MCPD de camundongos intensifica comportamentos relacionados à esquiva do predador. De maneira interessante, essas alterações produzidas pelo NMDA foram consistentemente revertidas pelo inibidor da NOSn, previamente microinjetado no mesmo sítio. Além disso, efeitos intrínsecos do NPLA atenuaram as respostas de esquiva e de avaliação de risco em camundongos submetidos ao RET. Por fim, os resultados da segunda etapa também apontaram para um efeito proaversivo (nas respostas de salto e de esquiva) do agonista de receptores CRF, indicando uma participação dos sistemas glutamatérgico, nitrérgico e CRFérgico, localizados na MCPD, na modulação de diferentes estratégias defensivas (ex. esquiva, avaliação de risco e saltos) de camundongos submetidos ao confronto com o predador. Em conjunto, nossos resultados corroboram a hipótese de que a MPCD está envolvida tanto na organização e expressão de comportamentos intempestivos do tipo fuga e luta como também no controle de comportamentos defensivos mais elaborados e orientados, tais como a avaliação de risco e a esquiva.
The midbrain dorsal periaqueductal grey (DPAG) is part of the brain defensive system involved in active defense reactions to threatening stimuli. Many lines of evidence suggest that besides fundamentally controlling fear-like responses (fight and flight) the DPAG also controls responses related to anxiety, such as avoidance and risk assessment. This study investigated the role of DPAG on different defensive strategies (i.e. flight, avoidance and risk assessment) elicited by artificial (chemical stimulation, Experiments 1-3) and naturalistic (exposure to predator, Experiments 4 and 5) paradigms in mice. Firstly, D,L-Homocysteic acid (DLH) was infused into the DPAG and behavioral responses of mice were evaluated in two different situations, a rectangular novel chamber and a large oval runway, the Mouse Defense Test Battery (MDTB) apparatus (Exp. 1). We also investigated the ability of mice to react to a threatening stimulus (ex. a predator) during (Exp. 3) and immediately after (Exp. 2) the hyperactive responses (ex. jumping and running) induced by DLH injection. Our results indicated that running as opposed to jumping is the primary response in mice injected with DLH into the DPAG when the environment enables flight. However, mice did not react the predator during the flight reaction induced by chemical stimulation, suggesting the behavioral profile induced by DLH infusion into the DPAG is not related to a normal antipredator flight. In the Experiments 4 and 5, we evaluated the effects of three different compounds, N-methyl-D-Aspartate (NMDA), a NMDA receptor agonist, N?-propyl-L-arginine (NPLA), an neuronal nitric oxide synthase (nNOS) inhibitor as well as ovine CRF (oCRF), a nonspecific corticotropin-releasing factor (CRF) receptor agonist, injected into the DPAG of mice, in two predator-stress situations, the Mouse Defense Test Battery (MDTB), and the Rat Exposure Test (RET). Firstly, our results demonstrated that NMDA receptor activation into the mouse DPAG enhances antipredator reactivity (avoidance), an effect that was attenuated by prior infusion of NPLA into the same site. Moreover, the results from the Experiment 4 indicated that the NPLA treatment per se induces consistent anti-aversive effects on defensive behaviors (avoidance and risk assessment) of mice confronted by predator. Finally, our results pointed out a proaversive effect (e.g. increased jump escapes and avoidance behaviors) following intra-DPAG infusion of oCRF, suggesting an important role of glutamatergic, nitrergic and CRFergic systems into the DPAG on the defensive behaviors (risk assessment, avoidance and jumps) elicited by the confront to the predator. Taken together, present results are compatible with previous studies which have emphasized the role of the periaqueductal gray in the modulation of behavioral responses related to anxiety such as risk assessment and avoidance besides fundamentally controlling fear-like responses.

L’œuf est un aliment riche en divers composés dont de nombreuses molécules antimicrobiennes qui sont les effectrices de son système de défense moléculaire innée et complètent l’action des immunoglobulines (IgYs) afin de protéger l’embryon. La composition en IgYs de l’œuf est sous la dépendance notamment des stimulations microbiennes de la poule. L’objectif de notre travail a été d’évaluer l’influence de ces mêmes stimulations sur le système de défense moléculaire innée du blanc d’œuf. Nous avons exploré cette hypothèse en utilisant deux approches expérimentales. La première était basée sur la comparaison de l’activité antibactérienne des blancs d’œufs de poules axéniques, de poules exemptes d’organismes pathogènes spécifiques (EOPS), et de poules conventionnelles. La seconde approche a testé les effets de deux types d’inductions du système immunitaire chez la poule: injection d’un immunostimulant, le lipopolysaccharide bactérien (LPS) et administration oral de souches vaccinales atténuées (virale, bactérienne et parasitaire). Nos résultats montrent que l’activité antibactérienne du blanc d’œuf est augmentée en fonction de la charge microbienne du milieu de vie de la poule, après stimulation de celle-ci par voie intraveineuse avec du LPS ou suite à la vaccination avec des souches atténuées virale ou bactérienne. Néanmoins cette augmentation est modérée de par son amplitude et son spectre antibactérien. Ces résultats suggèrent que les poules peuvent renforcer modérément l’activité antimicrobienne du blanc d’œuf en réponse à des stimuli microbiens de leur milieu et anticiper ainsi les besoins de l’embryon en termes de protection
The egg is a balanced source of different nutrients and contains a myriad of antibacterial peptides/proteins that ensure its chemical protection. These molecules are a part of its innate molecular defense and, in addition to the maternal immunoglobulins IgY, contribute to the protection of the forming embryo whose development occurs ex utero. It is well documented that yolk immunoglobulin deposition is induced by the environmental microbiome of the hen but no such evidence is available for antimicrobial peptides/proteins. Therefore the aim of this thesis was to assess whether the hen has the ability to stimulate the innate molecular immunity of the egg white when facing a higher environmental microbial load (commensal or pathogenic). To address these questions, we developed two main experimental approaches; the first assessed the impact of the hen environmental microbial load through the comparison of three groups of hens with different immune status:-Germ free, -Specific pathogen free (SPF), and -conventional. The second approach explored the effect of different types of immune stimulation in hens: non-infectious stimulation (systemic injection of bacterial lipopolysaccharide (LPS)); immune stimulation using attenuated live vaccines (Infectious bronchitis virus vaccine, Salmonella enterica Enteritidis vaccine and a complex of Eimeria vaccine). Our results show that the activity of egg white is increased in response to higher microbial environmental charge, after LPS systemic stimulation or after vaccinating hens with live attenuated viral and bacterial strains. However this response is moderate both in its amplitude and microbial spectrum. Altogether, it appears that hens when subjected to immune stimuli, have the ability to reinforce moderately the antibacterial activity of the egg white as an attempt to anticipate the need of protection of their embryos

In the frame of promoting sustainable vitiviniculture, the development of eco-friendly alternatives to synthetic chemical products for phytosanitary treatments against grapevine (Vitis vinifera) pests is gaining importance. One of the bio-control methods that can be proposed is the induction of plant immunity by using elicitors, also called plant defense stimulators (PDS), as these substances are biodegradable and, non-toxic to health and environment. A conferred resistance against various pathogens can be obtained with natural molecules acting most frequently through jasmonic acid (JA), salicylic acid (SA), and/or ethylene (ET) signaling pathways. These pathways are involved in the induction of defense-related genes such as those encoding enzymes responsible for the biosynthesis of stilbenes, which are the most important polyphenolic antimicrobial metabolites (phytoalexins) in Vitaceae. For vineyard protection, PDS can be applied as a complement for pesticides and not as a full replacement since their effectiveness is often variable according to pathogens and environmental conditions. In order to develop the strategies based on PDS use, more studies which could elucidate their mechanism of action are needed. The aim of this thesis was to examine the responses of grapevine to elicitors of different mode of action, as methyl jasmonate (MeJA), implicated in JA signaling pathway, 2,1,3-benzothiadiazole-7-carbothioic acid S-methyl ester (BTH), a synthetic analogue of SA, and phosphonates (PHOS), molecules of a double stimulator-fungicide action. Due to scarce information about steroids and pentacyclic triterpenoids in grapevine, their profile after PDS treatment were characterized in different grapevine experimental models using gas chromatography-mass spectrometry (GC-MS) analyses. Firstly, the effect of elicitation with MeJA was evaluated in cell suspension cultures (in vitro) of V. vinifera. An overproduction of bioactive pentacyclic triterpenoids occurred with differences according to the cultivar studied, i.e., acumulation of betulin and oleanolic acid or phytosterols was noted in respectively Petit Verdot, Gamay Teinturier and Cabernet Sauvignon cell suspension cultures. Then, elicitations were effectuated on the leaves of V. vinifera cv. Cabernet Sauvignon greenhouse cuttings. A stimulatory effect on the potentially defense-related pentacyclic triterpenoids at the expense of the biosynthesis of sterols, which are essential structural components of cell membranes, was shown. By the use of NeoVigen microarrays, and ultra-performance liquid chromatography-mass spectrometry (UHPLC-MS), the accumulation of defense-related transcripts and polyphenols (stilbenes, flavanols and flavonols) were noted after the three elicitors treatments. Grapevine protection conferred by these elicitors was confirmed on foliar discs against the biotrophic oomycete Plasmopara viticola, the causal agent of downy mildew. Furthermore, the impact of PDS on primary metabolism should be evaluated in order to ensure, in the longer term, the best trade-off between growth, yield and defense. Thus, a thorough metabolomic approach using proton nuclear magnetic resonance spectroscopy (1H-NMR) was performed. A reprogramming similar and/or specific to the elicitor applied was noted, particularly within carbohydrates, amino acids, and some of the Krebs cycle intermediates. The research presented in the current dissertation revealed that the thorough comprehension of the interaction between elicitor, plant molecular and metabolic responses and pathogen, is crucial for the development of effective protection strategies based on the use of PDS for grapevine diseases control.
Rozwój przyjaznych dla środowiska alternatywnych metod ochrony winorośli (Vitis vinifera) przeciw szkodnikom zyskuje na znaczeniu w kontekście zapobiegania nadmiernemu użyciu pestycydów i promowania zrównoważonego rolnictwa. Indukcja naturalnej obrony roślin poprzez działanie elicytorów (stymulatorów odporności roślin) jest jedną z najbardziej obiecujących metod kontroli biologicznej, jako że opiera się na zastosowaniu substancji biodegradowalnych i nietoksycznych dla środowiska i zdrowia ludzi. Elicytory zaaplikowane na roślinę wyzwalają w niej mechanizmy obronne, prowadząc do rozwijania odporności na kolejne ataki patogenów. Percepcja elicytora uruchamia szlaki sygnałowe, z których najbardziej kluczowe są te związane z kwasem salicylowym (SA), kwasem jasmonowym (JA) lub etylenem (ET). Konsekwencje aktywowania kaskady reakcji odpornościowych to m.in. indukcja ekspresji genów związanych z obroną, np. kodujących białka związane z patogenezą (PR), czy enzymy odpowiedzialne za biosyntezę wyspecjalizowanych metabolitów o właściwościach przeciwdrobnoustrojowych (przede wszystkim stilbenów u roślin z rodziny Vitaceae). Stymulatory odporności roślin są obecnie stosowane jako uzupełnienie pestycydów, a nie jako środek zastępczy, ponieważ ich skuteczność jest często zmienna w zależności od konkretnego patogena i warunków środowiskowych. W celu opracowania strategii ochrony winorośli przed chorobami całkowicie opartych na elicytorach, potrzeba jest wielu badań, które pozwolą m.in. wyjaśnić mechanizm działania tych środków. Celem niniejszej pracy było zbadanie odpowiedzi winorośli na elicytory o różnej aktywności biologicznej: jasmonian metylu (MeJA), związany ze szlakiem sygnałowym JA, benzotiadiazol (BTH), syntetyczny analog SA oraz sole fosforanowe (PHOS), o działaniu stymulującym, jak i grzybobójczym. Profil steroidów i triterpenoidów pentacyklicznych winorośli scharakteryzowano za pomocą chromatografii gazowej sprzężonej ze spektrometrią mas (GC-MS). Potencjalny wpływ MeJA na profil triterpenoidów oceniono w hodowlach zawiesin komórkowych in vitro V. vinifera. W zależności od badanej odmiany, zaobserwowano wzmożoną biosyntezę triterpenoidów pentacycklicznych, takich jak betulina (Petit Verdot) i kwas oleanolowy (Gamay Teinturier), a także fitosteroli (Cabernet Sauvignon). W elicytowanych liściach szklarniowych sadzonek Cabernet Sauvignon, wykazano konkurencyjność szlaków biosyntezy triterpenoidow, tzn. zwiększona akumulacja triterpenoidow pentacyklicznych, związanych z chemiczną obroną roślin, odbyła się kosztem biosyntezy steroli, związków niezbędnych dla budowy i funkcjonowania błon komórkowych. Dzięki analizom z wykorzystaniem mikromacierzy NeoVigen i ultrasprawnej chromatografii cieczowej ze spektrometrią mas (UHPLC-MS), w liściach poddanych elicytacji stwierdzono akumulację transkryptów związanych z obroną oraz zwiększoną zawartość polifenoli (stilbenów, flawanoli i flawonoli). Biotesty na krążkach liściowych wykazały, że badane elicytory nadały odporność liściom przeciwko Plasmopara viticola, tj. patogenowi wywołującemu mączniaka rzekomego. Ponadto, badanie wpływu elicytorów na metabolizm pierwotny jest istotny w celu zapewnienia roślinie swoistego kompromisu między aktywowaniem kosztownych energetycznie mechanizmów obronnych, a utrzymaniem prawidłowego funkcjonowania podstawowych procesów fizjologicznych. Badania metabolomiczne z wykorzystaniem spektroskopii protonowego jądrowego rezonansu magnetycznego (1H-NMR) wykazały szereg istotnych zmian w profilu węglowodanów, aminokwasów i niektórych substratów cyklu Krebsa w liściach poddanych elicytacji. Wyniki badań przedstawione w niniejszej rozprawie wykazują, iż dokładne zrozumienie interakcji między elicytorem, odpowiedzią molekularną i metaboliczną rośliny a patogenem, ma kluczowe znaczenie dla rozwoju skutecznych strategii obrony winorośli przed chorobami, opartych na stymulatorach odporności roślin.