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Parallélisation d'heuristiques d'optimisation sur les GPUs / Parallel optimization heuristics on GPUsBerrajaa, Achraf 27 December 2018 (has links)
Cette thèse, présente des contributions à la résolution (sur les GPUs) de problèmes d'optimisations réels de grandes tailles. Les problèmes de tournées de véhicules (VRP) et ceux de localisation des hubs (HLP) sont traités. Diverses approches et leur implémentions sur GPU pour résoudre des variantes du VRP sont présentées. Un algorithme génétique (GA) parallèle sur GPU est proposé pour résoudre différentes variantes du HLP. Le GA adapte son codage, sa solution initiale, ses opérateurs génétiques et son implémentation à chacune des variantes traitées. Enfin, nous avons utilisé le GA pour résoudre le HLP avec des incertitudes sur les données.Les tests numériques montrent que les approches proposées exploitent efficacement la puissance de calcul du GPU et ont permis de résoudre de larges instances jusqu'à 6000 nœuds. / This thesis presents contributions to the resolution (on GPUs) of real optimization problems of large sizes. The vehicle routing problems (VRP) and the hub location problems (HLP) are treated. Various approaches implemented on GPU to solve variants of the VRP. A parallel genetic algorithm (GA) on GPU is proposed to solve different variants of the HLP. The proposed GA adapts its encoding, initial solution, genetic operators and its implementation to each of the variants treated. Finally, we used the GA to solve the HLP with uncertainties on the data.The numerical tests show that the proposed approaches effectively exploit the computing power of the GPU and have made it possible to resolve large instances up to 6000 nodes.
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Social Innovation Hubs Supporting Social Entrepreneurs: Strategically Adopting the SDGs towards SustainabilityBhalerao, Akash, Louwerse, Sjaak, Quarmyne, Michael Tei, Ritchie, Dan January 2019 (has links)
The United Nations Sustainable Development Goals (SDGs) are a well-known and comprehensive framework for sustainable development. However due to the overlapping and interrelated nature of the goals, action towards one goal can positively or negatively contribute to another.Social innovation hubs including Impact Hub and Centre for Social Innovation use the SDGs to support social entrepreneurs to have a positive impact. Document analysis and interviews with 15 practitioners from these hubs informed the research on how the organizations perceive and contribute to sustainability, how they integrate the SDGs, and the challenges and benefits with using the SDGs. Based on that, this research has developed five recommendations for social innovation hubs to: 1) Define Sustainability; 2) Enhance Visioning; 3) Design co-creative programs; 4) Define Impact;and 5) Communicate Impact. While other elements of the Framework for Strategic Sustainable Development (FSSD) could be used to complement the SDGs, the Sustainability Principles (SPs) of the FSSD are recommended as a definition for sustainability.
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Neuropathies Périphériques Génétiques et Surdité : Etude des Relations Génétiques et Mécanistiques / Genetic Peripheral Neuropathies and Deafness : Study of Genetic and Mechanistic ConnectionsLerat, Justine 13 December 2018 (has links)
Les neuropathies périphériques héréditaires (NP) sont caractérisées par des phénotypes très divers et une hétérogénéité génétique importante. La maladie de Charcot-Marie-Tooth (CMT) représente la majeure partie des neuropathies périphériques sensitivo-motrices. D’autres symptômes peuvent être associés, telle que la surdité. A l’heure actuelle, aucune estimation précise de la surdité n’existe dans cette population et la pathogénicité est incertaine. L’objectif de cette thèse était de mieux comprendre la physiopathologie de la surdité chez les patients atteints de neuropathies périphériques. Pour cela plusieurs approches complémentaires ont été mises en œuvre : 1) Approche clinique sur une cohorte française de patients atteints à la fois de neuropathie périphérique et de surdité et tests de génétique moléculaire avec séquençage NGS (Panels NP, surdités et/ou exomes) ; 2) Approche biochimique sur des prélèvements de nerfs cochléaires murins et humains ; 3) Approche bioinformatique afin d’identifier des réseaux de protéines impliquées dans l’apparition de surdité liée à une neuropathie périphérique. Grâce à ce travail, nous avons pu caractériser les phénotypes variés des patients atteints de NP génétique et surdité, et ainsi constater que la surdité peut être endo, rétro ou endo et rétrocochléaire. Trente-six gènes ont été rapportés comme associées à NP et surdité. Le génotype de nos patients NP+Surdité a pu être établi dans 60% des cas, avec la découverte de sept nouveaux variants pathogènes dans cinq gènes différents. Nos travaux suggèrent également que PMP22, le gène le plus retrouvé dans les CMT, n’est probablement pas ou peu impliqué dans l’apparition de la surdité des patients NP. Chez deux de nos patients présentant un variant pathogène de PMP22, un deuxième gène impliqué a été trouvé avec respectivement COCH et MYH14. Des corrélations génotypes-phénotypes ont pu être mises en évidence avec les gènes ABHD12, SH3TC2, NEFL et PRPS1. Deuxièmement, l’étude préliminaire immunohistochimique sur des nerfs auditifs de rats sauvages a permis de mettre en évidence l’expression de pmp22, mpz, nefl et trpv4 au niveau du nerf cochléaire et de pister une différence d’expression chez les rats CMTpmp22/+. L’étude chez l’humain n’a pas été concluante. Dernièrement, la recherche in silico de voies communes aux différents gènes décrits comme impliqués dans NP+surdité a permis de confirmer le lien direct entre PMP22 et MPZ. Des liens indirects entre plusieurs autres protéines ont été pistés. Cette thèse montre également que la surdité est très certainement sous-diagnostiquée dans cette population de NP génétique. Nous proposons donc un suivi audiométrique systématique des patients atteints de NP héréditaire, et une évaluation neurologique pour les enfants diagnostiqués pour surdité. / Hereditary Peripheral Neuropathies (PN) are characterized by various phenotypes and great genetic heterogeneity. Charcot-Marie-Tooth disease (CMT) accounts for most sensori-motor peripheral neuropathies. Besides, other symptoms can be associated, such as deafness. No precise estimation of deafness within this population exist and its pathogenicity is uncertain. The aim of this PhD was to better understand the physiopathology of deafness in patients suffering from PN. Various complementary approaches were used; 1) a clinical approach on a French cohort of patients suffering from both PN and hearing loss and molecular genetic tests with NGS sequencing (PN, deafness panels, and/or exomes), 2) a biochemical approach on murine and human cochlear nerve samples and 3) a bioinformatic approach to identify protein hubs implicated in the onset of PN-associated deafness.This has enabled us to characterize the various phenotypes of patients suffering from both hereditary PN and deafness, and then notice that deafness can be endo-, retro- or endo- and retrocochlear. Thirty-six genes were reported to be associated with both PN and hearing impairment. Sixty percent of our patients were genotyped, highlighting seven novel pathogenic variants in five different genes. Our research also suggests that PMP22, the most frequent gene in CMT, is probably not or poorly implicated in deafness onset in PN patients. In two of our patients with PMP22 pathogenic variants, a second involved gene was found with COCH and MYH14 respectively. Genotype-phenotype correlations were found out with the ABHD12, SH3TC2, NEFL and PRPS1 genes. Secondly, the preliminary immunohistochemical study on wild-type rats auditory nerves highlighted the expression of pmp22, mpz, nefl and trpv4 on the cochlear nerve and tracked a different expression in CMTpmp22/+ rats. However, the study on humans was not conclusive. Recently, in silico research of pathways common to the different genes described to be involved in both PN and deafness, has found the direct link between PMP22 and MPZ. Indirect links between several other proteins have been tracked.This thesis also shows that hearing impairment is most probably under-diagnosed in this population of genetic PN sufferers. We suggest regular audiologic follow-up for PN patients and neurological assessment for deaf children.
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台灣地區資訊製造產業電子供應鏈策略管理之研究--以HP、MiTAC為個案研究案例 / Taiwan Area information Industry Supply Chain Management strategy郭淑玲, Sharon Kuo Unknown Date (has links)
網際網路(Internet)大量應用於商業化之後,企業經營模式產生了鉅大的改變,尤其是以應用於資訊傳遞的方法更是徹底。也由於Internet的普及,也使得構築於Internet的資訊應用科技系統更深化於企業經營之中。
台灣的資訊製造產業是全球化最深的產業,該產業本身從訂單到生產到交貨莫不與全球各地的企業習習相關,而速度、彈性與創新更是這些產業所追求的目標。為了達成這樣的目標,因此,企業資源規劃(Enterprise Resource Planning)、電子供應鏈管理(Supply Chain Management)與全球運籌管理(Global Logistics),對台灣資訊製造產業是相當重要的課題。由於這樣的產業特色,也使得如何應用Internet資訊技術整合供應鏈,促使台灣資訊製造產業的電子供應鏈體系能有效的經營,建構企業的核心能耐,亦是當前企業必須面對的商業策略議題。
本論文之研究方向是以台灣地區資訊製造產業間的電子供應鏈發展為對象,目的在於:1)瞭解電子供應鏈意涵及發展歷程;2)電子供應鏈對企業的效益;3)電子供應鏈的現況分析及其願景;同時以個案研究方式深入探討目前在企業體系的實際運作並由企業願景、企業能耐等構面來分析電子供應鏈應用於企業界的演進階段。
在個案研究部份,本論文是以政府的『產業自動化及電子化推動方案』(簡稱AB計畫)為主要基礎,再由『A、B計畫』中分別篩選足以作為示範應用系統的台灣惠普公司(HP)以及神達電腦公司(MiTAC)作為本論文的研究對象。並以1)電子供應鏈動態發展策略;2)使命願景;3)企業能耐;4)電子供應鏈管理制度;5)電子供應鏈與上下游廠商動態策略分期標準;5)電子供應鏈動態策略演進階段;等五個研究變項建立觀念模型進行實證研究,發展相關命題與建議。
經實證結果發現,電子供應鏈動態發展可歸納為四個階段,每一階段各有其不同之命題與概念發展,四大階段分別為:1)概念醞釀期,主要是建立成員間的共識;2)成員數量累積期,側重於企業實體資訊構架的建置;3)品質控制期,此一時期「品質」是命題的重點;4)策略聯盟期,著重在跨體系間的價值鏈運用。企業願景是貫穿每一階段的核心,企業能耐則是落實企業電子化的趨力。
本論文之研究發現可歸納為以下重要結論:台灣資訊製造業電子供應體系發展能否成功,關鍵是在於B計畫中的衛星體系能否電子化成功。因為B計畫的中心廠商對電子供應鏈的策略目標與組織共識是非常明確的,而且在企業能耐與實力上已經厚基了相當的實力。反而是衛星廠商由於經濟規模之限制需要投入更多的資源輔導。 / The operational approach of enterprises has been changed drastically after the vast commercialization “Internet” applications.
In particular, business management must be transformed to the new business model from the traditional design. We find the application of information technologies has become significantly important factors; especially, how to use related information technologies and methodology of the application implementation are the major points of transforming to the new business model.
The information industry in Taiwan area is a very profoundly globalizes. When a corporate obtains a international purchased order “PO”, we can find along the process from PO to manufacture and delivery to the buyers, the promotion on time efficiency availability commitment and flexibility are necessary for achieving the goal. Other words, the “e-Business” has become the most important topic on the corporate management of the new digital economy era.
The focus of this research is to e verify goals for supply chain management and the establishment of strategic alliance. The purpose of this paper builds a model for supply chain management (SCM) analysis based on the concept of the investigation on current situation of the SCM, the industrial development theories, the literature review on pervious studies, and the case study analysis.
The result of the study shows that the SCM dynamic development can be divided into four stages with each stage having its own topics and different conception development. The four stages are as follows:
(1) The stage for incubatory conception: the stage to form the consensus on SCM with the whole organization.
(2) The stage for the accumulation of physics: this stage constructs to the hardware and software capability.
(3) The stage for the quality control: this stage enforces the total quality management (TQM).
(4) The stage for the establishment of strategic alliance: this stage enhances of business partnership on SCM and the establishment of a learning organization, followed by the application of value chain, and setup of new strategies for new business model.
Generally speaking, the corporation’s vision is the core value that links and integrates all the every stage, and the corporate capability is core competence to implement e-Business and accomplish the performance of SCM.
The result of the study suggests the following conclusion: The key successful point of information industry is that its electronics supply chains depends on the implementation of computerize information system built in subsidiary manufactures in Plan B. The central manufacturers in Plan B have clear strategies and vision for their companies alredy. They have also invested a lot of resource, to build their core competence. Significantly, the subsidiary manufacturers need more assistances and resources from governments and central manufacturers due to their economic scale.
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Morphologies et dynamiques territoriales des services de messagerieBeyer, Antoine 15 January 1999 (has links) (PDF)
Le transport de messagerie a en charge les envois légers. Son caractère intersectoriel et multi-clients lui permet de jouer sur la complémentarité de flux hétérogènes. Il offre au géographe une lecture éclairante de l'interface entre territoire et industrie. En effet, l'éclatement croissant des structures et des espaces productifs n'a été envisageable que par l'amélioration constante de la fluidité des relations entre les divers acteurs économiques. Alors que l'organisation de la circulation des biens et de sa fiabilité est devenue une préoccupation majeure des industriels, la figure du réseau s'impose désormais comme un élément central dans l'articulation du nouvel espace industriel. Elle met en jeu un territoire dont la cohérence repose moins sur une approche fondée la notion de proximité spatiale, que sur la notion d'accessibilité (réduction des coûts et des temps de transports et de réapprovisionnement dans le strict respect des délais). Les contraintes<br />temporelles de plus en plus impérieuses conduisent alors selon l'expression de C. Raffestin à envisager le passage d'une territorialité régionalisée à une territorialité temporalisée dont les entreprises de messagerie offrent un beau modèle.<br />Dans un contexte de concurrence avivée, l'accroissement des performances logistiques exigé en termes de délais, de coûts et de qualité de service, pousse les prestataires de messagerie à une rapide restructuration spatiale et organisationnelle de leurs réseaux techniques. Prenant appui sur l'étude de cas concrets, la thèse examine les modalités du fonctionnement des réseaux de messagerie et les logiques de leurs dynamiques territoriales.<br />Elle s'attache à mettre en évidence les éléments géographiques structurant l'activité à différentes échelles spatiales et temporelles.
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Revenue and operational impacts of depeaking flights at hub airportsKatz, Donald Samuel 13 November 2012 (has links)
Post deregulation, many U.S. airlines created hubs with banked schedules, however, in the past decade these same airlines began to experiment with depeaking their schedules to reduce costs and improve operational performance. To date there has been little research that has investigated revenue and operational shifts associated with depeaked schedules; yet understanding the trade-offs among revenue, costs, and operational performance at a network level is critical before airlines will consider future depeaking and related congestion-management strategies. This study develops data cleaning and analysis methodologies based on publicly available data that are used to quantify airport-level and network-level revenue and operational changes associated with schedule depeaking. These methodologies are applied to six case studies of airline depeaking over the past decade. Results show that depeaking is associated with revenue per available seat mile (RASM) increasing slower than the rest of the network and the industry as a whole. Depeaking is associated with improved operations for both the depeaking airlines and competitors. Airports benefit from increases in non-aeronautical sales associated with connecting passengers spending more time in the terminal. The underlying reasons driving airlines' scheduling decisions during depeaking vary greatly by case. Results from the study provide insights for airlines that are considering depeaking and the airports which are affected. The results suggest that losses in RASM and no improvement in operations could potentially lead an airline to repeak, and that RASM is prone to fall when a strong competitive threat exists.
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Increasing the efficiency of multi-hub airline networks by means of flexible time-range tickets - An analysis of passenger acceptance, revenue potentials and implications on network designBadura, Felix 12 September 2011 (has links) (PDF)
After the complete liberalization of the airline industry during the 1990s the industry has faced a rapid growth in passenger numbers. This has mainly been caused by the emergence of so-called Low Cost Carrier (LCC) that offer a simplified product (i.e. point-to-point flights without any frills) at a lower cost than traditional Network Carriers. Furthermore LCC also introduced a less differentiated pricing structure (Restriction Free Pricing) which forced competing network carriers to reduce the degree of price discrimination which they were able to practice until then in order to defend their market shares. This has led to a decrease of average yields, which resulted in difficulties for (smaller) Network Carriers to cover their fixed costs, related to the operation of a hub & spoke network. In this environment network airlines are looking for new revenue sources as well as further sources of cost reduction. This development has amplified the consolidation trend of the airline industry and led to the emergence of several multi-hub networks (e.g. Lufthansa runs hub-operation in Frankfurt, Munich, Zurich and Vienna).
One way to leverage the fact that multi-hub networks allow several routings for one origin-destination city pair would be the introduction of flexible tickets, where the actual routing of the passenger is not defined at the moment of purchase but only a certain time prior to departure. This allows airlines to raise the load factor on their network by increasing the degree of overbooking which they currently practice by pooling the risk that more passengers arrive than there is capacity among several flights. Furthermore these tickets might allow network carriers to compete in the low-cost-airline segment without having to further reduce the price level of their regular product (with specified routing).
The present dissertation examined possible designs of such a ticket and their impact on the acceptance by passengers by means of a choice based conjoint study among 356 travelers. The findings suggest that while 77.5% of leisure travelers are willing to accept flexible time-range tickets in their relevant set, only 56% of business travelers are considering using this kind of ticket. More particular the results also showed that business travelers are not willing to compromise on travel duration and departure times, and are subsequently willing to pay a premium for specified tickets. A market share simulation showed that depending on the selected product layout flexible time-range tickets are able to gain up to 60% market share when offered at a discount of up to 33% relative to traditional tickets. When it comes to the actual layout, the largest lever to increase the acceptance is to exclude connection flights from the potential set of flights.
The results contribute to the young research area on flexible products by assessing the disutility which is experienced by customers with regard to particular product characteristics of flexible products. Furthermore the results aim at providing airline managers with a comprehensive overview of the possibilities which flexible time-range tickets bring along when it comes to increasing the load factor and thereby the revenues in a multi-hub network. (author's abstract)
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Design, Development And Manufacturing Of An All Terrain Modular Robot PlatformKul, Mustafa Cihangir 01 May 2010 (has links) (PDF)
The aim of this thesis is to create a flexible multi-purpose modular all terrain robot
platform, which has the potential to be used in commercial applications as well as in
education and research. In developing this robot platform, it is aimed to use readily
available commercial products as much as possible in order to keep the cost of the
product low, increase maintainability, and benefit from the improvements made to
these components in time. The modularity is attained by designing a two wheeled
base module which is autonomous on its own. This base module is composed of
two wheels where, the motors located inside these wheels. It is shown that the
proposed base module facilitates the configuration of various robots to suit the
needs of diverse applications. Detailed design and manufacturing of one of various
possible configurations is presented. Performance tests are conducted on this robot
configuration and effectiveness of the proposed modular approach is justified.
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Desafios na implementação da regionalização da saúde a partir da perspectiva de municípios não-polo: estudo de caso em regiões de saúde desenvolvidasSantos, Carolina Cavanha de Azeredo 23 February 2017 (has links)
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Previous issue date: 2017-02-23 / The research agenda on health regionalization seeks to identify challenges that undermine its effective implementation in a solidary model. In this regard, researches have been prioritizing certain analytical units, such as regulatory marks, States, health regions and Municipalities that are a hub for medium and high complexity health services. In this context, this research sheds light on the peculiarities of a pool of actors that appears marginally in the investigations, but that plays an important role within regional policy dynamic: non-hub municipalities, which are not reference for health services within their region. Therefore, this research has two objectives: first, to structure a taxonomy on the challenges concerning the implementation of health regionalization; secondly, to apply it to non-hub Municipalities, in order to validate them as an analytical unit. The selected methodology to reach these objectives was a case study with non-hub Municipalities of two health regions of the state of Sao Paulo: Metropolitan Region of Campinas and Marilia Region The taxonomy proved to be effective in its use as an analytical research tool: (i) it guided the fieldwork, (ii) supported the comparison of perception among the interviewees and (iii) was central to the broader understanding of regionalization from macro-categories. On the second objective, it became clear that non-hub Municipalities face particular challenges. Thus, this pool of players can be validated as specific analytical unit to understand cooperative regionalization, and deserve special attention when formulating and implementing public policies. Furthermore, we reach to the conclusion that the current regionalization model presents considerable limitations that negatively affect non-hub municipalities. / A agenda de pesquisa sobre regionalização da saúde busca, de modo geral, identificar os desafios que comprometem sua implementação efetiva e solidária. Para tanto, os autores têm priorizado algumas unidades analíticas, como os marcos-regulatórios, os estados, as regiões de saúde em si e os municípios-polo. Nesse contexto, a presente dissertação coloca luz sobre as particularidades de um grupo de atores que aparece de forma marginal nas pesquisas, mas que tem papel importante na dinâmica política regional: os municípios não-polo. Sendo assim, são dois os objetivos da dissertação: primeiro, estruturar uma taxonomia sobre os desafios à implementação da regionalização a partir da revisão da literatura; segundo, aplicá-la aos municípios não-polo, no intuito de validá-los enquanto categoria analítica. Como metodologia para alcançar tais objetivos foi desenvolvido um estudo de caso com municípios não-polo de duas regiões de saúde do estado de São Paulo: Região Metropolitana de Campinas e Região de Marília. A taxonomia mostrou-se efetiva no seu uso como instrumento de pesquisa analítica pois orientou o trabalho em campo, apoiou a comparação de percepção entre os diferentes entrevistados e foi central para o entendimento mais amplo da regionalização a partir de macro-categorias. Sobre o segundo objetivo, tornou-se evidente que os municípios não-polo enfrentam desafios particulares à sua natureza. Sendo assim, estes podem ser validados enquanto categoria analítica específica para entender a regionalização cooperativa, e merecem atenção especial quando da formulação e implementação de políticas públicas. No mais, conclui-se que o atual modelo de regionalização apresenta limitações consideráveis que prejudicam os municípios não-polo.
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Root-knot nematode effectors : key actors of parasitism : functional analysis and protein-protein interaction with host plants / Protéines effectrices de virulence des nématodes à galles : acteurs clés du parasitisme : Analyse fonctionnelle et interactions protéine-protéine avec la plante hôteGrossi De Sa, Maira 13 December 2016 (has links)
Les nématodes à galles (RKN), Meloidogyne spp. sont des petits vers parasites qui infectent les racines des plantes où ils induisent la formation de sites nourriciers. Les RKN sont des endoparasites à large gamme d'hôtes, englobant les principales espèces de plantes cultivées. Meloidogyne javanica, M. graminicola et M. incognita sont les principales espèces parasitant le riz (Oryza sativa). Le succès infectieux des RKN repose sur la production de protéines effecteurs de virulence, secrétées dans leurs glandes oesophagiennes et libérées dans les cellules de la plante hôte par leur stylet. La caractéristique principale des RKN est leur capacité à déréguler des cellules du parenchyme vasculaire pour induire la formation de cellules géantes multinucléées, à haute activité métabolique. Les processus moléculaires sous-jacents restent encore mal connus, alors que l’identification d’effecteurs de virulence et de leurs cibles végétales pourrait fournir de nouvelles perspectives pour le contrôle des RKN. Ainsi, les objectifs de cette étude étaient (1) d’évaluer le rôle de protéines de Meloidogyne sécrétées (MSP) au cours des interactions riz - RKN et (2) d'identifier des cibles des MSP parmi les principales protéines Hub d’Arabidopsis thaliana impliquées dans l'immunité des plantes, afin d'évaluer la fonction putative des MSP dans les cellules hôtes. Pour la première partie de notre étude, nous avons sélectionné trois MSP exprimées dans les glandes oesophagiennes et possiblement sécrétées. L’analyse de l’expression des gènes par RT-qPCR a montré que MSP2 est fortement exprimé dans les premiers stades du cycle du nématode, tandis que MSP18 et MSP19 sont activés au cours du parasitisme dans les racines du riz. Les essais de localisation subcellulaire dans les cellules d'oignon ont identifié le noyau (pour MSP2) et le cytoplasme (pour MSP7 et MSP18) comme compartiments cellulaires ciblés par les protéines du nématode. Des plants de riz (O. sativa Nipponbare) transgéniques ont été produits pour évaluer le rôle des MSP au cours des interactions riz-RKN. Des lignées de riz surexprimant MSP18 ont permis un taux de reproduction plus élevé de M. javanica et M. graminicola. Au contraire, des retards de développement et de reproduction de M. javanica ont été observés sur des lignées de riz exprimant des micro-RNAs capables de supprimer l’expression des gènes MSP2 ou MSP19. Ces données ont montré que MSP2, MSP18 et MSP19 peuvent être des gènes importants pour le parasitisme ou le développement du nématode. Les tests d'expression transitoire dans le tabac (Nicotiana benthamiana) ont montré que MSP18 peut interférer avec la mort cellulaire programmée déclenchée par INF1, ce qui suggère que MSP18 pourrait supprimer les voies de défense des plantes pour faciliter l’infection. Dans une deuxième partie de ce travail, des analyses systématiques en double-hybride chez la levure ont été menées pour vérifier les interactions protéine-protéine entre 6 MSP et 18 protéines Hub d’A. thaliana. Chez la levure, la protéine du nématode MSP400 interagit avec trois protéines Hub, l’Anaphase-Promoting-complex 8 (At-APC8) et les facteurs de transcription At-TCP14 et At-TCP15. L'interaction physique de MSP400 avec At-APC8, un régulateur clé du cycle cellulaire de la plante, a été confirmée in planta par complémentation bimoléculaire de fluorescence (BiFC). Ces résultats démontrent pour la première fois qu'un effecteur de nématode est capable d'interagir directement avec une protéine régulatrice du cycle cellulaire chez la plante, révélant un nouveau mécanisme utilisé par les RKN pour commander la machinerie du cycle de la cellule hôte et induire ainsi la formation du site d'alimentation. Les données obtenues dans cette étude élargissent considérablement notre connaissance des acteurs moléculaires qui contribuent à la pathogénicité des nématodes, mettant en évidence les différents mécanismes exploités par les RKN pour promouvoir la sensibilité des plantes. / Root-knot nematodes (RKN), Meloidogyne spp. are small parasitic worms that infect plant roots where they induce the formation of highly specialized nutrient feeding sites. RKN are endoparasites with a wide host range encompassing major plant crops, impairing effective specific control. Meloidogyne javanica, M. graminicola, and M. incognita are the principal RKN species responsible for rice (Oryza sativa) production losses. Successful plant infection is likely achieved by nematode effector proteins produced in their esophageal gland cells and released into the host plant cells through their stylet. In particular, one of the striking features of RKN is their ability to deregulate vascular parenchyma cells to induce the formation of multinucleated giant cells with a high metabolic activity in the roots. The molecular processes underlying plant-RKN interactions still remain poorly understood. Identification of nematode virulence effectors and their plant targets may provide new insights for developing control strategies towards RKN. Thus, the aims of this study were to (1) assess the role of Meloidogyne secreted proteins (MSP) in rice – RKN interactions and (2) identify MSP targets among the major Arabidopsis thaliana Hub proteins involved in plant immunity, to assess the putative MSP function into host cells. For the first part of our study, we selected three Meloidogyne-genus specific proteins expressed in esophageal glands and predicted to be secreted. Gene expression analysis by RT-qPCR showed that MSP2 is highly expressed in the early stages of the nematode cycle, while MSP18 and MSP19 are up-regulated during parasitism in rice roots. Subcellular localization assays in onion cells identified the nucleus (for MSP2) and cytoplasm (for MSP7 and MSP18) as the main cellular compartments targeted by nematode proteins. Transgenic rice (O. sativa Nipponbare) plants expressing the MSP cDNAs or artificial micro-RNAs (amiRNAs) able to silence MSP genes were used to assess the role of MSPs during rice-RKN interactions. Homozygous transgenic lines were inoculated with pre-parasitic juveniles (J2) and (i) the number and developmental stage of nematodes present in roots after 21 days, (ii) the number of egg masses laid after 28 days and, (iii) the number of next-generation hatched J2 after 45 days were assessed. Rice lines overexpressing MSP18 allowed a higher reproduction rate of M. javanica and M. graminicola. On the contrary, impaired M. javanica development and reproduction was observed in rice lines expressing amiRNAs against MSP2 or MSP19 genes. These data showed that MSP2, MSP18, and MSP19 genes might be important genes for nematode parasitism or development. Transient expression assays in tobacco (Nicotiana benthamiana) revealed that MSP18 interfered with the INF1-triggered programmed cell death, suggesting that MSP18 could suppress the plant defense pathways to facilitate nematode parasitism. In the second part of this work, systematic yeast-two-hybrid paired assays were conducted to check for protein-protein interactions between 6 MSP and 18 A. thaliana Hub proteins. In yeast, the nematode MSP400 protein interacts with three Hub proteins, the Anaphase-Promoting-Complex 8 (At-APC8) and the transcription factors At-TCP14 and At-TCP15. Physical interaction of MSP400 with At-APC8, a key plant cell cycle regulator, was confirmed in planta by bimolecular fluorescence complementation (BiFC) assays. These results demonstrated for the first time that a plant parasitic nematode effector is able to directly interact with a cell cycle regulatory protein, revealing a novel mechanism utilized by RKN to control the host cell cycle machinery and thereby induce feeding site formation. The data obtained in this study significantly broaden our knowledge of the molecular players contributing to nematode pathogenicity, highlighting the different mechanisms exploited by RKN to promote plant susceptibility.
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