Proteomic Analysis of Grape Berry Cell Cultures Reveals that Developmentally Regulated Ripening Related Processes Can Be Studied Using Cultured Cells

The original publication is available at http:/www.plosone.orgBackground: This work describes a proteomics profiling method, optimized and applied to berry cell suspensions to evaluate organ-specific cultures as a platform to study grape berry ripening. Variations in berry ripening within a cluster(s) on a vine and in a vineyard are a major impediment towards complete understanding of the functional processes that control ripening, specifically when a characterized and homogenous sample is required. Berry cell suspensions could overcome some of these problems, but their suitability as a model system for berry development and ripening needs to be established first. Methodology/Principal Findings: In this study we report on the proteomic evaluation of the cytosolic proteins obtained from synchronized cell suspension cultures that were established from callus lines originating from green, véraison and ripe Vitis vinifera berry explants. The proteins were separated using liquid phase IEF in a Microrotofor cell and SDS PAGE. This method proved superior to gel-based 2DE. Principal component analysis confirmed that biological and technical repeats grouped tightly and importantly, showed that the proteomes of berry cultures originating from the different growth/ripening stages were distinct. A total of twenty six common bands were selected after band matching between different growth stages and twenty two of these bands were positively identified. Thirty two % of the identified proteins are currently annotated as hypothetical. The differential expression profile of the identified proteins, when compared with published literature on grape berry ripening, suggested common trends in terms of relative abundance in the different developmental stages between real berries and cell suspensions. Conclusions: The advantages of having suspension cultures that accurately mimic specific developmental stages are profound and could significantly contribute to the study of the intricate regulatory and signaling networks responsible for berry development and ripening. © 2011 Sharathchandra et al.Publishers' Versio

Berry Flesh and Skin Ripening Features in Vitis vinifera as Assessed by Transcriptional Profiling

Background Ripening of fleshy fruit is a complex developmental process involving the differentiation of tissues with separate functions. During grapevine berry ripening important processes contributing to table and wine grape quality take place, some of them flesh- or skin-specific. In this study, transcriptional profiles throughout flesh and skin ripening were followed during two different seasons in a table grape cultivar ‘Muscat Hamburg’ to determine tissue-specific as well as common developmental programs. Methodology/Principal Findings Using an updated GrapeGen Affymetrix GeneChip® annotation based on grapevine 12×v1 gene predictions, 2188 differentially accumulated transcripts between flesh and skin and 2839 transcripts differentially accumulated throughout ripening in the same manner in both tissues were identified. Transcriptional profiles were dominated by changes at the beginning of veraison which affect both pericarp tissues, although frequently delayed or with lower intensity in the skin than in the flesh. Functional enrichment analysis identified the decay on biosynthetic processes, photosynthesis and transport as a major part of the program delayed in the skin. In addition, a higher number of functional categories, including several related to macromolecule transport and phenylpropanoid and lipid biosynthesis, were over-represented in transcripts accumulated to higher levels in the skin. Functional enrichment also indicated auxin, gibberellins and bHLH transcription factors to take part in the regulation of pre-veraison processes in the pericarp, whereas WRKY and C2H2 family transcription factors seems to more specifically participate in the regulation of skin and flesh ripening, respectively. Conclusions/Significance A transcriptomic analysis indicates that a large part of the ripening program is shared by both pericarp tissues despite some components are delayed in the skin. In addition, important tissue differences are present from early stages prior to the ripening onset including tissue-specific regulators. Altogether, these findings provide key elements to understand berry ripening and its differential regulation in flesh and skin.This study was financially supported by GrapeGen Project funded by Genoma España within a collaborative agreement with Genome Canada. The authors also thank The Ministerio de Ciencia e Innovacion for project BIO2008-03892 and a bilateral collaborative grant with Argentina (AR2009-0021). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Peer reviewe

Développer des alternatives à des systèmes de culture Maïs-Blé en polyculture-élevage sans irrigation : enseignements de trois expérimentations conduites dans différentes régions françaises.

Trois systèmes de culture expérimentaux, alternatifs à un système local dominant à base de Maïs-Blé non irrigué, dans un contexte de polyculture-élevage, partagent le même objectif principal de diminuer de 50% l’Indice de Fréquence de Traitement (IFT) par rapport au système local dominant tout en maintenant la rentabilité. Les résultats pluriannuels montrent une diminution de 22 à 50% de l’IFT total, de 55 à 80% de l’IFT hors herbicide, de 12 à 65% de l’IFT herbicide. Les systèmes présentent aussi des risques d’exposition toxique de l’agriculteur plus faibles et des risques de transfert de substances actives vers l’environnement plus faibles. Par ailleurs, leur rentabilité est plus élevée pour l’un, plutôt plus élevée pour le deuxième et plutôt plus faible pour le dernier. Deux voies ont été empruntées pour obtenir ces résultats positifs sur les herbicides et la rentabilité : soit une diversification moyenne de la succession Maïs-Blé combinée à un travail du sol intensif, soit peu de travail du sol associé à une couverture du sol et combinés à une diversification importante de la succession. La maîtrise des adventices qui en résulte est plus ou moins satisfaisante selon les systèmes de culture. L’analyse agronomique des réussites et des échecs de la maîtrise des adventices permet de tirer des enseignements sur les conditions de réussite de chacune des stratégies.Three innovative cropping systems were tested during several years. They aimed to reduce the use of pesticides by 50% compared to local reference cropping systems while being still profitable. Reference [br/] systems were maize-winter wheat rotations in mixed crop-livestock agricultural systems. The use of total pesticides was reduced by 22-50%, i.e. 12-65% for herbicides and 55-80% for other pesticides. Toxic exposure of farmers was lower for innovative systems ; risk of active substance transfer to the environment was also lower. Regarding profitability, results were different from one experiment to another, with higher economic margins for the innovative system for two of them, and lower margin for the last one. Two strategies were employed to achieve these positive results on pesticides use and impacts and on profitability : moderate diversification of crop sequences combined with intensive tillage on one hand versus minimum tillage associated with soil cover and high crop sequences diversification on the other hand. Resulting weed pressure was acceptable or rather acceptable in two out of the three experiments. Agronomic analysis of success and failure cases led to new insights on success conditions for both strategies of weed management

Expérimentation de systèmes de culture innovants : avancées méthodologiques et mise en réseau opérationnelle

L'expérimentation "système de culture" est une étape importante du processus de conceptionévaluation de systèmes de culture. Elle permet de tester au champ la faisabilité technique et la cohérence agronomique des systèmes de culture les plus prometteurs et d'évaluer leurs résultatsagronomiques et techniques, ainsi que leurs performances socio-économiques et environnementales. Elle représente également un lieu d'échanges interdisciplinaires intéressant, pouvant être source d'innovation. Dans cet article, nous décrivons une démarche d'expérimentation "système de culture" et ses principales spécificités méthodologiques dans le domaine des cultures assolées. Ensuite, nous présentons les acquis méthodologiques et les outils développés pour (i) faciliter la mise en oeuvre et la conduite de ce type d'expérimentation, (ii) permettre la mise en réseau opérationnelle d'expérimentations testant, dans des contextes de production divers, des systèmes de culture répondant à différents enjeux. Enfin, nous présentons la structuration et l'organisation de deux réseaux français d'expérimentations "système de culture" : celui du Réseau Mixte Technologique "Systèmes de Culture Innovants" pour les filières Grande Culture et Polyculture-Elevage et celui initié dans le cadre du projet RotAB pour les Grandes Cultures en agriculture biologique sans élevage. Des perspectives de travail et notamment l'analyse transversale des résultats entre expérimentations et la valorisation des résultats et performances des systèmes de culture testés sont abordées

Systèmes de culture innovants : une nouvelle génération de réseau expérimental et de réseau de compétences

International audienceLes conseillers ou les animateurs agricoles doivent de plus en plus accompagner les agriculteurs dans un changement de pratiques et l’innovation, en particulier dans les suites du Grenelle de l’Environnement. Pour leur permettre, ainsi qu’à leurs responsables hiérarchiques, de faire évoluer le métier, une nouvelle intelligibilité de la relation de conseil et d’accompagnement du changement a été produite. Elle est issue d’échanges entre conseillers ou animateurs et chercheurs autour d’expériences vécues et d’analyse de leur activité réalisée par les chercheurs. Elle repose sur la notion clé de « formats ». Ces formats peuvent être institués (le tour de plaine « à chaud », la formation avec intervention d’un expert), en cours d’institution (e.g. la co-construction d’un système de culture, le tour de plaine « à froid ») et leur combinaison dans l’accompagnement du changement en cours de construction (e.g. pour l’animation sur une aire d’alimentation de captage, ou l’accompagnement d’un groupe vers l’atteinte d’un cahier des charges « Grandes Cultures Économes »). Des outils sont proposés pour analyser ce qui perturbe le travail du conseiller ou de l’animateur dans des situations nouvelles et être opérationnels dans la mise en œuvre de nouveaux formats en vue d’aider les agriculteurs dans la transformation de leurs pratiques

Systèmes de culture innovants : une nouvelle génération de réseau expérimental et de réseau de compétences

International audienceLes conseillers ou les animateurs agricoles doivent de plus en plus accompagner les agriculteurs dans un changement de pratiques et l’innovation, en particulier dans les suites du Grenelle de l’Environnement. Pour leur permettre, ainsi qu’à leurs responsables hiérarchiques, de faire évoluer le métier, une nouvelle intelligibilité de la relation de conseil et d’accompagnement du changement a été produite. Elle est issue d’échanges entre conseillers ou animateurs et chercheurs autour d’expériences vécues et d’analyse de leur activité réalisée par les chercheurs. Elle repose sur la notion clé de « formats ». Ces formats peuvent être institués (le tour de plaine « à chaud », la formation avec intervention d’un expert), en cours d’institution (e.g. la co-construction d’un système de culture, le tour de plaine « à froid ») et leur combinaison dans l’accompagnement du changement en cours de construction (e.g. pour l’animation sur une aire d’alimentation de captage, ou l’accompagnement d’un groupe vers l’atteinte d’un cahier des charges « Grandes Cultures Économes »). Des outils sont proposés pour analyser ce qui perturbe le travail du conseiller ou de l’animateur dans des situations nouvelles et être opérationnels dans la mise en œuvre de nouveaux formats en vue d’aider les agriculteurs dans la transformation de leurs pratiques

Effects of water-deficit irrigation on hormonal content and nitrogen compounds in developing berries of Vitis vinifera L. cv. Tempranillo

13 Págs., 3 Figs., 4 Tabls.Water-deficit irrigation to grapevines reduces plant growth, yield, and berry growth, altering the ripening process, all of which may influence fruit composition and wine quality. Therefore, the goals of this study were (1) to investigate the influence of the main endogenous berry hormones, abscisic acid (ABA), indole-3-acetic acid (IAA), salicylic acid (SA), and jasmonic acid (JA), on berry growth and ripening under water-deficit conditions and (2) to analyze changes in fruit composition, specifically N compounds, under water deprivation. The study was carried out using container-grown Tempranillo grapevines grown under controlled conditions in a greenhouse. Two irrigation treatments were imposed: control (well-watered) and sustained deficit irrigation (SDI). Water deficit decreased leaf area and the source-to-sink ratio, reduced yield and berry size, and decreased concentrations of the main phenolic compounds. SDI also modified berry hormonal status. At the pea-size stage, SDI berries had lower IAA and higher JA and SA than nonstressed berries. At veraison (onset of ripening), accumulation of ABA was less accentuated in SDI than in control berries. At harvest, the content of amino acids and free ammonium was low in both treatments but SDI-treated berries showed a significant accumulation of amines. Results suggest that water restrictions to grapevines might be playing a physiological role in reducing berry growth through affecting hormone dynamics, phenolic synthesis, and the berry amino acid content and composition, which could compromise fruit quality. Possible roles of endogenous IAA controlling berry size and endogenous ABA and SA controlling levels of anthocyanins and flavonols at harvest are discussed. © 2013 Springer Science+Business Media New York.This work was supported by Fundación Universitaria de Navarra (2011) and Ministerio de Ciencia e Innovación (MCINN BFU2011-26989), Spain. F. Morales thanks Gobierno de Aragón (A03 research group) for financial support. M. Niculcea was the recipient of a grant from Asociación de Amigos de la Universidad de Navarra.Peer reviewe

Micorrização e indução de quitinases e β-1,3-glucanases e resistência à fusariose em porta-enxerto de videira Mycorrhizal inoculation and induction of chitinases and β-1,3-glucanases and fusarium resistance in grapevine rootstock

O objetivo deste trabalho foi avaliar os níveis de expressão de &#946;-1,3-glucanases e quitinases nos porta-enxertos de videira SO4 e R110, respectivamente suscetível e resistente a Fusarium oxysporum f. sp. herbemontis, bem como avaliar o efeito do fungo micorrízico arbuscular Glomus intraradices no crescimento, na expressão dessas enzimas e na supressão do patógeno no porta-enxerto suscetível. Foram quantificadas as atividades enzimáticas de &#946;-1,3-glucanases e quitinases nas raízes dos porta-enxertos. Mudas do porta-enxerto SO4 receberam inóculos de G. intraradices e F. oxysporum, e foram avaliadas quanto ao crescimento, atividade das duas enzimas e sintomas de doença. As atividades das enzimas nas raízes do porta-enxerto resistente aumentaram entre 0 e 5 dias após a inoculação do patógeno. A atividade de quitinases nas raízes do porta-enxerto suscetível aumentou com a inoculação do fungo micorrízico e do patógeno. A atividade de &#946;-1,3-glucanases foi maior somente com a presença do fungo micorrízico e do patógeno. Videiras com inoculação de G. intraradices apresentaram diminuição nos sintomas de infecção por Fusarium spp., o que indica que o fungo micorrízico promove a indução de quitinases e &#946;-1,3-glucanases especificamente na supressão ou inibição do patógeno.<br>The objective of this work was to evaluate the expression levels of &#946;-1,3-glucanases and chitinases in SO4 and 110 grapevine rootstocks, respectively susceptible and resistant to Fusarium oxysporum f. sp. herbemontis, as well as to evaluate the effect of the arbuscular mycorrhizal fungus Glomus intraradices on plant growth, on enzyme expression and on pathogen suppression in the susceptible rootstock. The enzyme activities of &#946;-1,3-glucanases and chitinases in the rootstocks roots were evaluated. Plant growth, enzyme activity, and disease symptoms were evaluated in SO4 plantlets inoculated with G. intraradices and F. oxysporum. Enzyme activities in the plant roots increased in the resistant rootstock, between 0 and 5 days after pathogen inoculation. In the susceptible rootstock, chitinase activity enhanced after the mycorrhizal fungus and the pathogen inoculations. &#946;-1,3-glucanase activity increased only in the presence of both the mycorrhizal and the pathogenic fungi. G. intraradices-inoculated SO4 grapevine plantlets showed decreased symptoms of Fusarium infection, which indicates that the mycorrhizal fungus promotes chitinase and &#946;-1,3-glucanase activities leading to pathogen suppression or inhibition