Day and night heat stress trigger different transcriptomic responses in green and ripening grapevine (vitis vinifera) fruit

Background: Global climate change will noticeably affect plant vegetative and reproductive development. The recent increase in temperatures has already impacted yields and composition of berries in many grapevine-growing regions. Physiological processes underlying temperature response and tolerance of the grapevine fruit have not been extensively investigated. To date, all studies investigating the molecular regulation of fleshly fruit response to abiotic stress were only conducted during the day, overlooking possible critical night-specific variations. The present study explores the night and day transcriptomic response of grapevine fruit to heat stress at several developmental stages. Short heat stresses (2 h) were applied at day and night to vines bearing clusters sequentially ordered according to the developmental stages along their vertical axes. The recently proposed microvine model (DRCF-Dwarf Rapid Cycling and Continuous Flowering) was grown in climatic chambers in order to circumvent common constraints and biases inevitable in field experiments with perennial macrovines. Post-véraison berry heterogeneity within clusters was avoided by constituting homogenous batches following organic acids and sugars measurements of individual berries. A whole genome transcriptomic approach was subsequently conducted using NimbleGen 090818 Vitis 12X (30 K) microarrays. Results: Present work reveals significant differences in heat stress responsive pathways according to day or night treatment, in particular regarding genes associated with acidity and phenylpropanoid metabolism. Precise distinction of ripening stages led to stage-specific detection of malic acid and anthocyanin-related transcripts modulated by heat stress. Important changes in cell wall modification related processes as well as indications for heat-induced delay of ripening and sugar accumulation were observed at véraison, an effect that was reversed at later stages. Conclusions: This first day - night study on heat stress adaption of the grapevine berry shows that the transcriptome of fleshy fruits is differentially affected by abiotic stress at night. The present results emphasize the necessity of including different developmental stages and especially several daytime points in transcriptomic studies

Is transcriptomic regulation of berry development more important at night than during the day?

Diurnal changes in gene expression occur in all living organisms and have been studied on model plants such as Arabidopsis thaliana. To our knowledge the impact of the nycthemeral cycle on the genetic program of fleshly fruit development has been hitherto overlooked. In order to circumvent environmental changes throughout fruit development, young and ripening berries were sampled simultaneously on continuously flowering microvines acclimated to controlled circadian light and temperature changes. Gene expression profiles along fruit development were monitored during both day and night with whole genome microarrays (Nimblegen® vitis 12x), yielding a total number of 9273 developmentally modulated probesets. All day-detected transcripts were modulated at night, whereas 1843 genes were night-specific. Very similar developmental patterns of gene expression were observed using independent hierarchical clustering of day and night data, whereas functional categories of allocated transcripts varied according to time of day. Many transcripts within pathways, known to be up-regulated during ripening, in particular those linked to secondary metabolism exhibited a clearer developmental regulation at night than during the day. Functional enrichment analysis also indicated that diurnally modulated genes considerably varied during fruit development, with a shift from cellular organization and photosynthesis in green berries to secondary metabolism and stress-related genes in ripening berries. These results reveal critical changes in gene expression during night development that differ from daytime development, which have not been observed in other transcriptomic studies on fruit development thus far. © 2014 Rienth et al.This work is part of the DURAVITIS program which is financially supported by the ANR (Agence national de la recherche) -Genopole (project ANR-2010-GENM-004-01) and the Jean Poupelain foundation (30 Rue Gâte Chien, 16100 Javrezac, France).Peer Reviewe

Identification of stable QTLs for vegetative and reproductive traits in the microvine (Vitis vinifera L.) using the 18 K Infinium chip

UMR AGAP - équipe DAAV - Diversité, adaptation et amélioration de la vigne[b]Background[/b] [br/]The increasing temperature associated with climate change impacts grapevine phenology and development with critical effects on grape yield and composition. Plant breeding has the potential to deliver new cultivars with stable yield and quality under warmer climate conditions, but this requires the identification of stable genetic determinants. This study tested the potentialities of the microvine to boost genetics in grapevine. A mapping population of 129 microvines derived from Picovine x Ugni Blanc flb, was genotyped with the Illumina® 18 K SNP (Single Nucleotide Polymorphism) chip. Forty-three vegetative and reproductive traits were phenotyped outdoors over four cropping cycles, and a subset of 22 traits over two cropping cycles in growth rooms with two contrasted temperatures, in order to map stable QTLs (Quantitative Trait Loci). [br/][b]Results[/b] [br/]Ten stable QTLs for berry development and quality or leaf area were identified on the parental maps. A new major QTL explaining up to 44 % of total variance of berry weight was identified on chromosome 7 in Ugni Blanc flb, and co-localized with QTLs for seed number (up to 76 % total variance), major berry acids at green lag phase (up to 35 %), and other yield components (up to 25 %). In addition, a minor QTL for leaf area was found on chromosome 4 of the same parent. In contrast, only minor QTLs for berry acidity and leaf area could be found as moderately stable in Picovine. None of the transporters recently identified as mutated in low acidity apples or Cucurbits were included in the several hundreds of candidate genes underlying the above berry QTLs, which could be reduced to a few dozen candidate genes when a priori pertinent biological functions and organ specific expression were considered. [br/][b]Conclusions[/b] [br/]This study combining the use of microvine and a high throughput genotyping technology was innovative for grapevine genetics. It allowed the identification of 10 stable QTLs, including the first berry acidity QTLs reported so far in a Vitis vinifera intra-specific cross. Robustness of a set of QTLs was assessed with respect to temperature variatio

Réponses développementales et physiologiques de la Microvigne aux températures élevées : caractérisation de l’effet sur le bilan carboné et son implication dans l’avortement précoce des organes reproducteurs.

Global warming is likely to be a major issue for grapevine yield sustainability in the next years. A common hypothesis is that long-term elevated temperatures may cause the failure of key phases of reproductive development, through their negative impact on carbon balance. However, testing the specific role of plant carbon status on yield elaboration under elevated temperatures is difficult on perennial crops, such as grapevine, and when environment fluctuates. To overcome these difficulties, the present work was conducted under fully controlled environment using a natural, giberrelic acid insensitive mutant of grapevine called Microvine, as a new model for grapevine genetics and physiological studies. This mutant present a dwarf stature and lack of juvenile characteristics leading to the continuous flowering and fruiting even during the first year after sowing. The present study was aimed at (i) developing a framework of analysis for Microvine; (ii) describing vegetative and reproductive developments responses to temperature elevation at whole plant level, and more precisely on early reproductive development abortions; (iii) characterizing grapevine carbon balance responses to temperature; and finally (iv) evaluating carbon balance implication in temperature-dependent early reproductive development abortions. This work shows that complete inflorescence abortions observed in response to temperature were related to carbon status, either expressed though total carbohydrate, or through the level of reserves at the whole plant level. This study opens the way to use Microvine as model to address the impact of climate warming on grapevine vegetative and reproductive developments.Dans le contexte de réchauffement climatique, le maintien des rendements est un enjeu majeur pour la filière viticole. L'augmentation des températures impacte négativement certaines phases clés du développement reproducteur en induisant notamment des phénomènes de coulure ou de pertes précoces d'inflorescence. L'altération du bilan carboné pourrait être un facteur majeur des diminutions de rendements en réponse aux températures élevées et l'évaluation de cette hypothèse permettrait d'ajuster les pratiques pour limiter les effets du réchauffement climatique. Si l'implication du bilan de carbone chez les plantes annuelles semble être à la base des baisses de rendement observées ces dernières années, les choses sont beaucoup moins claires chez des pérennes comme la vigne du fait des très nombreux co-facteurs possible ainsi que de la difficulté à expérimenter dans des conditions totalement contrôlées. Afin de s'affranchir de ces contraintes, notre étude a été réalisée en conditions contrôlées sur la Microvigne, un mutant naturel insensible à l'acide gibberelique et un nouveau modèle d'étude pour la génétique et la physiologie de la vigne. Ce mutant présente un phénotype nain et une floraison et fructification continues le long de l'axe proleptique. L'approche utilisée dans cette étude a consisté à (i) développer un cadre d'analyse adapté à la Microvigne; (ii) décrire l'impact de l'élévation de la température, sur les développements végétatif et reproducteur de la vigne, et plus particulièrement l'avortement des stades précoces du développement reproducteur; (iii) caractériser les changements du bilan de carbone de la Microvigne induits par la contrainte thermique, et enfin (iv) évaluer l'implication potentielle du bilan de carbone dans les phénomènes d'avortement précoces en réponse au stress thermique. Ce travail a permis de mettre en évidence des phénomènes d'avortement complet d'inflorescences en réponse à l'élévation des températures. Par ailleurs, nous avons pu montrer un rôle du statut carboné dans ce phénomène, soit via les teneurs totales en carbohydrates, soit via l'état global des réserves à l'échelle de la plante. Ce travail montre les potentialités su modèle Microvigne et ouvre de nouvelles perspectives pour l'analyse des effets des contraintes abiotiques sur le développement végétatif et reproducteur de la vigne

Exploring the Microvine developmental and physiological adaptation mechanisms to elevated temperature : characterization of the impact on carbon balance and its involvement in early reproductive development failure

Dans le contexte de réchauffement climatique, le maintien des rendements est un enjeu majeur pour la filière viticole. L'augmentation des températures impacte négativement certaines phases clés du développement reproducteur en induisant notamment des phénomènes de coulure ou de pertes précoces d'inflorescence. L'altération du bilan carboné pourrait être un facteur majeur des diminutions de rendements en réponse aux températures élevées et l'évaluation de cette hypothèse permettrait d'ajuster les pratiques pour limiter les effets du réchauffement climatique. Si l'implication du bilan de carbone chez les plantes annuelles semble être à la base des baisses de rendement observées ces dernières années, les choses sont beaucoup moins claires chez des pérennes comme la vigne du fait des très nombreux co-facteurs possible ainsi que de la difficulté à expérimenter dans des conditions totalement contrôlées. Afin de s'affranchir de ces contraintes, notre étude a été réalisée en conditions contrôlées sur la Microvigne, un mutant naturel insensible à l'acide gibberelique et un nouveau modèle d'étude pour la génétique et la physiologie de la vigne. Ce mutant présente un phénotype nain et une floraison et fructification continues le long de l'axe proleptique. L'approche utilisée dans cette étude a consisté à (i) développer un cadre d'analyse adapté à la Microvigne; (ii) décrire l'impact de l'élévation de la température, sur les développements végétatif et reproducteur de la vigne, et plus particulièrement l'avortement des stades précoces du développement reproducteur; (iii) caractériser les changements du bilan de carbone de la Microvigne induits par la contrainte thermique, et enfin (iv) évaluer l'implication potentielle du bilan de carbone dans les phénomènes d'avortement précoces en réponse au stress thermique. Ce travail a permis de mettre en évidence des phénomènes d'avortement complet d'inflorescences en réponse à l'élévation des températures. Par ailleurs, nous avons pu montrer un rôle du statut carboné dans ce phénomène, soit via les teneurs totales en carbohydrates, soit via l'état global des réserves à l'échelle de la plante. Ce travail montre les potentialités su modèle Microvigne et ouvre de nouvelles perspectives pour l'analyse des effets des contraintes abiotiques sur le développement végétatif et reproducteur de la vigne.Global warming is likely to be a major issue for grapevine yield sustainability in the next years. A common hypothesis is that long-term elevated temperatures may cause the failure of key phases of reproductive development, through their negative impact on carbon balance. However, testing the specific role of plant carbon status on yield elaboration under elevated temperatures is difficult on perennial crops, such as grapevine, and when environment fluctuates. To overcome these difficulties, the present work was conducted under fully controlled environment using a natural, giberrelic acid insensitive mutant of grapevine called Microvine, as a new model for grapevine genetics and physiological studies. This mutant present a dwarf stature and lack of juvenile characteristics leading to the continuous flowering and fruiting even during the first year after sowing. The present study was aimed at (i) developing a framework of analysis for Microvine; (ii) describing vegetative and reproductive developments responses to temperature elevation at whole plant level, and more precisely on early reproductive development abortions; (iii) characterizing grapevine carbon balance responses to temperature; and finally (iv) evaluating carbon balance implication in temperature-dependent early reproductive development abortions. This work shows that complete inflorescence abortions observed in response to temperature were related to carbon status, either expressed though total carbohydrate, or through the level of reserves at the whole plant level. This study opens the way to use Microvine as model to address the impact of climate warming on grapevine vegetative and reproductive developments

The microvine, a model to study the effect of temperature on grapevine latent bud development and fruitfulness

International audienceAim: The success of inflorescence primordia initiation and differentiation within latent buds (i.e. bud fruitfulness) is a critical issue for grapevine yield sustainability under climate change. The aim of the present study was to track the timing and rate of inflorescence development in latent buds along the cane and to quantify their responses to elevated day/night (D/N) temperatures. Methods and Results: The experiments were conducted under controlled conditions, using the microvine model, which is suitable for establishment in small areas. Two imagery methods for analyzing bud anatomy were assessed: light microscopy and x-ray microtomography. Light microscopy was laborious, but it was the most accurate method for investigating organogenesis in the primordial shoot of the latent bud. In plants grown in a greenhouse (D/N, 25°C/15°C), the number of phytomer primordia in latent buds increased linearly from the apical to the basal buds on the cane. A maximum of six phytomers and two inflorescence primordia were observed beneath the 20th bud position that is, slightly fewer than usually reported with macrovines. The first and second inflorescences started to differentiate at the 14th and 18th bud position, respectively. Temperature increases in the growth chamber (D/N, 20–30°C/15–25°C) only slightly changed the final number of preformed phytomers and the probability of inflorescence primordia differentiation per bud. However, elevated temperature sharply accelerated and thereby shortened development of the latent bud primordial shoot, resulting in differentiation of the first inflorescence primordia straight from the fifth bud position. Based on the spatiotemporal conversion of bud position into thermal time, the first inflorescence started to differentiate 332 growing degree days (°Cd) (or 41 days) after bud emergence at D/N 20°C/15°C, and only 98°Cd (or 5 days) after bud emergence at D/N 35°C/25°C. Finally, the number of preformed phytomers was shown to correlate with primary bud length and cane diameter, independent of temperature. These easily measured variables may be used as indicators of bud developmental stage and potential bud fruitfulness in further studies using the microvine. Conclusions: The microvine appears to be suitable for parameterizing a developmental model of grapevine latent buds under controlled environmental conditions and when evaluating the response to elevated D/N temperatures. Significance and impact of the study: The precise description of the timing and rate of differentiation of phytomers and inflorescences opens new perspectives for understanding the molecular processes underlying the response of bud fruitfulness to environmental constraints

In the microvine pericarp, critical transcriptome changes occur during the night

Póster presentado en la 2nd Annual ConferenceFleshy Fruit Development & Ripening (COST Action FA1106 QualityFruit), celebrada en Chania (Grecia) del 22 al 25 de septiembre de 2013.Many transcriptomic studies have been conducted on different vitis vinifera L. cultivars to better understand physiological changes occurring d uring berry development (Lijavetzky et al., 2012, Terrier et al., 2005, Pilati et al., 200 7). To our knowledge very few time course studies, assessing both day and night transcriptome , have been performed on any fleshy fruits until now. Still, it was demonstrated on mod el plants such as Arabidopsis thaliana (Schaffer et al., 2001, Chow and Kayb, 2013) that i mportant changes in gene expression following diurnal patterns occur in all living orga nisms and might influence organoleptic quality of the grapes. With the macrovine, which pr esents an annual reproductive cycle, studying day and night fruit development under cont rolled conditions is almost impossible. In our study, we useed the microvine (DRCF-mutant; Boss and Thomas, 2002) a recently proposed grapevine model (Chaib et al., 2010), whic h is a dwarf and present a rapid cycling and a continuous production of fruits. Gene express ion profile was studied on two pre - and two post-véraison stages of berry development using a whole genome microarray assay (Nimblegen® vitis 12x). Equivalent developmental st ages were sampled at day and night time. A total number of 9273 probesets were develop mentally modulated between all stages day and night with a high number of genes be ing modulated specifically at night (1755). Eight very similar clusters of gene express ion profiles could be obtained independently for day and night development. Our st udy allowed highlighting development stage specifics mechanisms of diurnal modulation of genes in the fruit that have been overlooked in other transcriptomic studies on fruit development so far. In the green berry functional enrichment analysis indicated major chan ges in cellular organization and photosynthesis whereas in the ripe berry mainly sec ondary metabolism was affected during the night.N

The carbon balance of (micro)vine plants is decreased under elevated temperature

UMR AGAP équipe DAAVClimate change results in crops being exposed to higher temperatures. Understanding how this affects production is key to designing resilient genotypes and adapted practices. Microvine, a mutant of grapevine, is a powerful model because of its small size and continuous fruiting phenotype. This study was thus performed to assess the impact of elevated temperatures on carbon balance using both photosynthesis and respiration measurement as well as growth estimations.One-year old potted Microvines were installed in a growth chamber. A multiplexed, whole plant gas-exchange chamber was designed and used in parallel with single leaf data cuvettes. Temperature steps of 48 hours were applied during the day (20-35°C) and night (15-30°C).Remarkably, neither night, nor day temperature had an effect on day photosynthesis and night respiration respectively. Higher temperature reduced photosynthesis with similar relationships both at the leaf and at the whole plant level, independently of the leaf age. By contrast, elevated temperature increased night respiration but with strong dependency on the leaf age and with lower values when compared to whole plant measurements. The respiration of stem and trunk were likely responsible for these discrepancies. Based on these data plus growth estimates from leaf development rate and leaf dry matter content, a carbon balance model was derived. It showed an average 4.6% and 3.3% reduction of the C balance every 2°C increase during the day and during the night respectively. Our results clearly highlight the deleterious effect of elevated temperature on grapevine C balance with potential negative impact on berry set and sugar accumulation