Biology subjects

Romieu, C.

Publications and source records attributed to Romieu, C..

9 recordsLinked to original sources

bioRxiv2024

Near-infrared real time non-destructive monitoring of sugar accumulation reveals that single berries ripen two times faster than previously documented on standard asynchronous samples

Understanding how climate change impacts berry ripening physiology is essential for selecting genotypes that balance sugars and acids under warming conditions. In this context, we used a portable near-infrared spectrometer in the vineyard, to monitor sugar and acid evolution in individual berries from 10 grapevine varieties over two years. Spectra were periodically acquired on the same berries all along ripening, and a subset of these berries was also collected for sugars and organic acids quantification by HPLC, to train partial least square regression models. Prediction models for glucose, fructose, and malic acid concentrations were characterized by validation R{superscript 2} of 0.71, 0.64, and 0.55, respectively. We further used these models to follow sugar accumulation in individual berries and observed that single berries ripen two times faster than found in samples composed of multiple berries. Our results pave avenues toward precise quantitative approaches on sugar and acid fluxes in berry ripening studies.

plant biology↗

bioRxiv2024

Time-resolved transcriptomic of single V. vinifera fruits: membrane transports as switches of the double sigmoidal growth

By revealing that the grape berry loses one H+ per accumulated sucrose at the inception of ripening, adopting a single fruit paradigm elucidates the fundamentals of the malate-sugar nexus, previously obscured by asynchrony in population-based models of ripening. More broadly, the development of the individual fruit was revisited from scratch to capture the simultaneous changes in gene expression and metabolic fluxes in a kinetically relevant way from flowering to overripening. Dynamics in water, tartrate, malate, hexoses, and K+ fluxes obtained by combining individual single fruit growth and concentration data allowed to define eleven sub-phases in fruit development, which distributed on a rigorous curve in RNAseq PCA. WGCNA achieved unprecedented time resolutions in exploring transcript level-metabolic rate associations. A comprehensive set of membrane transporters was found specifically expressed during the first growth phase related to vacuolar over-acidification. Unlike in slightly more acidic citrus, H+ V-PPase transcripts were predominantly expressed, followed by V-ATPase and PH5, clarifying the thermodynamic limit beyond which replacement by the PH1/PH5 complex turns compulsory. Puzzlingly, bona fide ALMT kept a low profile at this stage, possibly replaced by a predominating uncharacterized anion channel. Then, the switch role of HT6 in sugar accumulation was confirmed, electroneutralized by malate vacuolar leakage and H+ pumps activation. HighlightsTo alleviate asynchronicity biases, transcripts showing strict coincidental timing with pericarp physiological phases were disentangled on single berries, enlightening the tight multifaceted membrane developmental control of sugar and acid fluxes.

plant biology↗

bioRxiv2024

The single-berry metabolomic clock paradigm reveals new stages and metabolic switches during grapevine berry development

O_LIAsynchronous development of berries causes metabolic chimerism in usual samples. We thus revisited the developmental changes in the metabolome of the Vitis vinifera single berries from anthesis to over-ripening. C_LIO_LIA dataset of 9,256 ions obtained by non-targeted ultra-performance liquid chromatography coupled to high-resolution mass spectrometry was submitted to an analysis workflow combining classification and dimension reduction tools, to reveal the dynamics of metabolite composition without phenological a priori. C_LIO_LIThis approach led to a metabolome-based definition of developmental stages, as well as the clustering of metabolites into 12 specific kinetic patterns. The single berry intrinsic metabolomic clock alleviates constitutive asynchronicity biases in the usual combination of phenological scales and observer clock. Such increase in temporal resolution enabled the identification of metabolite clusters annunciative of the onset of ripening since the herbaceous plateau. In particular, these clusters included transient lipidic changes and the start of ABA accumulation. We also highlighted a cluster of stilbenes that accumulate after sugar loading stops, during fruit shriveling. C_LIO_LIThis non-targeted approach enables a more precise and unbiased characterization of grapevine berry development through the metabolomic clock paradigm. The discovery of new metabolic milestones of berry development paves the way towards an unbiased assessment of berry physiological stages. C_LI

plant biology↗

bioRxiv2023

Impact of water deficit on single grapevine berry ripening

The effect of water deficit on grapevine fruit ripening has most often been addressed under the assumption that individual berries behave identically to their blend in the future harvest, both kinetically and metabolically. However, mixing unsynchronized berries, whose water and sucrose import pathways critically change according to their own developmental stages, intrinsically blurs the physiological and phenological effects of stress. We investigated the consequences of water deprivation on berry growth and primary metabolites content (glucose, fructose, tartaric and malic acids) on sixteen genetically distant genotypes of Vitis vinifera and fungus-tolerant hybrids submitted to 10 watering regimes, from well-watered to partial leaf shedding. Then, six genotypes were selected for comprehensive single berry analyses. Own-rooted potted plants bearing berries at the late herbaceous plateau stage were subjected to the different water treatments for four weeks in a greenhouse with automated regulation of soil water content. Berry and cluster growth were monitored by image analysis, before performing a final destructive sampling to determine berry weight and composition. Grape phenology was highly dependent on water availability. In some cultivars, ripening was considerably delayed or even prevented under well-watered conditions. These cultivars required an intermediate water deficit to trigger the second berry growth period along with sugar accumulation and malate breakdown, typical of the ripening process. Ripening still occurred in all genotypes upon severe water deprivation, although sugar accumulation and concentration were dramatically impaired, and the second growth period was annihilated or even replaced by shrivelling. Water deficit increased malate breakdown, uncoupling it from sugar accumulation. Single berry analyses suggest that although asynchronicity in berry ripening was reduced upon stress, individual fruits within the same cluster may undergo heterogeneous water budgets, expansion or shrivelling.

physiology↗

bioRxiv2023

Enhancing grapevine breeding efficiency through genomic prediction and selection index

Grapevine (Vitis vinifera) breeding reaches a critical point. New cultivars are released every year with resistance to powdery and downy mildews. However, the traditional process remains time-consuming, taking 20 to 25 years, and demands the evaluation of new traits to enhance grapevine adaptation to climate change. Until now, the selection process has relied on phenotypic data and a limited number of molecular markers for simple genetic traits such as resistance to pathogens, without a clearly defined ideotype and was carried out on a large scale. To accelerate the breeding process and address these challenges, we investigated the use of genomic prediction, a methodology using molecular markers to predict genotypic values. In our study, we focused on two existing grapevine breeding programs: Rose wine and Cognac production. In these programs, several families were created through crosses of emblematic and inter-specific resistant varieties to powdery and downy mildews. 30 traits were evaluated for each program, using two genomic prediction methods: GBLUP (Genomic Best Linear Unbiased Predictor) and LASSO (Least Absolute Shrinkage Selection Operator). The results revealed substantial variability in predictive abilities across traits, ranging from 0 to 0.9. These discrepancies could be attributed to factors such as trait heritability and trait characteristics. Moreover, we explored the potential of across-population genomic prediction by leveraging other grapevine populations as training sets. Integrating genomic prediction allowed us to identify superior individuals for each program, using multivariate selection index method. The ideotype for each breeding program was defined collaboratively with representatives from the wine-growing sector.

genetics↗

bioRxiv2023

Ripening dynamics revisited: an automated method to track the development of asynchronous berries on time-lapse images

BackgroundGrapevine berries undergo asynchronous growth and ripening dynamics within the same bunch. Due to the lack of efficient methods to perform sequential non-destructive measurements on a representative number of individual berries, the genetic and environmental origins of this heterogeneity, as well as its impacts on both vine yield and wine quality, remain nearly unknown. To address these limitations, we propose to track the growth and coloration kinetics of individual berries on time-lapse images of grapevine bunches. ResultFirst, a deep-learning approach is used to detect berries with at least 50{+/-}10% of visible contours, and infer the shape they would have in the absence of occlusions. Second, a tracking algorithm was developed to assign a common label to shapes representing the same berry along the time-series. Training and validation of the methods were performed on challenging image datasets acquired in a robotised high-throughput phenotyping platform. Berries were detected on various genotypes with a F1-score of 91.8%, and segmented with a mean absolute error of 4.1% on their area. Tracking allowed to label and retrieve the temporal identity of more than half of the segmented berries, with an accuracy of 98.1%. This method was used to extract individual growth and colour kinetics of various berries from the same bunch, allowing us to propose the first statistically relevant analysis of berry ripening kinetics, with a time resolution lower than one day. ConclusionsWe successfully developed a fully-automated open-source method to detect, segment and track overlapping berries in time-series of grapevine bunch images. This makes it possible to quantify fine aspects of individual berry development, and to characterise the asynchrony within the bunch. The interest of such analysis was illustrated here for one genotype, but the method has the potential to be applied in a high throughput phenotyping context. This opens the way for revisiting the genetic and environmental variations of the ripening dynamics. Such variations could be considered both from the point of view of fruit development and the phenological structure of the population, which would constitute a paradigm shift.

plant biology↗

bioRxiv2022

The sugarless grape trait characterized by single berry phenotyping

For grape production, an important driver for the selection of varieties better adapted to climate fluctuations, especially warming, is the balance between fruit sugars and acidity. Since the past decades, temperature during ripening has constantly raised causing excessive sugars concentrations and insufficient acidity of the wine grapes in warmest regions. There is thus an increasing interest in breeding new cultivars, able to ripen at lower sugar concentration while preserving fruit acidity. However, the phenotyping of berry composition challenges both methodological and conceptual issues. Indeed, most authors predetermine either average harvest date, ripening duration, thermal time or even hexoses concentration threshold itself, to compare accessions at an hopefully similar ripe stage. Here, we have phenotyped the fruit development and composition of 6 genotypes, including 3 new disease-tolerant varieties known to produce wines with low alcoholic contents. The study was performed at single berry level from the end of green growth stage to the arrest of phloem unloading, when water and solute contents reach a maximum per berry. The results confirmed that sugarless genotypes achieve fruit ripening with 20-30% less hexoses than classical varieties, Grenache N and Merlot N, without impacting berry growth, total acidity or cations accumulation. Sugarless genotypes displayed a higher malic acid/tartaric acid balance than other genotypes with similar sucrose/H+ exchanges at the onset of ripening. Data suggest that sugarless phenotype results from a specific plasticity in the relationship between growth and the turgor imposed by organic acid accumulation and sugar loading. This opens interesting perspectives to understand the mechanism of grapevine berry growth and to breed varieties better coping with climate warming.

plant biology↗

bioRxiv2021

Transcripts repressed at the stop of phloem unloading highlight the energy efficiency of sugar import in the ripening V. vinifera fruit.

Transcriptomic changes at the cessation of sugar accumulation in the pericarp of Vitis vinifera were addressed on single berries re-synchronized according to their individual growth patterns. The net rates of water, sugars and K+ accumulation inferred from individual growth and solute concentration confirmed that these inflows stopped simultaneously in the ripe berry, while the small amount of malic acid remaining at this stage was still being oxidized at low rate. Re-synchronized individual berries displayed negligible variations in gene expression among triplicates. RNA-Seq studies revealed sharp reprogramming of cell wall enzymes and structural proteins at the stop of phloem unloading, associated with an 80% repression of multiple sugar transporters and aquaporins on the plasma or tonoplast membranes, with the noticeable exception of H+/sugar symporters, that were rather weakly and constitutively expressed. This was verified in three genotypes placed in contrasted thermo-hydric conditions. The prevalence of SWEET suggests that electrogenic transporters would play a minor role on the plasma membranes of SE/CC complex and the one of the flesh, while sucrose/H+ exchangers dominate on its tonoplast. Cis-regulatory elements present in their promoters allowed to sort these transporters in different groups, also including specific TIPs and PIPs paralogs, and cohorts of cell wall related genes. Together with simple thermodynamic considerations, these results lead to propose that H+/sugar exchangers at the tonoplast, associated with a considerably acidic vacuolar pH, may exhaust cytosolic sugars in the flesh and alleviate the need for supplementary energization of sugar transport at the plasma membrane.

plant biology↗

bioRxiv2020

Genome-wide association and prediction studies using a grapevine diversity panel give insights into the genetic architecture of several traits of interest

To cope with the challenges faced by agriculture, speeding-up breeding programs is a worthy endeavor, especially for perennials such as grapevine, but requires understanding the genetic architecture of target traits. To go beyond the mapping of quantitative trait locus (QTL) in bi-parental crosses, we exploited a diverse panel of 279 Vitis vinifera L. cultivars. This panel planted in five blocks in the vineyard was phenotyped over several years for 127 traits including yield components, organic acids, aroma precursors, polyphenols, and a water stress indicator. The panel was genotyped for 63k single nucleotide polymorphisms (SNPs) by combining an 18K microarray and genotyping-by-sequencing (GBS). The experimental design allowed to reliably assess the genotypic values for most traits. Marker densification via GBS markedly increased the proportion of genetic variance explained by SNPs, and two multi-SNP models identified QTLs not found by a SNP-by-SNP model. Overall, 489 reliable QTLs were detected for 41% more response variables than by a SNP-by-SNP model with microarray-only SNPs, many new ones compared to the results from bi-parental crosses. Prediction accuracy higher than 0.42 was obtained for 50% of the response variables. Our overall approach as well as QTL and prediction results provide insights into the genetic architecture of target traits. New candidate genes and the application in breeding are discussed.

genetics↗