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SummaryExperiments conducted under controlled conditions in vineyards and growth chambers demonstrated day- and night-specific responses of grape berry organic acid levels through altered TCA cycle and amino acid metabolism.

BackgroundVitis vinifera berry development is characterised by an initial phase where the fruit is small, hard and acidic, followed by a lag phase known as veraison. In the final phase, berries become larger, softer and sweeter and accumulate an array of organoleptic compounds. Since the physiological and biochemical makeup of grape berries at harvest has a profound impact on the characteristics of wine, there is great interest in characterising the molecular and biophysical changes that occur from flowering through veraison and ripening, including the coordination and temporal regulation of metabolic gene pathways. Advances in deep-sequencing technologies, combined with the availability of increasingly accurate V. vinifera genomic and transcriptomic data, have enabled us to carry out RNA-transcript expression analysis on a global scale at key points during berry development.ResultsA total of 162 million 100-base pair reads were generated from pooled Vitis vinifera (cv. Shiraz) berries sampled at 3-weeks post-anthesis, 10- and 11-weeks post-anthesis (corresponding to early and late veraison) and at 17-weeks post-anthesis (harvest). Mapping reads from each developmental stage (36-45 million) onto the NCBI RefSeq transcriptome of 23,720 V. vinifera mRNAs revealed that at least 75% of these transcripts were detected in each sample. RNA-Seq analysis uncovered 4,185 transcripts that were significantly upregulated at a single developmental stage, including 161 transcription factors. Clustering transcripts according to distinct patterns of transcription revealed coordination in metabolic pathways such as organic acid, stilbene and terpenoid metabolism. From the phenylpropanoid/stilbene biosynthetic pathway at least 46 transcripts were upregulated in ripe berries when compared to veraison and immature berries, and 12 terpene synthases were predominantly detected only in a single sample. Quantitative real-time PCR was used to validate the expression pattern of 12 differentially expressed genes from primary and secondary metabolic pathways.ConclusionsIn this study we report the global transcriptional profile of Shiraz grapes at key stages of development. We have undertaken a comprehensive analysis of gene families contributing to commercially important berry characteristics and present examples of co-regulation and differential gene expression. The data reported here will provide an invaluable resource for the on-going molecular investigation of wine grapes.

The growth and development of plants can be limited by environmental stresses such as salinity. It has been suggested that the non-phosphorylating alternative respiratory pathway in plants, mediated by the NAD(P)H dehydrogenase [NAD(P)H DH] and alternative oxidase (AOX), is important during environmental stresses. The involvement of this alternative pathway in a stress response may be linked to its capacity to uncouple carbon metabolism from adenylate control and/or the minimization of the formation of destructive reactive oxygen species (ROS). Salinity stress is a widespread, adverse environmental stress, which leads to an ionic imbalance, hyperosmotic stress and oxidative stress, the latter being the result of ROS formation. In this study, we show that salinity stress of Arabidopsis thaliana plants resulted in the formation of ROS, increased levels of Na+ in both the shoot and the root and an increase in transcription of Ataox1a, Atndb2 and Atndb4 genes, indicating the formation of an abridged non-phosphorylating electron transport chain in response to salinity stress. Furthermore, plants constitutively over-expressing Ataox1a, with increased AOX capacity, showed lower ROS formation, 30-40% improved growth rates and lower shoot Na+ content compared with controls, when grown under salinity stress conditions. Thus, more active AOX in roots and shoots can improve the salt tolerance of Arabidopsis as defined by its ability to grow more effectively in the presence of NaCl, and maintain lower shoot Na+ content. AOX does have an important role in stress adaptation in plants, and these results provide some validation of the hypothesis that AOX can play a critical role in cell re-programming under salinity stress.

Temperature, water, solar radiation, and atmospheric CO2 concentration are the main abiotic factors that are changing in the course of global warming. These abiotic factors govern the synthesis and degradation of primary (sugars, amino acids, organic acids, etc.) and secondary (phenolic and volatile flavor compounds and their precursors) metabolites directly, via the regulation of their biosynthetic pathways, or indirectly, via their effects on vine physiology and phenology. Several hundred secondary metabolites have been identified in the grape berry. Their biosynthesis and degradation have been characterized and have been shown to occur during different developmental stages of the berry. The understanding of how the different abiotic factors modulate secondary metabolism and thus berry quality is of crucial importance for breeders and growers to develop plant material and viticultural practices to maintain high-quality fruit and wine production in the context of global warming. Here, we review the main secondary metabolites of the grape berry, their biosynthesis, and how their accumulation and degradation is influenced by abiotic factors. The first part of the review provides an update on structure, biosynthesis, and degradation of phenolic compounds (flavonoids and non-flavonoids) and major aroma compounds (terpenes, thiols, methoxypyrazines, and C13 norisoprenoids). The second part gives an update on the influence of abiotic factors, such as water availability, temperature, radiation, and CO2 concentration, on berry secondary metabolism. At the end of the paper, we raise some critical questions regarding intracluster berry heterogeneity and dilution effects and how the sampling strategy can impact the outcome of studies on the grapevine berry response to abiotic factors.

HighlightVvGuaS, a novel allele of the VvTPS24 gene, is responsible for the production of the rotundone precursor α-guaiene. Two specific polymorphisms distinguish VvGuaS from its non-guaiene-producing homologue VvPNSeInt.

BackgroundGene expression datasets in model plants such as Arabidopsis have contributed to our understanding of gene function and how a single underlying biological process can be governed by a diverse network of genes. The accumulation of publicly available microarray data encompassing a wide range of biological and environmental conditions has enabled the development of additional capabilities including gene co-expression analysis (GCA). GCA is based on the understanding that genes encoding proteins involved in similar and/or related biological processes may exhibit comparable expression patterns over a range of experimental conditions, developmental stages and tissues. We present an open access database for the investigation of gene co-expression networks within the cultivated grapevine, Vitis vinifera.DescriptionThe new gene co-expression database, VTCdb (http://vtcdb.adelaide.edu.au/Home.aspx), offers an online platform for transcriptional regulatory inference in the cultivated grapevine. Using condition-independent and condition-dependent approaches, grapevine co-expression networks were constructed using the latest publicly available microarray datasets from diverse experimental series, utilising the Affymetrix Vitis vinifera GeneChip (16 K) and the NimbleGen Grape Whole-genome microarray chip (29 K), thus making it possible to profile approximately 29,000 genes (95% of the predicted grapevine transcriptome). Applications available with the online platform include the use of gene names, probesets, modules or biological processes to query the co-expression networks, with the option to choose between Affymetrix or Nimblegen datasets and between multiple co-expression measures. Alternatively, the user can browse existing network modules using interactive network visualisation and analysis via CytoscapeWeb. To demonstrate the utility of the database, we present examples from three fundamental biological processes (berry development, photosynthesis and flavonoid biosynthesis) whereby the recovered sub-networks reconfirm established plant gene functions and also identify novel associations.ConclusionsTogether, we present valuable insights into grapevine transcriptional regulation by developing network models applicable to researchers in their prioritisation of gene candidates, for on-going study of biological processes related to grapevine development, metabolism and stress responses.

In addition to the classical electron transport pathway coupled to ATP synthesis, plant mitochondria have an alternative pathway that involves type II NAD(P)H dehydrogenases (NDs) and alternative oxidase (AOX). This alternative pathway participates in thermogenesis in select organs of some species and is thought to help prevent cellular damage during exposure to environmental stress. Here, we investigated the function and role of one alternative path component, AtNDB2, using a transgenic approach in Arabidopsis (Arabidopsis thaliana). Disruption of AtNDB2 expression via T-DNA insertion led to a 90% decrease of external NADH oxidation in isolated mitochondria. Overexpression of AtNDB2 led to increased AtNDB2 protein abundance in mitochondria but did not enhance external NADH oxidation significantly unless AtAOX1A was concomitantly overexpressed and activated, demonstrating a functional link between these enzymes. Plants lacking either AtAOX1A or AtNDB2 were more sensitive to combined drought and elevated light treatments, whereas plants overexpressing these components showed increased tolerance and capacity for poststress recovery. We conclude that AtNDB2 is the predominant external NADH dehydrogenase in mitochondria and together with AtAOX1A forms a complete, functional, nonphosphorylating pathway of electron transport, whose operation enhances tolerance to environmental stress. This study demonstrates that at least one of the alternative NDs, as well as AOX, are important for the stress response.