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Lowering the storage temperature is an effective method to extend the postharvest and shelf life of fruits. Nevertheless, this technique often leads to physiological disorders, commonly known as chilling injuries. Apples and pears are susceptible to chilling injuries, among which superficial scald is the most economically relevant. Superficial scald is due to necrotic lesions of the first layers of hypodermis manifested through skin browning. In peaches and nectarines, chilling injuries are characterized by internal symptoms, such as mealiness. Fruits with these aesthetic or compositional/structural defects are not suitable for fresh consumption. Genetic variation is a key factor in determining fruit susceptibility to chilling injuries; however, physiological, or technical aspects such as harvest maturity and storage conditions also play a role. Multi-omics approaches have been used to provide an integrated explanation of chilling injury development. Metabolomics in pome fruits specifically targets the identification of ethylene, phenols, lipids, and oxidation products. Genomics and transcriptomics have revealed interesting connections with metabolomic datasets, pinpointing specific genes linked to cold stress, wax synthesis, farnesene metabolism, and the metabolic pathways of ascorbate and glutathione. When applied to Prunus species, these cutting-edge approaches have uncovered that the development of mealiness symptoms is linked to ethylene signaling, cell wall synthesis, lipid metabolism, cold stress genes, and increased DNA methylation levels. Emphasizing the findings from multi-omics studies, this review reports how the integration of omics datasets can provide new insights into understanding of chilling injury development. This new information is essential for successfully creating more resilient fruit varieties and developing novel postharvest strategies. [ABSTRACT FROM AUTHOR]

Dry yeast extracts (DYE) are applied to vineyards to improve aromatic and secondary metabolic compound content and wine quality; however, systematic information on the underpinning molecular mechanisms is lacking. This work aimed to unravel, through a systematic approach, the metabolic and molecular responses of Sauvignon Blanc berries to DYE treatments. To accomplish this, DYE spraying was performed in a commercial vineyard for two consecutive years. Berries were sampled at several time points after the treatment, and grapes were analyzed for sugars, acidity, free and bound aroma precursors, amino acids, and targeted and untargeted RNA-Seq transcriptional profiles. The results obtained indicated that the DYE treatment did not interfere with the technological ripening parameters of sugars and acidity. Some aroma precursors, including cys-3MH and GSH-3MH, responsible for the typical aromatic nuances of Sauvignon Blanc, were stimulated by the treatment during both vintages. The levels of amino acids and the global RNA-seq transcriptional profiles indicated that DYE spraying upregulated ROS homeostatic and thermotolerance genes, as well as ethylene and jasmonic acid biosynthetic genes, and activated abiotic and biotic stress responses. Overall, the data suggested that the DYE reduced berry oxidative stress through the regulation of specific subsets of metabolic and hormonal pathways. [ABSTRACT FROM AUTHOR]

Summary: In temperate zones, fruit trees regulate their annual growth cycle to seasonal environmental changes. During the cold season, growth is limited by both environmental and genetic factors. After the exposure to low temperature and fulfillment of chilling requirements, mild temperatures promote the growth and flowering. However, an insufficient chilling exposure may lead to nonuniform blooming, with a negative impact on fruit set.To gain insights into flower development in the fruit tree buds, peach is an interesting model, the flower and vegetative bud being distinct organs. To understand how flower bud development is regulated, we integrated cytological observations and epigenetic and chromatin genome‐wide data with transcriptional changes to identify the main regulatory factors involved in flower development during chilling accumulation.We demonstrated that growth cessation does not occur in peach flower buds during chilling accumulation, but that there are changes in transcript abundance of key genes of hormone metabolism and flower bud development, distribution of histone modifications (H3K4me3 and H3K27me3) and DNA methylation.Altogether, our findings indicate that during the cold season the flower bud is in a nondormant state and that the chilling experience allows flower differentiation to be completed. [ABSTRACT FROM AUTHOR]

The grapevine is subject to high number of fungal and viral diseases, which are responsible for important economic losses in the global wine sector every year. These pathogens deteriorate grapevine berry quality either directly via the modulation of fruit metabolic pathways and the production of endogenous compounds associated with bad taste and/or flavor, or indirectly via their impact on vine physiology. The most common and devastating fungal diseases in viticulture are gray mold, downy mildew (DM), and powdery mildew (PM), caused, respectively by Botrytis cinerea , Plasmopara viticola , and Erysiphe necator. Whereas B. cinerea mainly infects and deteriorates the ripening fruit directly, deteriorations by DM and PM are mostly indirect via a reduction of photosynthetic leaf area. Nevertheless, mildews can also infect berries at certain developmental stages and directly alter fruit quality via the biosynthesis of unpleasant flavor compounds that impair ultimate wine quality. The grapevine is furthermore host of a wide range of viruses that reduce vine longevity, productivity and berry quality in different ways. The most widespread virus-related diseases, that are known nowadays, are Grapevine Leafroll Disease (GLRD), Grapevine Fanleaf Disease (GFLD), and the more recently characterized grapevine red blotch disease (GRBD). Future climatic conditions are creating a more favorable environment for the proliferation of most virus-insect vectors, so the spread of virus-related diseases is expected to increase in most wine-growing regions. However, the impact of climate change on the evolution of fungal disease pressure will be variable and depending on region and pathogen, with mildews remaining certainly the major phytosanitary threat in most regions because their development rate is to a large extent temperature-driven. This paper aims to provide a review of published literature on most important grapevine fungal and viral pathogens and their impact on grape berry physiology and quality. Our overview of the published literature highlights gaps in our understanding of plant-pathogen interactions, which are valuable for conceiving future research programs dealing with the different pathogens and their impacts on grapevine berry quality and metabolism. [ABSTRACT FROM AUTHOR]

Abstract Background Auxins act as repressors of ripening inception in grape (véraison), while ethylene and abscisic acid (ABA) play a positive role as inducers of the syndrome. Despite the increasing amount of information made available on this topic, the complex network of interactions among these hormones remains elusive. In order to shed light on these aspects, a holistic approach was adopted to evaluate, at the transcriptomic level, the crosstalk between hormones in grape berries, whose ripening progression was delayed by applying naphtalenacetic acid (NAA) one week before véraison. Results The NAA treatment caused significant changes in the transcription rate of about 1,500 genes, indicating that auxin delayed grape berry ripening also at the transcriptional level, along with the recovery of a steady state of its intracellular concentration. Hormone indices analysis carried out with the HORMONOMETER tool suggests that biologically active concentrations of auxins were achieved throughout a homeostatic recovery. This occurred within 7 days after the treatment, during which the physiological response was mainly unspecific and due to a likely pharmacological effect of NAA. This hypothesis is strongly supported by the up-regulation of genes involved in auxin conjugation (GH3-like) and action (IAA4- and IAA31-like). A strong antagonistic effect between auxin and ethylene was also observed, along with a substantial ‘synergism’ between auxins and ABA, although to a lesser extent. Conclusions This study suggests that, in presence of altered levels of auxins, the crosstalk between hormones involves diverse mechanisms, acting at both the hormone response and biosynthesis levels, creating a complex response network.

Abstract Background Field observations and a few physiological studies have demonstrated that peach embryogenesis and fruit development are tightly coupled. In fact, attempts to stimulate parthenocarpic fruit development by means of external tools have failed. Moreover, physiological disturbances during early embryo development lead to seed abortion and fruitlet abscission. Later in embryo development, the interactions between seed and fruit development become less strict. As there is limited genetic and molecular information about seed-pericarp cross-talk and development in peach, a massive gene approach based on the use of the μPEACH 1.0 array platform and quantitative real time RT-PCR (qRT-PCR) was used to study this process. Results A comparative analysis of the transcription profiles conducted in seed and mesocarp (cv Fantasia) throughout different developmental stages (S1, S2, S3 and S4) evidenced that 455 genes are differentially expressed in seed and fruit. Among differentially expressed genes some were validated as markers in two subsequent years and in three different genotypes. Seed markers were a LTP1 (lipid transfer protein), a PR (pathogenesis-related) protein, a prunin and LEA (Late Embryogenesis Abundant) protein, for S1, S2, S3 and S4, respectively. Mesocarp markers were a RD22-like protein, a serin-carboxypeptidase, a senescence related protein and an Aux/IAA, for S1, S2, S3 and S4, respectively. The microarray data, analyzed by using the HORMONOMETER platform, allowed the identification of hormone-responsive genes, some of them putatively involved in seed-pericarp crosstalk. Results indicated that auxin, cytokinins, and gibberellins are good candidates, acting either directly (auxin) or indirectly as signals during early development, when the cross-talk is more active and vital for fruit set, whereas abscisic acid and ethylene may be involved later on. Conclusions In this research, genes were identified marking different phases of seed and mesocarp development. The selected genes behaved as good seed markers, while for mesocarp their reliability appeared to be dependent upon developmental and ripening traits. Regarding the cross-talk between seed and pericarp, possible candidate signals were identified among hormones. Further investigations relying upon the availability of whole genome platforms will allow the enrichment of a marker genes repertoire and the elucidation of players other than hormones that are involved in seed-pericarp cross-talk (i.e. hormone peptides and microRNAs).

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. [ABSTRACT FROM AUTHOR]

Tartaric acid (TA) is an obscure end point to the catabolism of ascorbic acid (Asc). Here, it is proposed as a "specialized primary metabolite", originating from carbohydrate metabolism but with restricted distribution within the plant kingdom and lack of known function in primary metabolic pathways. Grapes fall into the list of high TA-accumulators, with biosynthesis occurring in both leaf and berry. Very little is known of the TA biosynthetic pathway enzymes in any plant species, although recently some progress has been made in this space. New technologies in grapevine research such as the development of global co-expression network analysis tools and genome-wide association studies, should enable more rapid progress. There is also a lack of information regarding roles for this organic acid in plant metabolism. Therefore this review aims to briefly summarize current knowledge about the key intermediates and enzymes of TA biosynthesis in grapes and the regulation of its precursor, ascorbate, followed by speculative discussion around the potential roles of TA based on current knowledge of Asc metabolism, TA biosynthetic enzymes and other aspects of fruit metabolism. [ABSTRACT FROM AUTHOR]

The partitioning of assimilates in fruits, which are economically important sink organs, is ruled by different physiological processes and affected by both environmental and agronomical factors. The bulk of the water and solutes, required for growth, is imported into fruits and seeds through xylem and phloem. In the stone fruits, five vascular bundles enter the base of the fruit, then dividing to supply either the flesh or the seed. The main sugars accumulated in stone fruits include fructose, glucose, and sucrose, along with other minor saccharides. The mechanisms of phloem loading in these fruit species have not been fully elucidated yet, but the available data hint either an apoplastic or a symplastic type or possibly a combination of both, depending on the species and the sugar considered. Similarly, phloem unloading mechanisms, elucidated for a small number of species, depend on genotype and developmental stage. Remarkably, key enzymes and transporters involved in the main sugars-conversion and transport pathways have received considerable attention. In stone fruit trees, the presence of an elevated number of fruits alters the source-sink balance, with a consequent intensification of competition among them and between vegetative and reproductive growth. The main environmental factors affecting this balance and the agronomical/artificial manipulations of source-sink relationships to achieve adequate fruit production and quality are reviewed. [ABSTRACT FROM AUTHOR]

Non-structural carbohydrates are abundant constituents of the ripe flesh of all stone fruits. The bulk of their content comprises sucrose, glucose, fructose and sorbitol. However, the abundance of each of these carbohydrates in the flesh differs between species, and also with its stage of development. In this article the import, subcellular compartmentation, contents, metabolism and functions of non-structural carbohydrates in the flesh of commercially cultivated stone fruits of the family Rosaceae are reviewed. [ABSTRACT FROM AUTHOR]

Stone fruits of the Rosaceae family consist of several distinct parts, and these include the flesh, woody endocarp, and seed. To understand the metabolism of these fruits, it is necessary to have knowledge of both their structure and growth characteristics. The nitrogen metabolism of the different tissues of stone fruits is interlinked. For example, there is an import and storage of nitrogenous compounds in the endocarp that are then exported to the seed. Moreover, there are links between the metabolism of nitrogen and that of malic/citric acids. In this article, the structure and growth characteristics, together with the import/export, contents, metabolism, and functions of nitrogenous compounds and organic acids in the different parts of stone fruits and their seeds are reviewed. [ABSTRACT FROM AUTHOR]

Phytochemicals or secondary metabolites present in fruit are key components contributing to sensory attributes like aroma, taste, and color. In addition, these compounds improve human nutrition and health. Stone fruits are an important source of an array of secondary metabolites that may reduce the risk of different diseases. The first part of this review is dedicated to the description of the main secondary organic compounds found in plants which include (a) phenolic compounds, (b) terpenoids/isoprenoids, and (c) nitrogen or sulfur containing compounds, and their principal biosynthetic pathways and their regulation in stone fruit. Then, the type and levels of bioactive compounds in different stone fruits of the Rosaceae family such as peach (Prunus persica), plum (P. domestica , P. salicina and P. cerasifera) , sweet cherries (P. avium), almond kernels (P. dulcis , syn. P. amygdalus), and apricot (P. armeniaca) are presented. The last part of this review encompasses pre- and postharvest treatments affecting the phytochemical composition in stone fruit. Appropriate management of these factors during pre- and postharvest handling, along with further characterization of phytochemicals and the regulation of their synthesis in different cultivars, could help to increase the levels of these compounds, leading to the future improvement of stone fruit not only to enhance organoleptic characteristics but also to benefit human health. [ABSTRACT FROM AUTHOR]

Cell wall turnover and modification in its composition are key factors during stone fruit development and patterning. Changes in cell wall disassembly and reassembly are essential for fruit growth and ripening. Modifications in cell wall composition, resulting in the formation of secondary cell walls, are necessary for producing the most distinctive trait of drupes: the lignified endocarp. The contribution of primary metabolism to cell wall synthesis has been investigated in detail, while the knowledge on the contribution of the cell wall to primary metabolites and related processes is still fragmented. In this review, starting from peculiarities of cell wall of drupes cells (in mesocarp and endocarp layers), we discuss the structure and composition of cell wall, processes related to its modification and contribution to the synthesis of primary metabolites. In particular, our attention has been focused on the ascorbate synthesis cell wall-related and on the potential role of cyanogenic compounds in the deposition of the secondary cell wall. [ABSTRACT FROM AUTHOR]

Rosaceae is a family with an extraordinary spectrum of fruit types, including fleshy peach, apple, and strawberry that provide unique contributions to a healthy diet for consumers, and represent an excellent model for studying fruit patterning and development. In recent years, many efforts have been made to unravel regulatory mechanism underlying the hormonal, transcriptomic, proteomic and metabolomic changes occurring during Rosaceae fruit development. More recently, several studies on fleshy (tomato) and dry (Arabidopsis) fruit model have contributed to a better understanding of epigenetic mechanisms underlying important heritable crop traits, such as ripening and stress response. In this context and summing up the results obtained so far, this review aims to collect the available information on epigenetic mechanisms that may provide an additional level in gene transcription regulation, thus influencing and driving the entire Rosaceae fruit developmental process. The whole body of information suggests that Rosaceae fruit could become also a model for studying the epigenetic basis of economically important phenotypes, allowing for their more efficient exploitation in plant breeding. [ABSTRACT FROM AUTHOR]

Iron chlorosis is a serious deficiency that affects orchards and vineyards reducing quality and yield production. Chlorotic plants show abnormal photosynthesis and yellowing shoots. In grapevine iron uptake and homeostasis are most likely controlled by a mechanism known as "Strategy I," characteristic of non-graminaceous plants and based on a system of soil acidification, iron reduction and transporter-mediated uptake. Nowadays, grafting of varieties of economic interest on tolerant rootstocks is widely used practice against many biotic and abiotic stresses. Nevertheless, many interspecific rootstocks, and in particular those obtained by crossing exclusively non-vinifera genotypes, can show limited nutrient uptake and transport, in particular for what concerns iron. In the present study, 101.14, a commonly used rootstock characterized by susceptibility to iron chlorosis was subjected to both Fe-absence and Fe-limiting conditions. Grapevine plantlets were grown in control, Fe-deprived, and bicarbonate-supplemented hydroponic solutions. Whole transcriptome analyses, via mRNA-Seq, were performed on root apices of stressed and unstressed plants. Analysis of differentially expressed genes (DEGs) confirmed that Strategy I is the mechanism responsible for iron uptake in grapevine, since many orthologs genes to the Arabidopsis "ferrome" were differentially regulated in stressed plant. Molecular differences in the plant responses to Fe absence and presence of bicarbonate were also identified indicating the two treatments are able to induce response-mechanisms only partially overlapping. Finally, we measured the expression of a subset of genes differentially expressed in 101.14 (such as IRT1, FERRITIN1, bHLH38/39) or known to be fundamental in the "strategy I" mechanism (AHA2 and FRO2) also in a tolerant rootstock (M1) finding important differences which could be responsible for the different degrees of tolerance observed. [ABSTRACT FROM AUTHOR]

The NAP (NAC-like, activated by AP3/P1) transcription factor belongs to a subfamily of the NAC transcription factor family, and is believed to have an important role in regulating plant growth and development. However, there is very little information about this subfamily in Rosaceous plants. We identified seven NAP genes in the peach genome. PpNAP2 was categorized in the NAP I group, and contained a conserved transcription activation region. The other PpNAP genes belonged to the NAP II group. The expression patterns of the PpNAP genes differed in various organs and developmental stages. PpNAP1 and PpNAP2 were highly expressed in mature and senescing flowers, but not in leaves, fruits, and flower buds. PpNAP3 and PpNAP5 were only expressed in leaves. The PpNAP4 expression level was high in mature and senescing fruits, while PpNAP6 and PpNAP7 expression was up-regulated in mature and senescent leaves and flowers. During the fruit development period, the PpNAP4 and PpNAP6 expression levels rapidly increased during the S1 and S4 stages, which suggests these genes are involved in the first exponential growth phase and fruit ripening. During the fruit ripening and softening period, the PpNAP1, PpNAP4, and PpNAP6 expression levels were high during the early storage period, which was accompanied by a rapid increase in ethylene production. PpNAP1, PpNAP4, and PpNAP6 expression slowly increased during the middle or late storage periods, and peaked at the end of the storage period. Additionally, abscisic acid (ABA)-treated fruits were softer and produced more ethylene than the controls. Furthermore, the PpNAP1, PpNAP4, and PpNAP6 expression levels were higher in ABA-treated fruits. These results suggest that PpNAP1, PpNAP4, and PpNAP6 are responsive to ABA and may regulate peach fruit ripening. [ABSTRACT FROM AUTHOR]

In modern viticulture, grafting commercial grapevine varieties on interspecific rootstocks is a common practice required for conferring resistance to many biotic and abiotic stresses. Nevertheless, the use of rootstocks to gain these essential traits is also known to impact grape berry development and quality, although the underlying mechanisms are still poorly understood. In grape berries, the onset of ripening (véraison) is regulated by a complex network of mobile signals including hormones such as auxins, ethylene, abscisic acid, and brassinosteroids. Recently, a new rootstock, designated M4, was selected based on its enhanced tolerance to water stress and medium vigor. This study investigates the effect of M4 on Cabernet Sauvignon (CS) berry development in comparison to the commercial 1103P rootstock. Physical and biochemical parameters showed that the ripening rate of CS berries is faster when grafted onto M4. A multifactorial analysis performed on mRNA-Seq data obtained from skin and pulp of berries grown in both graft combinations revealed that genes controlling auxin action (ARF and Aux/IAA) represent one of main categories affected by the rootstock genotype. Considering that the level of auxin tightly regulates the transcription of these genes, we investigated the behavior of the main gene families involved in auxin biosynthesis and conjugation. Molecular and biochemical analyses confirmed a link between the rate of berry development and the modulation of auxin metabolism. Moreover, the data indicate that this phenomenon appears to be particularly pronounced in skin tissue in comparison to the flesh. [ABSTRACT FROM AUTHOR]

Many plant processes depend on differential gene expression, which is generally controlled by complex proteins called transcription factors (TFs). In peach, 1533 TFs have been identified, accounting for about 5.5% of the 27,852 protein-coding genes. These TFs are the reference for the rest of the Prunus species. TF studies in Prunus have been performed on the gene expression analysis of different agronomic traits, including control of the flowering process, fruit quality, and biotic and abiotic stress resistance. These studies, using quantitative RT-PCR, have mainly been performed in peach, and to a lesser extent in other species, including almond, apricot, black cherry, Fuji cherry, Japanese apricot, plum, and sour and sweet cherry. Other tools have also been used in TF studies, including cDNA-AFLP, LC-ESI-MS, RNA, and DNA blotting or mapping. More recently, new tools assayed include microarray and high-throughput DNA sequencing (DNA-Seq) and RNA sequencing (RNA-Seq). New functional genomics opportunities include genome resequencing and the well-known synteny among Prunus genomes and transcriptomes. These new functional studies should be applied in breeding programs in the development of molecular markers. With the genome sequences available, some strategies that have been used in model systems (such as SNP genotyping assays and genotyping-by-sequencing) may be applicable in the functional analysis of Prunus TFs as well. In addition, the knowledge of the gene functions and position in the peach reference genome of the TFs represents an additional advantage. These facts could greatly facilitate the isolation of genes via QTL (quantitative trait loci) map-based cloning in the different Prunus species, following the association of these TFs with the identified QTLs using the peach reference genome. [ABSTRACT FROM AUTHOR]

Differences in gene expression were studied after Plum pox virus ( PPV, sharka disease) infection in peach GF305 leaves with and without sharka symptoms using RNA-Seq. For each sample, more than 80% of 100-nucleotide paired-end ( PE) Illumina reads were aligned on the peach reference genome. In the symptomatic sample, a significant proportion of reads were mapped to PPV reference genomes (1.04% compared with 0.00002% in non-symptomatic leaves), allowing for the ultra-deep assembly of the complete genome of the PPV isolate used (9775 nucleotides, missing only 11 nucleotides at the 5′ genome end). In addition, significant alternative splicing events were detected in 359 genes and 12 990 single nucleotide polymorphisms ( SNPs) were identified, 425 of which could be annotated. Gene ontology annotation revealed that the high-ranking mRNA target genes associated with the expression of sharka symptoms are mainly related to the response to biotic stimuli, to lipid and carbohydrate metabolism and to the negative regulation of catalytic activity. A greater number of differentially expressed genes were observed in the early asymptomatic phase of PPV infection in comparison with the symptomatic phase. These early infection events were associated with the induction of genes related to pathogen resistance, such as jasmonic acid, chitinases, cytokinin glucosyl transferases and Lys- M proteins. Once the virus had accumulated, the overexpression of Dicer protein 2a genes suggested a gene silencing plant response that was suppressed by the virus HCPro and P1 proteins. These results illustrate the dynamic nature of the peach- PPV interaction at the transcriptome level and confirm that sharka symptom expression is a complex process that can be understood on the basis of changes in plant gene expression. [ABSTRACT FROM AUTHOR]

Background Alternative splicing (AS) significantly enhances transcriptome complexity. It is differentially regulated in a wide variety of cell types and plays a role in several cellular processes. Here we describe a detailed survey of alternative splicing in grape based on 124 SOLiD RNAseq analyses from different tissues, stress conditions and genotypes. Results We used the RNAseq data to update the existing grape gene prediction with 2,258 new coding genes and 3,336 putative long non-coding RNAs. Several gene structures have been improved and alternative splicing was described for about 30% of the genes. A link between AS and miRNAs was shown in 139 genes where we found that AS affects the miRNA target site. A quantitative analysis of the isoforms indicated that most of the spliced genes have one major isoform and tend to simultaneously co-express a low number of isoforms, typically two, with intron retention being the most frequent alternative splicing event. Conclusions As described in Arabidopsis, also grape displays a marked AS tissue-specificity, while stress conditions produce splicing changes to a minor extent. Surprisingly, some distinctive splicing features were also observed between genotypes. This was further supported by the observation that the panel of Serine/Arginine-rich splicing factors show a few, but very marked differences between genotypes. The finding that a part the splicing machinery can change in closely related organisms can lead to some interesting hypotheses for evolutionary adaptation, that could be particularly relevant in the response to sudden and strong selective pressures. [ABSTRACT FROM AUTHOR]

Peach ( Prunus persica var. laevis Gray) was chosen to unravel the molecular basis underlying the ability of spermidine (Sd) to influence fruit development and ripening. Field applications of 1 m M Sd on peach fruit at an early developmental stage, 41 days after full bloom (dAFB), i.e. at late stage S1, led to a slowing down of fruit ripening. At commercial harvest (125 dAFB, S4II) Sd-treated fruits showed a reduced ethylene production and flesh softening. The endogenous concentration of free and insoluble conjugated polyamines (PAs) increased (0.3-2.6-fold) 1 day after treatment (short-term response) butsoon it declined to control levels; starting from S3/S4, when soluble conjugated forms increased (up to five-fold relative to controls at ripening), PA levels became more abundant in treated fruits, (long-term response). Real-time reverse transcription-polymerase chain reaction analyses revealed that peaks in transcript levels of fruit developmental marker genes were shifted ahead in accord with a developmental slowing down. At ripening (S4I-S4II) the upregulation of the ethylene biosynthetic genes ACO1 and ACS1 was dramatically counteracted by Sd and this led to a strong downregulation of genes responsible for fruit softening, such as PG and PMEI. Auxin-related gene expression was also altered both in the short term ( TRPB) and in the long term ( GH3, TIR1 and PIN1), indicating that auxin plays different roles during development and ripening processes. Messenger RNA amounts of other hormone-related ripening-regulated genes, such as NCED and GA2-OX, were strongly downregulated at maturity. Results suggest that Sd interferes with fruit development/ripening by interacting with multiple hormonal pathways. [ABSTRACT FROM AUTHOR]

Abstract: Specific polyphenol compound concentrations and gene expression patterns were determined by both microarray and qRT-PCR analyses in the epicarp of red-skinned grape berries (Vitis vinifera L. cv ‘Raboso Piave’) dehydrated, after harvest, at slow (S) and rapid (R) rates of up to 10 and 30% weight loss (WL). Increases in flavonols (quercetin) and trans-resveratrol concentrations were observed in the skins of all dehydrated samples, whereas flavan-3-ols concentrations showed a decreasing trend, which was more pronounced in S samples. The decrease in flavan-3-ol concentrations was paralleled by a reduction in procyanidin B1 and, particularly B2. Computational analysis of microarray data revealed that several key genes of the flavonoid pathways were unaffected or down-regulated during berry dehydration, with the exception of flavonol synthase, which was induced as well as one MybB transcription factor. Chalcone synthase (CHS), flavanone 3-hydroxylase (F3H), leucoanthocyanidin dioxygenase (LDOX) and MybA were markedly down-regulated, particularly in relation to 30% WL, whereas UDP-glucose:flavonoid 3-O-glucosyltransferase (UFGT) was unaffected in all the samples considered. Specific genes involved in the lignin biosynthetic pathway, including laccase, were induced in the most dehydrated samples. Considering the accumulation pattern and gene expression profiles, stilbenes and flavonols may represent useful biomarkers to monitor postharvest dehydration. [Copyright &y& Elsevier]

Background: Auxins act as repressors of ripening inception in grape (véraison), while ethylene and abscisic acid (ABA) play a positive role as inducers of the syndrome. Despite the increasing amount of information made available on this topic, the complex network of interactions among these hormones remains elusive. In order to shed light on these aspects, a holistic approach was adopted to evaluate, at the transcriptomic level, the crosstalk between hormones in grape berries, whose ripening progression was delayed by applying naphtalenacetic acid (NAA) one week before véraison. Results: The NAA treatment caused significant changes in the transcription rate of about 1,500 genes, indicating that auxin delayed grape berry ripening also at the transcriptional level, along with the recovery of a steady state of its intracellular concentration. Hormone indices analysis carried out with the HORMONOMETER tool suggests that biologically active concentrations of auxins were achieved throughout a homeostatic recovery. This occurred within 7 days after the treatment, during which the physiological response was mainly unspecific and due to a likely pharmacological effect of NAA. This hypothesis is strongly supported by the up-regulation of genes involved in auxin conjugation (GH3-like) and action (IAA4- and IAA31-like). A strong antagonistic effect between auxin and ethylene was also observed, along with a substantial 'synergism' between auxins and ABA, although to a lesser extent. Conclusions: This study suggests that, in presence of altered levels of auxins, the crosstalk between hormones involves diverse mechanisms, acting at both the hormone response and biosynthesis levels, creating a complex response network. [ABSTRACT FROM AUTHOR]

The recent sequencing of the complete genome of the peach offers new opportunities for further transcriptomic studies in Prunus species in the called post-genomics era . First works on transcriptome analysis in Prunus species started in the early 2000s with the development of ESTs (expressed sequence tags) and the analysis of several candidate genes. Later, new strategies of massive analysis (high throughput) of transcriptomes have been applied, producing larger amounts of data in terms of expression of a large number of genes in a single experiment. One of these systems is massive transcriptome analysis using cDNA biochips (microarrays) to analyze thousands of genes by hybridization of mRNA labelled with fluorescence. However, the recent emergence of a massive sequencing methodology ('deep-sequencing') of the transcriptome (RNA-Seq), based on lowering the costs of DNA (in this cases complementary, cDNA) sequencing, could be more suitable than the application of microarrays. Recent papers have described the tremendous power of this technology, both in terms of profiling coverage and quantitative accuracy in transcriptomic studies. Now this technology is being applied to plant species, including Prunus. In this work, we analyze the potential in using this RNA-Seq technology in the study of Prunus transcriptomes and the development of genomic tools. In addition, the strengths and limitations of RNA-Seq relative to microarray profiling have been discussed. [ABSTRACT FROM AUTHOR]

Background: Field observations and a few physiological studies have demonstrated that peach embryogenesis and fruit development are tightly coupled. In fact, attempts to stimulate parthenocarpic fruit development by means of external tools have failed. Moreover, physiological disturbances during early embryo development lead to seed abortion and fruitlet abscission. Later in embryo development, the interactions between seed and fruit development become less strict. As there is limited genetic and molecular information about seed-pericarp crosstalk and development in peach, a massive gene approach based on the use of the μPEACH 1.0 array platform and quantitative real time RT-PCR (qRT-PCR) was used to study this process. Results: A comparative analysis of the transcription profiles conducted in seed and mesocarp (cv Fantasia) throughout different developmental stages (S1, S2, S3 and S4) evidenced that 455 genes are differentially expressed in seed and fruit. Among differentially expressed genes some were validated as markers in two subsequent years and in three different genotypes. Seed markers were a LTP1 (lipid transfer protein), a PR (pathogenesis-related) protein, a prunin and LEA (Late Embryogenesis Abundant) protein, for S1, S2, S3 and S4, respectively. Mesocarp markers were a RD22-like protein, a serin-carboxypeptidase, a senescence related protein and an Aux/IAA, for S1, S2, S3 and S4, respectively. The microarray data, analyzed by using the HORMONOMETER platform, allowed the identification of hormoneresponsive genes, some of them putatively involved in seed-pericarp crosstalk. Results indicated that auxin, cytokinins, and gibberellins are good candidates, acting either directly (auxin) or indirectly as signals during early development, when the cross-talk is more active and vital for fruit set, whereas abscisic acid and ethylene may be involved later on. Conclusions: In this research, genes were identified marking different phases of seed and mesocarp development. The selected genes behaved as good seed markers, while for mesocarp their reliability appeared to be dependent upon developmental and ripening traits. Regarding the cross-talk between seed and pericarp, possible candidate signals were identified among hormones. Further investigations relying upon the availability of whole genome platforms will allow the enrichment of a marker genes repertoire and the elucidation of players other than hormones that are involved in seed-pericarp cross-talk (i.e. hormone peptides and microRNAs). [ABSTRACT FROM AUTHOR]

A large-scale transcriptome analysis was conducted using an oligo-based microarray (14,562 probes) on skins of wine grape (Vitis vinifera L.) berries dehydrated at different rates (slow, S and rapid, R) after harvest up to 10 and 30% of weight loss (WL). At 10% WL, a total of 84 and 68 probes were differentially expressed following S and R dehydration, respectively. At 30% WL, 309 and 262 differentially expressed probes were detected in S and R samples, respectively, indicating that grape berries are still reactive at advanced stages of postharvest dehydration. Bioinformatic analysis revealed that about 70% of the differentially expressed probes could be annotated and putative functions were assigned. Functional characterization highlighted that, independently of the rate and intensity of dehydration, differential expression occurred in particular for genes associated with general metabolism, regulatory processes, and responses to biotic and abiotic stimuli. A total of 16 (induced) and 10 (repressed) probes, common to all four dehydrated samples, were associated with hormone (ethylene) metabolism, transcription factors, carbohydrate and secondary (polyphenols) metabolism, transport and stress responses. Together with the total number of differentially expressed probes, enhancing the dehydration level from 10 to 30% WL also affected the distribution of genes within functional categories: this behaviour was observed in particular for R samples. A higher level of water stress in grapes appears to be associated with modification to the expression of genes mainly involved in hormone and sugar metabolism, and defence mechanisms. Besides the intensity of dehydration, a significant effect on gene expression was also associated with rate of water loss: an increase in the percentage incidence of differentially expressed probes was present for categories involved in defence and environmental stress when comparing R and S samples. The microarray data, validated by RT-PCR analyses, represent robust evidence for the marked effects of postharvest water loss on metabolic processes in fruit tissues. [Copyright &y& Elsevier]

• The time course of ethylene biosynthesis and perception was investigated in ripening peach fruit ( Prunus persica) following treatments with the polyamines putrescine (Pu) and spermidine (Sd), and with aminoethoxyvinylglycine (AVG). • Fruit treatments were performed in planta. Ethylene production was measured by gas chromatography, and polyamine content by high-performance liquid chromatography; expression analyses were performed by Northern blot or real-time polymerase chain reaction. • Differential increases in the endogenous polyamine pool in the epicarp and mesocarp were induced by treatments; in both cases, ethylene production, fruit softening and abscission were greatly inhibited. The rise in 1-aminocyclopropane-1-carboxylate oxidase ( PpACO1) mRNA was counteracted and delayed in polyamine-treated fruit, whereas transcript abundance of ethylene receptors PpETR1 (ethylene receptor 1) and PpERS1 (ethylene sensor 1) was enhanced at harvest. Transcript abundance of arginine decarboxylase (ADC) and S-adenosylmethionine decarboxylase (SAMDC) was transiently reduced in both the epicarp and mesocarp. AVG, here taken as a positive control, exerted highly comparable effects to those of Pu and Sd. • Thus, in peach fruit, increasing the endogenous polyamine pool in the epicarp or in the mesocarp strongly interfered, both at a biochemical and at a biomolecular level, with the temporal evolution of the ripening syndrome. [ABSTRACT FROM AUTHOR]

Abstract: The transition from pre-climacteric to climacteric phase is a critical step during fruit development. A holistic approach to study this transition has been undertaken using the first available peach microarray (μPEACH1.0) containing about 4800 oligonucleotide probes corresponding to a set of unigenes most of them expressed during the last stages of fruit development. Microarray hybridizations indicated that among the genes present in the microarray slide, 267 and 109 genes are up- and down-regulated, respectively. Genes have been classified according to the TAIR Gene Ontology into three main categories based on cellular localization, molecular function and biological process. Considering the cellular localization, the most significant up- and down-regulated gene products belong to cell wall and chloroplast compartments. Within the molecular function and biological process categories, a dramatic up-regulation has been detected for genes encoding transcription factors and enzymes involved in ethylene biosynthesis and action. A new member of ETR peach family (Pp-ETR2) has been characterized: this gene shows high similarity to Arabidopsis EIN4, tomato Le-ETR4, and strawberry Fa-ETR2. Transition from S3 to S4 is paralleled by changes in expression of 19 genes encoding transcription factors (TFs) belonging to several families including MADS-box, AUX/IAA, bZIP, bHLH, HD, and Myb. Differential expression of genes involved in specific quality traits has also been observed: besides confirming previous data on cell wall-related gene expression, a new pectin-methyl esterase and two new expansins have been identified. Several genes encoding enzymes acting in the isoprenoid biosynthetic pathway appeared to be strongly induced at S3/S4 transition. Among those involved in carotenoid biosynthesis we found also a β-carotene hydroxylase, responsible for the formation of β-cryptoxanthin, the most abundant carotenoid of ripe yellow peaches. [Copyright &y& Elsevier]

We report the isolation of a new peach gene, Pp-MLP1, that shows significant similarity to a family of fruit- and flower-specific genes, designated as major latex protein (MLP) homologues. Transcript of Pp-MLP1 highly accumulated in cells of fruit pedicel, similar to lacticifers, adjacent to the abscission zone (non-abscission zone) and, to a lesser extent, in epicotyls, stems and roots, while no accumulation was detected in leaves. In contrast to the MLP homologues isolated so far, the Pp-MLP1 transcript was detected during fruit cells expansion, though its expression appeared unrelated to fruit ripening. Propylene treatment caused a decrease in mRNA accumulation of Pp-MLP1 in all tested tissues. The function of Pp-MLP1, as with all previously described MLP homologues, is unknown. MLPs are associated with fruit and flower development in addition to plant pathogenesis responses. Expression in tissues associated with abscission would be consistent with a role in implementing this aspect of floral development or possibly protective responses to plant pathogens which may infect post-abscission wounds. In addition, the high similarity between proteins encoding by Pp-MLP1 and Csf2, an MLP gene associated with the early development of cucumber fruit, could suggest an alternative developmental role such as cell and tissue expansion. [Copyright &y& Elsevier]

The characterization and expression of PP-ACO1 and PP-ACO2, two members of the peach 1-aminocyclopropane-1-carboxylate (ACC) oxidase (ACO) gene family, are reported. PP-ACO1 is organized in 4 exons interrupted by 3 introns, whereas PP-ACO2 has only 2 of the 3 introns present in PP-ACO1. Comparison of the deduced amino acid sequences of PP-ACO1 and PP-ACO2 reveals a 77.7% identity. PP-ACO1 and PP-ACO2 show highest degree of similarity with petunia (PH-ACO3; 84.1%) and apple (85.4%) ACO genes, respectively. PP-ACO1 is expressed in flowers, fruitlet abscission zones, mesocarp and in young fully expanded leaves. PP-ACO1 transcript accumulation strongly increases during fruitlet abscission, in ripe mesocarp and senescing leaves, and is enhanced by propylene. PP-ACO2 mRNA accumulation is detected in fruits only during early development and is unaffected by propylene treatment. Both ACO genes are expressed in epicotyl and roots of growing seedlings, although a stronger accumulation of PP-ACO2 mRNA is observed. [ABSTRACT FROM AUTHOR]

Superficial scald is a post-harvest chilling storage injury leading to browning of the surface of the susceptible cv Granny Smith apples. Wounding of skins has been reported to play a preventive role on scald development however its underlying molecular factors are unknown. We have artificially wounded the epidermal and sub-epidermal layers of apple skins consistently obtaining the prevention of superficial scald in the surroundings of the wounds during two independent vintages. Time course RNA-Seq analyses of the transcriptional changes in wounded versus unwounded skins revealed that two transcriptional waves occurred. An early wave included genes up-regulated by wounding already after 6 h, highlighting a specific transcriptional rearrangement of genes connected to the biosynthesis and signalling of JA, ethylene and ABA. A later transcriptional wave, occurring after three months of cold storage, included genes up-regulated exclusively in unwounded skins and was prevented from its occurrence in wounded skins. A significant portion of these genes was related to decay of tissues and to the senescence hormones ABA, JA and ethylene. Such changes suggest a wound-inducible reversed hormonal balance during post-harvest storage which may explain the local inhibition of scald in wounded tissues, an aspect that will need further studies for its mechanistic explanation. [ABSTRACT FROM AUTHOR]

In grapevines, as in other plants, sucrose and its constituents glucose and fructose are fundamentally important and carry out a multitude of roles. The aims of this review are three-fold. First, to provide a summary of the metabolism and transport of sucrose in grapevines, together with new insights and interpretations. Second, to stress the importance of considering the compartmentation of metabolism. Third, to outline the key role of acid invertase in osmoregulation associated with sucrose metabolism and transport in plants. [ABSTRACT FROM AUTHOR]

During peach (Prunus persica [L.] Batsch) fruit development and ripening the cell wall undergoes several structural and biochemical changes driven by several hydrolases. Among these, the endo-β-1,4-D-glucanase (EGase, EC 3.2.1.4), or cellulase, may play a crucial role. Involvement of EGase throughout development and ripening of the fruit of cv. Redhaven was assessed by monitoring enzyme activity, specific polypeptide accumulation and gene transcription. During the four stages of growth EGase activity was high during S1 and in the early S2, declined during S3, and increased with the onset of ripening (S4). Two isoforms with isolectric points of 6.5 and 9.5 were identified. The pI 6.5 EGase was the only form present during the early stages of growth. whereas the pI 9.5 EGase was most abundant during ripening. The same isoforms were present in leaf and fruit abscission zones. The antibody raised against the pI 9.5 EGase, purified from leaf abscission zones, cross-reacted with a protein of 54 kDa. A cDNA clone of 753 bp encoding peach EGase was obtained by RT-PCR. EGase transcripts, detectable only after amplification of total RNA by RT-PCR, were observed during S1, and at the preclimacteric and climacteric stages. However, the strongest hybridisation occurred at ripening, in correspondence with the maximal enzyme activity and polypeptide accumulation, which took place before the ethylene climacteric and in the early stage of flesh softening. Propylene treatments reduced EGase activity during the early stage of fruit growth but dramatically enhanced enzyme activity and the related transcript accumulation at ripening, and accelerated the loss of firmness. In fruit treated with 2,5-norbornadiene the softening process was strongly inhibited and the rise in EGase transcripts and activity did not take place. The results point to the EGases being involved in early fruit growth and the initial phases of softening. The presence of two isoforms and the dual effect of propylene on enzyme activity suggest that different EGase genes operate during the early and late developmental stages in peach fruit. [ABSTRACT FROM AUTHOR]

Consumers' choices are mainly based on fruit external characteristics such as the final size, weight, and shape. The majority of edible fruit are by tree fruit species, among which peach is the genomic and genetic reference for Prunus. In this research, we used a peach with a slow ripening (SR) phenotype, identified in the Fantasia (FAN) nectarine, associated with misregulation of genes involved in mesocarp identity and showing a reduction of final fruit size. By investigating the ploidy level, we observed a progressive increase in endoreduplication in mesocarp, which occurred in the late phases of FAN fruit development, but not in SR fruit. During fruit growth, we also detected that genes involved in endoreduplication were differentially modulated in FAN compared to SR. The differential transcriptional outputs were consistent with different chromatin states at loci of endoreduplication genes. The impaired expression of genes controlling cell cycle and endocycle as well as those claimed to play a role in fruit tissue identity result in the small final size of SR fruit. [ABSTRACT FROM AUTHOR]