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A "box-in-box" cocultivation system was used to investigate plant responses to microbial volatile compounds (VCs) and to evaluate the contributions of organic and inorganic VCs (VOCs and VICs, respectively) to these responses. Arabidopsis plants were exposed to VCs emitted by adjacent Alternaria alternata and Penicillium aurantiogriseum cultures, with and without charcoal filtration. No VOCs were detected in the headspace of growth chambers containing fungal cultures with charcoal filters. However, these growth chambers exhibited elevated CO 2 and bioactive CO and NO headspace concentrations. Independently of charcoal filtration, VCs from both fungal phytopathogens promoted growth and distinct developmental changes. Plants cultured at CO 2 levels observed in growth boxes containing fungal cultures were identical to those cultured at ambient CO 2 . Plants exposed to charcoal-filtered fungal VCs, nonfiltered VCs, or superelevated CO 2 levels exhibited transcriptional changes resembling those induced by increased irradiance. Thus, in the "box-in-box" system, (a) fungal VICs other than CO 2 and/or VOCs not detected by our analytical systems strongly influence the plants' responses to fungal VCs, (b) different microorganisms release VCs with distinct action potentials, (c) transcriptional changes in VC-exposed plants are mainly due to enhanced photosynthesis signaling, and (d) regulation of some plant responses to fungal VCs is primarily posttranscriptional., (© 2018 John Wiley & Sons Ltd.)

The zucchini (Cucurbita pepo) is an important food crop, the transcriptomics of which are a fundamental tool to accelerate the development of new varieties by breeders. However, the suitability of reference genes for data normalization in zucchini has not yet been studied. The aim of this study was to assess the suitability of 13 genes for their potential use as reference genes in quantitative real-time PCR. Assays were performed on 34 cDNA samples representing plants under different stresses and at different developmental stages. The application of geNorm and NormFinder software revealed that the use of a combination of UFP, EF-1A, RPL36aA, PP2A, and CAC genes for the different experimental sets was the best strategy for reliable normalization. In contrast, 18S rRNA and TUA were less stable and unsuitable for use as internal controls. These results provide the possibility to allow more accurate use of qPCR in this horticultural crop.

The main objective of this study was to monitor apricot development and ripening through gene expression analysis of key candidate genes using the RT-qPCR technique. Eight apricot cultivars were selected to analyze phenological and genetic patterns from pre-ripening stages through to postharvest. In addition, 19 selected genes were analyzed in the contrasting cultivars 'Cebas Red' and 'Rojo Pasión' in different stages (two preharvest stages S1 and S2, one harvest stage S3, and two postharvest stages S4 and S5). This pool of genes included genes related to fruit growth and ripening, genes associated with fruit color, and genes linked to the fruit's nutraceutical aspects. Among the studied genes, Polygalacturonase (PG), Pectin methylesterase (PME), Aminocyclopropane-1-carboxylate synthase (ACS), and Myo-inositol-1-phosphate synthase (INO1) were directly related to fruit maturation and quality. Significant differential expression was observed between the cultivars, which correlated with variations in firmness, shelf life, and sensory characteristics of the apricots. 'Rojo Pasión' displayed high levels of PG, associated with rapid maturation and shorter postharvest shelf life, whereas 'Cebas Red' exhibited lower levels of this gene, resulting in greater firmness and extended shelf life. Genes CCD4, CRTZ, and ZDS, related to carotenoids, showed varied expression patterns during growth and postharvest stages, with higher levels in 'Rojo Pasión'. On the other hand, Sucrose synthase (SUSY) and Lipoxygenase (LOX2) were prominent during the postharvest and growth stages, respectively. Additionally, GDP-L-galactose phosphorylase (VTC2_5) was linked to better postharvest performance. This research provides valuable insights for future breeding initiatives aimed at enhancing the quality and sustainability of apricot cultivation. [ABSTRACT FROM AUTHOR]

This study aimed at the monitoring of the apricot (Prunus armeniaca L.) ripening progression through the expression analysis of 25 genes related to fruit quality traits in nine cultivars with great differences in fruit color and ripening date. The level of pigment compounds, such as anthocyanins and carotenoids, is a key factor in food taste, and is responsible for the reddish blush color or orange skin and flesh color in apricot fruit, which are desirable quality traits in apricot breeding programs. The construction of multiple linear regression models to predict anthocyanins and carotenoids content from gene expression allows us to evaluate which genes have the strongest influence over fruit color, as these candidate genes are key during biosynthetic pathways or gene expression regulation, and are responsible for the final fruit phenotype. We propose the gene CHS as the main predictor for anthocyanins content, CCD4 and ZDS for carotenoids content, and LOX2 and MADS-box for the beginning and end of the ripening process in apricot fruit. All these genes could be applied as RNA markers to monitoring the ripening stage and estimate the anthocyanins and carotenoids content in apricot fruit during the ripening process. [ABSTRACT FROM AUTHOR]

The study of fruit development in zucchini via gene expression has proven to be applicable in breeding programs. Phenotypic and transcriptomic studies of fruit set and parthenocarpy have been previously developed and some relevant genes have been reported. From these studies, three genotypes (MUCU-16, Whitaker, and Cavili) and six genes (CpAUX22, CpIAA4, CpIAMT-1, CpPIN5, CpCYCD6-1, and CpEXPLB1) were selected. The expression of these genes was analyzed in each genotype under three different treatments (pollination, auxin-treatment and non-treatment) during one week post anthesis. Also, a phenotyping analysis was conducted. The different nature of the samples and the genes selected allowed associations between different fruit traits and fruit development stages. There was a rapid response of CpAUX22 and CpIAA4 to the auxin treatment. Also, these genes and the CpIAMT-1 became more overexpressed in pollinated samples over time. The CpPIN5 gene increased its expression over time in all genotypes while CpCYCD6-1 was overexpressed in the early stages of fruit development in all samples. The CpEXPLB1 was highly up-regulated in non-treated samples, suggesting a relationship with fruit abortion. The overexpression of CpAUX22 and the non-overexpression of CpEXPLB1 in early stages may be associated with fruit growth in zucchini. [ABSTRACT FROM AUTHOR]

Cucurbita species can be affected by soil-borne pathogens, such as Phytopthora capsici and Fusarium solani f. sp. cucurbitae (Fsc). Diverse commercial and conserved lines of Cucurbita spp. were tested. C. pepo subsp. pepo genotypes showed the highest susceptibility to both pathogens. The tolerance to P. capsici and Fsc was then screened in a zucchini mutant population. Two M3 mutant lines (Cp107 and Cp116) with a high occurrence of tolerant individuals to Phytophthora capsici were obtained from a screening of 160 M2 mutant lines. The M3 lines presented higher tolerance than the background MUCU-16. Furthermore, in the inoculated samples, both mutants overexpressed CpDEF and expressed more CpPAL and CpChiIV than the susceptible control. It has been previously shown that this expression pattern could be associated with tolerance in the P. capsici - Cucurbita spp. pathosystem. The M3 lines obtained could be applied in breeding programs, as they are likely to be compatible with the highly susceptible C. pepo subsp. pepo genotype. [ABSTRACT FROM AUTHOR]

Apricot (Prunus armeniaca L.) is a valuable worldwide agronomical crop, with a delicious fruit highlighted as a functional food with both nutritional and bioactive properties, remarkably beneficial to human health. Apricot fruit ripening is a coordinated developmental process which requires change in the expression of hundreds to thousands of genes to modify many biochemical and physiological processes arising from quality characteristics in ripe fruit. In addition, enhancing fruit and nutraceutical quality is one of the central objectives to be improved in the new varieties developed by breeding programs. In this study we analyzed the contents of main metabolites linked to the nutraceutical value of apricot fruits, together with the most important pomological characteristics and biochemical contents of fruit during the ripening process in two contrasted apricot genotypes. Additionally, the gene expression changes were analyzed using RNA-Seq and real time qPCR. Results showed that genes with differential expression in the biosynthetic pathways, such as phenylpropanoids, flavonoids, starch and sucrose and carotenoid metabolism, could be possible candidates as molecular markers of fruit quality characteristics for fruit color and soluble solid content. The gene involves in carotenoid metabolism carotenoid cleavage dioxygenase 4 , and the gene sucrose synthase in starch and sucrose metabolism were identified as candidate genes in the ripening process for white skin ground color and flesh color and high soluble sugar content. The application of these candidate genes on marker-assisted selection in apricot breeding programs may contribute to the early selection of high-quality fruit genotypes with suitable nutraceutical values. [ABSTRACT FROM AUTHOR]

Capsaicin is a natural compound present in chili and red peppers and the responsible of their spicy flavor. It has recently provoked interest because of its antitumoral effects in many cell types although its action mechanism is not clearly understood. As metabolic dysregulation is one of the hallmarks of cancer cells and the key metabolic sensor in the AMP-activated kinase (AMPK), in this study we explored the ability of capsaicin to modulate AMPK activity. We found that capsaicin activated AMPK in HepG2 cells by increasing AMPK phosphorylation and its downstream target ACC. Mechanistically, we determined that capsaicin activated AMPK through the calcium/calmodulin-dependent protein kinase kinase β, CaMKKβ as either the CaMKK inhibitor STO-609 or CaMKK knock down with siRNA abrogated the activation of AMPK. Moreover, capsaicin decreased cell viability, inhibited Akt/mTOR pathway and increased reactive oxygen species (ROS) in HepG2 cells. AMPK activation was involved in the underpinning mechanism of capsaicin-induced cell death. [ABSTRACT FROM AUTHOR]

Plant responses to drought stress are regulated at the transcriptional and post-transcriptional levels through noncoding endogenous microRNAs. These microRNAs play key roles in gene expression, mainly by down-regulating target mRNAs. In this work, an in silico search and validation for microRNAs related to drought response in peach ('G.H. Hill'), almond ('Sefied') and an interspecific peach-almond hybrid ('GN 15') has been performed. We used qPCR to analyse the gene expression of several miRNAs described as being related to drought response in peach, including miR156, miR159, miR160, miR167, miR171, miR172, miR398, miR403, miR408, miR842 and miR2275 under mild and severe water deficit. These miRNAs were in silico selected on the basis of previous works, their conservation in plants and their drought response. qPCR analysis confirmed the implication of these miRNAs in the dehydration stress response in the three assayed genotypes. Comparison of miRNA expression patterns in the three evaluated genotypes indicated that the hybrid GN 15 showed higher expression levels of specific miRNAs which should be related to the observed drought tolerance. mRNA target transcripts of the miRNAs studied were predicted using the Rose database, which includes transcription factors that regulate plant growth and development. In addition, results showed that the promoter region contains responsive elements to hormone-mediated regulatory elements. Network analysis not only unravelled the interaction between miRNAs and their predicted gene targets but also highlighted the roles of miRNAs in response to drought stress. [ABSTRACT FROM AUTHOR]

Background: Simple sequence repeats (SSRs) are defined as sequence repeat units between 1 and 6 bp that occur in both coding and non-coding regions abundant in eukaryotic genomes, which may affect the expression of genes. In this study, expressed sequence tags (ESTs) of eight Prunus species were analyzed for in silico mining of EST-SSRs, protein annotation, and open reading frames (ORFs), and the identification of codon repetitions. Results: A total of 316 SSRs were identified using MISA software. Dinucleotide SSR motifs (26.31 %) were found to be the most abundant type of repeats, followed by tri- (14.58 %), tetra- (0.53 %), and penta- (0.27 %) nucleotide motifs. An attempt was made to design primer pairs for 316 identified SSRs but these were successful for only 175 SSR sequences. The positions of SSRs with respect to ORFs were detected, and annotation of sequences containing SSRs was performed to assign function to each sequence. SSRs were also characterized (in terms of position in the reference genome and associated gene) using the two available Prunus reference genomes (mei and peach). Finally, 38 SSR markers were validated across peach, almond, plum, and apricot genotypes. This validation showed a higher transferability level of EST-SSR developed in P. mume (mei) in comparison with the rest of species analyzed. Conclusions: Findings will aid analysis of functionally important molecular markers and facilitate the analysis of genetic diversity. [ABSTRACT FROM AUTHOR]

Phytophtora capsici causes major diseases in cucurbit crops worldwide. In this study, we inoculated this pathogen into Cucurbita pepo subsp. pepo susceptible MUCU-16 and C. moschata tolerant M63. The gene expression of plant pathogenesis-related proteins chitinase (CpChiIV), lignin-forming peroxidase (CpLPOX), and defensin (CpDEF) and hormone-related enzymes salicylic acid (CpPAL) and ethylene (CpACO) was analyzed for two weeks post-inoculation in root and crown tissues. Differentially expressed genes were found between genotypes, tissues, days post-inoculation, and inoculated/non-inoculated samples. After inoculation, CpPAL and CpChiIV (crown) were downregulated in MUCU-16, while CpLPOX and CpDEF were upregulated in M63. In inoculated samples, higher expression changes were presented on days 10–14 than on day 3 for CpACO, CpLPOX, and CpDEF genes. Overexpression was higher for CpDEF compared to the other tested genes, indicating good suitability as a marker of biotic stress. The overexpression of CpDEF was higher in crown than in roots for both inoculated genotypes. The basal expression of CpPAL and CpDEF was higher in MUCU-16, but after inoculation, CpPAL and CpDEF gene expression were higher in M63. These changes suggest an association between CpDEF upregulation and tolerance, and between CpPAL downregulation and susceptibility. [ABSTRACT FROM AUTHOR]

Summary: Characterization of the tomato falsiflora mutant shows that fa mutation mainly alters the development of the inflorescence resulting in the replacement of flowers by secondary shoots, but also produces a late-flowering phenotype with an increased number of leaves below first and successive inflorescences. This pattern suggests that the FALSIFLORA (FA) locus regulates both floral meristem identity and flowering time in tomato in a similar way to the floral identity genes FLORICAULA (FLO) of Antirrhinum and LEAFY (LFY) of Arabidopsis. To analyse whether the fa phenotype is the result of a mutation in the tomato FLO/LFY gene, we have cloned and analysed the tomato FLO/LFY homologue (TOFL) in both wild-type and fa plants following a candidate gene strategy. The wild-type gene is predicted to encode a protein sharing 90% identity with NFL1 and ALF, the FLO/LFY-like proteins in Nicotiana and Petunia, and about 80 and 70% identity with either FLO or LFY. In the fa mutant, however, the gene showed a 16 bp deletion that results in a frameshift mutation and in a truncated protein. The co-segregation of this deletion with the fa phenotype in a total of 240 F[sub 2] plants analysed supports the idea that FA is the tomato orthologue to FLO and LFY. The gene is expressed in both vegetative and floral meristems, in leaf primordia and leaves, and in the four floral organs. The function of this gene in comparison with other FLO/LFY orthologues is analysed in tomato, a plant with a sympodial growth habit and a cymose inflorescence development. [ABSTRACT FROM AUTHOR]

No natural sources of resistance to Plum pox virus (PPV, sharka disease) have been identified in peach. However, previous studies have demonstrated that grafting a "Garrigues" almond scion onto "GF305" peach rootstock seedlings heavily infected with PPV can progressively reduce disease symptoms and virus accumulation. Furthermore, grafting a "Garrigues" scion onto the "GF305" rootstock has been shown to completely prevent virus infection. This study aims to analyse the rewiring of gene expression associated with this resistance to PPV transmitted by grafting through the phloem using RNA-Seq and RT-qPCR analysis. A total of 18 candidate genes were differentially expressed after grafting "Garrigues" almond onto healthy "GF305" peach. Among the up-regulated genes, a HEN1 homolog stands out, which, together with the differential expression of RDR- and DCL2-homologs, suggests that the RNA silencing machinery is activated by PPV infection and can contribute to the resistance induced by "Garrigues" almond. Glucan endo-1,3-beta D-glucosidase could be also relevant for the "Garrigues"-induced response, since its expression is much higher in "Garrigues" than in "GF305". We also discuss the potential relevance of the following in PPV infection and "Garrigues"-induced resistance: several pathogenesis-related proteins; no apical meristem proteins; the transcription initiation factor, TFIIB; the speckle-type POZ protein; in addition to a number of proteins involved in phytohormone signalling. [ABSTRACT FROM AUTHOR]

Apricot (Prunus armeniaca L.) is globally the third most cultivated stone fruit, prized for its flavourful and nutritious fruit. However, its climacteric nature impacts postharvest behaviour, shortening shelf-life. While the role and interaction of key genes and proteins involved in the ripening process of this species have been extensively studied over the years, research on ethylene suppressors like 1-methylcyclopropene (1-MCP) and cold storage is still limited. Fruit samples from 10 genotypes were collected at commercial maturity from the CEBAS-CSIC research centre and divided into three groups: control (T0), one application of 1-MCP at harvest (T1), and two applications of 1-MCP at harvest and after seven days of cold storage (T2). The study aimed to assess the effects of these treatments on fruit quality traits, such as skin color, soluble solids, acidity, firmness, and ethylene production, after various cold storage periods (10, 20, and 30 days) at 1°C, followed by 48 hours at 20°C. Cold storage combined with 1-MCP effectively reduced fruit softening and ethylene production, with varying impacts on fruit quality traits depending on the genotype. '906–12' and 'Goldrich' were identified as the most contrasting genotypes in terms of ethylene production and fruit softening. Additionally, the application of 1-MCP reduced the expression of genes responsible for fruit softening, coloration and some organoleptic characteristics while enhancing those promoting antioxidant activity during ripening. This study sheds light on the molecular processes involved in apricot fruit ripening and highlights the potential of this approach for enhancing postharvest preservation. • Deeper exploration of fruit quality, postharvest traits, and key ripening genes. • 1-MCP significantly reduced ethylene production, maintaining fruit firmness. • Genotype-dependent response to 1-MCP treatment in delaying ripening. • Suppression of PME, PG, and ACS genes in fruit softening by 1-MCP. • Potential impact on aroma and antioxidant activity due to 1-MCP. [ABSTRACT FROM AUTHOR]

Nine important fruit quality traits—including fruit weight, stone weight, fruit diameter, skin ground colour, flesh colour, blush colour, firmness, soluble solids content and acidity content—were studied for two consecutive years in two F1 apricot progeny derived from the crosses 'Bergeron' × 'Currot' (B×C) and 'Goldrich' × 'Currot' (G×C). Results showed great segregation variability between populations, which was expected because of the polygenic nature and quantitative inheritance of all the studied traits. In addition, some correlations were observed among the fruit quality traits studied. QTL (quantitative trait loci) analysis was carried out using the phenotypic data and genetic linkages maps of 'B×C' and 'G×C' obtained with SSR and SNP markers. The most significant QTLs were localised in LG4 for soluble solids content and in LG3 for skin and flesh colour. In LG4, we can highlight the presence of candidate genes involved in D-glucose and D-mannose binding, while in LG3, we identified MYB genes previously linked to skin colour by other authors. In order to clearly identify the candidate genes responsible for the analysed traits, we converted the QTLs into expression QTLs and analysed the abundance of transcripts in the segregating genotypes 'GC 2–11' and 'GC 3–7' from the G×C population. Using qPCR, we analysed the gene expression of nine candidate genes associated with the QTLs identified, including transcription factors (MYB 10), carotenoid biosynthesis genes (LOX 2, CCD1 and CCD4), anthocyanin biosynthesis genes (ANS, UFGT and F3'5'H), organic acid biosynthesis genes (NAD ME) and ripening date genes (NAC). Results showed variable expression patterns throughout fruit development and between contrasted genotypes, with a correlation between validated genes and linked QTLs. The MYB10 gene was the best candidate gene for skin colour. In addition, we found that monitoring NAC expression is a good RNA marker for evaluating ripening progression. [ABSTRACT FROM AUTHOR]

Highlights: [•] cDNA-AFLP analysis has been used to identify genes responsible for drought tolerance in wild almond P. scoparia identifying twenty differentially expressed transcripts. [•] Five differentially expressed genes were annotated to known function genes including AFC2 kinase. [•] AFC2 kinase interacts with nuclear mRNA splicing proteins as well as epigenetic response proteins during drought stress. [•] The AFC2 derived network has a key regulatory role in P. scoparia drought response. [•] Promoter analysis showed that differentially expressed genes harbour binding sites of MYB1 and MYB2 transcription factors which are involved in dehydration response through ABA signalling pathway. [Copyright &y& Elsevier]