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Plums are affected by a cancerous disease called “black knot disease” caused by the fungus Apiosporina morbosa. It affects both Japanese (Prunus salicina) and European (Prunus domestica) plums equally. To understand the spread of the disease, histological analysis was performed in two different European plum cultivars (susceptible and tolerant). Light and scanning electron microscope (SEM) analyses confirmed the presence of the growing hyphae in the internal tissues of the susceptible trees. By using stereoscopic analysis with a fluorescence filter, we were able to detect the hyphae in the visible lesion area. At about 2 inches from above and below the knots, no spore or hypha were visible with the light microscope. However, SEM images showed strong evidence that the fungus is capable of migrating to adjacent vessels in the susceptible plum genotype. In fact, at that distance below and above the knots, conidia were detected inside xylem vessels suggesting a systemic movement of the fungus that has not been shown so far. No symptoms were observed in the resistant genotype. Starch granules, vessel occlusions, and lipid droplets were the main distinguishable characteristics between susceptible and tolerant varieties.

Somatic mutations may alter important traits in tree fruits, such as fruit color, size and maturation date. Autumn Gala (AGala), a somatic mutation from apple cultivar Gala, matures 4 weeks later than Gala. To understand the mechanisms underlying the delayed maturation, RNA-seq analyses were conducted with fruit sampled at 13 (Gala) and 16 (AGala) time-points during their growth and development. Weighted gene co-expression network analysis (WGCNA) of 23 372 differentially expressed genes resulted in 25 WGCNA modules. Of these, modules 1 (r = -0.98, P = 2E-21) and 2 (r = -0.52, P = 0.004), which were suppressed in AGala, were correlated with fruit maturation date. Surprisingly, 77 of the 152 member genes in module 1 were harbored in a 2.8-Mb genomic region on chromosome 6 that was deleted and replaced by a 10.7-kb gypsy-like retrotransposon (Gy-36) from chromosome 7 in AGala. Among the 77 member genes, MdACT7 was the most suppressed (by 10.5-fold) in AGala due to a disruptive 2.5-kb insertion in coding sequence. Moreover, MdACT7 is the exclusive apple counterpart of Arabidopsis ACT7 known of essential roles in plant development, and the functional allele MdACT7, which was lost to the deletion in AGala, was associated with early fruit maturation in 268 apple accessions. Overexpressing alleles MdACT7 and Mdact7 in an Arabidopsis act7 line showed that MdACT7 largely rescued its stunted growth and delayed initial flowering while Mdact7 did not. Therefore, the 2.8-Mb hemizygous deletion is largely genetically causal for fruit maturation delay in AGala, and the total loss of MdACT7 might have contributed to the phenotype.

Fruit development is orchestrated by a complex network of interactions between hormone signaling pathways. The phytohormone gibberellin (GA) is known to regulate a diverse range of developmental processes; however, the mechanisms of GA action in perennial fruit species are yet to be elucidated. In the current study, a GA signaling gene PslSLY1, encoding a putative F-box protein that belongs to the SLY1 (SLEEPY1)/GID2 (gibberellin-insensitive dwarf2) gene family, was isolated from Japanese plum (Prunus salicina). PslSLY1 transcript abundance declined as fruit development progressed, along with potential negative feedback regulation of PslSLY1 by GA. Subcellular localization and protein–protein interaction assays suggested that PslSLY1 functions as an active GA signaling component that interacts with the ASK1 (Arabidopsis SKP1) subunit of an SCF–ubiquitin ligase complex and with PslDELLA repressors, in a GA-independent manner. By using a domain omission strategy, we illustrated that the F-box and C-terminal domains of PslSLY1 are essential for its interactions with the downstream GA signaling components. PslSLY1 overexpression in wild-type and Arabidopsissly1.10 mutant backgrounds resulted in a dramatic enhancement in overall plant growth, presumably due to triggered GA signaling. This includes germination characteristics, stem elongation, flower structure, and fertility. Overall, our findings shed new light on the GA strategy and signaling network in commercially important perennial crops.

Parthenocarpy and stenospermocarpy are the two mechanisms underlying the seedless fruit set program. Seedless fruit occurs naturally and can be produced using hormone application, crossbreeding, or ploidy breeding. However, the two types of breeding are time-consuming and sometimes ineffective due to interspecies hybridization barriers or the absence of appropriate parental genotypes to use in the breeding process. The genetic engineering approach provides a better prospect, which can be explored based on an understanding of the genetic causes underlying the seedlessness trait. For instance, CRISPR/Cas is a comprehensive and precise technology. The prerequisite for using the strategy to induce seedlessness is identifying the crucial master gene or transcription factor liable for seed formation/development. In this review, we primarily explored the seedlessness mechanisms and identified the potential candidate genes underlying seed development. We also discussed the CRISPR/Cas-mediated genome editing approaches and their improvements.

Muscadine grapes accumulate higher amounts of bioactive phenolics compared with other grape species. To identify the molecular events associated with polyphenolic accumulation that influence antioxidant capacity, two contrasting muscadine genotypes (C5 and C6) with varied phenolic/flavonoid content and antioxidant activity were investigatedviaRNA-sequencing during berry development. The results showed that berry development is concomitant with transcriptome profile changes, which was more pronounced at the véraison (V) stage. Despite that the downregulation pattern of gene expression dominated the upregulation through berry development, the C5 genotype maintained higher expression levels. Comparative transcript profiling allowed the identification of 94 differentially expressed genes with potential relevance in regulating fruit secondary metabolism, including 18 transcription factors and 76 structural genes. The genes underlying the critical enzymes in the modification reactions of polyphenolics biosynthetic pathway, including hydroxylation, methylation, and glycosylation were more pronounced during the immature stages of prevéraison (PrV), V, and postvéraison (PoV) in the C5 genotype, resulting in more accumulation of biologically active phenolic/flavonoid derivatives. The results suggested that muscadine grapes, as in bunch grapes (Vitissp.); possess a similar mechanism that organizes polyphenolics accumulation; however, the set of total flavonoids (TFs) and structural genes coordinating the pathway varies between the two species.

Potato (Solanum tuberosum L.) is an important crop in Egypt with great trade value for the export market. The addition of agrochemicals with possibly containing heavy metals, such as cadmium (Cd), decreases the quality of the final product. Generally, little is known about the factors that influence Cd content in this crop. The current study estimated the Cd levels in different organs of three potato cultivars grown in four commercial regions across Egypt. Further, we investigated the soil properties that affected Cd uptake during two growing seasons. With the exception of the Cd content of the soils, no relationships were detected between the tested soil properties (i.e., pH, conductivity, organic matter, and clay content) and Cd content in potato organs, because the soil from different regions showed comparable levels for these parameters. The average Cd content in the peeled tubers among different cultivars (0.145 mg Kg−1 DW) was below the maximum limit (0.5 mg Kg−1 DW). The patterns of Cd accumulation in potato organs were constant among cultivars, with the highest levels detected in leaves (~82%), followed by stems (~16.5%), and the lowest content observed in tubers (~1.5%). The study showed that the tested potato cultivars exhibited diversity in the accumulation levels of Cd in the tubers (~2.6-fold). The cultivar Suntana displayed the lowest Cd levels among different field sites for the two growing seasons, suggesting the potential involvement of genetic factors.

Malate accumulation in the vacuole largely determines apple (Malus domestica) fruit acidity, and low fruit acidity is strongly associated with truncation of Ma1, an ortholog of ALUMINUM-ACTIVATED MALATE TRANSPORTER9 (ALMT9) in Arabidopsis (Arabidopsis thaliana). A mutation at base 1,455 in the open reading frame of Ma1 leads to a premature stop codon that truncates the protein by 84 amino acids at its C-terminal end. Here, we report that both the full-length protein, Ma1, and its naturally occurring truncated protein, ma1, localize to the tonoplast; when expressed in Xenopus laevis oocytes and Nicotiana benthamiana cells, Ma1 mediates a malate-dependent inward-rectifying current, whereas the ma1-mediated transmembrane current is much weaker, indicating that ma1 has significantly lower malate transport activity than Ma1. RNA interference suppression of Ma1 expression in ‘McIntosh’ apple leaves, ‘Empire’ apple fruit, and ‘Orin’ apple calli results in a significant decrease in malate level. Genotyping and phenotyping of 186 apple accessions from a diverse genetic background of 17 Malus species combined with the functional analyses described above indicate that Ma1 plays a key role in determining fruit acidity and that the truncation of Ma1 to ma1 is genetically responsible for low fruit acidity in apple. Furthermore, we identified a C-terminal domain conserved in all tonoplast-localized ALMTs essential for Ma1 function; protein truncations into this conserved domain significantly lower Ma1 transport activity. We conclude that the truncation of Ma1 to ma1 reduces its malate transport function by removing a conserved C-terminal domain, leading to low fruit acidity in apple.

Background/aim Triple-negative breast cancer (TNBC) is the most aggressive subtype, predominant in African American women. In this study, the antioxidant/anticancer activity of muscadine grape extracts and the role of their phenolic and flavonoid contents in exerting these properties were investigated in TNBC cells. Materials and methods Berry extracts from muscadine genotypes were investigated for total phenolic content (TPC), total flavonoid content (TFC), antioxidant capacity, and anticancer effects using breast cancer cell lines, representing Caucasians and African Americans. Results The antioxidant activity was associated with high TPC content. Extracts showed cytotoxicity up to 78.6% in Caucasians and 90.7% in African American cells, with an association with high antioxidant capacity. There was a strong correlation between TPC and anticancer/antioxidant activities. Conclusion The anticancer and antioxidant effects of muscadine grapes are attributed to the TPC of extracts, which showed a stronger positive correlation with growth inhibition of African American breast cancer cells compared to Caucasians.

Vitis has two subgenera: Euvitis, which includes commercially important Vitis vinifera and interspecific hybrid cultivars, and Muscadinia. Of note, the market for Muscadinia grapes remains small, and only Muscadinia rotundifolia is cultivated as a commercial crop. To establish a basis for the study of Muscadinia species, we generated chromosome-level whole-genome sequences of Muscadinia rotundifolia cv. Noble. A total of 393.8 Mb of sequences were assembled from 20 haploid chromosomes, and 26 394 coding genes were identified from the sequences. Comparative analysis with the genome sequence of V. vinifera revealed a smaller size of the M. rotundifolia genome but highly conserved gene synteny. A genome-wide association study of 12 Muscadinia berry-related traits was performed among 356 individuals from breeding populations of M. rotundifolia. For the transferability of markers between Euvitis and Muscadinia, we used 2000 core genome rhAmpSeq markers developed to allow marker transferability across Euvitis species. A total of 1599 (80%) rhAmpSeq markers returned data in Muscadinia. From the GWAS analyses, we identified a total of 52 quantitative trait nucleotides (QTNs) associated with the 12 berry-related traits. The transferable markers enabled the direct comparison of the QTNs with previously reported results. The whole-genome sequences along with the GWAS results provide a new basis for the extensive study of Muscadinia species.

Three muscadine grape genotypes (Muscadinia rotundifolia (Michx.) Small) were evaluated for their metabolite profiling and antioxidant activities at different berry developmental stages. A total of 329 metabolites were identified using UPLC-TOF-MS analysis (Ultimate 3000LC combined with Q Exactive MS and screened with ESI-MS) in muscadine genotypes throughout different developmental stages. Untargeted metabolomics study revealed the dominant chemical groups as amino acids, organic acids, sugars, and phenolics. Principal component analysis indicated that developmental stages rather than genotypes could explain the variations among the metabolic profiles of muscadine berries. For instance, catechin, epicatechin-3-gallate, and gallic acid were more accumulated in ripening seeds (RIP-S). However, tartaric acid and malonic acid were more abundant during the fruit-set (FS) stage, and malic acid was more abundant in the veraison (V) stage. The variable importance in the projection (VIP &gt, 0.5) in partial least-squares–discriminant analysis described 27 biomarker compounds, representing the muscadine berry metabolome profiles. A heatmap of Pearson’s correlation analysis between the 27 biomarker compounds and antioxidant activities was able to identify nine antioxidant determinants, among them, gallic acid, 4-acetamidobutanoic acid, trehalose, catechine, and epicatechin-3-gallate displayed the highest correlations with different types of antioxidant activities. For instance, DPPH and FRAP conferred a similar antioxidant activity pattern and were highly correlated with gallic acid and 4-acetamidobutanoic acid. This comprehensive study of the metabolomics and antioxidant activities of muscadine berries at different developmental stages is of great reference value for the plant, food, pharmaceutical, and nutraceutical sectors.

Biochemical juice attributes and color-related traits of muscadine grape genotypes have been investigated. For this study, 90 muscadine genotypes, including 21 standard cultivars, 60 breeding lines, and 9 Vitis x Muscadinia hybrids (VM), were evaluated. The biochemical properties of total soluble solids (TSS), titratable acidity, and TSS/Acid (T/A) ratio showed modest diversity among genotypes with a range of 10.3 °Brix, 2.1 mg tartaric acid/L, and 4.6, respectively. Nonetheless, the pH trait exhibited a tight range of 0.74 among the population with a minimum and maximum pH of 3.11 ± 0.12 and 3.85 ± 0.12. Color-related traits showed more deviation between individuals. Total anthocyanin content (TAC), luminosity index (L*), hue angle (h°), and chroma index (C*) displayed a range of 398 µg/g DW, 33.2, 352.1, and 24, respectively. The hierarchical clustering map classified the population into two large groups of colored and non-colored grapes based on L* and h°, suggesting the predominance of these two characters among the population. The colored berries genotypes clade was further divided into several sub-clades depending on C*, TAC, and TSS levels. The principal component analysis (PCA) separated the four-color characteristics into two groups with a negative correlation between them, L* and C* versus TAC and h°. Further, PCA suggested the positive influence of acidity in enhancing the different nutraceutical components. Despite the nature of anthocyanins as a member of phenolic compounds, a lack of significant correlation between TAC and nutraceutical-related traits was detected. The dissimilatory matrix analysis highlighted the muscadine individuals C11-2-2, E16-9-1, O21-13-1, and Noble as particular genotypes among the population due to enhanced color characteristics.

Eight blueberry cultivars at three developmental stages were investigated for metabolite profiling, antioxidant, and anticancer activities. Cultivars- and developmental stages-variations were determined in total phenolic, flavonoid, DPPH, and FRAP antioxidant assays. The anticancer capacity was equal against A549, HepG2, and Caco-2 cancer cells, whereas the inhibition rate was dose-, incubation period-, cultivar-, and developmental stages-dependent. The untargeted metabolite profiling by UPLC-TOF-MS analysis of two contrast cultivars, 'Vernon' and 'Star', throughout the developmental stages revealed 328 metabolites; the majority of them were amino acids, organic acids, and flavonoids. The multivariate statistical analysis identified five metabolites, including quinic acid, methyl succinic acid, chlorogenic acid, oxoadipic acid, and malic acid, with positively higher correlations with all anticancer activities. This comprehensive database of blueberry metabolites along with anticancer activities could be targeted as natural anticancer potentials. This study would be of great value for food, nutraceutical, and pharmaceutical industries as well as plant biotechnologists.

Plant stress is a real dilemma; it puzzles plant biologists and is a global problem that negatively affects people’s daily lives. Of particular interest is salinity, because it represents one of the major water-related stress types. We aimed to determine the signals that guide the cellular-related events where various adaptation mechanisms cross-talk to cope with salinity-related water stress in plants. In an attempt to unravel these mechanisms and introduce cellular events in the right context, we expansively discussed how salt-related signals are sensed, with particular emphasis on aquaporins, nonselective cation channels (NSCCs), and glycosyl inositol phosphorylceramide (GIPC). We also elaborated on the critical role Ca2+, H+, and ROS in mediating signal transduction pathways associated with the response and tolerance to salt stress. In addition, the fragmentary results from the literature were compiled to develop a harmonized, informational, and contemplative model that is intended to improve our perception of these adaptative mechanisms and set a common platform for plant biologists to identify intriguing research questions in this area. Published version

While horizontally transferred transposable elements (TEs) have been reported in several groups of plants, their importance for genome evolution remains poorly understood. To understand how horizontally transferred TEs contribute to plant genome evolution, we investigated the composition and activity of horizontally transferred TEs in the genomes of four Vitis species. A total of 35 horizontal transfer (HT) events were identified between the four Vitis species and 21 other plant species belonging to 14 different families. We determined the donor and recipient species for 28 of these HTs, with the Vitis species being recipients of 15 of them. As a result of HTs, 8–10 LTR retrotransposon clusters were newly formed in the genomes of the four Vitis species. The activities of the horizontally acquired LTR retrotransposons differed among Vitis species, showing that the consequences of HTs vary during the diversification of the recipient lineage. Our study provides the first evidence that the HT of TEs contributes to the diversification of plant genomes by generating additional TE subfamilies and causing their differential proliferation in host genomes.

In this study, fertility-related traits of 90 muscadine grape genotypes were evaluated. Selected genotypes included 21 standard cultivars, 60 breeding lines, and nine Vitis × Muscadinia hybrids (VM hybrids). The first fruiting bud (FFB), bud fertility (BF), bud fertility coefficient (BFC), number of flowers/flower cluster (N.F/FC), fruit-set efficiency (FSE), number of clusters/vine (N.C/V), and yield/vine (Y/V) traits were evaluated. The FFB trait did not show significant differences among genotypes. The muscadine genotype O28-4-2-2 (1.6 ± 0.2) displayed the FFB closest to the base, however, O17-16-2-1, O18-2-1, and VM A12-10-2 genotypes had the most distant FFB (3.6 ± 0.3). All the other fertility-related traits varied widely among the population. The BF, BFC, N.F/FC, FSE, N.C/V, and Y/V exhibited a range estimated at 35.1%, 81.5%, 259.7, 63.3%, 177 C/V, and 22.3 kg/V, respectively. The muscadine genotypes O42-3-1 (36.7% ± 1.3) and Majesty (34% ± 1.2) exhibited the highest BF, however, the VM A12-10-2 (1.6% ± 0.1) recorded the lowest BF. The VM genotype O15-16-1 (82.8% ± 4.1) displayed the highest BFC, however, the VM A12-10-2 (1.3% ± 0.1) showed the lowest BFC. The muscadine genotypes D7-1-1 (280.3 F/FC ± 21.7) and O17-17-1 (20.7 F/FC ± 5.5) showed the highest and lowest N.F/FC, respectively. The maximum and minimum FSE was observed for the Rosa cultivar (65.7% ± 2.4) and muscadine genotype D7-1-1 (2.4% ± 0.2), respectively. The minimum N.C/V was recorded for VM genotype A12-10-2 (6 C/V ± 0.2) and maximum noted for muscadine genotypes B20-18-2 (183 C/V ± 7.5) and O44-14-1 (176 C/V ± 7.3). Muscadine genotype O23-11-2 (22.6 kg ± 1.1) produced the highest Y/V, however, the lowest yield was recorded for O15-17-1, Fry Seedless, Sugargate, and the VM genotypes and A12-10-2, with an average yield among them estimated at 0.4 kg ± 0.2.

In this study, biometrics assessment of flower structure, cluster-, and berry-related traits were evaluated in a population of 90 muscadine grape genotypes for three consecutive years. This population consisted of 21 standard cultivars, 60 breeding lines, and 9 Vitis x Muscadinia hybrids (VM hybrids). Cluster length (CL) and width (CWI) characteristics exhibited slight differences among the population, with a range estimated at 7.1 and 4.6 cm, respectively. However, cluster weight (CWE), number of berries/cluster (N.B/C), and cluster compactness (CC) traits showed more diversity between individuals with a calculated range of 205.6 g, 32.6 B/C, and 24.1, respectively. Interestingly, all berry-related traits greatly varied between individuals, excluding the number of seeds/berry (N.S/B) character. The N.S/B trait displayed a narrow range of 5.6 seeds within the population. However, characters of berry length (BL), width (BWI), weight (BWE), the weight of seeds/berry (W.S/B), firmness (FF), and dry scar pattern (SP) demonstrated a wide estimated range of 21.2 mm, 21.7 mm, 25.4 g, 0.71 g, 0.21 N, and 82%, respectively. Normal distribution analysis for each trait suggested different distribution patterns extended between unimodal to multimodal behavior. Hierarchical mapping analysis was able to classify the population into several clades based on physical cluster- and berry-related attributes. The PCA suggested that hermaphroditic (perfect) flower structure is associated with compact clusters exhibiting small berries in size and weight (i.e., muscadine genotypes suitable for wine production). However, female flower structure is associated with clusters displaying large berries in size and weight (i.e., muscadine genotypes appropriate for fresh consumption). These patterns occurred independently of cluster size and weight characters. This research is the first study evaluating muscadine biometrics characters at a population level, providing valuable information for market demand and muscadine breeding programs.

European plum fruit (Prunus domestica) are normally blue-black to dark purple. However, some genotypes remain green/yellow after ripening. We hypothesized that in such genotypes anthocyanin biosynthesis is genetically disturbed. To examine this hypothesis, six european plum genotypes with diverse fruit colors were investigated for the expression pattern of several anthocyanin biosynthetic genes (ABGs)—e.g., phenylalanine ammonia-lyase, chalcone synthase (CHS), dihydroflavonol 4-reductase (DFR), anthocyanin synthase (ANS), and UDP-glucose:flavonoid 3-O-glucosyltransferase 1 and 2 (UFGT 1 and 2). Expression profiles indicated that ABGs, especially Pd-CHS and UFGT 2, were significantly downregulated in the green/yellow fruit compared with the dark-purple fruit. Furthermore, the quantification of total polyphenols and individual flavonoid compounds showed substantial differences between the off-colored and the purple genotype. To further examine the contribution of each of the ABGs in color development, the open reading frame (ORP) of Pd-CHS, Pd-DFR, Pd-ANS, and Pd-UFGT 2 was ectopically expressed in tobacco (Nicotiana tabacum). The characterization of transgenic plants showed that the petals of plants expressing Pd-CHS were darker in color and had higher anthocyanin content than control or even other transgenic types, suggesting the significant contribution of CHS in determining anthocyanin production levels and hence fruit coloration. The results of this study provides better understanding of color development in european plum, which can be rewarding in developing european plum cultivars with desired colors through classical or modern breeding tools.

Ripening is a highly coordinated, irreversible event involving a series of physiological and biochemical changes, leading to the development of a soft fruit. One of the limiting factors that influence the economic value of fruit is the relatively short ripening period and limited shelf-life. In climacteric fruit, ethylene is a key regulator of ripening; however, recent research has shown that auxin also plays an important role during the event. To understand the contributions of ethylene, auxin and their interaction in ripening, two plum cultivars with widely varying fruit ripening behaviors were compared. The early cultivar, EG, exhibited a brief ripening process in association with rapid decline in firmness. The late cultivar, V9, displayed slow ripening behavior accompanying by notable extension in fruit firmness, resulting in prolonged shelf-life along with preserved fruit quality traits. Auxin has been suggested to play an indirect role in promoting fruit ripening via stimulating the transcription of several ethylene components, resulting in ethylene-induced fruit ripening and softening. This study shows further that there is a direct involvement of auxin in advancing ripening events independent of ethylene action through stimulating the transcription of several genes that encode cell-wall metabolism-related proteins critical for determining the fruit softening rate and potential shelf-life. These results support the hypothesis that the autonomous role played by auxin is as important as that of ethylene in determining not only fruit ripening behavior, but also in mediating other fruit quality traits including shelf-life.

The role of hormones during fruit development and ripening in strawberry (Fragaria × ananassa) is poorly understood. In this study, two strawberry cultivars (‘Jewel’ and ‘Wendy’) were chosen based on their shelf life quality, which were intermediate and excellent, respectively. Hexanal and growth regulators were applied to the fruit as two preharvest sprays. Fruits treated with “Enhanced Freshness Formulation” (a formulation with hexanal as a key ingredient) showed improvement in the shelf life of both cultivars. Auxin application reduced loss of fruit firmness and abscisic acid was able to accelerate the ripening process without having a significant effect on the relevant fruit quality parameters. The regulation of gene expression during ripening in relation to hexanal and hormones was examined in 21 genes potentially involved in cell wall degradation. Gene expression profiles showed similar patterns in the two cultivars, with more prominent amplitude in ‘Wendy’. The expression of hormone-responsive gene...

In order to obtain nutrients from plants for survival and reproduction, phytopathogenic bacteria interfere with and disrupt many plant functions. Interactions between plants and bacteria and their final outcomes are tightly controlled at the molecular level by both organisms. During their coevolution, pathogenic bacteria and plants have evolved various mechanisms to increase their pathogenicity or resistance, respectively. The purpose of this review is to shed light on the different stages where interactions between plants and bacteria take place, how plants respond at each stage, and how bacteria can evade plant responses. Advances in molecular and cellular biology, genomics, and bioinformatics have revealed different elements in this continuous interplay. The successful use of molecular elements to produce plants resistant to a particular disease or showing broad-spectrum resistance to different bacterial strains/species has already been demonstrated in many horticultural and field crops.

Peach ERF3b is a potent transcriptional repressor for defense-related genes even in the presence of similar levels of transcriptional activators and can interfere with plant development through pathways independent of the EAR motif. Ethylene response factors (ERFs) are a major group of plant transcription factors with either activation or repression capabilities on gene transcription. Repressor-type ERFs are characterised by an intrinsic motif, namely the ERF-associated amphiphilic repression motif (EAR). Here we report the identification of three genes from peach (Prunus persica), PpERF12, PpERF3a and PpERF3b, encoding for ERF repressors. The transcription kinetics of these genes was investigated by qRT-PCR after inoculation of peach leaves with Xanthomonas campestris pv. pruni. All three genes showed higher induction in the susceptible 'BabyGold 5', than in the resistant 'Venture' peach varieties suggesting a negative role for these genes in disease resistance. The functional potency of PpERF3b has been confirmed in vivo by its ability to repress the expression of GUS-reporter gene. To better understand the functional role of PpERF3b, the full-length and the EAR-truncated (PpERF3b∆EAR) genes were overexpressed in tobacco (Nicotiana tabacum). Both transgenic plants (PpERF3b and PpERF3b∆EAR) uniformly exhibited precocious side branching, which suggests the interference of PpERF3b with auxin-mediated dormancy of lateral shoots. Consistent with that the expression of auxin-response factors (Nt-ARF1, Nt-ARF6 and Nt-ARF8) was significantly downregulated in transgenic plants compared to the wild type (WT). Although side branching was independent of EAR motif, the response of transgenic plants to inoculation by Pseudomonas syringae pv. tabaci was EAR dependent. Transgenic plants overexpressing PpERF3b∆EAR showed less disease symptoms than those overexpressing the full-length gene or WT plants. Resistance of PpERF3b∆EAR plants was associated with enhanced induction of pathogenesis-related (PR) genes. Our results indicate that repressor-type ERFs might act through pathways that are dependent or independent of the EAR motif.

International audience; The apical hook of dark-grown dicotyledonous plants results from asymmetric growth of the inner and outer sides of the upper part of the hypocotyl. This protective structure prevents damage of the shoot apical meristem and the young leaves as the seedling pushes through the soil. HOOKLESS (HLS1) of Arabidopsis was recognised as an ethylene response gene whose product is required for hook formation. We cloned two cDNAs from tomato, Sl-HLS1 and Sl-HLS2, and showed through the complementation of the Arabidopsis hls1 mutant that the encoded proteins are functional orthologs of At-HLS1. The genomic clones of Sl-HLS1 and Sl-HLS2 showed similar structure with two introns and three exons. While the data indicate complete functional redundancy between the two tomato HLS genes, only the expression of Sl-HLS2 is regulated by ethylene and auxin and the ethylene-induced expression of Sl-HLS2 is localised in the outer side of the hook while the auxin-responsive expression is not restricted to a specific side of the hook. 1-MCP, an inhibitor of ethylene signalling, inhibited auxin-enhanced accumulation of Sl-HLS2 mRNA in the apical hook suggesting that regulation of Sl-HLS2 by auxin is ethylene-dependent. The overexpression of tomato Sl-HLS1 and Sl-HLS2 in Arabidopsis confers hypersensitivity to ethylene. The data presented bring further insight into the complex ethylene-auxin interplay in hook formation.

Traditional Charentais melons have a typical climacteric behavior with ethylene playing a major role in the regulation of the ripening process. Genetic studies using climacteric and non-climacteric types of Cucumis melo demonstrated that the climacteric character is dominant and conferred by 2 duplicated loci only which are of great importance for the regulation of storability and sensory quality. Commercial varieties of Charentais melon with long shelf-life have been generated, some of them by crossing with a non-ripening Charentais genotype (Vauclusien). The introduction of the long shelf-life character resulted in undesirable loss of aroma volatiles production. The inhibition of ethylene synthesis by knocking-down ACC oxidase gene expression has been achieved in Charentais melon. It results is a strong inhibition of the synthesis of aroma volatiles while the accumulation of sugars is not affected or is even improved and the softening of the flesh is strongly affected but not abolished. It was also demonstrated that ethylene-inhibited fruit exhibited better resistance to chilling injury. Due to the importance of aroma volatiles in sensory quality and to the strong negative correlation between aroma production and ethylene synthesis, we have developed a research program aimed at isolating genes involved in the synthesis of volatile esters, compounds that are essential for the flavor of Cantaloupe melons. We report here on the recent advances in the field with special emphasis on the characterization of two families of genes encoding aldehyde reductases and alcohol acyl transferases.

Background TIR1-like proteins are F-box auxin receptors. Auxin binding to the F-box receptor proteins promotes the formation of SCFTIR1 ubiquitin ligase complex that targets the auxin repressors, Aux/IAAs, for degradation via the ubiquitin/26S proteasome pathway. The release of auxin response factors (ARFs) from their Aux/IAA partners allows ARFs to mediate auxin-responsive changes in downstream gene transcription. In an attempt to understand the potential role of auxin during fruit development, a plum auxin receptor, PslTIR1, has previously been characterized at the cellular, biochemical and molecular levels, but the biological significance of this protein is still lacking. In the present study, tomato (Solanum lycopersicum) was used as a model to investigate the phenotypic and molecular changes associated with the overexpression of PslTIR1. Results The findings of the present study highlighted the critical role of PslTIR1 as positive regulator of auxin-signalling in coordinating the development of leaves and fruits. This was manifested by the entire leaf morphology of transgenic tomato plants compared to the wild-type compound leaf patterning. Moreover, transgenic plants produced parthenocarpic fruits, a characteristic property of auxin hypersensitivity. The autocatalytic ethylene production associated with the ripening of climacteric fruits was not significantly altered in transgenic tomato fruits. Nevertheless, the fruit shelf-life characteristics were affected by transgene presence, mainly through enhancing fruit softening rate. The short shelf-life of transgenic tomatoes was associated with dramatic upregulation of several genes encoding proteins involved in cell-wall degradation, which determine fruit softening and subsequent fruit shelf-life. Conclusions The present study sheds light into the involvement of PslTIR1 in regulating leaf morphology, fruit development and fruit softening-associated ripening, but not autocatalytic ethylene production. The results demonstrate that auxin accelerates fruit softening independently of ethylene action and this is probably mediated through the upregulation of many cell-wall metabolism genes. Electronic supplementary material The online version of this article (doi:10.1186/s12870-016-0746-z) contains supplementary material, which is available to authorized users.

The sensory quality of fruit has become a major criterion in making the purchasing decision by consumers. Breeding programs have mainly been directed, from the post-harvest stand point, towards improving shelf-life. Chance seedlings or mutants with improved agronomic traits and/or extended shelf-life have been used for introgressing the long shelf-life character and eventually improved sensory quality traits in commercial genotypes of apple, melon or tomato. Because the plant hormone ethylene plays a central role in both storability and ripening of climacteric fruit, the generation by biotechnology of ethylene-inhibited fruit has offered a powerful tool to better understand, at the molecular and genetic level, the interrelations between storability and sensory quality. In the melon, inhibition of ethylene synthesis results is a strong inhibition of the synthesis of aroma volatiles while the accumulation of sugars is not affected or is even improved. The softening of the flesh is strongly affected but not abolished. Mid or long shelf-life melons generated by classical breeding present the same behavior. The generation of recombinant inbred lines by crossing a typical climacteric melon (Cantaloupe Charentais of the cantalupensis group) with a non climacteric melon (PI161375 of the agrestis chinensis group) allowed to demonstrate that the climacteric character is conferred by 2 duplicated loci only, which are of great importance for the regulation of storability and sensory quality. Due to the importance of aroma volatiles in sensory quality and to the strong negative correlation between aroma production and ethylene synthesis, we have developed a research program aimed at isolating genes involved in the synthesis of aroma volatiles. We will report on the recent advances in the field with special emphasis on the characterization of genes responsible for the synthesis of esters, a family of compounds crucial for the flavor of many fruit.

The differential regulation of ACC synthase genes has been studied in pear cultivars that either require a long chilling treatment before they are capable of ripening (‘Passe-Crassane’, PC) or not (‘Old-Home’, OH) and in OH x PC hybrids having no (A16) or intermediate (A50) cold requirement. Among the seven Pc-ACS cDNAs isolated, four of them (Pc-ACS1a/b and Pc-ACS2a/b) showed differential expression in relation with cold requirement. Pc-ACS1a transcripts accumulated specifically during chilling and ripening of cold-dependent cultivars while Pc-ACS1b transcripts were detected only during ripening of cold-independent genotypes. Pc- ACS2a mRNA was expressed specifically in cold-dependent genotypes and negatively regulated by ethylene while Pc-ACS2b transcripts accumulated only in cold-independent genotypes and positively regulated by ethylene. Pc-ACS3, 4 and 5 transcripts accumulation was similar in all genotypes, independently of coldrequirements.

Fruit growth depends on highly coordinated hormonal activities. The phytohormone gibberellin (GA) promotes growth by triggering degradation of the growth-repressing DELLA proteins; however, the extent to which such proteins contribute to GA-mediated fruit development remains to be clarified. Three new plum genes encoding DELLA proteins, PslGAI, PslRGL and PslRGA were isolated and functionally characterized. Analysis of expression profile during fruit development suggested that PslDELLA are transcriptionally regulated during flower and fruit ontogeny with potential positive regulation by GA and ethylene, depending on organ and developmental stage. PslGAI and PslRGL deduced proteins contain all domains present in typical DELLA proteins. However, PslRGA exhibited a degenerated DELLA domain and subsequently lacks in GID1-DELLA interaction property. PslDELLA-overexpression in WT Arabidopsis caused dramatic disruption in overall growth including root length, stem elongation, plant architecture, flower structure, fertility, and considerable retardation in development due to dramatic distortion in GA-metabolic pathway. GA treatment enhanced PslGAI/PslRGL interaction with PslGID1 receptors, causing protein destabilization and relief of growth-restraining effect. By contrast, PslRGA protein was not degraded by GA due to its inability to interact with PslGID1. Relative to other PslDELLA-mutants, PslRGA-plants displayed stronger constitutive repressive growth that was irreversible by GA application. The present results describe additional complexities in GA-signalling during plum fruit development, which may be particularly important to optimize successful reproductive growth.

Highlight text A mutation in the plum auxin-receptor encoded by PslAFB5, which acts as a negative regulator of auxin responses, results in inactive Pslafb5 protein causing accelerated fruit-ontogeny events associated with auxin hypersensitivity., Ethylene has long been considered the key regulator of ripening in climacteric fruit. Recent evidence showed that auxin also plays an important role during fruit ripening, but the nature of the interaction between the two hormones has remained unclear. To understand the differences in ethylene- and auxin-related behaviours that might reveal how the two hormones interact, we compared two plum (Prunus salicina L.) cultivars with widely varying fruit development and ripening ontogeny. The early-ripening cultivar, Early Golden (EG), exhibited high endogenous auxin levels and auxin hypersensitivity during fruit development, while the late-ripening cultivar, V98041 (V9), displayed reduced auxin content and sensitivity. We show that exogenous auxin is capable of dramatically accelerating fruit development and ripening in plum, indicating that this hormone is actively involved in the ripening process. Further, we demonstrate that the variations in auxin sensitivity between plum cultivars could be partially due to PslAFB5, which encodes a TIR1-like auxin receptor. Two different PslAFB5 alleles were identified, one (Pslafb5) inactive due to substitution of the conserved F-box amino acid residue Pro61 to Ser. The early-ripening cultivar, EG, exhibited homozygosity for the inactive allele; however, the late cultivar, V9, displayed a PslAFB5/afb5 heterozygous genotype. Our results highlight the impact of auxin in stimulating fruit development, especially the ripening process and the potential for differential auxin sensitivity to alter important fruit developmental processes.

Publication Inra prise en compte dans l'analyse bibliométrique des publications scientifiques mondiales sur les Fruits, les Légumes et la Pomme de terre. Période 2000-2012. http://prodinra.inra.fr/record/256699; International audience; Several dwarf plum genotypes (Prunus salicina L.), due to deficiency of unknown gibberellin (GA) signalling, were identified. A cDNA encoding GA 2-oxidase (PslGA2ox), the major gibberellin catabolic enzyme in plants, was cloned and used to screen the GA-deficient hybrids. This resulted in the identification of a dwarf plum hybrid, designated as DGO24, that exhibits a markedly elevated PslGA2ox signal. Grafting 'Early Golden' (EG), a commercial plum cultivar, on DGO24 (EG/D) enhanced PslGA2ox accumulation in the scion part and generated trees of compact stature. Assessment of active GAs in such trees revealed that DGO24 and EG/D accumulated relatively much lower quantities of main bioactive GAs (GA(1) and GA(4)) than control trees (EG/M). Moreover, the physiological function of PslGA2ox was studied by determining the molecular and developmental consequences due to ectopic expression in Arabidopsis. Among several lines, two groups of homozygous transgenics that exhibited contrasting phenotypes were identified. Group-1 displayed a dwarf growth pattern typical of mutants with a GA deficiency including smaller leaves, shorter stems, and delay in the development of reproductive events. In contrast, Group-2 exhibited a 'GA overdose' phenotype as all the plants showed elongated growth, a typical response to GA application, even under limited GA conditions, potentially due to co-suppression of closely related Arabidopsis homologous. The studies reveal the possibility of utilizing PslGA2ox as a marker for developing size-controlling rootstocks in Prunus.

Publication Inra prise en compte dans l'analyse bibliométrique des publications scientifiques mondiales sur les Fruits, les Légumes et la Pomme de terre. Période 2000-2012. http://prodinra.inra.fr/record/256699; International audience; Auxin-binding protein1 (ABP1) is an active element involved in auxin signaling and plays critical roles in auxin-mediated plant development. Here, we report the isolation and characterization of a putative sequence from Prunus salicina L., designated PslABP1. The expected protein exhibits a similar molecular structure to that of well-characterized maize-ABP1; however, PslABP1 displays more sequence polarity in the active-binding site due to substitution of some crucial amino-acid residues predicted to be involved in auxin-binding. Further, PslABP1 expression was assessed throughout fruit ontogeny to determine its role in fruit development. Comparing the expression data with the physiological aspects that characterize fruit-development stages indicates that PslABP1 up-regulation is usually associated with the signature events that are triggered in an auxin-dependent manner such as floral induction, fruit initiation, embryogenesis, and cell division and elongation. However, the diversity in PslABP1 expression profile during the ripening process of early and late plum cultivars seems to be due to the variability of endogenous auxin levels among the two cultivars, which consequently can change the levels of autocatalytic ethylene available for the fruit to co-ordinate ripening. The effect of auxin on stimulating ethylene production and in regulating PslABP1 was investigated. Our data suggest that auxin is involved in the transition of the mature green fruit into the ripening phase and in enhancing the ripening process in both auxin- and ethylene-dependent manners thereafter.