MYB transcription factor family in sweet cherry (Prunus avium L.): genome-wide investigation, evolution, structure, characterization and expression patterns.

BACK GROUND: MYB Transcription factors (TFs) are most imperative and largest gene family in plants, which participate in development, metabolism, defense, differentiation and stress response. The MYB TFs has been studied in various plant species. However, comprehensive studies of MYB gene family in the sweet cherry (Prunus avium L.) are still unknown. RESULTS: In the current study, a total of 69 MYB genes were investigated from sweet cherry genome and classified into 28 subfamilies (C1-C28 based on phylogenetic and structural analysis). Microcollinearity analysis revealed that dispersed duplication (DSD) events might play an important role in the MYB genes family expansion. Chromosomal localization, the synonymous (Ks) and nonsynonymous (Ka) analysis, molecular characteristics (pI, weight and length of amino acids) and subcellular localization were accomplished using several bioinformatics tools. Furthermore, the members of distinct subfamilies have diverse cis-acting regions, conserved motifs, and intron-exon architectures, indicating functional heterogeneity in the MYB family. Moreover, the transcriptomic data exposed that MYB genes might play vital role in bud dormancy. The quantitative real-time qRT-PCR was carried out and the expression pattern indicated that MYB genes significantly expressed in floral bud as compared to flower and fruit. CONCLUSION: Our comprehensive findings provide supportive insights into the evolutions, expansion complexity and functionality of PavMYB genes. These PavMYB genes should be further investigated as they seem to be brilliant candidates for dormancy manipulation in sweet cherry.

Summer pruning of sweet cherry: a way to control sugar content in different organs.

BACKGROUND: Sweet cherry trees (Prunus avium L.) of the cultivar Grace Star were pruned either in dormancy or in summer. The response was studied by analyzing the sugar content in different organs (flower bud, leaf, and fruit) at three sections of the canopy (inner, outer, and upper) using high-performance liquid chromatography. The effect of summer pruning was evaluated by measuring photosynthetic photon flux density (PPFD) and leaf chlorophyll content (SPAD). RESULTS: In this study, the timing of pruning had a significant effect on sugar content in flower buds, leaves, and fruit. Trees pruned in summer had higher glucose, fructose, sorbitol, and sucrose content in flower buds, higher glucose and fructose contents in leaves, and lower fructose, sorbitol, and total sugar content in fruit than in trees pruned at dormancy. Higher average PPFD and lower SPAD values were measured in the inner canopy of trees pruned in summer. All measured parameters were influenced by position in the canopy. The lowest fructose and sorbitol contents in the flower bud, the lowest content of glucose, fructose, sorbitol, total sugars and the highest SPAD values in the leaf, while less dark and lighter fruit were measured in the inner part of the canopy. CONCLUSION: Summer pruning affects sugar distribution in the tree by altering irradiation conditions within the canopy. Our results suggest that summer pruning is an effective technological measure to improve sugar content in the buds. A strong, well nourished flower bud is a good indication of high fruit production next season. © 2021 Society of Chemical Industry.

Cold induced genes (CIGs) regulate flower development and dormancy in Prunus avium L.

The flower buds continue to develop during the whole winter in tree fruit species, which is affected by environmental factors and hormones. However, little is known about the molecular mechanism of flower development during dormancy phase of sweet cherry in response to light, temperature and ABA. Therefore, we identified two cold induced gene (CIG) PavCIG1 and PavCIG2 from sweet cherry, which were closely to PpCBF and PyDREB from Prunus persica and Prunus yedoensis by using phylogenetic analysis, suggesting conserved functions with these evolutionarily closer DREB subfamily genes. Subcellular localization analysis indicated that, PavCIG1 and PavCIG2 were both localized in the nucleus. The seasonal expression levels of PavCIG1 and PavCIG2 were higher at the stage of endodormancy in winter, and induced by low temperature. Ectopic expression of PavCIG1 and PavCIG2 resulted in a delayed flowering in Arabidopsis. Furthermore, PavCIG2 increased light-responsive gene PavHY5 transcriptional activity by binding to its promoter, meanwhile, PavHY5-mediated positive feedback regulated PavCIG2. Moreover, ABA-responsive protein PavABI5-like could also increase transcriptional activity of PavCIG and PavCIG2. In addition, PavCIG and PavCIG2 target gene PavCAL-like was involved in floral initiation, demonstrated by ectopic expression in Arabidopsis. These findings provide evidences to better understand the molecular mechanism of CIG-mediated flower development and dormancy in fruit species, including sweet cherry.

Genome-Wide Identification of ARF Gene Family Suggests a Functional Expression Pattern during Fruitlet Abscission in Prunus avium L.

Auxin response factors (ARFs) play a vital role in plant growth and development. In the current study, 16 ARF members have been identified in the sweet cherry (Prunus avium L.) genome. These genes are all located in the nucleus. Sequence analysis showed that genes in the same subgroup have similar exon-intron structures. A phylogenetic tree has been divided into five groups. The promoter sequence includes six kinds of plant hormone-related elements, as well as abiotic stress response elements such as low temperature or drought. The expression patterns of PavARF in different tissues, fruitlet abscission, cold and drought treatment were comprehensively analyzed. PavARF10/13 was up-regulated and PavARF4/7/11/12/15 was down-regulated in fruitlet abscising. These genes may be involved in the regulation of fruit drop in sweet cherry fruits. This study comprehensively analyzed the bioinformatics and expression pattern of PavARF, which can lay the foundation for further understanding the PavARF family in plant growth development and fruit abscission.

Auxin Treatment Enhances Anthocyanin Production in the Non-Climacteric Sweet Cherry (Prunus avium L.).

Abscisic acid (ABA) is a key signaling molecule promoting ripening of non-climacteric fruits such as sweet cherry (Prunus avium L.). To shed light on the role of other hormones on fruit development, ripening and anthocyanin production, the synthetic auxin 1-naphthaleneacetic acid (NAA) was applied to sweet cherry trees during the straw-color stage of fruit development. NAA-treated fruits exhibited higher concentrations of 1-aminocyclopropane-1-carboxylic acid (ACC) and ABA-glucose ester (ABA-GE), which are a precursor of ethylene and a primary storage form of ABA, respectively. Consistent with these observations, transcript levels of genes encoding ACC synthase and ACC oxidase, both involved in ethylene biosynthesis, were increased after 6 days of NAA treatment, and both ABA concentration and expression of the regulator gene of ABA biosynthesis (NCED1 encoding 9-cis-epoxycarotenoid dioxygenase) were highest during early fruit ripening. In addition, transcript levels of key anthocyanin regulatory, biosynthetic and transport genes were significantly upregulated upon fruit exposure to NAA. This was accompanied by an increased anthocyanin concentration and fruit weight whilst fruit firmness and cracking index decreased. Altogether our data suggest that NAA treatment alters ethylene production, which in turn induces ripening in sweet cherry and enhanced anthocyanin production, possibly through ABA metabolism. The results from our study highlight the potential to use a single NAA treatment for manipulation of cherry ripening.

Meta-analysis of RNA-Seq studies reveals genes with dominant functions during flower bud endo- to eco-dormancy transition in Prunus species.

In deciduous fruit trees, entrance into dormancy occurs in later summer/fall, concomitantly with the shortening of day length and decrease in temperature. Dormancy can be divided into endodormancy, ecodormancy and paradormancy. In Prunus species flower buds, entrance into the dormant stage occurs when the apical meristem is partially differentiated; during dormancy, flower verticils continue their growth and differentiation. Each species and/or cultivar requires exposure to low winter temperature followed by warm temperatures, quantified as chilling and heat requirements, to remove the physiological blocks that inhibit budburst. A comprehensive meta-analysis of transcriptomic studies on flower buds of sweet cherry, apricot and peach was conducted, by investigating the gene expression profiles during bud endo- to ecodormancy transition in genotypes differing in chilling requirements. Conserved and distinctive expression patterns were observed, allowing the identification of gene specifically associated with endodormancy or ecodormancy. In addition to the MADS-box transcription factor family, hormone-related genes, chromatin modifiers, macro- and micro-gametogenesis related genes and environmental integrators, were identified as novel biomarker candidates for flower bud development during winter in stone fruits. In parallel, flower bud differentiation processes were associated to dormancy progression and termination and to environmental factors triggering dormancy phase-specific gene expression.

Cross-talk between transcriptome, phytohormone and HD-ZIP gene family analysis illuminates the molecular mechanism underlying fruitlet abscission in sweet cherry (Prunus avium L).

BACKGROUND: The shedding of premature sweet cherry (Prunus avium L) fruitlet has significantly impacted production, which in turn has a consequential effect on economic benefits. RESULT: To better understand the molecular mechanism of sweet cherry fruitlet abscission, pollen viability and structure had been observed from the pollination trees. Subsequently, the morphological characters of the shedding fruitlet, the plant hormone titers of dropping carpopodium, the transcriptome of the abscising carpopodium, as well as the HD-ZIP gene family were investigated. These findings showed that the pollens giving rise to heavy fruitlet abscission were malformed in structure, and their viability was lower than the average level. The abscising fruitlet and carpopodium were characterized in red color, and embryos of abscising fruitlet were aborted, which was highly ascribed to the low pollen viability and malformation. Transcriptome analysis showed 6462 were significantly differentially expressed, of which 2456 genes were up-regulated and 4006 down-regulated in the abscising carpopodium. Among these genes, the auxin biosynthesis and signal transduction genes (α-Trp, AUX1), were down-regulated, while the 1-aminocyclopropane-1-carboxylate oxidase gene (ACO) affected in ethylene biosynthesis, was up-regulated in abscising carpopodium. About genes related to cell wall remodeling (CEL, PAL, PG EXP, XTH), were up-regulated in carpopodium abscission, which reflecting the key roles in regulating the abscission process. The results of transcriptome analysis considerably conformed with those of proteome analysis as documented previously. In comparison with those of the retention fruitlet, the auxin contents in abscising carpopodium were significantly low, which presumably increased the ethylene sensitivity of the abscission zone, conversely, the abscisic acid (ABA) accumulation was considerably higher in abscising carpopodium. Furthermore, the ratio of (TZ + IAA + GA3) / ABA also obviously lower in abscising carpopodium. Besides, the HD-ZIP gene family analysis showed that PavHB16 and PavHB18 were up-regulated in abscising organs. CONCLUSION: Our findings combine morphology, cytology and transcriptional regulation to reveal the molecular mechanism of sweet cherry fruitlet abscission. It provides a new perspective for further study of plant organ shedding.

Titer and distribution of little cherry virus 2 in Prunus avium.

Little cherry virus 2 (LChV-2) is a causal agent of little cherry disease, which produces small, misshapen fruit with poor color and taste. As LChV-2 symptoms are only present near harvest, molecular detection is essential for effective control. Therefore, we determined the titer and distribution of this virus in infected trees over time. While initial infections were found to be basipetal, in field trees, early-stage infection was characterized by uneven distribution and low titer, concentrated in woody stems. In contrast, established infections were systemic, and detection was consistent across tissues. These data provide improved sampling recommendations for the detection of LChV-2.

Detection and monitoring of Drosophila suzukii in raspberry and cherry orchards with volatile organic compounds in the USA and Europe.

Spotted wing drosophila (SWD) causes significant economic loss in fruit crops to growers worldwide. There is immediate need for efficacious and selective monitoring tools that can detect infestations early. Previously, volatile organic compounds derived from apple were studied and a quinary chemical component blend (QB) was identified as the key SWD attractant in a blueberry orchard in the United States. This study's aim was to determine whether previously observed QB efficacy, selectivity, and early detection levels could be attained within raspberry and cherry fields in the USA and Europe. Results demonstrated that sticky trap baited QB dispenser provided earlier SWD detection potential than the usually adopted apple cider vinegar (ACV) trap. The number of SWD captured/trap by QB baited trapping systems was significantly lower than that of the ACV trap. However, percent SWD/trap of QB baited traps was same within cherry. Lower non-target capture will save farmer/grower's labor and time allocated to traps installation and drosophila species identification. Within the USA, SWD selectivity of QB baited liquid traps was consistently greater than sticky trap in raspberry field, suggesting that the QB dispenser can be an alternative to the standard ACV lure and that trap design could improve selectivity further.

Genome-Wide Identification of the B-BOX Genes that Respond to Multiple Ripening Related Signals in Sweet Cherry Fruit.

B-BOX proteins are zinc finger transcription factors that play important roles in plant growth, development, and abiotic stress responses. In this study, we identified 15 PavBBX genes in the genome database of sweet cherry. We systematically analyzed the gene structures, clustering characteristics, and expression patterns of these genes during fruit development and in response to light and various hormones. The PavBBX genes were divided into five subgroups. The promoter regions of the PavBBX genes contain cis-acting elements related to plant development, hormones, and stress. qRT-PCR revealed five upregulated and eight downregulated PavBBX genes during fruit development. In addition, PavBBX6, PavBBX9, and PavBBX11 were upregulated in response to light induction. We also found that ABA, BR, and GA3 contents significantly increased in response to light induction. Furthermore, the expression of several PavBBX genes was highly correlated with the expression of anthocyanin biosynthesis genes, light-responsive genes, and genes that function in multiple hormone signaling pathways. Some PavBBX genes were strongly induced by ABA, GA, and BR treatment. Notably, PavBBX6 and PavBBX9 responded to all three hormones. Taken together, BBX proteins likely play major roles in regulating anthocyanin biosynthesis in sweet cherry fruit by integrating light, ABA, GA, and BR signaling pathways.

Could Causal Discovery in Proteogenomics Assist in Understanding Gene-Protein Relations? A Perennial Fruit Tree Case Study Using Sweet Cherry as a Model.

Genome-wide transcriptome analysis is a method that produces important data on plant biology at a systemic level. The lack of understanding of the relationships between proteins and genes in plants necessitates a further thorough analysis at the proteogenomic level. Recently, our group generated a quantitative proteogenomic atlas of 15 sweet cherry (Prunus avium L.) cv. 'Tragana Edessis' tissues represented by 29,247 genes and 7584 proteins. The aim of the current study was to perform a targeted analysis at the gene/protein level to assess the structure of their relation, and the biological implications. Weighted correlation network analysis and causal modeling were employed to, respectively, cluster the gene/protein pairs, and reveal their cause-effect relations, aiming to assess the associated biological functions. To the best of our knowledge, this is the first time that causal modeling has been employed within the proteogenomics concept in plants. The analysis revealed the complex nature of causal relations among genes/proteins that are important for traits of interest in perennial fruit trees, particularly regarding the fruit softening and ripening process in sweet cherry. Causal discovery could be used to highlight persistent relations at the gene/protein level, stimulating biological interpretation and facilitating further study of the proteogenomic atlas in plants.

PaMADS7, a MADS-box transcription factor, regulates sweet cherry fruit ripening and softening.

E-class MADS-box transcription factors, SEPALLATA (SEP) genes have an important role in floral organ initiation and development and fruit ripening. In this study, four sweet cherry SEP-like genes (PaMADS2, PaMADS4, PaMADS5, and PaMADS7) were cloned and functionally characterized. Gene expression analysis showed that the differential expression levels of PaMADS4 and PaMADS7 coincided with fruit ripening, and expression of PaMADS2 and PaMADS5 did not. Expression of PaMADS7 was affected by ABA and IAA. Subcellular localization assay demonstrated that four sweet cherry SEP-like proteins were all localized inside the nucleus. Silencing PaMADS7 using TRV-mediated virus-induced gene silencing inhibited fruit ripening and influenced major ripening-related physiological processes, such as ABA content, soluble sugar contents, fruit firmness, and anthocyanin content, as well as expression of ripening-related genes. In addition, silencing of PaMADS7 induced phenotype defects that suppressed fruit ripening, which were rescued by exogenous ABA. Furthermore, yeast one-hybrid assay (Y1H) and transient expression analyses revealed that PaMADS7 directly binds to the promoter of PaPG1, which is involved in sweet cherry fruit softening, and positively activated PaPG1expression. These results showed that PaMADS7 is an indispensable positive regulator of sweet cherry fruit ripening and softening.

Decreased deposition and increased swelling of cell walls contribute to increased cracking susceptibility of developing sweet cherry fruit.

MAIN CONCLUSION: During fruit development, cell wall deposition rate decreases and cell wall swelling increases. The cell wall swelling pressure is very low relative to the fruit's highly negative osmotic potential. Rain cracking of sweet cherry fruit is preceded by the swelling of the cell walls. Cell wall swelling decreases both the cell: cell adhesion and the cell wall fracture force. Rain cracking susceptibility increases during fruit development. The objectives were to relate developmental changes in cell wall swelling to compositional changes taking place in the cell wall. During fruit development, total mass of cell wall, of pectins and of hemicelluloses increases, but total mass of cellulose remains constant. The mass of these cell wall fractions increases at a lower rate than the fruit fresh mass-particularly during stage II and early stage III. During stage III, on a whole-fruit basis, the HCl-soluble pectin fraction, followed by the water-soluble pectin fraction, the NaOH-soluble pectin fraction and the oxalate-soluble pectin fraction all increase. At maturity, just the HCl-soluble pectin decreases. Cell wall swelling increases during stages I and II of fruit development, with little change thereafter. This was indexed by light microscopy of skin sections following turgor release, and by determinations of the swelling capacity, water holding capacity and water retention capacity. The increase in cell wall swelling during development was due primarily to increases in NaOH-soluble pectins. The in vitro swelling of cell wall extracts depends on the applied pressure. The swelling pressure of the alcohol-insoluble residue is low throughout development and surprisingly similar across different cell wall fractions. Thus, swelling pressure does not contribute significantly to fruit water potential.

Novel insights into the calcium action in cherry fruit development revealed by high-throughput mapping.

KEY MESSAGE: This work provides the first system-wide datasets concerning metabolic changes in calcium-treated fruits, which reveal that exogenously applied calcium may specifically reprogram sweet cherry development and ripening physiognomy. Calcium modulates a wide range of plant developmental processes; however, the regulation of fruit ripening by calcium remains largely uncharacterized. In this study, transcriptome, proteome and metabolome profiling was used to document the responses of sweet cherry fruit to external calcium application (0.5% CaCl2) at 15, 27 and 37 days after full blossom. Endogenous calcium loading in fruit across development following external calcium feeding was accompanied by a reduction in respiration rate. Calcium treatment strongly impaired water-induced fruit cracking tested by two different assays, and this effect depended on the fruit size, water temperature and light/dark conditions. Substantial changes in the levels of numerous polar/non-polar primary and secondary metabolites, including malic acid, glucose, cysteine, epicatechin and neochlorogenic acid were noticed in fruits exposed to calcium. At the onset of ripening, we identified various calcium-affected genes, including those involved in ubiquitin and cysteine signaling, that had not been associated previously with calcium function in fruit biology. Calcium specifically increased the abundance of a significant number of proteins that classified as oxidoreductases, transferases, hydrolases, lyases, and ligases. The overview of temporal changes in gene expression and corresponding protein abundance provided by interlinked analysis revealed that oxidative phosphorylation, hypersensitive response, DNA repair, stomata closure, biosynthesis of secondary metabolites, and proton-pump activity were mainly affected by calcium. This report provides the fullest characterization of expression patterns in calcium-responsive genes, proteins and metabolites currently available in fruit ripening and will serve as a blueprint for future biological endeavors.

Transfer of endogenous small RNAs between branches of scions and rootstocks in grafted sweet cherry trees.

Grafting is a well-established agricultural practice in cherry production for clonal propagation, altered plant vigor and architecture, increased tolerance to biotic and abiotic stresses, precocity, and higher yield. Mobile molecules, such as water, hormones, nutrients, DNAs, RNAs, and proteins play essential roles in rootstock-scion interactions. Small RNAs (sRNAs) are 19 to 30-nucleotides (nt) RNA molecules that are a group of mobile signals in plants. Rootstock-to-scion transfer of transgene-derived small interfering RNAs enabled virus resistance in nontransgenic sweet cherry scion. To determine whether there was long-distance scion-to-rootstock transfer of endogenous sRNAs, we compared sRNAs profiles in bud tissues of an ungrafted 'Gisela 6' rootstock, two sweet cherry 'Emperor Francis' scions as well as their 'Gisela 6' rootstocks. Over two million sRNAs were detected in each sweet cherry scion, where 21-nt sRNA (56.1% and 55.8%) being the most abundant, followed by 24-nt sRNAs (13.1% and 12.5%). Furthermore, we identified over three thousand sRNAs that were potentially transferred from the sweet cherry scions to their corresponding rootstocks. In contrast to the sRNAs in scions, among the transferred sRNAs in rootstocks, the most abundant were 24-nt sRNAs (46.3% and 34.8%) followed by 21-nt sRNAs (14.6% and 19.3%). In other words, 21-nt sRNAs had the least transferred proportion out of the total sRNAs in sources (scions) while 24-nt had the largest proportion. The transferred sRNAs were from 574 cherry transcripts, of which 350 had a match from the Arabidopsis thaliana standard protein set. The finding that "DNA or RNA binding activity" was enriched in the transcripts producing transferred sRNAs indicated that they may affect the biological processes of the rootstocks at different regulatory levels. Overall, the profiles of the transported sRNAs and their annotations revealed in this study facilitate a better understanding of the role of the long-distance transported sRNAs in sweet cherry rootstock-scion interactions as well as in branch-to-branch interactions in a tree.

Oxidation processes related to seed storage and seedling growth of Malus sylvestris, Prunus avium and Prunus padus.

Seeds stored in controlled conditions in gene banks, faster or slower lose their viability. The effects of seed moisture content levels (ca. 5, 8, 11%) combined with storage temperatures (-3°, -18°, -196°C) were investigated in terms of the description of seeds defined as orthodox under oxidative stress after seed storage, during germination, and initial seedling growth. Hydrogen peroxide (H2O2), thiobarbituric acid reactive substances (TBARS) and ascorbate (Asc) were analyzed in relation to seed germinability and seedlings emergence in three species: Malus sylvestris L., Prunus avium L. and Prunus padus L. The effect of seed storage conditions on H2O2 levels appeared in germinated seeds after the third year of storage in each species. The H2O2 levels were negatively correlated with the germination and seedling emergence of P. avium seeds after three years of storage under all examined combinations. The emergence of P. padus seedlings was not linked to any of the stress markers tested. The P. padus seed biochemical traits were least altered by storage conditions, and the seeds produced tolerant seedlings of relatively high levels of H2O2 and TBARS. To cope with different H2O2 levels, TBARS levels, and Asc levels in seeds of three species varying storage conditions different molecular responses, i.e. repairing mechanisms, were applied during stratification to compensate for the storage conditions and, as a result, seeds remained viable and seedlings were successfully established.

Comparative transcriptomic analysis reveals novel roles of transcription factors and hormones during the flowering induction and floral bud differentiation in sweet cherry trees (Prunus avium L. cv. Bing).

In sweet cherry trees, flowering is commercially important because the flowers, after fertilization, will generate the fruits. In P. avium, the flowering induction and flower organogensis are the first developmental steps towards flower formation and they occur within specialized organs known as floral buds during the summer, nine months before blooming. During this period the number of floral buds per tree and the bud fruitfulness (number of flowers per bud) are stablished affecting the potential yield of orchards and the plant architecture. The floral bud development is sensitive to any type of stress and the hotter and drier summers will interfere with this process and are calling for new adapted cultivars. A better understanding of the underlying molecular and hormonal mechanisms would be of help, but unlike the model plant Arabidopsis, very little is known about floral induction in sweet cherry. To explore the molecular mechanism of floral bud differentiation, high-throughput RNA sequencing was used to detect differences in the gene expression of P. avium floral buds at five differentiation stages. We found 2,982 differentially expressed genes during floral bud development. We identified genes associated with floral initiation or floral organ identity that appear to be useful biomarkers of floral development and several transcription factor families (ERF, MYB, bHLH, MADS-box and NAC gene family) with novel potential roles during floral transition in this species. We analyzed in deep the MADS-box gene family and we shed light about their key role during floral bud and organs development in P. avium. Furthermore, the hormonal-related signatures in the gene regulatory networks and the dynamic changes of absicic acid, zeatin and indolacetic acid contents in buds suggest an important role for these hormones during floral bud differentiation in sweet cherry. These data provide a rich source of novel informacion for functional and evolutionary studies about floral bud development in sweet cherry and new tools for biotechnology and breeding.

Proteomic and metabolic analysis reveals novel sweet cherry fruit development regulatory points influenced by girdling.

Despite the application of girdling technique for several centuries, its impact on the metabolic shifts that underly fruit biology remains fragmentary. To characterize the influence of girdling on sweet cherry (Prunus avium L.) fruit development and ripening, second-year-old shoots of the cultivars 'Lapins' and 'Skeena' were girdled before full blossom. Fruit characteristics were evaluated across six developmental stages (S), from green-small fruit (stage S1) to full ripe stage (stage S6). In both cultivars, girdling significantly altered the fruit ripening physiognomy. Time course fruit metabolomic along with proteomic approaches unraveled common and cultivar-specific responses to girdling. Notably, several primary and secondary metabolites, such as soluble sugars (glucose, trehalose), alcohol (mannitol), phenolic compounds (rutin, naringenin-7-O-glucoside), including anthocyanins (cyanidin-3-O-rutinoside, cyanidin-3-O-galactoside, cyanidin-3.5-O-diglucoside) were accumulated by girdling, while various amino acids (glycine, threonine, asparagine) were decreased in both cultivars. Proteins predominantly associated with ribosome, DNA repair and recombination, chromosome, membrane trafficking, RNA transport, oxidative phosphorylation, and redox homeostasis were depressed in fruits of both girdled cultivars. This study provides the first system-wide datasets concerning metabolomic and proteomic changes in girdled fruits, which reveal that shoot girdling may induce long-term changes in sweet cherry metabolism.

Melatonin Accumulation in Sweet Cherry and Its Influence on Fruit Quality and Antioxidant Properties.

Although the effects of melatonin on plant abiotic and biotic stress resistance have been explored in recent decades, the accumulation of endogenous melatonin in plants and its influence on fruit quality remains unclear. In the present study, melatonin accumulation levels and the expression profiles of five synthesis genes were investigated during fruit and leaf development in sweet cherry (Prunus avium L.). Melatonin was strongly accumulated in young fruits and leaves, then decreased steadily with maturation. Transcript levels of PacTDC and PacSNAT were highly correlated with melatonin content in both fruit and leaves, indicating their importance in melatonin accumulation. Furthermore, application of 50 and 100 µmol·L-1 of melatonin to leaves had a greater influence on fruit quality than treatments applied to fruits, by significantly improving fruit weight, soluble solids content, and phenolic content including total phenols, flavanols, total anthocyanins, and ascorbic acid. Meanwhile, melatonin application promoted the antioxidant capacity of fruit assayed by 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azinobis (3-ethylben zothiazoline-6-sulfonic acid) (ABTS), and ferric reducing antioxidant power (FRAP). These results provide insights into the physiological and molecular mechanisms underlying melatonin metabolism of sweet cherry.

Dormancy-Associated MADS-Box (DAM) Genes Influence Chilling Requirement of Sweet Cherries and Co-Regulate Flower Development with SOC1 Gene.

Floral bud dormancy release of fruit tree species is greatly influenced by climate change. The lack of chilling accumulation often results in the occurrence of abnormal flower and low yields of sweet cherries (Prunus avium L.) in warm regions. To investigate the regulation of dormancy in sweet cherries, six DAM genes with homology to peach DAM, designated PavDAM1-6, have been identified and characterized. Phylogenetic analysis indicate that these genes are similar to DAMs in peach, apple and pear. The expression patterns of the PavDAMs in the low-chill cultivar 'Royal Lee' were different from that in the high-chill cultivar 'Hongdeng'. 'Royal Lee' exhibits lower transcriptional level of PavDAM1 compared to 'Hongdeng', especially at the stage of chilling accumulation, and transcriptional levels of PavDAM4/5 were high in both cultivars during the endodormancy. Ectopic expression of PavDAM1 and PavDAM5 in Arabidopsis resulted in plants with abnormal flower and seed development, especially the PavDAM5. Higher transcriptional levels of SOC1 were observed in transgenic PavDAM1/5 lines, and ectopic expression of PavSOC1 had the similar floral phenotype. Further, protein interaction analysis demonstrated that PavDAM1/5 could interact with PavSOC1 in vivo and in vitro, which will help clarify the molecular mechanism of the flower development in sweet cherry or other fruit trees.