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Background Bud dormancy is a crucial stage in perennial trees and allows survival over winter to ensure optimal flowering and fruit production. Recent work highlighted physiological and molecular events occurring during bud dormancy in trees. However, they usually examined bud development or bud dormancy in isolation. In this work, we aimed to further explore the global transcriptional changes happening throughout bud development and dormancy onset, progression and release. Results Using next-generation sequencing and modelling, we conducted an in-depth transcriptomic analysis for all stages of flower buds in several sweet cherry ( Prunus avium L.) cultivars that are characterized for their contrasted dates of dormancy release. We find that buds in organogenesis, paradormancy, endodormancy and ecodormancy stages are defined by the expression of genes involved in specific pathways, and these are conserved between different sweet cherry cultivars. In particular, we found that DORMANCY ASSOCIATED MADS-box ( DAM ), floral identity and organogenesis genes are up-regulated during the pre-dormancy stages while endodormancy is characterized by a complex array of signalling pathways, including cold response genes, ABA and oxidation-reduction processes. After dormancy release, genes associated with global cell activity, division and differentiation are activated during ecodormancy and growth resumption. We then went a step beyond the global transcriptomic analysis and we developed a model based on the transcriptional profiles of just seven genes to accurately predict the main bud dormancy stages. Conclusions Overall, this study has allowed us to better understand the transcriptional changes occurring throughout the different phases of flower bud development, from bud formation in the summer to flowering in the following spring. Our work sets the stage for the development of fast and cost effective diagnostic tools to molecularly define the dormancy stages. Such integrative approaches will therefore be extremely useful for a better comprehension of complex phenological processes in many species.

Background. Due to the increasing demand for organic food by consumers, agricultural systems had to change towards a more sustainable and environmental approach. One of the practices aligned with this purpose is the application of efficient biostimulants as a tool to improve food production with the smallest possible ecological footprint. For this reason, the aim of this work was to evaluate the effectiveness and comparison of an organic system composed of a Lombrico® SET biostimulant formulated from seaweed extracts, amino acids and carbohydrates from plant-based sources on two different varieties of cherries, Nimba and Red Pacific, against a conventional system composed by Maxi-Grow Excel®, Equilibrium® and Exelgrow® products according to the usual biostimulant treatments of the farmer for sweet cherry crop. Both options were applied at 12 times throughout the flowering and ripening stages under real field conditions in a 68-day study. Results. Although the program effectiveness differed by variety, both biostimulants treatments improved bud breakage, enhanced flowering, and advanced ripening, while improving the fruit quality and the organoleptic properties. Remarkable findings included increased levels of major carbohydrates and increased oxalic acid. Additionally, significant changes in the phenolic and flavonoid profile were observed. The organic program led to higher levels of chlorogenic acid as well as increased naringin across both varieties. Conversely, cyanidin-3-O-rutinoside levels were increased in the organic program for Red Pacific but were higher in the conventional method for Nimba. Conclusions. The increase in marketable first category fruit and the reduction in unmarketable fruit due to both programs contribute to improved farmer profitability and demonstrate the clear effectiveness of the application of biostimulant systems. In addition, the organic treatment produced an enhanced ripening, enabling an early harvest, with a positive impact on the quality and organoleptic parameters of the cherries. In particular, the organic treatment where a single product composed of seaweed extracts, amino acids and carbohydrates from plant-based origin is applied instead of the three products required in the conventional system is particularly valuable in organic farming where external inputs are limited.

To avoid the negative impacts of winter unfavorable conditions for plant development, temperate trees enter a rest period called dormancy. Winter dormancy is a complex process that involves multiple signaling pathways and previous studies have suggested that transport capacity between cells and between the buds and the twig may regulate the progression throughout dormancy stages. However, the dynamics and molecular actors involved in this regulation are still poorly described in fruit trees. Here, in order to validate the hypothesis that transport capacity regulates dormancy progression in fruit trees, we combined physiological, imaging and transcriptomic approaches to characterize molecular pathways and transport capacity during dormancy in sweet cherry (Prunus avium L.) flower buds. Our results show that transport capacity is reduced during dormancy and could be regulated by environmental signals. Moreover, we demonstrate that dormancy release is not synchronized with the transport capacity resumption but occurs when the bud is capable of growth under the influence of warmer temperatures. We highlight key genes involved in transport capacity during dormancy. Based on long-term observations conducted during six winter seasons, we propose hypotheses on the environmental and molecular regulation of transport capacity, in relation to dormancy and growth resumption in sweet cherry.

Sweet cherry ( L.) is an important fruit crop in which fruit size is strongly associated with commercial value; few genes associated with fruit size have, however, been identified in sweet cherry. Members of the CYP78A subfamily, a group of important cytochrome P450s, have been found to be involved in controlling seed size and development in , rice, soybean, and tomato. However, the influence of CYP78A members in controlling organ size and the underlying molecular mechanisms in sweet cherry and other fruit trees remains unclear. Here, we characterized a CYP78A gene that is thought to be involved in the regulation of fruit size and organ development using overexpression and silencing approaches. was significantly expressed in the flowers and fruit of sweet cherry. RNAi silencing of produced small cherry fruits and was found to affect fruit size by mediating mesocarp cell proliferation and expansion during fruit growth and development. Overexpression of in resulted in increased silique and seed size and was found to be highly expressed in the inflorescences and siliques of transgenic plants. Genes related to cell cycling and proliferation were downregulated in fruit from sweet cherry -silencing lines, suggesting that is likely to be an important upstream regulator of cell cycle processes. Together, our findings indicate that plays an essential role in the regulation of cherry fruit size and provide insights into the molecular basis of the mechanisms regulating traits such as fruit size in .

Sweet cherry is an important non-climacteric fruit with a high commercial interest, but exploitation of sweet cherry trees (Prunus avium L.) in orchards is usually subject to important economic losses due to fruit decay by pathogenic fungi and other microorganisms. Sweet cherries development and ripening are characterized by profound physiological changes in the fruit, among which the phytohormone abscisic acid (ABA) plays a pivotal role. In addition, sweet cherries are usually affected by fruit decay pathogens, and the role of other stress-related hormones such as jasmonic acid (JA) and salicylic acid (SA) may also be of paramount importance, not only from a developmental point of view, but also from a fruit-microbe interaction perspective. Here, a tissue-specific hormone quantification by LC-MS/MS, including the contents of JA, SA, and ABA, in the fruit exocarp and mesocarp of sweet cherries during fruit development from trees growing in a commercial orchard was carried out. Additionally, this study was complemented with the characterization of the culturable epiphytic and endophytic microbial communities of sweet cherries at various stages of fruit development and during cracking lesion formation. Our results revealed a completely differential behavior of phytohormones between both tissues (the exocarp and mesocarp), with a more dynamic exocarp in front of a more stable mesocarp, and with marked variations during fruit development. Microbial epiphytic community was mainly composed by yeasts, although rot-causing fungi like Alternaria spp. were always also present throughout fruit development. Endophytic colonization was poor, but it increased throughout fruit development. Furthermore, when the exocarp was naturally disrupted in sweet cherries suffering from cracking, the colonization by Alternaria spp. markedly increased. Altogether, results suggest that the fruit exocarp and mesocarp are very dynamic tissues in which endogenous phytohormones not only modulate fruit development and ripening but also fruit-microbe interactions.

Sweet cherry fruits harvested at commercial maturity stage were treated with an edible coating based on sodium alginate at several concentrations (1%, 3% or 5% w/v ). The coatings were effective on delaying the evolution of the parameters related to postharvest ripening, such as colour, softening and loss of acidity, and reducing respiration rate. In addition, the edible coatings showed a positive effect on maintaining higher concentration of total phenolics and total antioxidant activity, which decreased in control fruits associated with the over-ripening and senescence processes. Results from quality parameters and antioxidant activity suggested that the maximum storability period for control fruits was 8 days at 2 °C plus 2 days at 20 °C, while alginate-coated cherries could be stored with optimal quality and enhanced antioxidant activity up to 16 days at 2 °C plus 2 days at 20 °C.

The phenology of wild cherry in the Northwest of Tunisia was studied over the year 2013. The phenological events reserved are the leafing, the flowering and the fruiting. The observation of the various phenological phases of this species (bud burst, blossoming or fruiting) reveals a variation inter-site and intra-site. The precocity reveals for the sites of Tabarka (Hamdia and Kroufa) can give some explanation of differences of edaphic and topographic characteristics. The analyses of variances show that the duration of the various phenological phases of the wild cherry is significantly connected with the geographical localization of sites. The differences observed within the same site can give some explanation by the genetic variability of trees. These results open perspectives for the valuation and the genetic preservation of this commercial species in the northeast of Tunisia.

Sweet cherry is a high-value crop, and strategies to enhance production and sustainability are at the forefront of research linked to this crop. The improvement of plant status is key to achieving optimum yield. Biostimulants, such as glycine betaine (GB) or seaweed-based biostimulants [e.g., Ecklonia maxima (EM)], can represent a sustainable approach to improving plant conditions, even under adverse environmental circumstances. Despite their potential, few studies have focused on the effects of GB or EM exogenous application on sweet cherry tree physiology. To address this lack of research, a study was conducted in a Portuguese sweet cherry commercial orchard, using Lapins and Early Bigi cultivars. Trees were treated with products based on GB and EM at two different concentrations [GB 0.25% (v/v) and GB 0.40% (v/v); EM 0.30% (v/v) and EM 0.15% (v/v)], a combination of the lowest concentrations of both biostimulants (Mix —GB 0.25% and EM 0.15%), and a control group (C) treated with water. Applications were performed over three consecutive years (2019, 2020, and 2021) at three different phenological stages, according to the BBCH scale: 77, 81, and 86 BBCH. Results showed, in general, that the application of biostimulants led to improvements in water status as well as significantly lower values of electrolyte leakage and thiobarbituric acid reactive substances compared to C samples. Additionally, biostimulants reduced pigment loss in the leaves and enhanced their biosynthesis. The Chlorophyll a /Chlorophyll b ratio, ranging from 2 to 4, indicated a greater capacity for light absorption and lower stress levels in treated leaves. Soluble sugar and starch content decreased during fruit development in both cultivars and years; however, biostimulants increased these contents, with increments of approximately 15% to 30% in leaves treated with EM. Soluble protein content also showed the same pattern for treated leaves. Biostimulants, especially EM, demonstrated a significant positive effect ( p ≤ 0.001) on total phenolic content, with increases of approximately 25% to 50% in treated leaves. In conclusion, the application of biostimulants, especially algae-based, significantly improved tree performance by enhancing physiological parameters and stress resilience and could represent a novel approach in fruit production systems.

Anthocyanins play a crucial role in shaping the visual appeal and nutritional quality of fruits. Previous research on anthocyanin biosynthesis in sweet cherry (Prunus avium L.) has primarily relied on single-omics approaches or focused on a limited range of metabolites, leaving the regulatory mechanisms and dynamic metabolism of anthocyanins during ripening inadequately characterized. This study integrated anthocyanin-targeted metabolomics and transcriptomics to identify key anthocyanins in sweet cherry and construct a transcriptional regulatory network for anthocyanin biosynthesis. A novel bHLH transcription factor, Prunus avium bHLH transcription factor 102 (PavbHLH102), was identified, and its role in regulating cyanidin levels was validated through overexpression and silencing experiments. Both in vitro and in vivo assays demonstrated that PavbHLH102 activates key anthocyanin biosynthetic genes, including PavF3H, PavDFR, and PavUFGT, thereby enhancing fruit coloration. Notably, PavF3′H upregulation significantly increased cyanidin accumulation. This study provides new insights into anthocyanin regulation in sweet cherry and offers valuable resources for improving fruit quality.

Chromatin immunoprecipitation-sequencing (ChIP-seq) is a robust technique to study interactions between proteins, such as histones or transcription factors and DNA. This technique in combination with RNA-sequencing (RNA-seq) is a powerful tool to better understand biological processes in eukaryotes. We developed a combined ChIP-seq and RNA-seq protocol for tree buds (Prunus avium L., Prunus persica L Batch, Malus x domestica Borkh.) that has also been successfully tested on Arabidopsis thaliana and Saccharomyces cerevisiae. Tree buds contain phenolic compounds that negatively interfere with ChIP and RNA extraction. In addition to solving this problem, our protocol is optimised to work on small amounts of material. Furthermore, one of the advantages of this protocol is that samples for ChIP-seq are cross-linked after flash freezing, making it possible to work on trees growing in the field and to perform ChIP-seq and RNA-seq on the same starting material. Focusing on dormant buds in sweet cherry, we explored the link between expression level and H3K4me3 enrichment for all genes, including a strong correlation between H3K4me3 enrichment at the DORMANCY-ASSOCIATED MADS-BOX 5 (PavDAM5) loci and its expression pattern. This protocol will allow analysis of chromatin and transcriptomic dynamics in tree buds, notably during its development and response to the environment.