Osmia lignaria (Hymenoptera: Megachilidae) increase pollination of Washington sweet cherry and pear crops.

Apis mellifera Linnaeus (Hymenoptera: Apis), honey bees, are the most widely used managed crop pollinators. However, their high rental cost and uncertain availability for North American orchard crops have motivated growers to explore alternative pollination options. We examined whether adding solitary, spring-flying Osmia lignaria Say (Hymenoptera: Megachilidae), blue orchard bees, as co-pollinators with A. mellifera in Washington sweet cherry and pear orchards enhances fruit set and yield compared to the use of A. mellifera alone. We added managed O. lignaria to orchard sites where A. mellifera hives were already present. Fruit set, fruit yield, and O. lignaria reproduction at O. lignaria-supplemented sites were compared to nearby, paired sites pollinated only by A. mellifera (3 paired cherry and 3 paired pear sites). For both crops, the addition of O. lignaria significantly increased fruit set but did not yield at harvest. Microscopic inspection of pollen grains from O. lignaria nest cell provisions confirmed that O. lignaria primarily visited orchard flowers. Mean retention of O. lignaria in cherry orchards was slightly higher (65%) than O. lignaria retention reported in other orchard crops (30%-60%). However, retention in pear orchards was much lower (≤20%). These results show that supplementing hives with O. lignaria in Washington spring orchard crops can increase overall pollination, but that trees fail to bear developing fruit to maturity. The strategy of using co-pollinators, O. lignaria and A. mellifera, in US orchards may act as "pollination insurance" when A. mellifera hives are in low supply or when the weather is not amenable for A. mellifera flight during the bloom period.

Effect of preharvest biofilm application regimes on cracking and fruit quality traits in '0900 Ziraat' sweet cherry cultivar.

BACKGROUND: Fruit cracking impacts the quality of sweet cherry, significantly affecting its marketability due to increased susceptibility to injury, aesthetic flaws, and susceptibility to pathogens. The effect of 1% biofilm (Parka™) application regimes on fruit cracking and other quality parameters in the '0900 Ziraat' cherry cultivar was investigated in this study. Fruit sprayed with water were served as control (U1). Fruit treated only once with biofilm three, two and one week before the commercial harvest were considered as U2, U3 and U4, respectively. Fruit treated with biofilm three, two, and one week before harvest were considered as U5; three and two week before harvest as U6; two and one week before harvest as U7; and fruit treated three and one week before harvest as U8. RESULTS: In both measurement periods, the lower cracking index was obtained in biofilm-treated sweet cherry fruit. However, the firmness of biofilm-treated fruit was higher than that of the control fruit. The lowest respiration rate was observed in U7, while the highest weight was recorded in U4 and U5 than the control. The biofilm application decreased fruit coloration. The biofilm application also increased the soluble solids content of the fruit. The U2, U3 and U4 applications at harvest showed higher titratable acidity than the control. In both measurement periods, the vitamin C content of the U2, U5, U6, U7 and U8 applications was found to be higher than that of the control. The total monomeric anthocyanin of the U3 and U8 applications was higher than that of the control. Furthermore, the antioxidant activity of the U2, U3 and U5 in the DPPH, and the U7 and U8 in FRAP were measured higher thanthat of the control. CONCLUSIONS: The application of biofilms has the potential to mitigate fruit cracking, prolong postharvest life of sweet cherries, and enhance fruit firmness.

Overexpression of PavHIPP16 from Prunus avium enhances cold stress tolerance in transgenic tobacco.

BACKGROUND: The heavy metal-associated isoprenylated plant protein (HIPP) is an important regulatory element in response to abiotic stresses, especially playing a key role in low-temperature response. RESULTS: This study investigated the potential function of PavHIPP16 up-regulated in sweet cherry under cold stress by heterologous overexpression in tobacco. The results showed that the overexpression (OE) lines' growth state was better than wild type (WT), and the germination rate, root length, and fresh weight of OE lines were significantly higher than those of WT. In addition, the relative conductivity and malondialdehyde (MDA) content of the OE of tobacco under low-temperature treatment were substantially lower than those of WT. In contrast, peroxidase (POD), superoxide dismutase (SOD), catalase (CAT) activities, hydrogen peroxide (H2O2), proline, soluble protein, and soluble sugar contents were significantly higher than those of WT. Yeast two-hybrid assay (Y2H) and luciferase complementation assay verified the interactions between PavbHLH106 and PavHIPP16, suggesting that these two proteins co-regulated the cold tolerance mechanism in plants. The research results indicated that the transgenic lines could perform better under low-temperature stress by increasing the antioxidant enzyme activity and osmoregulatory substance content of the transgenic plants. CONCLUSIONS: This study provides genetic resources for analyzing the biological functions of PavHIPPs, which is important for elucidating the mechanisms of cold resistance in sweet cherry.

Genetic factors acting prior to dormancy in sour cherry influence bloom time the following spring.

Understanding the process of Prunus species floral development is crucial for developing strategies to manipulate bloom time and prevent crop loss due to climate change. Here, we present a detailed examination of flower development from initiation until bloom for early- and late-blooming sour cherries (Prunus cerasus) from a population segregating for a major bloom time QTL on chromosome 4. Using a new staging system, we show floral buds from early-blooming trees were persistently more advanced than those from late-blooming siblings. A genomic DNA coverage analysis revealed the late-blooming haplotype of this QTL, k, is located on a subgenome originating from the late-blooming P. fruticosa progenitor. Transcriptome analyses identified many genes within this QTL as differentially expressed between early- and late-blooming trees during the vegetative-to-floral transition. From these, we identified candidate genes for the late bloom phenotype, including multiple transcription factors homologous to Reproductive Meristem B3 domain-containing proteins. Additionally, we determined that the basis of k in sour cherry is likely separate from candidate genes found in sweet cherry-suggesting several major regulators of bloom time are located on Prunus chromosome 4.

Integrative analyses of metabolome and transcriptome reveals metabolomic variations and candidate genes involved in sweet cherry (Prunus avium L.) fruit quality during development and ripening.

Sweet cherry (Prunus avium L.), one of the most appreciated and most important commercial temperate fruits, has high sensory quality and nutritional value. Investigating its metabolic variations provides valuable information on the formation of fruit quality. In this study, widely targeted LC-MS/MS based metabolomics was used to identify and quantify metabolic changes during 'Black Pearl' sweet cherry development and ripening. A total of 263 significant differentially expressed metabolites (DEMs) were detected during the four fruit-development stages. Significant differences were observed in the composition and content of compounds in the four stages of cherry development, especially sugars, organic acids, and flavonoids. Moreover, transcriptome analysis provided a molecular basis for metabolic variations during fruit development. A total of 6724 significant differentially expressed genes (DEGs) were identified. Further correlation analysis of major DEMs and DEGs showed that 19 key DEGs were involved in sugar metabolism, 23 key DEGs in organic acid metabolism, and 13 key DEGs in flavonoid metabolism. The upregulated genes involved in the flavonoid pathway probably play an important role in regulating the rapid increase of anthocyanin content during fruit development. These comprehensive analysis data provide a better understanding to improve fruit quality traits based on molecular and metabolic levels.

Preanthesis changes in freeze resistance, relative water content, and ovary growth preempt bud phenology and signify dormancy release of sour cherry floral buds.

MAIN CONCLUSION: Growing degree hours (GDH) predicted floral bud development of 'Montmorency' sour cherry and explained changes in lethal temperatures (LT50) that preempted any visible changes in bud phenology. The gradual warming during late winter and early spring promotes floral bud development and, concomitantly, the de-acclimation of Prunus sp. flowers. In fact, once ecodormancy releases, an approximate 20 °C loss of hardiness occurs prior to any distinguishable changes in external bud phenology. The aim of the following work was to characterize the physiological changes of 'Montmorency' sour cherry floral buds as they transition from endo- and ecodormancy and resume growth, and to determine whether physiological and anatomical characteristics within the buds preempt or signify dormancy release to enable a better prediction of freeze susceptibility. Here, we present a developmental timeline of the preanthesis changes of 'Montmorency' floral buds, ovaries and anthers over 2 years following their completion of chilling and relate these changes to growing degree hours (GDH) and the lethal temperature (LT50) of flowers. Changes in bud dry weight (DW), fresh weight (FW), volume, and external phenology stage including the percentage of green color development of bud scales were predicted by heat accumulation but were not early predictors of the increasing freeze susceptibility of pistils. Between endodormancy and green tip stage, ovary volume increased nearly threefold and relative water content (RWC) increased from ~ 45 to 70% in both years. A linear mixed regression model indicated that RWC and the interaction between RWC and ovary growth were significant predictors of LT50. Importantly, the loss of ~ 20 °C of freeze resistance occurred between 45 and 57% RWC and preceded any detectable changes in bud phenology. Microsporogenesis was observed after dormancy release when measurable changes in the ovary and bud RWC had already occurred. A GDH model estimated freeze sensitivity of pistils and explained 93% of the variation in LT50 during preanthesis development. A simple GDH model to predict critical freeze temperature of pistils should aid producers to manage frost protection.

Complex Analysis of Antioxidant Activity, Abscisic Acid Level, and Accumulation of Osmotica in Apple and Cherry In Vitro Cultures under Osmotic Stress.

Plant response to osmotic stress is a complex issue and includes a wide range of physiological and biochemical processes. Extensive studies of known cultivars and their reaction to drought or salinity stress are very important for future breeding of new and tolerant cultivars. Our study focused on the antioxidant activity, accumulations of osmotica, and the content of abscisic acid in apple (cv. "Malinové holovouské", "Fragrance", "Rubinstep", "Idared", "Car Alexander") and cherry (cv. "Regina", "Napoleonova", "Kastánka", "Sunburst", "P-HL-C") cultivated in vitro on media containing different levels of polyethylene glycol PEG-6000. Our results indicated that the studied genotypes responded differently to osmotic stress manifested as reduction in the leaf relative water content (RWC) and increment in the activities of antioxidant enzymes, proline, sugars, and abscisic acid content. Overall, cherry cultivars showed a smaller decrease in percentage RWC and enzymatic activities, but enhanced proline content compared to the apple plants cultivars. Cultivars "Rubinstep", "Napoleonova", and "Kastánka" exhibited higher antioxidant capacity and accumulation of osmoprotectants like proline and sorbitol that can be associated with the drought-tolerance system.

Response of chloroplast pigments, sugars and phenolics of sweet cherry leaves to chilling.

The aim of the present study was to evaluate the effect of post-flowering chilling of sweet cherry (Prunus avium L.) on the content of biochemical parameters in the leaf (chloroplast pigments, sugars and phenolics). The effect of chilling was investigated in two experiments. Potted 2-year-old trees of cv. 'Grace Star' and 'Schneiders' were exposed to one, two or three consecutive overnight chillings at an average air temperature of 4.7 °C (Experiment I), but in the following year only trees of 'Grace Star' were chilled at 2.2 °C (Experiment II), 3 to 7 weeks after flowering. The analysis of the biochemical parameters was performed by high performance liquid chromatography combined with electrospray ionization mass spectrometry. Chilling at 4.7 °C caused little or no stress, while 2.2 °C induced more intense stress with increased zeaxanthin, sugar and phenolic content in leaves, while exposure of trees to higher temperatures and closer to flowering showed no changes. Two or three consecutive overnight chilling periods increased the phenolic content and enhanced the accumulation of zeaxanthin in the leaves. Sucrose, sorbitol, fructose, total sugar, and total flavonoid content in leaves increased within 48 h after chilling. Zeaxanthin epoxidized within 24 h after one and 48 h after one and two consecutive overnight chillings.

Impact of Background Fruit Odors on Attraction of Drosophila suzukii (Diptera: Drosophilidae) to Its Symbiotic Yeast.

Background odors produced by plants in the environment can interfere with the response of insects to a point-releasing attractant, especially when their compositions overlap. In this study, a series of binary choice tests was conducted in a wind tunnel to investigate whether background odors emitted from cherry, blueberry, blackberry, or raspberry fruits would affect the level of Drosophila suzukii (Matsumura) attraction to its symbiotic yeast, Hanseniaspora uvarum (Niehaus) (Saccharomycetales: Saccharomycetaceae). Whether an increase in the intensity of background odors would affect the attractiveness of H. uvarum to D. suzukii was also investigated, either by increasing the number of cherry or raspberry fruit per cup or by increasing the number of fruit cups surrounding the cup baited with the yeast. In wind tunnel assays, background fruit odors interfering with D. suzukii attraction to the yeast varied among fruit types. Raspberry odor inhibited the attractiveness of H. uvarum to the fly the most, followed by blackberry odor, whereas cherry and blueberry odors had no significant impact on the attraction. An increase in the intensity of odors by adding more cherry or raspberry fruit per cup did not increase the impact of fruit odor on the attraction; however, adding more raspberry cups around H. uvarum linearly decreased its attractiveness, suggesting that background host fruit abundance and likely increase in host odor may influence D. suzukii attraction to yeast odor depending on host species.

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.

Ubiquitination of S4-RNase by S-LOCUS F-BOX LIKE2 Contributes to Self-Compatibility of Sweet Cherry 'Lapins'.

Recent studies have shown that loss of pollen-S function in S4' pollen from sweet cherry (Prunus avium) is associated with a mutation in an S haplotype-specific F-box4 (SFB4) gene. However, how this mutation leads to self-compatibility is unclear. Here, we examined this mechanism by analyzing several self-compatible sweet cherry varieties. We determined that mutated SFB4 (SFB4') in S4' pollen (pollen harboring the SFB4' gene) is approximately 6 kD shorter than wild-type SFB4 due to a premature termination caused by a four-nucleotide deletion. SFB4' did not interact with S-RNase. However, a protein in S4' pollen ubiquitinated S-RNase, resulting in its degradation via the 26S proteasome pathway, indicating that factors in S4' pollen other than SFB4 participate in S-RNase recognition and degradation. To identify these factors, we used S4-RNase as a bait to screen S4' pollen proteins. Our screen identified the protein encoded by S 4 -SLFL2, a low-polymorphic gene that is closely linked to the S-locus. Further investigations indicate that SLFL2 ubiquitinates S-RNase, leading to its degradation. Subcellular localization analysis showed that SFB4 is primarily localized to the pollen tube tip, whereas SLFL2 is not. When S 4 -SLFL2 expression was suppressed by antisense oligonucleotide treatment in wild-type pollen tubes, pollen still had the capacity to ubiquitinate S-RNase; however, this ubiquitin-labeled S-RNase was not degraded via the 26S proteasome pathway, suggesting that SFB4 does not participate in the degradation of S-RNase. When SFB4 loses its function, S4-SLFL2 might mediate the ubiquitination and degradation of S-RNase, which is consistent with the self-compatibility of S4' pollen.

Sweet Cherry (Prunus avium L.) PaPIP1;4 Is a Functional Aquaporin Upregulated by Pre-Harvest Calcium Treatments that Prevent Cracking.

The involvement of aquaporins in rain-induced sweet cherry (Prunus avium L.) fruit cracking is an important research topic with potential agricultural applications. In the present study, we performed the functional characterization of PaPIP1;4, the most expressed aquaporin in sweet cherry fruit. Field experiments focused on the pre-harvest exogenous application to sweet cherry trees, cultivar Skeena, with a solution of 0.5% CaCl2, which is the most common treatment to prevent cracking. Results show that PaPIP1;4 was mostly expressed in the fruit peduncle, but its steady-state transcript levels were higher in fruits from CaCl2-treated plants than in controls. The transient expression of PaPIP1;4-GFP in tobacco epidermal cells and the overexpression of PaPIP1;4 in YSH1172 yeast mutation showed that PaPIP1;4 is a plasma membrane protein able to transport water and hydrogen peroxide. In this study, we characterized for the first time a plasma membrane sweet cherry aquaporin able to transport water and H2O2 that is upregulated by the pre-harvest exogenous application of CaCl2 supplements.

Effects of exogenous compound sprays on cherry cracking: skin properties and gene expression.

BACKGROUND: Cherry fruit cracking is a costly problem for cherry growers. The effect of repeated sprayings (gibberellic acid - GA3 ; abscisic acid - ABA; salicylic acid - SA; glycine betaine - GB, and Ascophyllum nodosum - AN) combined with CaCl2 , on 'Sweetheart' cherry fruit-cracking characteristics was investigated. Cracking was quantified in terms of cracking incidence, crack morphology, confocal scanning laser microscopy, cuticular wax content, cell-wall modification, and cuticular wax gene expression. RESULTS: All spray treatments reduced cracking compared with an untreated control (H2 O), with fewer cheek cracks. The least cracking incidence was observed for ABA + CaCl2 - and GB + CaCl2 -treated fruits, indicating an added benefit compared to spraying with CaCl2 alone. In addition, GB + CaCl2 -treated fruits showed higher fruit diameter. ABA + CaCl2 and GB + CaCl2 sprays showed higher wax content and higher cuticle and epidermal thickness compared with the control, including increased expression of wax synthase (ABA + CaCl2 ) and expansin 1 (GB + CaCl2 ). CONCLUSION: In general, factors that improve the cuticle thickness appear to be important at the fruit-coloring stage. At the fruit-ripening stage, larger cell sizes of the epidermis, hypodermis, and parenchyma cells lower cracking incidence, indicating the importance of flexibility and elasticity of the epidermis. © 2020 Society of Chemical Industry.

From bud formation to flowering: transcriptomic state defines the cherry developmental phases of sweet cherry bud dormancy.

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.

Determination of the bruise degree for cherry using Vis-NIR reflection spectroscopy coupled with multivariate analysis.

Determination and classification of the bruise degree for cherry can improve consumer satisfaction with cherry quality and enhance the industry's competiveness and profitability. In this study, visible and near infrared (Vis-NIR) reflection spectroscopy was used for identifying bruise degree of cherry in 350-2500 nm. Sampling spectral data were extracted from normal, slight and severe bruise samples. Principal component analysis (PCA) was implemented to determine the first few principal components (PCs) for cluster analysis among samples. Optimal wavelengths were selected by loadings of PCs from PCA and successive projection algorithm (SPA) method, respectively. Afterwards, these optimal wavelengths were empolyed to establish the classification models as inputs of least square-support vector machine (LS-SVM). Better performance for qualitative discrimination of the bruise degree for cherry was emerged in LS-SVM model based on five optimal wavelengths (603, 633, 679, 1083, and 1803 nm) selected directly by SPA, which showed acceptable results with the classification accuracy of 93.3%. Confusion matrix illustrated misclassification generally occurred in normal and slight bruise samples. Furthermore, the latent relation between spectral property of cherries in varying bruise degree and its firmness and soluble solids content (SSC) was analyzed. The result showed both colour, firmness and SSC were consistent with the Vis-NIR reflectance of cherries. Overall, this study revealed that Vis-NIR reflection spectroscopy integrated with multivariate analysis can be used as a rapid, intact method to determine the bruise degree of cherry, laying a foundation for cherry sorting and postharvest quality control.

Exogenous melatonin delays postharvest fruit senescence and maintains the quality of sweet cherries.

We assessed the effects of postharvest exogenous melatonin (50,100 and 150 µmol L-1) on the senescence and quality of sweet cherries during storage at 0 ±â€¯0.5 °C. Melatonin treatment decreased decay incidence, respiration rate, and weight loss. It delayed the degradation of firmness, lightness, saturation, hue angle, titratable acidity, and total soluble solids content, thus maintaining better fruit quality. Melatonin treatment inhibited increases in O2-, H2O2, malondialdehyde content, and relative membrane permeability, while maintaining higher endogenous melatonin levels and increasing superoxide dismutase and catalase activity. Additionally, melatonin treatment enhanced the activity of antioxidant enzymes, increased the levels of ascorbic acid, and reduced glutathione levels, which are related to the ascorbate-glutathione cycle, as well as increasing the AsA:DHA and GSH:GSSG ratios. Delayed senescence in sweet cherries after exogenous melatonin treatment may be associated with high endogenous melatonin levels and increased antioxidant activity and content.

Sweet cherry water relations and fruit production efficiency are affected by rootstock vigor.

Rootstock vigor is well known to affect yield and productive performance in many fruit crops and the dwarfing trait is often the preferred choice for modern orchard systems thanks to its improved productivity and reduced canopy volume. This work investigates the different physiological responses induced by rootstock vigor on cherry, by comparing shoot and fruit growth, water relations, leaf gas exchanges as well as fruit vascular and transpiration in/outflows of "Black Star" trees grafted on semi-vigorous (CAB6 P) and on semi-dwarfing (Gisela™6) rootstocks. The daily patterns of stem (Ψstem), leaf (Ψleaf) and fruit (Ψfruit) water potential, leaf photosynthesis, stomatal conductance and transpiration, shoot and fruit growth, fruit phloem, xylem and transpiration flows were assessed both in pre- and post-veraison, while productivity and fruit quality were determined at harvest. At both stages, no significant differences were found on Ψleaf, photosynthesis, fruit daily growth rates as well as fruit vascular and transpiration flows, while trees on Gisela™6 showed lower shoot growth rates and lower Ψstem and Ψfruit than trees on CAB6 P. The resulting decrease in stem-to-leaf Ψ gradient on Gisela™6 trees determined a reduction in shoot growth by decreasing shoot strength as sinks for water and carbohydrates. On the other hand, Gisela™6 fruit lowered their Ψfruit thanks to a higher osmotic accumulation and increased their competitiveness towards shoots, as confirmed by the higher productivity and fruit soluble solid content found at harvest for these trees. These results indicate that rootstock vigor alters resource competition between vegetative and reproductive growth, which can affect water use efficiency, yield, and fruit quality.

Sweet cherry fruit miRNAs and effect of high CO2 on the profile associated with ripening.

MAIN CONCLUSION: 157 known and 55 novel miRNAs were found in sweet cherry fruit. MiRNA target genes involved in fruit ripening and the differentially expressed miRNAs under CO2 treatment were identified. MicroRNAs (miRNAs) are short non-coding RNAs and play important functions in many biological processes, including fruit ripening and senescence. In the current study, the high-throughput sequencing and bioinformatics methods were implemented to decipher the miRNAs landscape in sweet cherry fruit. A total of 157 known miRNAs belonging to 50 families and 55 putative novel miRNAs were found. Target genes of the miRNAs were predicted and genes involved in fruit ripening were found, including F-box proteins and TFs such as SPL, TCP, NAC, MYB, ARF and AP2/ERF. And these target genes were further confirmed by degradome sequencing. A regulatory network model was constructed to uncover the miRNAs and their targets involved in fruit ripening and senescence. Importantly, elevated carbon dioxide can significantly postpone the ripening and senescence of sweet cherry fruit and the differentially expressed miRNAs exposed to CO2 were identified. These miRNAs included miR482j, miR6275, miR164, miR166, miR171, miR393, miR858, miR3627a, miR6284, miR6289 and miR7122b, and some of their functions were linked to fruit ripening. This study was the first report to profile miRNAs in sweet cherry fruit and it would provide more information for further study of miRNA roles in the ripening processes and their regulation mechanism underlying the effects of high carbon dioxide treatment on fruit ripening.

Yield potential definition of the chilling requirement reveals likely underestimation of the risk of climate change on winter chill accumulation.

Evaluation of chilling requirements of cultivars of temperate fruit trees provides key information to assess regional suitability, according to winter chill, for both industry expansion and ongoing profitability as climate change progresses. Traditional methods for calculating chilling requirements use climate-controlled chambers and define chilling requirements (CR) using a fixed bud burst percentage, usually close to 50% (CR-50%). However, this CR-50% definition may estimate chilling requirements that lead to flowering percentages that are lower than required for orchards to be commercially viable. We used sweet cherry to analyse the traditional method for calculating chilling requirements (CR-50%) and compared the results with a more restrictive method, where the chilling requirement was defined by a 90% bud break level (CRm-90%). For sweet cherry, this higher requirement of flowering success (90% as opposed to 50%) better represents grower production needs as a greater number of flowers leads to greater potential yield. To investigate the future risk of insufficient chill based on alternate calculations of the chilling requirement, climate projections of winter chill suitability across Europe were calculated using CR-50% and CRm-90%. Regional suitability across the landscape was highly dependent on the method used to define chilling requirements, and differences were found for both cold and mild winter areas. Our results suggest that bud break percentage levels used in the assessment of chilling requirements for sweet cherry influence production risks of current and future production areas. The use of traditional methods to determine chilling requirements can result in an underestimation of productivity chilling requirements for tree crops like sweet cherry which rely on a high conversion of flowers to mature fruit to obtain profitable yields. This underestimation may have negative consequences for the fruit industry as climate change advances with climate risk underestimated.

Foliage motion under wind, from leaf flutter to branch buffeting.

The wind-induced motion of the foliage in a tree is an important phenomenon both for biological issues (photosynthesis, pathogens development or herbivory) and for more subtle effects such as on wi-fi transmission or animal communication. Such foliage motion results from a combination of the motion of the branches that support the leaves, and of the motion of the leaves relative to the branches. Individual leaf dynamics relative to the branch, and branch dynamics have usually been studied separately. Here, in an experimental study on a whole tree in a large-scale wind tunnel, we present the first empirical evidence that foliage motion is actually dominated by individual leaf flutter at low wind velocities, and by branch turbulence buffeting responses at higher velocities. The transition between the two regimes is related to a weak dependence of leaf flutter on wind velocity, while branch turbulent buffeting is strongly dependent on it. Quantitative comparisons with existing engineering-based models of leaf and branch motion confirm the prevalence of these two mechanisms. Simultaneous measurements of the wind-induced drag on the tree and of the light interception by the foliage show the role of an additional mechanism, reconfiguration, whereby leaves bend and overlap, limiting individual leaf flutter. We then discuss the consequences of these findings on the role of wind-mediated phenomena.