Analysis of flavonoid metabolism of compounds in succulent fruits and leaves of three different colors of Rosaceae.

Red flesh apple (Malus pumila var. medzwetzkyana Dieck), purple leaf plum (Prunus cerasifera Ehrhar f), and purple leaf peach (Prunus persica 'Atropurpurea') are significant ornamental plants within the Rosaceae family. The coloration of their fruits and leaves is crucial in their appearance and nutritional quality. However, qualitative and quantitative studies on flavonoids in the succulent fruits and leaves of multicolored Rosaceae plants are lacking. To unveil the diversity and variety-specificity of flavonoids in these three varieties, we conducted a comparative analysis of flavonoid metabolic components using ultra-high-performance liquid phase mass spectrometry (UPLC-MS/MS). The results revealed the detection of 311 metabolites, including 47 flavonoids, 105 flavonols, 16 chalcones, 37 dihydroflavonoids, 8 dihydroflavonols, 30 anthocyanins, 14 flavonoid carbon glycosides, 23 flavanols, 8 isoflavones, 11 tannins, and 12 proanthocyanidins. Notably, although the purple plum and peach leaves exhibited distinct anthocyanin compounds, paeoniflorin and corythrin glycosides were common but displayed varying glycosylation levels. While the green purple leaf peach fruit (PEF) and red flesh apple leaf (AL) possessed the lowest anthocyanin content, they exhibited the highest total flavonoid content. Conversely, the red flesh apple fruit (AF) displayed the highest anthocyanin content and a diverse range of anthocyanin glycosylation modifications, indicating that anthocyanins predominantly influenced the fruit's color. Purple PLF, PLL, and PEL showcased varying concentrations of anthocyanins, suggesting that their colors result from the co-color interaction between specific types of anthocyanins and secondary metabolites, such as flavonols, flavonoids, and dihydroflavonoids. This study provides novel insights into the variations in tissue metabolites among Rosaceae plants with distinct fruit and leaf colors.

Transcription factor LBD16 targets cell wall modification/ion transport genes in peach lateral root formation.

LATERAL ORGAN BOUNDARIES DOMAIN/ASYMMETRIC LEAVES2-LIKEs (LBDs/ASLs) are plant-specific transcription factors that function downstream of auxin-regulated lateral root (LR) formation. Our previous research found that PpLBD16 positively regulates peach (Prunus persica) LR formation. However, the downstream regulatory network and target genes of PpLBD16 are still largely unknown. Here, we constructed a PpLBD16 homologous overexpression line and a PpLBD16 silenced line. We found that overexpressing PpLBD16 promoted peach root initiation, while silencing PpLBD16 inhibited peach root formation. Through RNA sequencing (RNA-seq) analysis of roots from PpLBD16 overexpression and silenced lines, we discovered that genes positively regulated by PpLBD16 were closely related to cell wall synthesis and degradation, ion/substance transport, and ion binding and homeostasis. To further detect the binding motifs and potential target genes of PpLBD16, we performed DNA-affinity purification sequencing (DAP-seq) analysis in vitro. PpLBD16 preferentially bound to CCNGAAANNNNGG (MEME-1), [C/T]TTCT[C/T][T/C] (MEME-2), and GCGGCGG (ABR1) motifs. By combined analysis of RNA-seq and DAP-seq data, we screened candidate target genes for PpLBD16. We demonstrated that PpLBD16 bound and activated the cell wall modification-related genes EXPANSIN-B2 (PpEXPB2) and SUBTILISIN-LIKE PROTEASE 1.7 (PpSBT1.7), the ion transport-related gene CYCLIC NUCLEOTIDE-GATED ION CHANNEL 1 (PpCNGC1) and the polyphenol oxidase (PPO)-encoding gene PpPPO, thereby controlling peach root organogenesis and promoting LR formation. Moreover, our results displayed that PpLBD16 and its target genes are involved in peach LR primordia development. Overall, this work reveals the downstream regulatory network and target genes of PpLBD16, providing insights into the molecular network of LBD16-mediated LR development.

The PpMYB75-PpDFR module reveals the difference between 'SR' and its bud variant 'RMHC' in peach red flesh.

'Red Meat Honey Crisp (RMHC)' has been widely cultivated by growers in recent years due to its early maturity, and red meat type characteristics. As a bud variant of 'Super Red (SR)' peach, red flesh is the most distinctive characteristic of 'Red Meat Honey Crisp (RMHC)'. However, the mechanism of red flesh formation in 'RMHC' remains unclear. In this study, 79 differentially produced metabolites were identified by metabolomics analysis. The anthocyanin content in 'RMHC' was significantly higher than that in 'SR' during the same period, such as cyanidin O-syringic acid and cyanidin 3-O-glucoside. Other flavonoids also increased during the formation of red flesh, including flavonols (6-hydroxykaempferol-7-O-glucoside, hyperin), flavanols (protocatechuic acid, (+)-gallocatechin), and flavonoids (chrysoeriol 5-O-hexoside, tricetin). In addition, transcriptomic analysis and RT-qPCR showed that the expression levels of the flavonoid synthesis pathway transcription factor MYB75 and some structural genes, such as PpDFR, PpCHS, PpC4H, and PpLDOX increased significantly in 'RMHC'. Subcellular localization analysis revealed that MYB75 was localized to the nucleus. Yeast single hybridization assays showed that MYB75 bound to the cis-acting element CCGTTG of the PpDFR promoter region. The MYB75-PpDFR regulatory network was identified to be a key pathway in the reddening of 'RMHC' flesh. Moreover, this is the first study to describe the cause for red meat reddening in 'RMHC' compared to 'SR' peaches using transcriptomics, metabolomics and molecular methods. Our study identified a key transcription factor involved in the regulation of the flavonoid synthetic pathway and contributes to peach breeding-related efforts as well as the identification of genes involved in color formation in other species.

Formation of key aroma-active and off-flavor components in concentrated peach puree.

Non-volatiles offer some insight into the formation of aroma-active components in peach puree (PP), but more depth investigation is still needed. Formation pathways of key aroma-active and off-flavor components in PP during thermal concentration (PP + C) and sterilization (PP + C + S) are unclear. Therefore, GC-O-MS combined with UPLC-MS/MS was used to identify the volatile and nonvolatile components and their formation pathways. Among the 36 aroma-active compounds, the contents of γ-decalactone, hexyl acetate, leaf acetate, hexanal, and 1-hexanol (odor activity value ≥ 1) decreased by 46 %, 100 %, 100 %, 92 %, and 100 % between PP and PP + C + S, causing the weakening of "green" and "fruity" attributes. Off-flavor components including 1-octen-3-one, isobutyric acid, isothiazole, and isovaleric acid were identified during thermal processing. 1-Octen-3-one content increased by 75 % from PP to PP + C + S through linolenic acid metabolism, which contributed to "cooked"; the formation of isobutyric and isovaleric acids, isothiazole, resulted in the enhancement of "sour/rancid" via serine and leucine metabolism.

Water extract of Fagopyrum dibotrys (D. Don) Hara straw increases selenium accumulation in peach seedlings under selenium-contaminated soil.

To promote the selenium (Se) uptakes in fruit trees under Se-contaminated soil, the effects of water extract of Fagopyrum dibotrys (D. Don) Hara straw on the Se accumulation in peach seedlings under selenium-contaminated soil were studied. The results showed that the root biomass, chlorophyll content, activities of antioxidant enzymes, and soluble protein content of peach seedlings were increased by the F. dibotrys straw extract. The different forms of Se (total Se, inorganic Se, and organic Se) were also increased in peach seedlings following treatment with the F. dibotrys straw extract. The highest total shoot Se content was treated by the 300-fold dilution of F. dibotrys straw, which was 30.87% higher than the control. The F. dibotrys straw extract also increased the activities of adenosine triphosphate sulfurase (ATPS), and adenosine 5'-phosphosulfate reductase (APR) in peach seedlings, but decreased the activity of serine acetyltransferase (SAT). Additionally, correlation and grey relational analyses revealed that chlorophyll a content, APR activity, and root biomass were closely associated with the total shoot Se content. Overall, this study shows that the water extract of F. dibotrys straw can promote Se uptake in peach seedlings, and 300-fold dilution is the most suitable concentration.

The water extract of Fagopyrum dibotrys (D. Don) Hara straw promoted the selenium (Se) uptake in peach seedlings under selenium-contaminated soil. The concentration of F. dibotrys straw extract showed a quadratic polynomial regression relationship with the total root and shoot Se. Furthermore, chlorophyll a content, APR activity, and root biomass were closely associated with the total shoot Se. This study shows that water extract of F. dibotrys straw can promote Se uptake in peach seedlings, and 300-fold dilution is the most suitable concentration.

Melatonin Promotes Iron Reactivation and Reutilization in Peach Plants under Iron Deficiency.

The yellowing of leaves due to iron deficiency is a prevalent issue in peach production. Although the capacity of exogenous melatonin (MT) to promote iron uptake in peach plants has been demonstrated, its underlying mechanism remains ambiguous. This investigation was carried out to further study the effects of exogenous MT on the iron absorption and transport mechanisms of peach (Prunus persica) plants under iron-deficient conditions through transcriptome sequencing. Under both iron-deficient and iron-supplied conditions, MT increased the content of photosynthetic pigments in peach leaves and decreased the concentrations of pectin, hemicellulose, cell wall iron, pectin iron, and hemicellulose iron in peach plants to a certain extent. These effects stemmed from the inhibitory effect of MT on the polygalacturonase (PG), cellulase (Cx), phenylalanine ammonia-lyase (PAL), and cinnamoyl-coenzyme A reductase (CCR) activities, as well as the promotional effect of MT on the cinnamic acid-4-hydroxylase (C4H) activity, facilitating the reactivation of cell wall component iron. Additionally, MT increased the ferric-chelate reductase (FCR) activity and the contents of total and active iron in various organs of peach plants under iron-deficient and iron-supplied conditions. Transcriptome analysis revealed that the differentially expressed genes (DEGs) linked to iron metabolism in MT-treated peach plants were primarily enriched in the aminoacyl-tRNA biosynthesis pathway under iron-deficient conditions. Furthermore, MT influenced the expression levels of these DEGs, regulating cell wall metabolism, lignin metabolism, and iron translocation within peach plants. Overall, the application of exogenous MT promotes the reactivation and reutilization of iron in peach plants.

Crucial roles of sorbitol metabolism and energy status in the chilling tolerance of yellow peach.

In this study, we compared sorbitol metabolism, energy metabolism, and CI development in yellow peach fruit at 1 °C (less susceptible to CI) and 8 °C (more susceptible to CI) storage to elucidate potential connections between them. The results indicated that storage at 1 °C effectively maintained the textural quality of yellow peach fruit and delayed the onset of CI by 12 days compared to 8 °C. This positive effect might be attributable to 1 °C storage maintaining higher sorbitol content throughout the storage duration, thus sustaining the higher adenosine triphosphate (ATP) level and energy charge. The regulation of sorbitol accumulation by 1 °C storage was closely linked to the metabolic activity of sorbitol, which stimulated sorbitol synthesis by enhancing sorbitol-6-phosphate dehydrogenase (S6PDH) activity after 12 days while suppressing sorbitol degradation via decreased sorbitol oxidase (SOX) and NAD+-sorbitol dehydrogenase (NAD+-SDH) activities before 24 days. In addition, the notable up-regulation in the NAD+-SDH activity in the late storage period promoted the conversion of sorbitol to fructose and glucose under 1 °C storage, thereby providing ample energy substrate for ATP generation. Moreover, sorbitol acts as a vital signaling molecule, and substantially up-regulated expressions of sorbitol transporters genes (PpeSOT3, PpeSOT5, and PpeSOT7) were observed in fruit stored at 1 °C, which might promote sorbitol transport and improve cold tolerance in peach fruit. Taken together, these findings suggested that 1 °C storage delayed CI by enhancing sorbitol metabolism and transporter activity, promoting sorbitol accumulation, and finally elevating the energy status in yellow peach fruit.

Integration of metabolomics and transcriptomics analyses reveals the mechanism of nano-selenium treated to activate phenylpropanoid metabolism and enhance the antioxidant activity of peach.

Foliar spraying to improve the quality of fruits is a general approach nowadays. In this study, 10 ppm nano-selenium (nano-Se) diluted with distilled water was sprayed on peach leaves every 10 days for a total of 7 sprays during the fruit set period. And then their fruit quality was compared with that of control group. It was found that the firmness, soluble solid concentration, total phenol, and proanthocyanidin content of the peaches were raised after the nano-Se treatment. Moreover, the ascorbic acid glutathione loop (ASA-GSH loop) was fully activated in the nano-Se treated group, and the associated antioxidant capacity and enzyme activity were significantly increased. Metabolomics revealed that nano-Se could upregulate some metabolites, such as phenylalanine, naringenin, and pinocembrin, to fully activate the metabolism of phenylpropanoids. Further, based on transcriptomics, nano-Se treatment was found to affect fruit quality by regulating genes related to phenylpropanoid metabolism, such as arogenate/prephenate dehydratase (ADT), genes related to abscisic acid metabolism such as (+)-abscisic acid 8'-hydroxylase (CYP707A), and some transcription factors such as MYB. Based on the comprehensive analysis of physicochemical indicators, metabolomics, and transcriptomics, it was found that nano-Se improved fruit quality by activating phenylpropanoid metabolism and enhancing antioxidant capacity. This work provides insights into the mechanism of the effect of nano-Se fertilizer on peach fruit quality. PRACTICAL APPLICATION: The firmness and soluble solid concentration of peaches are higher after nano-Se treatment, which is more in line with people's demand for hard soluble peaches like "Yingzui." The antioxidant capacity, antioxidant substance content, and antioxidant enzyme activity of nano-Se-treated peaches are higher, with potential storage resistance and health effects on human body. The mechanism of nano-Se affecting peach quality was analyzed by metabolomics and transcriptomics, which is a reference and guide for the research and application of nano-Se.

ß-Cyclocitric acid enhances drought tolerance in peach (Prunus persica) seedlings.

The ß-cyclocitric acid (ß-CCA) is a bioactive apocarotenoid previously shown to improve drought tolerance in annual plants. However, the underlying molecular mechanism of this process remains largely elusive. Moreover, the question about the activity of ß-CCA in perennial fruit crops is still open. Here, we found that treatment of ß-CCA enhances drought tolerance in peach seedlings. The application of ß-CCA significantly increased the relative water content and root activity and reduced the electrolyte leakage of peach seedlings under drought stress. Moreover, treatment with ß-CCA under drought stress increased chlorophyll fluorescence, indicating a positive effect on photosynthesis, while also enhancing superoxide dismutase and peroxidase activity and reducing reactive oxygen species (ROS) levels. Consistent with these alterations, transcriptome analysis revealed an up-regulation of photosynthesis and antioxidant-related genes upon the application of ß-CCA under drought stress. We also detected an induction in genes related to detoxification, environmental adaptation, primary metabolism, phytohormone, phenylpropanoid and the biosynthesis of cutin, suberine and wax, which might contribute to the induction of drought resistance. Altogether, our study reveals that ß-CCA positively modulates peach drought tolerance, which is mainly mediated by enhancing photosynthesis and reducing ROS, indicating the potential of utilizing ß-CCA for drought control in peach and perhaps other fruit crops.

Low temperature inhibits pectin degradation by PpCBFs to prolong peach storage time.

Low-temperature storage is a widely used method for peach fruit storage. However, the impact of PpCBFs on pectin degradation during low-temperature storage is unclear. As such, in this study, we stored the melting-flesh peach cultivar "Fuli" at low temperature (LT, 6°C) and room temperature (RT, 25°C) to determine the effect of different temperatures on its physiological and biochemical changes. Low-temperature storage can inhibit the softening of "Fuli" peaches by maintaining the stability of the cell wall. It was found that the contents of water-soluble pectin and ionic-soluble pectin in peach fruit stored at RT were higher than those stored at LT. The enzyme activities of polygalacturonase (PG), pectate lyase (PL), and pectin methylesterase (PME) were all inhibited by LT. The expressions of PpPME3, PpPL2, and PpPG were closely related to fruit firmness, but PpCBF2 and PpCBF3 showed higher expression levels at LT than RT. The promoters of PpPL2 and PpPG contain the DER motif, which suggested that PpCBF2 and PpCBF3 might negatively regulate their expression by directly binding to their promoters. These results indicated that LT may maintain firmness by activating PpCBFs to repress pectin-degradation-related enzyme genes during storage.

Transcriptomic and metabolite analyses provided a new sight of 1-MCP on organic acid metabolism in peach during storage.

In this study, "Xiahui 6" peaches were treated with 10 µL/L 1-methylcyclopropene (1-MCP) for 12 h and then stored at 20°C for 9 days; the regulation of 1-MCP on organic acids during storage was investigated through transcriptomic and metabolite analyses. Results showed that 1-MCP maintained higher gene expression of malate synthesis (PpPEPC1, PpPEPC2, and PpNAD-cytMDH) at the end of storage but extremely inhibited the gene expression of malate degradation (PpNADP-cytME) during storage, resulting that malate content in treated peaches was twice that of control group at day 7. Besides, the increasement of citrate synthesis and degradation-related genes (PpmitCS, PpcytACO, PpNAD-mitIDH, and PpNADP-cytIDH) at days 3 and 5 was postponed by 1-MCP treatment, accompanied by 0.5 times higher citrate content at day 7. Our results suggested that 1-MCP has inhibitory effects on both the synthesis and degradation of organic acids; however, the inhibitory effect of 1-MCP on organic acid degradation may be greater than that on organic acid synthesis. Practical Application: This study provides a theoretical basis for the application of 1-methylcyclopropene (1-MCP) in fruit preservation.

Evaluation of fungal dynamics during sheep manure composting employing peach shell biochar.

In this study, explored the influence of different proportion (0%, 2.5%, 5%, 7.5%, and 10%) peach shell biochar (PSB) with microbial agents (EM) on the carbon transformation, humification process and fungal community dynamics during sheep manure (SM) composting. And no additives were used as control. The results manifested that the CO2 and CH4 emissions were effectively reduced 8.23%∼13.10% and 17.92%∼33.71%. The degradation rate of fulvic acid increased by 17.12%∼23.08% and the humic acid contents were enhanced by 27.27%∼33.97% so that accelerated the composting. Besides, the dominant fungal phylum was Ascomycota (31.43%∼52.54%), Basidiomycota (3.12%∼13.85%), Mucoromycota (0.40%∼7.61%) and Mortierellomycota (0.97%∼2.39%). Pearson correlation analysis and network indicated that there were different correlations between physicochemical indexes and fungal community under different additive concentrations. In brief, the two modifiers application promoted the SM degradation and affected the fungal community structure.

The Mechanism of Peach Gum Polysaccharide Preventing UVB-Induced Skin Photoaging by Regulating Matrix Metalloproteinanse and Oxidative Factors.

Exposure to ultraviolet light can cause oxidative damage and accelerate skin aging and is one of the main causes of skin aging. Peach gum polysaccharide (PG) is a natural edible plant component that has many biological activities, such as regulating blood glucose and blood lipids and improving colitis, as well as antioxidant and anticancer properties. However, there are few reports on the antiphotoaging effect of peach gum polysaccharide. Therefore, in this paper, we study the basic composition of the raw material peach gum polysaccharide and its ability to improve UVB-induced skin photoaging damage in vivo and in vitro. The results show that peach gum polysaccharide is mainly composed of mannose, glucuronic acid, galactose, xylose, and arabinose, and its molecular weight (Mw) is 4.10 × 106 g/mol. The results of the in vitro cell experiments show that PG could significantly alleviate UVB-induced apoptosis of human skin keratinocytes, promote cell growth repair, reduce the expression of intracellular oxidative factors and matrix metal collagenase, and improve the extent of oxidative stress repair. Moreover, the results from the in vivo animal experiments showed that PG could not only effectively improve the phenotype of UVB-induced photoaged skin in model mice but also significantly improve their oxidative stress status, regulate the contents of ROS and the levels of SOD and CAT, and repair the oxidative skin damage induced by UVB in vivo. In addition, PG improved UVB-induced photoaging-mediated collagen degradation in mice by inhibiting the secretion of matrix metalloproteinases. The above results indicate that peach gum polysaccharide has the ability to repair UVB-induced photoaging and may be used as a potential drug and antioxidant functional food to resist photoaging in the future.

Two interacting ethylene response factors negatively regulate peach resistance to Lasiodiplodia theobromae.

Gummosis is 1 of the most common and destructive diseases threatening global peach (Prunus persica) production. Our previous studies have revealed that ethylene and methyl jasmonate enhance peach susceptibility to Lasiodiplodia theobromae, a virulent pathogen inducing gummosis; however, the underlying molecular mechanisms remain obscure. Here, 2 ethylene response factors (ERFs), PpERF98 and PpERF1, were identified as negative regulators in peach response to L. theobromae infection. Expression of 2 putative paralogs, PpERF98-1/2, was dramatically induced by ethylene and L. theobromae treatments and accumulated highly in the gummosis-sensitive cultivar. Silencing of PpERF98-1/2 increased salicylic acid (SA) content and pathogenesis-related genes PpPR1 and PpPR2 transcripts, conferring peach resistance to L. theobromae, whereas peach and tomato (Solanum lycopersicum) plants overexpressing either of PpERF98-1/2 showed opposite changes. Also, jasmonic acid markedly accumulated in PpERF98-1/2-silenced plants, but reduction in PpPR3, PpPR4, and PpCHI (Chitinase) transcripts indicated a blocked signaling pathway. PpERF98-1 and 2 were further demonstrated to directly bind the promoters of 2 putative paralogous PpERF1 genes and to activate the ERF branch of the jasmonate/ethylene signaling pathway, thus attenuating SA-dependent defenses. The lesion phenotypes of peach seedlings overexpressing PpERF1-1/2 and PpERF98-1/2 were similar. Furthermore, PpERF98-1/2 formed homodimers/heterodimers and interacted with the 2 PpERF1 proteins to amplify the jasmonate/ethylene signaling pathway, as larger lesions were observed in peach plants cooverexpressing PpERF98 with PpERF1 relative to individual PpERF98 overexpression. Overall, our work deciphers an important regulatory network of ethylene-mediated peach susceptibility to L. theobromae based on a PpERF98-PpERF1 transcriptional cascade, which could be utilized as a potential target for genetic engineering to augment protection against L. theobromae-mediated diseases in crops and trees.

Effect of ultrahigh-pressure treatment on the structure and allergenicity of peach allergenic proteins.

Peach is a common plant-derived allergenic food and ultrahigh-pressure treatment is often used in peach products. In our study, an in-depth analysis of the structural and allergenicity changes of peach allergenic proteins after UHP treatment was performed by spectroscopy, mass spectrometry combined with serology and cytology. The results indicated that UHP treatment could reduce the content of peach soluble proteins and cause changes in secondary and tertiary structures. In addition, more hydrophobic residues were exposed and proteins tended to polymerize after UHP-treatment. The results of immunological assays showed that UHP treatment could reduce the IgE binding capacity of peach proteins and affect the ability of basophil degranulation, the upregulation of some cytokines may contribute to the reduction of peach protein allergenicity. Notably, UHP treatment may lead to the masking of some digestion sites in Pru p 3 epitopes, thus impeding human digestion and increasing the potential risk of allergenicity.

MADS-box protein PpDAM6 regulates chilling requirement-mediated dormancy and bud break in peach.

Bud dormancy is crucial for winter survival and is characterized by the inability of the bud meristem to respond to growth-promotive signals before the chilling requirement (CR) is met. However, our understanding of the genetic mechanism regulating CR and bud dormancy remains limited. This study identified PpDAM6 (DORMANCY-ASSOCIATED MADS-box) as a key gene for CR using a genome-wide association study analysis based on structural variations in 345 peach (Prunus persica (L.) Batsch) accessions. The function of PpDAM6 in CR regulation was demonstrated by transiently silencing the gene in peach buds and stably overexpressing the gene in transgenic apple (Malus × domestica) plants. The results showed an evolutionarily conserved function of PpDAM6 in regulating bud dormancy release, followed by vegetative growth and flowering, in peach and apple. The 30-bp deletion in the PpDAM6 promoter was substantially associated with reducing PpDAM6 expression in low-CR accessions. A PCR marker based on the 30-bp indel was developed to distinguish peach plants with non-low and low CR. Modification of the H3K27me3 marker at the PpDAM6 locus showed no apparent change across the dormancy process in low- and non-low- CR cultivars. Additionally, H3K27me3 modification occurred earlier in low-CR cultivars on a genome-wide scale. PpDAM6 could mediate cell-cell communication by inducing the expression of the downstream genes PpNCED1 (9-cis-epoxycarotenoid dioxygenase 1), encoding a key enzyme for ABA biosynthesis, and CALS (CALLOSE SYNTHASE), encoding callose synthase. We shed light on a gene regulatory network formed by PpDAM6-containing complexes that mediate CR underlying dormancy and bud break in peach. A better understanding of the genetic basis for natural variations of CR can help breeders develop cultivars with different CR for growing in different geographical regions.

Silencing of PpNRAMP5 improves manganese toxicity tolerance in peach (Prunus persica) seedlings.

The natural resistance-associated macrophage protein (NRAMP) gene family assists in the transport of metal ions in plants. However, the role and underlying physiological mechanism of NRAMP genes under heavy metal toxicity in perennial trees remain to be elucidated. In Prunus persica, five NRAMP family genes were identified and named according to their predicted phylogenetic relationships. The expression profiling analysis indicated that PpNRAMPs were significantly induced by excess manganese (Mn), iron, zinc, and cadmium treatments, suggesting their potential role in heavy metal uptake and transportation. Notably, the expression of PpNRAMP5 was tremendously increased under Mn toxicity stress. Heterologous expression of PpNRAMP5 in yeast cells also confirmed Mn transport. Suppression of PpNRAMP5 through virus-induced gene silencing enhanced Mn tolerance, which was compromised when PpNRAMP5 was overexpressed in peach. The silencing of PpNRAMP5 mitigated Mn toxicity by dramatically reducing Mn contents in roots, and effectively reduced the chlorophyll degradation and improved the photosynthetic apparatus under Mn toxicity stress. Therefore, PpNRAMP5-silenced plants were less damaged by oxidative stress, as signified by lowered H2O2 contents and O2•- staining intensity, also altered the reactive oxygen species (ROS) homeostasis by activating enzymatic antioxidants. Consistently, these physiological changes showed an opposite trend in the PpNRAMP5-overexpressed peach plants. Altogether, our findings suggest that downregulation of PpNRAMP5 markedly reduces the uptake and transportation of Mn, thus activating enzymatic antioxidants to strengthen ROS scavenging capacity and photosynthesis activity, thereby mitigating Mn toxicity in peach plants.

Waterlogging resistance and evaluation of physiological mechanism of three peach (Prunus persica) rootstocks.

Waterlogging occurs due to poor soil drainage or excessive rainfall. It is a serious abiotic stress factor that negatively affects crop growth. Waterlogging often causes plants to shed leaves, fruits, and, ultimately, to die. Peach (Prunus persica) trees are generally intolerant to waterlogging, and the primary peach rootstock used in Chinais "Maotao," which has very poor resistance to sensitivity. Therefore, waterlogging has become a restriction on the development of the peach industry in many regions. In this experiment, we tested the waterlogging resistance of "Maotao (Prunus persica (L.) Batsch)" (MT), "Shannong1 (GF677 × Cadaman)" (SN1), and "Mirabolano 29C (Prunus cerasifera)" (M29C) rootstocks. Using a simulated waterlogging method, the effects of waterlogging on the photosynthetic system, leaf pigments, osmotic adjustment, lipid membrane peroxidation, and antioxidant system of these three peach rootstocks were studied, and the changes of chlorophyll fluorescence parameters and fluorescence imaging were observed. The results showed that, with prolonged waterlogging, the photosynthetic pigment content and photosynthesis of the three peach rootstocks decreased rapidly, but the decomposition rate of SN1 and M29C chlorophyll was slower, and it still had high light energy absorption and energy transfer capabilities under waterlogging stress, which reduced the damage caused by waterlogging stress; under the stress of flooding, the osmoregulatory substances of the three rootstocks increased to varying degrees compared with normal conditions. At the same time, the enzyme activity of superoxide dismutase (SOD) activity, peroxidase (POD) activity, and catalase (CAT) activity in the leaves of the three rootstocks under flooding stress all increased and then decreased; during this period, malondialdehyde (MDA) continued to increase, and SN1 and M29C were significantly lower than MT; and chlorophyll fluorescence parameters, including the maximum photochemical efficiency (Fv/Fm), actual photochemical efficiency (ΦPSII), photochemical quenching coefficient (qP), non-photochemical quenching (NPQ), and electron transfer rate (ETR) decreased significantly. The tolerance of SN1 and M29C to waterlogging was significantly better than that of MT rootstocks. The rootstock and grafted seedlings of SN1 have good waterlogging tolerance.

Peach DELLA Protein PpeDGYLA Is Not Degraded in the Presence of Active GA and Causes Dwarfism When Overexpressed in Poplar and Arabidopsis.

Controlling the tree size of fruit species such as peach can reduce the amount of labor and input needed for orchard management. The phytohormone gibberellin (GA) positively regulates tree size by inducing degradation of the GA signaling repressor DELLA. The N-terminal DELLA domain in this protein is critical for its GA-dependent interaction with the GA receptor GID1 and the resulting degradation of the DELLA protein, which allows for growth-promoting GA signaling. In this study, a DELLA family member, PpeDGYLA, contains a DELLA domain but has amino acid changes in three conserved motifs (DELLA into DGYLA, LEQLE into LERLE, and TVHYNP into AVLYNP). In the absence or presence of GA3, the PpeDGYLA protein did not interact with PpeGID1c and was stable in 35S-PpeDGYLA peach transgenic callus. The overexpression of PpeDGYLA in both polar and Arabidopsis showed an extremely dwarfed phenotype, and these transgenic plants were insensitive to GA3 treatment. PpeDGYLA could interact with PpeARF6-1 and -2, supposed growth-promoting factors. It is suggested that the changes in the DELLA domain of PpeDGYLA may, to some extent, account for the severe dwarf phenotype of poplar and Arabidopsis transgenic plants. In addition, our study showed that the DELLA family contained three clades (DELLA-like, DELLA, and DGLLA). PpeDGYLA clustered into the DGLLA clade and was expressed in all of the analyzed tissues. These results lay the foundation for the further study of the repression of tree size by PpeDGYLA.

The microRNA ppe-miR393 mediates auxin-induced peach fruit softening by promoting ethylene production.

Auxin can inhibit or promote fruit ripening, depending on the species. Melting flesh (MF) peach fruit (Prunus persica L. Batsch) cultivars produce high levels of ethylene caused by high concentrations of indole-3-acetic acid (IAA), which leads to rapid fruit softening at the late stage of development. In contrast, due to the low concentrations of IAA, the fruit of stony hard (SH) peach cultivars does not soften and produces little ethylene. Auxin seems necessary to trigger the biosynthesis of ethylene in peach fruit; however, the mechanism is not well understood. In this study, we identified miRNA gene family members ppe-miR393a and ppe-miR393b that are differentially expressed in SH and MF fruits. RNA ligase-mediated 5' rapid amplification of cDNA ends and transient transformation of Nicotiana benthamiana revealed TRANSPORT INHIBITOR RESPONSE 1 (PpTIR1), part of the auxin perception and response system, as a target of ppe-miR393a and b. Yeast 2-hybrid assay and bimolecular fluorescence complementation assay revealed that PpTIR1 physically interacts with an Aux/IAA protein PpIAA13. The results of yeast 1-hybrid assay, electrophoretic mobility shift assay, and dual-luciferase assay indicated that PpIAA13 could directly bind to and trans-activate the promoter of 1-aminocyclopropane-1-carboxylic acid synthase 1 (PpACS1), required for ethylene biosynthesis. Transient overexpression and suppression of ppe-miR393a and PpIAA13 in peach fruit induced and repressed the expression of PpACS1, confirming their regulatory role in ethylene synthesis. Gene expression analysis in developing MF and SH fruits, combined with postharvest α-naphthalene acetic acid (NAA) treatment, supports a role for a ppe-miR393-PpTIR1-PpIAA13-PpACS1 module in regulating auxin-related differences in ethylene production and softening extent in different types of peach.