Advancing understanding of Ficus carica: a comprehensive genomic analysis reveals evolutionary patterns and metabolic pathway insights.

Ficus carica L. (dioecious), the most significant commercial species in the genus Ficus, which has been cultivated for more than 11,000 years and was one of the first species to be domesticated. Herein, we reported the most comprehensive F. carica genome currently. The contig N50 of the Orphan fig was 9.78 Mb, and genome size was 366.34 Mb with 13 chromosomes. Based on the high-quality genome, we discovered that F. carica diverged from Ficus microcarpa ~34 MYA, and a WGD event took place about 2─3 MYA. Throughout the evolutionary history of F. carica, chromosomes 2, 8, and 10 had experienced chromosome recombination, while chromosome 3 saw a fusion and fission. It is worth proposing that the chromosome 9 experienced both inversion and translocation, which facilitated the emergence of the F. carica as a new species. And the selections of F. carica for the genes of recombination chromosomal fragment are compatible with their goal of domestication. In addition, we found that the F. carica has the FhAG2 gene, but there are structural deletions and positional jumps. This gene is thought to replace the one needed for female common type F. carica to be pollinated. Subsequently, we conducted genomic, transcriptomic, and metabolomic analysis to demonstrate significant differences in the expression of CHS among different varieties of F. carica. The CHS playing an important role in the anthocyanin metabolism pathway of F. carica. Moreover, the CHS gene of F. carica has a different evolutionary trend compared to other Ficus species. These high-quality genome assembly, transcriptomic, and metabolomic resources further enrich F. carica genomics and provide insights for studying the chromosomes evolution, sexual system, and color characteristics of Ficus.

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.

Multiple-statistical genome-wide association analysis and genomic prediction of fruit aroma and agronomic traits in peaches.

'Chinese Cling' is an important founder in peach breeding history due to the pleasant flavor. Genome-wide association studies (GWAS) combined with genomic selection are promising tools in fruit tree breeding, as there is a considerable time lapse between crossing and release of a cultivar. In this study, 242 peaches from Shanghai germplasm were genotyped with 145 456 single-nucleotide polymorphisms (SNPs). The six agronomic traits of fruit flesh color, fruit shape, fruit hairiness, flower type, pollen sterility, and soluble solids content, along with 14 key volatile odor compounds (VOCs), were recorded for multiple-statistical GWAS. Except the reported candidate genes, six novel genes were identified as associated with these traits. Thirty-nine significant SNPs were associated with eight VOCs. The putative candidate genes were confirmed for VOCs by RNA-seq, including three genes in the biosynthesis pathway found to be associated with linalool, soluble solids content, and cis-3-hexenyl acetate. Multiple-trait genomic prediction enhanced the predictive ability for γ-decalactone to 0.7415 compared with the single-trait model value of 0.1017. One PTS1-SSR marker was designed to predict the linalool content, and the favorable genotype 187/187 was confirmed, mainly existing in the 'Shanghai Shuimi' landrace. Overall, our findings will be helpful in determining peach accessions with the ideal phenotype and show the potential of multiple-trait genomic prediction to improve accuracy for highly correlated genetic traits. The diagnostic marker will be valuable for the breeder to bridge the gap between quantitative trait loci and marker-assisted selection for developing strong-aroma cultivars.

Transcriptome Co-Expression Network Analysis of Peach Fruit with Different Sugar Concentrations Reveals Key Regulators in Sugar Metabolism Involved in Cold Tolerance.

Peach fruits are known to be highly susceptible to chilling injury (CI) during low-temperature storage, which has been linked to the level of sugar concentration in the fruit. In order to better understand the relationship between sugar metabolism and CI, we conducted a study examining the concentration of sucrose, fructose, and glucose in peach fruit with different sugar concentrations and examined their relationship with CI. Through transcriptome sequencing, we screened the functional genes and transcription factors (TFs) involved in the sugar metabolism pathway that may cause CI in peach fruit. Our results identified five key functional genes (PpSS, PpINV, PpMGAM, PpFRK, and PpHXK) and eight TFs (PpMYB1/3, PpMYB-related1, PpWRKY4, PpbZIP1/2/3, and PpbHLH2) that are associated with sugar metabolism and CI development. The analysis of co-expression network mapping and binding site prediction identified the most likely associations between these TFs and functional genes. This study provides insights into the metabolic and molecular mechanisms regulating sugar changes in peach fruit with different sugar concentrations and presents potential targets for breeding high-sugar and cold-tolerant peach varieties.

Profiling of naturally occurring proanthocyanidins and other phenolic compounds in a diverse peach germplasm by LC-MS/MS.

Peach fruits are rich in phenolic compounds and have considerable health benefits. In this study, 19 proanthocyanidins (PACs) and 37 other phenolic compounds are identified and evaluated in the mature fruits of 217 peach accessions using LC-QTRAP-MS/MS and LC-QTOF-MS analyses. Total PAC quantities ranged from 18.93 to 697.52 µg/g fresh weight with a variance of 36.8-fold, and accounted for 11.2 % - 85.6 % of total phenolics content. Kaempferol-3-O-rutinoside (0.001-0.968 µg/g), isorhamnetin-3-O-rutinoside (0.001-0.300 µg/g), taxifolin (0.006-0.078 µg/g), luteoloside (0.002-0.068 µg/g), prunin (0.043-33.333 µg/g), phlorizin (0.018-1.100 µg/g), and trans-piceid (0.013-0.472 µg/g) were also highly diverse. The fruit ripening period, breeding background and fruit type significantly influenced the PACs and phenolic glycosides. This study presents a complete profile of PACs and other major phenolics in 217 peach germplasms, and is expected to aid future peach breeding procedures targeted at producing plants rich in specific phenolics.

Genome-wide analysis of terpene synthase gene family to explore candidate genes related to disease resistance in Prunus persica.

In plants, a family of terpene synthases (TPSs) is responsible for the biosynthesis of terpenes and contributes to species-specific diversity of volatile organic compounds, which play essential roles in fitness of plants. However, little is known about the TPS gene family in peach and/or nectarine (Prunus persica L.). In this study, we identified 40 PpTPS genes in peach genome v2.0. Although these PpTPSs could be clustered into five classes, they distribute in several gene clusters of three chromosomes, share conserved exon-intron organizations, and code similar protein motifs. Thirty-five PpTPSs, especially PpTPS2, PpTPS23, PpTPS17, PpTPS18, and PpTPS19, altered their transcript levels after inoculation with Botryosphaeria dothidea, a cause of peach gummosis, compared to the mock treatments, which might further affect the contents of 133 terpenoids at 48 hours and/or 84 hours post inoculations in the current-year shoots of 'Huyou018', a highly susceptible nectarine cultivar. Moreover, about fifteen PpTPSs, such as PpTPS1, PpTPS2, PpTPS3, and PpTPS5, showed distinct expression patterns during fruit development and ripening in two peach cultivars, yellow-fleshed 'Jinchun' and white-fleshed 'Hikawa Hakuho'. Among them, the transcription level of chloroplast-localized PpTPS3 was obviously related to the content of linalool in fruit pulps. In addition, elevated concentrations (0.1 g/L to 1.0 g/L) of linalool showed antifungal activities in PDA medium. These results improve our understanding of peach PpTPS genes and their potential roles in defense responses against pathogens.

Controlled atmosphere storage alleviates internal browning in flat peach fruit by regulating energy and sugar metabolisms.

Flat peach fruit are cold-sensitive and vulnerable to chilling injury (CI), particularly internal browning (IB) during cold storage, which limits the consumer acceptance and market value of the fruit. Controlled atmosphere (CA) has been used to alleviate IB in fruit. However, the mechanisms of CA on IB in peach remains unknown. This study investigated the effects of CA (3-3.5% Oxygen, 3-3.5% Carbon dioxide, and 93-94% nitrogen) treatment on IB development, sugar metabolism, and energy metabolism in cold-stored (1 ± 0.5 °C) peach. The CA treatment effectively inhibited the development of IB and markedly inhibited the reduction of sugar contents and energy charge. The protein expression of the V-type proton ATPase subunit was significantly inhibited by the CA treatment, accompanied by higher adenosine triphosphate (ATP) content, and energy charge than the control fruit. Notably, the expressions of the pyruvate kinase family of proteins, pyruvate decarboxylases, and sucrose synthase were induced by CA treatment that had complex protein interactions with the ATPase and the energy metabolism pathway. These results indicated that CA treatment enhanced the chilling tolerance attributed to maintaining higher levels of energy status and sugar contents by regulating the expression of key proteins involved in energy metabolism during cold storage and shelf life. Taken together, our study can provide theoretical support for the research and development of fresh-keeping and cold-chain logistics technology.

Peach allergen Pru p 1 content is generally low in fruit but with large variation in different varieties.

BACKGROUND: Pru p 1 is a major allergen in peach and nectarine, and the different content in varieties may affect the degree of allergic reactions. This study aimed to quantify Pru p 1 levels in representative peach varieties and select hypoallergenic Pru p 1 varieties. METHODS: To obtain monoclonal and polyclonal antibodies, mice and rabbits, respectively, were immunized with recombinant Pru p 1.01 and Pru p 1.02. The Pru p 1 levels in fruits from 83 representative peach varieties was quantified by sandwich enzyme-linked immunosorbent assay (sELISA). nPru p 1 was obtained through specific monoclonal antibody affinity purification and confirmed by Western blot and mass spectrometry. The variable Pru p 1 content of selected varieties was evaluated by Western blot and the expression level of encoding Pru p 1 genes by quantitative polymerase chain reaction. RESULTS: A sELISA method with monoclonal and polyclonal antibodies was built for quantifying Pru p 1 levels in peach. Pru p 1 was mainly concentrated in the peel (0.20-73.44 µg/g, fresh weight), being very low in the pulp (0.05-9.62 µg/g) and not detected in wild peach. For the 78 peach and nectarine varieties, Pru p 1 content varied widely from 0.12 to 6.45 µg/g in whole fruit. We verified that natural Pru p 1 is composed of 1.01 and 1.02 isoallergens, and the Pru p 1 expression level and Pru p 1 band intensity in the immunoblots were in agreement with protein quantity determined by ELISA for some tested varieties. In some cases, the reduced levels of Pru p 1 did not coincide with low Pru p 3 in the same variety in whole fruit, while some ancient wild peach and nectarines contained low levels of both allergens, and late-ripening yellow flesh varieties were usually highly allergenic. CONCLUSION: Pru p 1 content is generally low in peach compared to Pru p 3. Several hypoallergenic Pru p 1 and Pru p 3 varieties, "Zi Xue Tao," "Wu Yue Xian," and "May Fire," were identified, which could be useful in trials for peach allergy patients.

Single Nucleotide Polymorphism Detection for Peach Gummosis Disease Resistance by Genome-Wide Association Study.

Peach gummosis is one of the most widespread and destructive diseases. It causes growth stunting, yield loss, branch, trunk, and tree death, and is becoming a restrictive factor in healthy and sustainable development of peach production. Although a locus has been identified based on bi-parental quantitative trait locus (QTL) mapping, selection of gummosis-resistant cultivars remains challenging due to the lack of resistant parents and of the complexity of an inducing factor. In this study, an integrated approach of genome-wide association study (GWAS) and comparative transcriptome was used to elucidate the genetic architecture associated with the disease using 195 accessions and 145,456 genome-wide single nucleotide polymorphisms (SNPs). The broad-sense and narrow-sense heritabilities were estimated using 2-year phenotypic data and genotypic data, which gave high values of 70 and 73%, respectively. Evaluation of population structure by neighbor-joining and principal components analysis (PCA) clustered all accessions into three major groups and six subgroups, mainly according to fruit shape, hairy vs. glabrous fruit skin, pedigree, geographic origin, and domestication history. Five SNPs were found to be significantly associated with gummosis disease resistance, of which SNPrs285957, located on chromosome6 across 28 Mb, was detected by both the BLINK and the FarmCPU model. Six candidate genes flanked by or harboring the significant SNPs, previously implicated in biotic stress tolerance, were significantly associated with this resistance. Two highly resistant accessions were identified with low disease severity, which could be potential sources of resistance genes for breeding. Our results provide a fresh insight into the genetic control of peach gummosis disease.

A comparative UPLC-Q-TOF/MS-based metabolomics approach for distinguishing peach (Prunus persica (L.) Batsch) fruit cultivars with varying antioxidant activity.

Peaches (Prunus persica (L.) Batsch) are nutritionally and economically important and they are one of the most popular fruits consumed worldwide. Understanding metabolite-caused bioactivity differences among cultivars is essential for designing a peach with enhanced nutritional traits. In this study, we report an untargeted UPLC-Q-TOF/MS-based metabolomics approach for comprehensively discriminating between peaches with different antioxidant activities. Mature fruit from 40 peach cultivars were distinguished using principal component analysis and orthogonal partial least squares discrimination analysis. Seventeen differential metabolites were tentatively identified between peach cultivars with high and low antioxidant potency composite indices, and eight metabolites, including procyanidin C1, procyanidin trimer isomer 1, procyanidin trimer isomer 2, procyanidin B1, procyanidin B2, procyanidin B3, prunus inhibitor b, and phloridzin, were identified as marker compounds responsible for the discrimination of the cultivars base on potential antioxidant activity. Our study highlights the essence and predictive power of metabolomics for detecting small differences and for identifying potential marker metabolites based on their levels and composition in plants exhibiting varying bioactivities. Overall, the variations in the metabolites in peach pulp reflected the diversity in the peach germplasm, and these eight compounds are good candidate markers for future genetic breeding of peach fruit with enhanced antioxidant activity.

Comparison of Phytochemical Differences of the Pulp of Different Peach [Prunus persica (L.) Batsch] Cultivars with Alpha-Glucosidase Inhibitory Activity Variations in China Using UPLC-Q-TOF/MS.

In order to fully understand the variation of the fruit alpha-glucosidase inhibitory activity-related phytochemical basis in the Chinese peach [Prunus persica (L.) Batsch], mature fruit from 33 cultivars was used for the investigation of fruit phenolic phytochemical attributes, including total phenolics, flavonoids, anthocyanins, and procyanidins, as well as the alpha-glucosidase inhibitory activity in vitro. Alpha-glucosidase inhibitory activity varied significantly among tested peach cultivars and was strongly correlated with total phenolics, total procyanidins, and total flavonoids. Untargeted UPLC-Q-TOF/MS-based metabolomics were used to comprehensively discriminate between peaches with different inhibitory activity on alpha-glucosidase. Principal component analysis (PCA) and orthogonal partial least squares discrimination analysis (OPLS-DA) were used for this process. Twenty-three differential compounds were identified between peach cultivars with high and low alpha-glucosidase inhibitory activity, and nine, including procyanidin C1, procyanidin trimer isomer 1, procyanidin trimer isomer 2, procyanidin B1, procyanidin dimer, epicatechin-epicatechin-epicatechin, phloridzin, kaempferol 3-(2'',6''-di-(E)-p-coumarylglucoside), and luteolin 3'-methyl ether 7-malonylglucoside, were identified as marker compounds responsible for the discrimination. Overall, variations in metabolites in peach pulp reflect the diversity in peach germplasm, and these nine compounds are good candidate markers for future genetic breeding of peach fruit with high alpha-glucosidase inhibitory activity.

Auxin biosynthesis by the YUCCA6 flavin monooxygenase gene in woodland strawberry.

Auxin has been regarded as the main signal molecule coordinating the growth and ripening of fruits in strawberry, the reference genomic system for Rosaceae. The mechanisms regulating auxin biosynthesis in strawberry are largely elusive. Recently, we demonstrated that two YUCCA genes are involved in flower and fruit development in cultivated strawberry. Here, we show that the woodland strawberry (Fragaria vesca L.) genome harbors nine loci for YUCCA genes and eight of them encode functional proteins. Transcription pattern in different plant organs was different for all eight FvYUCs. Functionality of the FvYUC6 gene was studied in transgenic strawberry overexpressing FvYUC6, which showed typical high-auxin phenotypes. Overexpression of FvYUC6 also delayed flowering and led to complete male sterility in F. vesca. Additionally, specific repression of FvYUC6 expression by RNA interference significantly inhibited vegetative growth and reduced plant fertility. The development of leaves, roots, flowers, and fruits was greatly affected in FvYUC6-repressed plants. Expression of a subset of auxin-responsive genes was well correlated with the changes of FvYUC6 transcript levels and free indole-3-acetic acid levels in transgenic strawberry. These observations are consistent with an important role of FvYUC6 in auxin synthesis, and support a main role of the gene product in vegetative and reproductive development in woodland strawberry.

Genome-wide identification and comparative expression analysis of NBS-LRR-encoding genes upon Colletotrichum gloeosporioides infection in two ecotypes of Fragaria vesca.

Anthracnose caused by Colletotrichum spp. is one of the most destructive diseases of cultivated strawberry (Fragaria×ananassa Duchesne) worldwide. The correlation between NBS-LRR genes, the largest class of known resistance genes, and strawberry anthracnose resistance has been elusive. BLAST search in NCBI identified 94 FvNBSs in the diploid genome of strawberry Fragaria vesca, with 67 of the TIR-NBS-LRR type. At least 36 FvNBSs were expressed, with 25% being non-coding genes. Two F. vesca ecotypes, HLJ and YW, showed great variations in both morphological and physiological responses upon C. gloeosporioides infection. qRT-PCR revealed that 5 of the 12 leaf-expressed FvNBSs displaying opposite transcription responses to C. gloeosporioides infection in two ecotypes. These results showed that the transcriptional responses of several FvNBSs were involved in the ecotype-specific responses to C. gloeosporioides in F. vesca. These FvNBSs hold potential in characterizing molecular components and developing novel markers associated with anthracnose resistance in strawberry.

Physical changes and physiological characteristics of red and green peel during nectarine (cv. Hu018) maturation.

BACKGROUND: The aim of this study was to determine physical changes in nectarine and distinctive physiological characteristics related to red and green peel under stresses occurring during fruit maturation, information on which is currently not available. RESULTS: Fruit firmness increased from 4 to 6 weeks after blooming (WAB) then decreased from 6 WAB until ripening. Anthocyanins in red and green peel during nectarine maturation were identified by high-performance liquid chromatography as cyanidin 3-glucoside together and, at a much lower level, cyanidin 3-rutinoside. Cyanidins in red and green peel decreased from 4 to 8 WAB then increased from 8 to 12 WAB. Anthocyanin contents were positively correlated with PAL, POD, A*, MDA and O2(·-) values and inversely correlated with L* and B* values. Red and green peel during maturation could be separated by hierarchical cluster analysis of the tested data. CONCLUSION: This study has provided an overview of red and green peel characteristics during nectarine (cv. Hu018) maturation. Values of A*, anthocyanins, O2(·-), MDA, PAL, PPO and POD in red peel were higher than those in green peel, while values of L*, B* and chroma in red peel were lower than those in green peel throughout fruit maturation.