Alternative splicing of the PECTINESTERASE gene encoding a cell wall-degrading enzyme affects postharvest softening in grape

The firmness of table grape berries is a crucial quality parameter. Despite extensive research on postharvest fruit softening, its precise molecular mechanisms remain elusive. To enhance our comprehension of the underlying molecular factors, we initially identified differentially expressed genes(DEGs) by comparing the transcriptomes of folic acid(FA)-treated and water-treated(CK) berries at different time points. We then analyzed the sequences to detect alternatively spliced(AS) genes associated with postharvest softening. A total of 2,559 DEGs were identified and categorized into four subclusters based on their expression patterns, with subcluster-4 genes exhibiting higher expression in the CK group compared with the FA treatment group. There were 1,045 AS-associated genes specific to FA-treated berries and 1,042 in the CK-treated berries, respectively. Gene Ontology(GO) annotation indicated that the AS-associated genes in CK-treated berries were predominantly enriched in cell wall metabolic processes,particularly cell wall degradation processes. Through a comparison between treatment-associated AS genes and subcluster-4 DEGs, we identified eight genes, including Pectinesterase 2(VvPE2, Vitvi15g00704), which encodes a cell wall-degrading enzyme and was predicted to undergo an A3SS event. The reverse transcription polymerase chain reaction further confirmed the presence of a truncated transcript variant of VvPE2 in the FA-treated berries.Our study provides a comprehensive analysis of AS events in postharvest grape berries using transcriptome sequencing and underscores the pivotal role of VvPE2 during the postharvest storage of grape berries.

High-Throughput MicroRNA and mRNA Sequencing Reveals that MicroRNAs May Be Involved in Pectinesterase-Mediated Cold Resistance in Potato

Since potato cultivars are sensitive to low temperature,cold injury severely affects the geographical distribution and yield of potato.Although some miRNAs have been identified in response to cold stress in plants,there is no report about the role of miRNAs in the response to cold stress in potato.Here,via high throughput sequencing,we described the profiling of cold stress response to miRNA and mRNA in potato.Two small RNA and six mRNA libraries were constructed and sequenced.296 known and 211 novel miRNAs were identified,in which 34 miRNAs in Cold Group(CG)had the higher expression quantity than which in Normal Group(NG)and 32 in CG had lower expression quantity than which in NG.3068 differentially expressed genes were detected between NG and CG,in which 1400 genes were up-regulated and 1668 genes were down-regulated.The metabolism pathway of starch and sucrose(ko00500)is the common KEGG pathway in differentially expressed miRNA and mRNA.In this pathway,StuPME21575 and StuPME42971 are pectinesterase which mainly catalyzes the pectin-forming pectate,which are controlled by stu-miR6023 and stu-novel-miR42365.As the potato suffering cold stress,these two miRNAs expression levels became higher,but their target genes expression levels were just opposite and this result is the same with qRT-PCR.

Biochemical Changes of Cubiu Fruits (Solanum sessiliflorum Dunal, Solanaceae) According to Different Tissue Portions and Ripening Stages

Cubiu fruits (Solanum sessiliflorum Dunal, Solanaceae) are known for their high nutritional value and low caloric content. This work aimed at evaluating biochemical indices of cubiu fruits according to different tissues (peel, pulp and placenta) and ripening stages (green, turning, ripe and fully ripe). The fruits were randomly harvested to investigate sensory aspects (colorimetry, blanching effect, pigments) and biochemical indices (moisture and dry matter, Total Soluble Solids (TSS), pH, Titratable Acidity (TA), TSS/TA ratio, ascorbic acid, Alcohol-Insoluble Solids (AIS), pectinesterase activity and pectin content). The analyses were performed at the laboratory of Instituto Nacional de Pesquisas da Amazonia (INPA), Manaus, Brazil. The blanching process preserved pulp colors at all ripening stages. Chlorophylls were higher in green peels;flavonoids and carotenoids, in fully ripe peels. Anthocyanins were entirely absent. Pulp (turning fruits) showed the highest moisture content (91.05), followed by ripe pulp (90.70) and fully ripe pulp (90.62). Pulp TSS changed little whereas placenta TSS declined and were associated with increased TA and pH reduction (fully ripe fruits). Pulp pH and TA varied little whereas placenta pH was notably low and TA was remarkably high (fully ripe fruits). Pulp TSS/TA ratio showed predominance of TSS while in placenta, there was predominance of organic acids. AIS, precursors of pectin, were stable during ripening. The statistical analysis of dietary fiber content showed one modal value in the AIS. In pulp, pectinesterase activity correlated inversely with pectin content. Cubiu fruits were significant sources of bioactive compounds, e.g. chlorophylls, flavonoids and carotenoids, predominantly in the peel and the pulp;soluble functional fibers, e.g. pectin (g/100 g fresh weight), particularly in the green peel (1.00) and the fully ripe pulp (1.12), and other versatile molecules, e.g. ascorbic acid (mg/100 g fresh weight), especially in the fully ripe peel (32.45) and placenta (24.84) and the turning placenta (21.27). Cubiu fruits are rich in ascorbic acid and should be included in the human diet.