A new graphene-based electrochemical biosensor was constructed and applied for the simple, rapid and highly selective determination of glucose. The modified glassy carbon electrode (GCE) was prepared using dopamine (DA) as a reducing agent for graphene oxide (GO) and as a capping agent to stabilize and coat the resulting reduced graphene oxide (RGO) surface as it can polymerize to form polydopamine (PDA). PDA-RGO composites were dropped onto a glassy carbon electrode surface, and glucose oxidase (GOx) was immobilized on the surface of the composites by Michael addition. Finally, Pt nanoparticles (PtNPs) were electrodeposited on the modified electrode. Such modified electrodes (denoted as GCE/PDA-RGO/GOx/PtNPs) were characterized by UV-visible spectroscopy, atomic force microscopy, X-ray power diffractometry, Fourier-transform infrared spectroscopy, and scanning electron microscopy. Electrochemical behaviors were investigated by cyclic voltammetry and electrochemical impedance spectroscopy. The fabricated biosensor exhibited a high sensitivity of 33 μA mM⁻¹ cm⁻² over a wide linear range of 0.2 mM to 1 mM, good stability, excellent repeatability, and a detection limit of 0.10 mM.

To date, there has been no high-quality sequence for genomes of the East Asian grape species, hindering biological and breeding efforts to improve grape cultivars. This study presents ~522 Mb of the Vitis amurensis (Va) genome sequence containing 27 635 coding genes. Phylogenetic analysis indicated that Vitis riparia (Vr) may have first split from the other two species, Va and Vitis vinifera (Vv). Divergent numbers of duplicated genes reserved among grapes suggests that the core eudicot-common hexaploidy (ECH) and the subsequent genome instability still play a non-negligible role in species divergence and biological innovation. Prominent accumulation of sequence variants might have improved cold resistance in Va, resulting in a more robust network of regulatory cold resistance genes, explaining why it is extremely cold-tolerant compared with Vv and Vr. In contrast, Va has preserved many fewer nucleotide binding site (NBS) disease resistance genes than the other grapes. Notably, multi-omics analysis identified one trans-cinnamate 4-monooxygenase gene positively correlated to the resveratrol accumulated during Va berry development. A selective sweep analysis revealed a hypothetical Va sex-determination region (SDR). Besides, a PPR-containing protein-coding gene in the hypothetical SDR may be related to sex determination in Va. The content and arrangement order of genes in the putative SDR of female Va were similar to those of female Vv. However, the putative SDR of female Va has lost one flavin-containing monooxygenase (FMO) gene and contains one extra protein-coding gene uncharacterized so far. These findings will improve the understanding of Vitis biology and contribute to the improvement of grape breeding.

In plant cells, multiple paralogs from ribosomal protein (RP) families are always synchronously expressed, which is likely contributing to ribosome heterogeneity or functional specialization. However, previous studies have shown that most RP mutants share common phenotypes. Consequently, it is difficult to distinguish whether the phenotypes of the mutants have resulted from the loss of specific genes or a global ribosome deficiency. Here, to investigate the role of a specific RP gene, we employed a gene overexpression strategy. We found that Arabidopsis lines overexpressing RPL16D (L16D-OEs) display short and curled rosette leaves. Microscopic observations reveal that both the cell size and cell arrangement are affected in L16D-OEs. The severity of the defect is positively correlated with RPL16D dosage. By combining transcriptomic and proteomic profiling, we found that overexpressing RPL16D decreases the expression of genes involved in plant growth, but increases the expression of genes involved in immune response. Overall, our results suggest that RPL16D is involved in the balance between plant growth and immune response.

Grapevine (Vitis vinifera) is an important and popular perennial fruit tree cultivated worldwide. Grapevine ripening is affected by flowering time, and although members of the MADS-box protein family play vital roles in regulating flowering in plants, the functions of MADS-box proteins in grapevine remain largely unknown. AGAMOUS-LIKE 11 (VvAGL11), a MADS-box gene in grapevine, was reported to be a regulator of seed morphogenesis. In this study, heterologous overexpression of VvAGL11 was found to significantly promote flowering in Arabidopsis, suggesting that its active expression in grapevine may induce early flowering and ripening. Transcriptome analysis showed that VvAGL11 overexpression affected the expression of genes involved in stress responses, hormonal signaling responses, and flowering regulation. Notably, VvAGL11 significantly increased the expression of key flowering genes such as FLOWERING LOCUS T (FT), APETALA3 (AP3), and SQUAMOSA PROMOTER BINDING PROTEIN-LIKE 5 (SPL5), which might have contributed to the early flowering in Arabidopsis. In summary, we characterized a novel flowering regulator, VvAGL11, which could be a potential target for early ripening breeding in grapevine.

Soil salinity severely restricts grapevine growth and development. Here, we integrated whole-genome bisulfite sequencing (WGBS) and RNA sequencing (RNA-seq) to investigate DNA methylation changes and their relationship with gene expression under salt stress in Vitis vinifera L. ‘Shine Muscat’ grapevine. Salt stress altered genome-wide DNA methylation patterns, reducing methylation levels in CG, CHG, and CHH contexts following NaCl treatment. We identified 8606 differentially methylated regions (DMRs) and 3106 DMR-associated genes (DMGs) under salt stress. RNA-seq analysis revealed 2691 differentially expressed genes (DEGs), including multiple stress-related transcription factors (e.g., MYB, NAC, WRKY) and hormone-related genes strongly induced by salinity. Integrative analysis identified 171 genes that were both differentially methylated and differentially expressed, primarily enriched in metabolic pathways, fructose and mannose metabolism, and fatty acid biosynthesis. Notably, several key stress-responsive genes (e.g., VvNAC72, VvBAK1, VvMYBS3) showed coordinated changes between methylation status and transcript abundance. Collectively, this study provides a comprehensive integrative analysis of DNA methylation and transcriptome reprogramming in ‘Shine Muscat’ grapevine under salt stress, revealing potential epigenetic mechanisms involved in transcriptional regulation and salt adaptation. The identified candidate genes provide valuable targets for further functional validation and genetic improvement of grapevine salt tolerance.

Soil salinity severely limits grapevine yield and fruit quality; however, the mechanisms underlying salt tolerance remain poorly understood. Small secreted peptides, particularly rapid alkalinization factor (RALF) peptides, are key regulators of plant responses to environmental stress. However, a systematic analysis of the RALF gene family in grapevine (Vitis vinifera L.) has not been conducted. In this study, we identified 11 VvRALFs in the grapevine genome (PN_T2T) and characterized their physicochemical properties, structural features, and evolutionary relationships. Expression profiling revealed that VvRALF9 was consistently upregulated in the roots of the salt-sensitive rootstock ‘Beta’ but downregulated in the salt-tolerant rootstock ‘3309C’. Silencing VvRALF9 in the roots of grapevine rootstocks significantly enhanced their salt tolerance. Protein interaction assays, including yeast two-hybrid, split-LUC, and pull-down assays, identified VvFER2 as a potential receptor of VvRALF9. Overexpression of VvFER2 in the roots of grapevine rootstocks also improved salt tolerance. Transcriptomic analysis of VvRALF9-RNAi roots revealed significant enrichment of the plant hormone signal transduction pathway, together with extensive changes in the expression of transcription factors and genes associated with multiple hormone signaling pathways. Together, these findings suggest that the VvRALF9–VvFER2 module is involved in the regulation of grapevine salt stress responses.