The red coloring of pear fruits is mainly caused by anthocyanin accumulation.  Red sport, represented by the green pear cultivar ‘Bartlett’ (BL) and the red-skinned derivative ‘Max Red Bartlett’ (MRB), is an ideal material for studying the molecular mechanism of anthocyanin accumulation in pear.  Genetic analysis has previously revealed a quantitative trait locus (QTL) associated with red skin color in MRB.  However, the key gene in the QTL and the associated regulatory mechanism remain unknown.  In the present study, transcriptomic and methylomic analyses were performed using pear skin for comparisons between BL and MRB.  These analyses revealed differential PcHY5 DNA methylation levels between the two cultivars; MRB had lower PcHY5 methylation than BL during fruit development, and PcHY5 was more highly expressed in MRB than in BL.  These results indicated that PcHY5 is involved in the variations in skin color between BL and MRB.  We further used dual luciferase assays to verify that PcHY5 activates the promoters of the anthocyanin biosynthesis and transport genes PcUFGT, PcGST, PcMYB10 and PcMYB114, confirming that PcHY5 not only regulates anthocyanin biosynthesis but also anthocyanin transport.  Furthermore, we analyzed a key differentially methylated site between MRB and BL, and found that it was located in an intronic region of PcHY5.  The lower methylation levels in this PcHY5 intron in MRB were associated with red fruit color during development, whereas the higher methylation levels at the same site in BL were associated with green fruit color.  Based on the differential expression and methylation patterns in PcHY5 and gene functional verification, we hypothesize that PcHY5, which is regulated by methylation levels, affects anthocyanin biosynthesis and transport to cause the variations in skin color between BL and MRB.

In this study, we tested the ability of sodium dehydroacetate (SD) to extend the shelf-life of ‘Kyoho’ grape.  Among the different concentrations of SD tested (0, 0.01, 0.1 and 1.0 mmol L^–1), 0.01 mmol L^–1 SD was the most effective in prolonging the shelf-life of ‘Kyoho’ grape.  Compared with the control, the weight loss rate, browning index and hydrogen peroxide (H₂O₂) and malonaldehyde contents were significantly lower in the 0.01 mmol L^–1 SD treatment, whereas the healthy berry rate, berry firmness, total soluble solids (TSS) content, ascorbic acid content and superoxide dismutase (SOD) activity were significantly higher.  In addition, an analysis of ‘Kyoho’ grape DNA using methylation sensitive amplification polymorphism (MSAP) markers showed that the average DNA methylation level was significantly higher in the 0.01 mmol L^–1 SD treatment than in the control.  Together, these results indicate that 0.01 mmol L^–1 SD could be used to extend the shelf-life of ‘Kyoho’ grape.  Moreover, a strong connection between reactive oxygen species (ROS) metabolism and DNA methylation change during storage was revealed.

Grapevine growing areas are increasingly affected by drought, which has greatly limited global wine production and quality.  DEAD-box is one of the largest subfamilies of the RNA helicase family, and its members play key roles in the growth and development of plants and their stress responses.  Previous studies have shown the potential of DEAD-box genes in the drought stress responses of Arabidopsis and tomato, rice, and other crop species.  However, information about DEAD-box genes in grapevine remains limited.  In this report, a total of 40 DEAD-box genes were identified in grapevine and their protein sequence characteristics and gene structures were analyzed.  By comparing the expression profiles of VviDEADRHs in response to drought stress in different grapevine varieties, nine candidate genes (VviDEADRH10c, -13, -22, -25a, -25b, -33, -34, -36, and -39) were screened based on expression profiling data.  Combined with qRT-PCR results, VviDEADRH25a was selected for functional verification.  Heterologous overexpression of VviDEADRH25a in Arabidopsis showed the transgenic plants were more sensitive to drought stress than the control.  Both electrolyte permeability and malondialdehyde content were significantly increased in transgenic plants, whereas the chlorophyll content and superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and ascorbate peroxidase (APX) enzyme activities were significantly decreased.  Furthermore, VviDEADRH25a-overexpressing plants showed down-regulated expression levels of several drought stress-related marker genes, namely AtCOR15a, AtRD29A, AtERD15, and AtP5CS1, which indicated that they participated in the drought stress response.  In summary, this study provides new insights into the structure, evolution, and participation of DEAD-box RNA helicase genes in the response to drought stress in grapevines.