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Low temperature stress is an important abiotic stress for garden roses in northern regions. Two garden rose cultivars ('Dagmar Hastrup' and 'Chandos Beauty') were selected to study the role of dehydrin and of carbohydrate metabolism during cold acclimation and deacclimation under the controlled daylength and temperature. The presence of bud dormancy was also observed as this could prevent budburst during warm spells. Both cultivars showed a similar changing pattern of cold acclimation and deacclimation and did not differ in their lowest LT 50 values. Dehydrin (RhDHN5) was upregulated by low temperatures and not by dehydration stress as the stem water content remained stable during the treatments. Total soluble sugars accumulated with a transient up-regulation of RhBAM3 (a key gene in starch hydrolysis) for 'Dagmar Hastrup' at 2 • C and a strong expression under both 2 and −3 • C for 'Chandos Beauty'. At 2 and −3 • C, raffinose and stachyose strongly accumulated though the up-regulation of RhRS6 and RhGK differed in the cultivars. Although similar cold hardiness levels were reached, carbohydrate metabolism in response to cold stress is different in the two cultivars. Increasing the temperature after a cold period resulted in fast deacclimation as found by the downregulation of RhDHN5 and RhBAM3, the decrease of raffinose and stachyose. Bud endodormancy was hardly present in both cultivars.

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Basic helix-loop-helix (bHLH) proteins play pivotal roles in plant growth, development, and stress responses. However, the molecular and functional properties of bHLHs have not been fully characterized. In this study, a novel XI subgroup of the bHLH protein gene, RcbHLH59, was isolated and identified in rose (Rosa spp.). This gene was induced by salinity stress in both rose leaves and roots, and functioned as a transactivator. Accordingly, silencing RcbHLH59 affected the antioxidant system, Na+/K+ balance, and photosynthetic system, thereby reducing salt tolerance, while the transient overexpression of RcbHLH59 improved salinity stress tolerance. Additionally, RcbLHLH59 was found to regulate the expression of sets of pathogenesis-related (PR) genes in RcbHLH59-silenced (TRV-RcbHLH59) and RcbHLH59-overexpressing (RcbHLH59-OE) rose plants. The RcPR4/1 and RcPR5/1 transcript levels showed opposite changes in the TRV-RcbHLH59 and RcbHLH59-OE lines, respectively, suggesting that these two genes are regulated by RcbHLH59. Further analysis revealed that RcbHLH59 binds to the promoters of RcPR4/1 and RcPR5/1, and that the silencing of RcPR4/1 or RcPR5/1 led to decreased tolerance to salinity stress. Moreover, callose degradation- and deposition-related genes were impaired in RcPR4/1 or RcPR5/1 silenced plants, which displayed a salt tolerance phenotype by balancing the Na+/K+ ratio through callose deposition. Collectively, our data highlight a new RcbLHLH59-RcPRs module that positively regulates salinity stress tolerance by balancing Na+/K+ and through callose deposition in rose plants.