Flower development is central to angiosperm reproduction and is regulated by a broad range of endogenous and exogenous stimuli. It has been well documented that ambient temperature plays a key role in controlling flowering time; however, the mechanisms by which temperature regulates floral organ differentiation remain largely unknown. In this study, we show that low temperature treatment significantly increases petal number in rose (Rosa hybrida) through the promotion of stamen petaloidy. Quantitative RT-PCR analysis revealed that the expression pattern of RhAG, a rose homolog of the Arabidopsis thaliana AGAMOUS C-function gene, is associated with low temperature regulated flower development. Silencing of RhAG mimicked the impact of low temperature treatments on petal development by significantly increasing petal number through an increased production of petaloid stamens. In situ hybridization studies further revealed that low temperature restricts its spatial expression area. Analysis of DNA methylation level showed that low temperature treatment enhances the methylation level of the RhAG promoter, and a specific promoter region that was hypermethylated at CHH loci under low temperature conditions, was identified by bisulfite sequencing. This suggests that epigenetic DNA methylation contributes to the ambient temperature modulation of RhAG expression. Our results provide highlights in the role of RhAG gene in petal number determination and add a new layer of complexity in the regulation of floral organ development. We propose that RhAG plays an essential role in rose flower patterning by regulating petal development, and that low temperatures increase petal number, at least in part, by suppressing RhAG expression via enhancing DNA CHH hypermethylation of the RhAG promoter.

Gerbera hybrida is one of the most popular cut flowers in the world; however, stem bending, which always happens when gerbera flower harvested from the field, greatly limits its vase life. To date the molecular mechanisms underlying stem bending remain poorly understood. In this study, we performed high-throughput transcriptome sequencing of gerbera during stem bending using the Illumina sequencing technology. Three cDNA libraries constructed from mRNAs of gerbera stem at stem bending stage 0, 2 and 4 were sequenced. More than 300 million high-quality reads were generated and assembled into 96,492 unigenes. Among them, 34,166 unigenes were functionally annotated based on similarity search with known protein. Sequences derived from plants at different stem bending stages were mapped to the assembled transcriptome, and 9,406 differentially expressed genes (DEGs) were identified. Through Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, specific pathways were identified during the stem bending process, such as phenylpropanoid biosynthesis pathway, phenylalanine metabolism pathway, starch and sucrose metabolism pathway, and plant hormone signal transduction pathway. A total of 211 transcription factors (TFs), including TF families involved in plant senescence, such as NAC, MYB, WRKY, and AP2/ERF members, as well as TFs related to water stress tolerance, were shown to be regulated during stem bending. Gene Onotology (GO) functional enrichment analysis indicated that key genes involved in responses to osmotic and oxidative stresses were also varied in expression during this process. Furthermore, analysis of DEGs involved in the hormone signaling pathways and determination of endogenous abscisic acid (ABA) content showed that stem bending may be an ethylene-independent process, but regulated by ABA. In short, our findings suggested that the stem bending of cut gerbera may be caused by the involvement of water stress and regulation of ABA during the postharvest life. The transcriptome sequences provide a valuable resource in revealing the molecular mechanism underlying stem bending of cut flower and offer novel genes that can be used to guide future studies for ornamental plant breeding.

Flower opening is a process primarily caused by water uptake-driven petal cell expansion. while which is easily affected by water deficit during transportation of cut flowers, resulting in abnormal flower opening. The knowledge of important players during this process remains limited. We previously reported that the aquaporin RhPIP1;1 plays an important role in ethylene-regulated petal cell expansion in rose flower. Here, we identified RhRab5ip as a new interactor of RhPIP1;1. RhRab5ip belongs to the Rab5-interacting protein (Rab5ip) family and may function in vesicle trafficking pathway. By using split ubiquitin yeast two-hybrid (SUY2H) system, bimolecular fluorescence complementation (BiFC) and subcellular colocalization we confirmed the existence of physical interaction between RhPIP1;1 and RhRab5ip in yeast and plant cell. The interaction of these two proteins happened at the small punctate structures in the cytoplasm. Expression of RhRab5ip in petals increased substantially at the initial stage of flower opening and maintained at high level until flower wilting. The transcripts of both RhRab5ip and RhPIP1;1 were greatly up-regulated by ABA and GA₃ treatments, while only RhPIP1;1 was down-regulated by ethylene. Moreover, both RhRab5ip and RhPIP1;1 were significantly induced by water deficit treatment after 12 h-treatment, when flowers started to wilt and showed neck bending. Taken together, these findings suggested that RhRab5ip might functionally coordinate with RhPIP1;1 in response to water deficit stress in rose flower, expanding our understanding of the possible involvement of Rab5ip protein in the regulatory network of flower opening during water deficit.

Mediator is a multiprotein complex integral to the transcription machinery, mediated by RNA polymerase II. Some Mediator subunits have been found to have critical functions in plants’ responses to abiotic stresses. However, the role of plant Mediator subunits in drought responses remains largely enigmatic. Here, we identified a Mediator subunit, RhMED15a, in roses (Rosa hybrida). Its expression was greatly and swiftly induced by dehydration treatment in the root. The promoter sequence of RhMED15a contains cis-acting elements that respond to abscisic acid (ABA) and methyl jasmonate (MeJA). In addition, the expression of RhMED15a was significantly up-regulated with ABA treatment and inversely down-regulated with MeJA treatment. Silencing RhMED15a using virus-induced gene silencing (VIGS) in roses significantly reduced drought tolerance in rose plants. This resulted in a significant increase in the malondialdehyde (MDA) level and a decreased survival rate in comparison to TRV controls. Moreover, we found that the expression of five drought-related genes, including dehydration responsive element binding factor 1B (DREB1B), responsive to desiccation stress 29A (RD29A), responsive to desiccation stress 29B (RD29B), early response to dehydration 14 (ERD14), and 9-cis-epoxycarotenoid dioxygenase 1 (NCED1), was considerably suppressed in RhMED15a-silenced plants during drought stress. Taken together, our results present that the Mediator tail module subunit RhMED15a serves as an enhancer of drought tolerance in rose, probably through the modulation of the expression of some drought-related genes.

Background: Rose (Rosa hybrida) is a globally recognized ornamental plant whose growth and distribution are strongly limited by drought stress. The role of Mediator, a multiprotein complex crucial for RNA polymerase II-driven transcription, has been elucidated in drought stress responses in plants. However, its physiological function and regulatory mechanism in horticultural crop species remain elusive. Results: In this study, we identified a Tail module subunit of Mediator, RhMED15a-like, in rose. Drought stress, as well as treatment with methyl jasmonate (MeJA) and abscisic acid (ABA), significantly suppressed the transcript level of RhMED15a-like. Overexpressing RhMED15a-like markedly bolstered the osmotic stress tolerance of Arabidopsis, as evidenced by increased germination rate, root length, and fresh weight. In contrast, the silencing of RhMED15a-like through virus induced gene silencing in rose resulted in elevated malondialdehyde accumulation, exacerbated leaf wilting, reduced survival rate, and downregulated expression of drought-responsive genes during drought stress. Additionally, using RNA-seq, we identified 972 differentially expressed genes (DEGs) between tobacco rattle virus (TRV)-RhMED15a-like plants and TRV controls. Gene Ontology (GO) analysis revealed that some DEGs were predominantly associated with terms related to the oxidative stress response, such as ‘response to reactive oxygen species’ and ‘peroxisome’. Furthermore, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment highlighted pathways related to ‘plant hormone signal transduction’, in which the majority of DEGs in the jasmonate (JA) and ABA signalling pathways were induced in TRV-RhMED15a-like plants. Conclusion: Our findings underscore the pivotal role of the Mediator subunit RhMED15a-like in the ability of rose to withstand drought stress, probably by controlling the transcript levels of drought-responsive genes and signalling pathway elements of stress-related hormones, providing a solid foundation for future research into the molecular mechanisms underlying drought tolerance in rose.