Phytohormone abscisic acid (ABA) induces anthocyanin biosynthesis; however, the underlying molecular mechanism is less known. In this study, we found that the apple MYB transcription factor MdMYB1 activated anthocyanin biosynthesis in response to ABA. Using a yeast screening technique, we isolated MdbZIP44, an ABA-induced bZIP transcription factor in apple, as a co-partner with MdMYB1. MdbZIP44 promoted anthocyanin accumulation in response to ABA by enhancing the binding of MdMYB1 to the promoters of downstream target genes. Furthermore, we identified MdBT2, a BTB protein, as an MdbZIP44-interacting protein. A series of molecular, biochemical, and genetic analysis suggested that MdBT2 degraded MdbZIP44 protein through the Ubiquitin-26S proteasome system, thus inhibiting MdbZIP44-modulated anthocyanin biosynthesis. Taken together, we reveal a novel working mechanism of MdbZIP44-mediated anthocyanin biosynthesis in response to ABA.
The basic leucine zipper (bZIP) transcription factor HY5 plays a multifaceted role in plant growth and development. Here the apple MdHY5 gene was cloned based on its homology with Arabidopsis HY5. Expression analysis demonstrated that MdHY5 transcription was induced by light and abscisic acid treatments. Electrophoretic mobility shift assays and transient expression assays subsequently showed that MdHY5 positively regulated both its own transcription and that of MdMYB10 by binding to E-box and G-box motifs, respectively. Furthermore, we obtained transgenic apple calli that overexpressed the MdHY5 gene, and apple calli coloration assays showed that MdHY5 promoted anthocyanin accumulation by regulating expression of the MdMYB10 gene and downstream anthocyanin biosynthesis genes. In addition, the transcript levels of a series of nitrate reductase genes and nitrate uptake genes in both wild-type and transgenic apple calli were detected. In association with increased nitrate reductase activities and nitrate contents, the results indicated that MdHY5 might be an important regulator in nutrient assimilation. Taken together, these results indicate that MdHY5 plays a vital role in anthocyanin accumulation and nitrate assimilation in apple.
Ethylene regulates climacteric fruit ripening, and EIN3-LIKE1 (EIL1) plays an important role in this process. In apple (), fruit coloration is accompanied by ethylene release during fruit ripening, but the molecular mechanism that underlies these two physiological processes is unknown. In this study, we found that ethylene treatment markedly induced fruit coloration as well as the expression of , a positive regulator of anthocyanin biosynthesis and fruit coloration. In addition, we found that MdEIL1 directly bound to the promoter of and transcriptionally activated its expression, which resulted in anthocyanin biosynthesis and fruit coloration. Furthermore, MdMYB1 interacted with the promoter of , a key regulator of ethylene biosynthesis, thereby providing a positive feedback for ethylene biosynthesis regulation. Overall, our findings provide insight into a mechanism involving the synergistic interaction of the ethylene signal with the MdMYB1 transcription factor to regulate ethylene biosynthesis and fruit coloration in apple.
SUMMARY
Drought stress induces anthocyanin biosynthesis in many plant species, but the underlying molecular mechanism remains unclear. Ethylene response factors (ERFs) play key roles in plant growth and various stress responses, including affecting anthocyanin biosynthesis. Here, we characterized an ERF protein, MdERF38, which is involved in drought stress‐induced anthocyanin biosynthesis. Biochemical and molecular analyses showed that MdERF38 interacted with MdMYB1, a positive modulator of anthocyanin biosynthesis, and facilitated the binding of MdMYB1 to its target genes. Therefore, MdERF38 promoted anthocyanin biosynthesis in response to drought stress. Furthermore, we found that MdBT2, a negative modulator of anthocyanin biosynthesis, decreased MdERF38‐promoted anthocyanin biosynthesis by accelerating the degradation of the MdERF38 protein. In summary, our data provide a mechanism for drought stress‐induced anthocyanin biosynthesis that involves dynamic modulation of MdERF38 at both transcriptional and post‐translational levels.
Cold stress severely affects plant growth and yield. C-repeat binding factors (CBFs) play important roles in the response to cold stress. In the present study, we identified an R2R3-MYB transcription factor (TF) MdMYB23 from apple (Malus × domestic) using transcriptome analyses, which was notably induced in response to cold stress. Transgenic apple calli and Arabidopsis with overexpression of MdMYB23 exhibited increased cold tolerance. Electrophoretic mobility shift assay (EMSA) and transient expression assays indicated that MdMYB23 directly bound to the promoters of MdCBF1 and MdCBF2 and activated their expression. MdMYB23 interacted with the promoter of MdANR, a key modulator of proanthocyanidin biosynthesis, and activated its expression to promote proanthocyanidin accumulation and reactive oxygen species (ROS) scavenging. MdBT2 was identified as an MdMYB23-interacting protein using yeast two-hybrid (Y2H), pull-down, and bimolecular fluorescence complementation (BiFC) assays. MdBT2 repressed cold tolerance and proanthocyanidin accumulation by promoting the degradation of MdMYB23 protein. Our findings shed light on the functions of MYB TFs and underlying mechanism in the modulation of plant cold tolerance.
Glucose induces anthocyanin accumulation in many plant species; however, the molecular mechanism involved in this process remains largely unknown. Here, we found that apple hexokinase MdHXK1, a glucose sensor, was involved in sensing exogenous glucose and regulating anthocyanin biosynthesis. In vitro and in vivo assays suggested that MdHXK1 interacted directly with and phosphorylated an anthocyanin-associated bHLH transcription factor (TF) MdbHLH3 at its Ser361 site in response to glucose. Furthermore, both the hexokinase_2 domain and signal peptide are crucial for the MdHXK1-mediated phosphorylation of MdbHLH3. Moreover, phosphorylation modification stabilized MdbHLH3 protein and enhanced its transcription of the anthocyanin biosynthesis genes, thereby increasing anthocyanin biosynthesis. Finally, a series of transgenic analyses in apple calli and fruits demonstrated that MdHXK1 controlled glucose-induced anthocyanin accumulation at least partially, if not completely, via regulating MdbHLH3. Overall, our findings provide new insights into the mechanism of the glucose sensor HXK1 modulation of anthocyanin accumulation, which occur by directly regulating the anthocyanin-related bHLH TFs in response to a glucose signal in plants.
Due to the exponentially increased demands of mobile data traffic, e.g., a 1000-fold increase in traffic demand from 4G to 5G, network densification is considered as a key mechanism in the evolution of cellular networks, and ultra-dense heterogeneous network (UDHN) is a promising technique to meet the requirements of explosive data traffic in 5G networks. In the UDHN, base station is brought closer and closer to users through densely deploying small cells, which would result in extremely high spectral efficiency and energy efficiency. In this article, we first present a potential network architecture for the UDHN, and then propose a generalized orthogonal/non-orthogonal random access scheme to improve the network efficiency while reducing the signaling overhead. Simulation results demonstrate the effectiveness of the proposed scheme. Finally, we present some of the key challenges of the UDHN.
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