WRKYs play important roles in plant growth, defense regulation, and stress response. However, the mechanisms through which WRKYs are involved in drought and salt tolerance have been rarely characterized in sweetpotato [Ipomoea batatas (L.) Lam.]. In this study, we cloned a WRKY gene, IbWRKY2, from sweetpotato and its expression was induced with PEG6000, NaCl, and abscisic acid (ABA). The IbWRKY2 was localized in the nucleus. The full-length protein exhibited transactivation activity, and its active domain was located in the N-terminal region. IbWRKY2-overexpressing Arabidopsis showed enhanced drought and salt tolerance. After drought and salt treatments, the contents of ABA and proline as well as the activity of superoxide dismutase (SOD) were higher in transgenic plants, while the malondialdehyde (MDA) and H₂O₂ contents were lower. In addition, several genes related to the ABA signaling pathway, proline biosynthesis, and the reactive oxygen species (ROS)-scavenging system, were significantly up-regulated in transgenic lines. These results demonstrate that IbWRKY2 confers drought and salt tolerance in Arabidopsis. Furthermore, IbWRKY2 was able to interact with IbVQ4, and the expression of IbVQ4 was induced by drought and salt treatments. These results provide clues regarding the mechanism by which IbWRKY2 contributes to the regulation of abiotic stress tolerance.

NAC (NAM, ATAF1,2, and CUC2) transcription factors (TFs) play critical roles in controlling plant growth, development, and abiotic stress responses. However, few studies have examined NAC proteins related to drought stress tolerance in rose (Rosa chinensis). Here, we identified a drought and abscisic acid (ABA)-induced NAC TF, RcNAC091, that localizes to the nucleus and has transcriptional activation activity. Virus-induced silencing of RcNAC091 resulted in decreased drought stress tolerance, and RcNAC091 overexpression had the opposite effect. Specifically, ABA mediated RcNAC091-regulated drought tolerance. A transcriptomic comparison showed altered expression of genes involved in ABA signaling and oxidase metabolism in RcNAC091-silenced plants. We further confirmed that RcNAC091 directly targets the promoter of RcWRKY71 in vivo and in vitro. Moreover, RcWRKY71-slienced rose plants were not sensitive to both ABA and drought stress, whereas RcWRKY71-overexpressing plants were hypersensitive to ABA, which resulted in drought-tolerant phenotypes. The expression of ABA biosynthesis- and signaling-related genes was impaired in RcWRKY71-slienced plants, suggesting that RcWRKY71 might facilitate the ABA-dependent pathway. Therefore, our results show that RcWRKY71 is transcriptionally activated by RcNAC091, which positively modulates ABA signaling and drought responses. The results of this study provide insights into the roles of TFs as functional links between RcNAC091 and RcWRKY71 in priming resistance; our findings also have implications for the approaches to enhance the drought resistance of roses.

Plant Myeloblastosis (MYB) proteins function crucially roles upon variegated abiotic stresses. Nonetheless, their effects and mechanisms in rose (Rosa chinensis) are not fully clarified. In this study, we characterized the effects of rose RcMYB8 under salt and drought tolerances. For induction of the RcMYB8 expression, NaCl and drought stress treatment were adopted. Rose plants overexpressing RcMYB8 displayed enhanced tolerance to salinity and drought stress, while silencing RcMYB8 resulted in decreased tolerance, as evidenced by lowered intra-leaf electrolyte leakage and callose deposition, as well as photosynthetic sustainment under stressed conditions. Here, we further show that RcMYB8 binds similarly to the promoters of RcPR5/1 and RcP5C51 in vivo and in vitro. Inhibiting RcP5CS1 by virus-induced gene silencing led to decreased drought tolerance through the reactive oxygen species (ROS) homeostatic regulation. RcP5CS1-silenced plants showed an increase in ion leakage and reduce of proline content, together with the content of malondialdehyde (MDA) increased, lowered activities of Catalase (CAT), peroxidase (POD) and superoxide dismutase (SOD). Our study highlights the transcriptional modulator role of RcMYB8 in drought and salinity tolerances, which bridges RcPR5/1 and RcP5CS1 by promoting ROS scavenging. Besides, it is probably applicable to the rose plant engineering for enhancing their abiotic stress tolerances. Graphical Abstract

DIRIGENT (DIR) genes are key players in environmental stress responses that have been identified in many vascular plant species. However, few studies have examined the VrDIR genes in mungbean. In this study, we characterized 37 VrDIR genes in mungbean using a genome-wide identification method. VrDIRs were distributed on seven of the 11 mungbean chromosomes, and chromosome three contained the most VrDIR genes, with seven members. Thirty-two of the 37 VrDIRs contained a typical DIR gene structure, with one exon; the conserved DIR domain (i.e., Pfam domain) occupied most of the protein in 33 of the 37 VrDIRs. The gene structures of VrDIR genes were analyzed, and a total of 19 distinct motifs were detected. VrDIR genes were classified into five groups based on their phylogenetic relationships, and 13 duplicated gene pairs were identified. In addition, a total of 92 cis-acting elements were detected in all 37 VrDIR promoter regions, and VrDIR genes contained different numbers and types of cis-acting elements. As a result, VrDIR genes showed distinct expression patterns in different tissues and in response to salt and drought stress.

The K⁺ transporter KT/HAK/KUP (K⁺ transporter/high-affinity K⁺/K⁺ uptake) family has a critical effect on K⁺ uptake and translocation in plants under different environmental conditions. However, the functional analysis of KT/HAK/KUP members in sweet potatoes is still limited. The present work reported the physiological activity of a new gene, IbHAK11, in the KT/HAK/KUP family in sweet potatoes. IbHAK11 expression increased significantly in the low K⁺-tolerant line compared with the low K⁺-sensitive line following treatment with low K⁺ concentrations. IbHAK11 upregulation promoted root growth in Arabidopsis under low K⁺ conditions. Under high saline stress, transgenic lines had superior growth and photosynthetic characteristics compared with the wild-type (WT). As for IbHAK11-overexpressing plants, activation of both the non-enzymatic and enzymatic reactive oxygen species (ROS) scavenging systems was observed. Therefore, IbHAK11-overexpressing plants had lower malondialdehyde (MDA) and ROS levels (including H₂O₂ and O²⁻) compared with WT under salt-induced stress. We also found that under both low K⁺ and high salinity conditions, overexpression of IbHAK11 enhanced K⁺ translocation from the root to the shoot and decreased Na⁺ absorption in Arabidopsis. Consequently, IbHAK11 positively regulated K⁺ deficiency and high salinity stresses by regulating K⁺ translocation and Na⁺ uptake, thus maintaining K⁺/Na⁺ homeostasis in plants.

Background: CIRCADIAN CLOCK ASSOCIATED 1 like (CCA1L) proteins are important components that participate in plant growth and development, and now have been characterized in multiple plant species. However, information on mungbeanCCA1Lgenes is limited.Results: In this study, we identified 27VrCCA1Lgenes from the mungbean genome.VrCCA1Lgenes were unevenly distributed on 10 of the 11 chromosomes and showed one tandem and two interchromosomal duplication events. Two distinct kinds of conserved MYB domains, MYB 1 and MYB 2, were found, and the conserved SHAQK(Y/F) F sequence was found at the C terminus of each MYB 2 domain. TheVrCCA1Lsdisplayed a variety of exon-intron organizations, and 24 distinct motifs were found among these genes. Based on phylogenetic analysis, VrCCA1L proteins were classified into five groups; group I contained the most members, with 11VrCCA1Ls.VrCCA1Lpromoters contained different types and numbers ofcis-acting elements, andVrCCA1Lsshowed different expression levels in different tissues. TheVrCCA1Lsalso displayed distinct expression patterns under different photoperiod conditions throughout the day in leaves.VrCCA1L26shared greatest homology toArabidopsis CCA1andLATE ELONGATED HYPOCOTYL(LHY).It delayed the flowering time inArabidopsisby affecting the expression levels ofCONSTANS(CO),FLOWERING LOCUS T(FT), andSUPPRESSOR OF OVEREXPRESSION OF CONSTANS 1(SOC1).Conclusion: We identified and characterized 27VrCCA1Lgenes from mungbean genome, and investigated their spatio-temporal expression patterns. Further analysis revealed thatVrCCA1L26delayed flowering time in transgenicArabidopsisplants. Our results provide useful information for further functional characterization of theVrCCA1Lgenes.