Yan Li

The WRKY transcription factors (TFs) are key regulators of plant responses to biotic and abiotic stresses. However, their roles in Amaranthus palmeri remain unexplored. In this study, 32 ApWRKYs were identified through bioinformatics and gene expression analyses. Subcellular localization predictions placed ApWRKYs in the nucleus, and transient expression assays of ApWRKY2 and ApWRKY5 confirmed nuclear targeting, supporting their role as transcriptional regulators. ApWRKYs are distributed across 15 genomic scaffolds, and phylogenetic analysis grouped them into three subfamilies, with conserved motifs identified within specific clades. Interaction analysis suggested potential post-transcriptional regulation by miRNAs. Gene expression profiling of ApWRKYs under glufosinate ammonium, NaCl, and PEG-induced osmotic stress treatments revealed potential distinct regulatory roles. Furthermore, transient overexpression in Arabidopsis thaliana found that ApWRKY1, ApWRKY2, and ApWRKY5 potentially regulate chlorophyll fluorescence and photosynthetic efficiency under glufosinate treatment. These findings establish ApWRKYs as central regulators of stress adaptation in A. palmeri, provide novel insights into WRKY-mediated regulation, and lay a foundation for future functional investigations aimed at enhancing stress resilience and herbicide management in horticultural systems.

Glyoxalase I (GLYI) is the key regulatory enzyme in the glyoxalase pathway. This pathway enables plants to neutralize methylglyoxal (MG) using glutathione (GSH), a mechanism significant for their acclimation to environmental stress. While functionally significant, the specific functions of GLYI genes in Amaranthus palmeri remain unexplored. In this study, integrated bioinformatics and expression analysis was used to identify five GLYI genes in A. palmeri. The results indicate that ApGLYI proteins are hydrophilic and slightly acidic, localized to scaffolds 1, 11, 13, and 16 of the A. palmeri genome. Phylogenetic analysis grouped ApGLYIs with other plant GLYI proteins into three distinct clades, each exhibiting conserved motif patterns. Expression analyses demonstrate that ApGLYI genes participate in both early and late regulatory phases of MG detoxification and signaling, responding to diverse stimuli including high temperature, NaCl, osmotic stress, exogenous methylglyoxal, abscisic acid (ABA), and methyl jasmonate (MeJA). Conversely, glufosinate ammonium treatment appears to compromise this cellular detoxification system. These results offer the evolutionary trajectory and functional significance of the ApGLYI gene. They establish a foundation for subsequent studies toward managing A. palmeri infestation and using these genes to improve stress resilience in cultivated crops through breeding strategies.

Sucrose non-fermenting-1-related protein kinases (SnRKs) have been implicated in plant growth and stress responses. Although SnRK3.23 is known to be involved in drought stress, the underlying mechanism of resistance differs between Arabidopsis and rice, and little is known about its function in wheat. In the current work, TaSnRK3.23B was detected on the cell membrane and in the nucleus. TaSnRK3.23B overexpression in Arabidopsis promoted reactive oxygen species (ROS) scavenging via the accumulation of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) and then conferred significant tolerance to drought. The prediction analysis, yeast two-hybrid, and bimolecular fluorescence complementation (BiFC) assays revealed that TaSnRK3.23B interacted with TaCBL2B and TaCBL6B, which are calcineurin B-like (CBL) proteins. The predicted model demonstrated that TaSnRK3 subfamily proteins participate in Ca2+ signaling mediated by TaCBL2B/TaCBL6B and subsequently provide drought stress tolerance by promoting ROS scavenging in wheat. Altogether, the obtained findings contribute to a better understanding of the functions of SnRK3.23 in wheat and offer genetic suggestions for improving drought resistance of wheat.

Aspartic proteases (APs), hydrolases with aspartic acid residues as catalytic active sites, are closely associated with processes such as plant growth and development and fungal and bacterial pathogenesis. F. graminearum is the dominant pathogenic fungus that causes Fusarium head blight (FHB) in wheat. However, the relationship of APs to the growth, development, and pathogenesis of F. graminearum is not clear. Therefore, we selected the FGSG_09558 gene, whose function annotation is aspartate protease, for further study. In this study, FGSG_09558 was found to contain a conserved structural domain and signal peptide sequence of aspartic acid protease and was therefore named FgAsp. The function of FgAsp in F. graminearum was investigated by constructing the knockout and complementation mutants of this gene. The results showed that with respect to the wild type (PH-1), the knockout mutant showed a significant reduction in mycelial growth, asexual spore production, and sexual spore formation, highlighting the key role of FgAsp in the growth and development of F. graminearum. In addition, the mutants showed a significant reduction in the virulence and accumulation level of deoxynivalenol (DON) content on maize whiskers, wheat germ sheaths, and wheat ears. DON, as a key factor of virulence, plays an important role in the F. graminearum infection of wheat ears, suggesting that FgAsp is involved in the regulation of F. graminearum pathogenicity by affecting the accumulation of the DON toxin. FgAsp had a significant effect on the ability of F. graminearum to utilize various sugars, especially arabinose. In response to the stress, hydrogen peroxide inhibited the growth of the mutant most significantly, indicating the important function of FgAsp in the strain’s response to environmental stress. Finally, FgAsp plays a key role in the regulation of F. graminearum growth and development, pathogenicity, and environmental stress response.