Glucose-6-phosphate dehydrogenase (G6PDH) is a key enzyme in the pentose phosphate pathway responsible for the generation of nicotinamide adenine dinucleotide phosphate (NADPH), thereby playing a central role in facilitating cellular responses to stress and maintaining redox homeostasis. This study aimed to characterize five G6PDH gene family members in maize. The classification of these ZmG6PDHs into plastidic and cytosolic isoforms was enabled by phylogenetic and transit peptide predictive analyses and confirmed by subcellular localization imaging analyses using maize mesophyll protoplasts. These ZmG6PDH genes exhibited distinctive expression patterns across tissues and developmental stages. Exposure to stressors, including cold, osmotic stress, salinity, and alkaline conditions, also significantly affected the expression and activity of the ZmG6PDHs, with particularly high expression of a cytosolic isoform (ZmG6PDH1) in response to cold stress and closely correlated with G6PDH enzymatic activity, suggesting that it may play a central role in shaping responses to cold conditions. CRISPR/Cas9-mediated knockout of ZmG6PDH1 on the B73 background led to enhanced cold stress sensitivity. Significant changes in the redox status of the NADPH, ascorbic acid (ASA), and glutathione (GSH) pools were observed after exposure of the zmg6pdh1 mutants to cold stress, with this disrupted redox balance contributing to increased production of reactive oxygen species and resultant cellular damage and death. Overall, these results highlight the importance of cytosolic ZmG6PDH1 in supporting maize resistance to cold stress, at least in part by producing NADPH that can be used by the ASA-GSH cycle to mitigate cold-induced oxidative damage.

Phlox subulata, a perennial herbaceous flower, can survive during the winter of northeast China, where the temperature can drop to −30 °C, suggesting that P. subulata is an ideal model for studying the molecular mechanisms of cold acclimation in plants. However, little is known about the gene expression profile of P. subulata under cold stress. Here, we examined changes in cold stress-related genes in P. subulata. We sequenced three cold-treated (CT) and control (CK) samples of P. subulata. After de novo assembly and quantitative assessment of the obtained reads, 99,174 unigenes were generated. Based on similarity searches with known proteins in public protein databases, 59,994 unigenes were functionally annotated. Among all differentially expressed genes (DEGs), 8302, 10,638 and 11,021 up-regulated genes and 9898, 17,876, and 12,358 down-regulated genes were identified after treatment at 4, 0, and −10 °C, respectively. Furthermore, 3417 up-regulated unigenes were expressed only in CT samples. Twenty major cold-related genes, including transcription factors, antioxidant enzymes, osmoregulation proteins, and Ca²⁺ and ABA signaling components, were identified, and their expression levels were estimated. Overall, this is the first transcriptome sequencing of this plant species under cold stress. Studies of DEGs involved in cold-related metabolic pathways may facilitate the discovery of cold-resistance genes.

The objective of the study was to identify suitable reference genes that can be used for quantitative real-time PCR (qPCR) analysis in mung bean (Vigna radiata). Therefore, 10 potential reference genes were selected and the results showed that ubiquitin-conjugating enzyme was suitable as reference under drought and pathogen infection stress; elongation factor 1-á was the most stable gene under waterlogging; and actin performed the best under saline stress. These selected reference genes were further confirmed by analysis of the expression profiles of catalase and peroxidase under waterlogging. Our results will contribute to the improvement of the accuracy of gene expression evaluation in mung bean.

Adzuki bean (Vigna angularis) rust, caused by the fungus Uromyces vignae, is an important disease affecting adzuki bean yield and quality. Previously, several NAC transcription factors (TFs) were induced by rust infection in a resistant adzuki bean variety, suggesting that NAC TF members may play important roles in rust resistance. To further explore the functions of NAC TFs in rust resistance and to provide a reference for resistant varietal breeding, 101 NAC TFs were identified from the adzuki bean genome. The synteny analysis revealed 25 pairs of VaNACs in the genome, which exhibited whole-genome/segmental duplication. Based on the phylogenetic relationships and conserved motif characteristics, the NAC TFs of V. angularis can be divided into 16 subfamilies. Previous transcriptome data showed that nine VaNACs are significantly induced by rust infection. Here, a cis-acting element analysis of these nine genes revealed that most contain hormone responsive elements, such as abscisic acid and methyl jasmonate (MeJA). The expression levels of these nine VaNACs were dynamically regulated in response to exogenous MeJA treatment, as revealed by quantitative real-time PCR analysis. Among them, seven VaNACs exhibited significantly upregulated expression, peaking at 12 h post treatment (hpt) and remaining significantly higher than that of the untreated control group for 48 hpt. These results suggest that these VaNACs are responsive to MeJA signaling and may play roles in the early and sustained transcriptional regulation of stress-related pathways. The exogenous MeJA decreased rust severity on adzuki bean leaves by 45.68%. Additionally, the expression levels of these nine genes in adzuki bean leaves in response to rust infection after pretreatment with MeJA were investigated. The expression of VaNAC002 rapidly peaked at 24 h post inoculation (hpi) and remained significantly higher than the control from 120 to 192 hpi. Subsequently, transient overexpression of VaNAC002 significantly enhanced the resistance of tobacco to Botrytis cinerea, indicating that VaNAC002 positively regulates plant disease resistance. These findings suggest that adzuki bean NAC family members may play important roles in disease resistance through JA signaling, with VaNAC002 having a positive regulatory role in plant immunity.