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Maize (Zea mays) is an important cereal crop worldwide. However, its yield and quality are adversely affected by salt stress resulting from soil hypersalinity. Exploring the regulatory mechanisms of stress responses is of vital importance to increase maize seed production. In the present study, we screened ethyl methanesulfonate-induced maize mutants and identified a salt-tolerant mutant. A single base was mutated in ZmWRKY20, leading to the formation of a truncated protein variant. A detailed phenotypic analysis revealed that this mutant had significantly higher resistance to wilting and lower reactive oxygen species levels than the inbred line B73. ZmWRKY20 showed transcriptional activity in yeast and specifically bound W-boxes according to the results of our yeast one-hybrid, electrophoretic mobility shift, and dual-luciferase assays. Overexpression of ZmWRKY20 decreased salt tolerance in maize. Transcriptome profiling revealed that ZmWRKY20 overexpression extensively reprogrammed genes involved in regulating defense and oxidation-reduction responses. The results substantiate that ZmWRKY20 is directly targeted to the basic leucine zipper (bZIP) motif in the transcription factor ZmbZIP111. It was also verified that ZmWRKY20 interacts with ZmWRKY115 and both proteins act jointly to enhance ZmbZIP111 repression. The results indicate that the ZmWRKY20 and ZmWRKY115 transcription factors interact in the nucleus, leading to repression of ZmbZIP111 expression by directly binding its promoter, and increase the sensitivity of maize seedlings to salt stress. The current study improves our understanding of the complicated responses of maize to salt stress., (© 2022 Society for Experimental Biology and John Wiley & Sons Ltd.)

WRKYs represent an important family of transcription factors that are widely involved in plant development, defense regulation and stress response. Transgenic rice that constitutively expressed ZmWRKY114 had shorter plant height and showed less sensitivity to gibberellic acid (GA 3 ). Further investigation proved that transgenic rice accumulated lower levels of bioactive GAs than that in wild-type plants. Application of exogenous GA 3 fully rescued the semi-dwarf phenotype of ZmWRKY114 transgenic plants. Transcriptome and qRT-PCR analyses indicated that the expression of OsGA2ox4 , encoding the repressor of GA biosynthesis, was markedly increased. Electrophoretic mobility shift assay and dual-luciferase reporter assay indicated that ZmWRKY114 directly binds to a W-box motif in the OsGA2ox4 promoter. Taken together, these results confirm that ZmWRKY114 is a GA-responsive gene and is participated in the regulation of plant height in rice.

As one of the largest families of transcription factors in plants, the WRKY proteins play crucial roles in plant growth and development, defense regulation and stress responses. In this research, ZmWRKY86 encoding a WRKY transcription factor was cloned from maize (Zea mays L.). ZmWRKY86 expression was up-regulated by salt stress. ZmWRKY86 is a nuclear protein and has no transcriptional activation ability in yeast. ZmWRKY86 can specifically bind to W-box (TTGACC), which was confirmed by electrophoretic mobility shift assay (EMSA) and dual-LUC experiments. As compared with control, the wrky86 mutants showed enhanced plant tolerance to salt stress with higher survival rate, catalase activity and K + content, but lower malondialdehyde accumulation, relative electrical leakage level and Na + content under salt-stress condition. Transcriptome and quantitative real-time PCR analyses indicated that the mutation of ZmWRKY86 led to significant changes in the expression of stress-related genes in maize. Further assays showed that ZmWRKY86 directly interacted with the promoter of two salt stress-related genes Zm00001d020840 and Zm00001d046813. In summary, we identified a WRKY transcription factor ZmWRKY86 that participates in salt stress regulation through controlling the expression of stress-related genes., (Copyright © 2021 Elsevier Masson SAS. All rights reserved.)