The papain-like cysteine proteases (PLCPs) is a subfamily of cysteine proteases that plays an important role in leaf senescence, and some of its members are involved in the regulation of plant growth and development under stress. In this study, we cloned a new gene, ZmSAG39, from maize. Expression profile analysis showed that ZmSAG39 was induced by darkness and drought treatments. In addition, the ZmSAG39 overexpression in maize accelerated the senescence of maize leaves under darkness and drought treatments. However, the knockout of ZmSAG39 in maize enhanced the resistance of maize to darkness and drought stresses and reduced the degree of senescence of maize leaves. Under drought stress, compared with WT plants, the knockout lines had a higher seed germination rate, seedling survival rate and chlorophyll content, and lower reactive oxygen species (ROS) level and malondialdehyde (MDA) content. In addition, quantitative real-time PCR (qRT-PCR) analysis showed that ZmSAG39 negatively regulated some stress-related genes but positively regulated senescence-related genes under darkness and drought stress conditions. To summarize, these results indicate that ZmSAG39 is a senescence-related gene and plays a negative role in response to darkness and drought stresses. This study laid a theoretical foundation for the innovation of maize germplasm resources with high quality, high yield and strong stress resistance.

Flowering is an essential process in plant development, and there are six major flowering pathways: the photoperiodic pathway, gibberellin pathway, vernalization pathway, age pathway, autonomous pathway, and temperature pathway. In this study, we screened the transcriptome sequencing of early flowering mutants from the laboratory for the significantly differentially expressed ZmGRAS46, which belongs to the DELLA subfamily of the GRAS family. DELLA is involved in the gibberellin pathway to regulate plant flowering. However, it is not clear whether ZmGRAS46 is involved in the gibberellin pathway which regulates plant flowering; therefore, in this experiment, we investigated the regulatory role of this gene in Arabidopsis flowering by overexpressing ZmGRAS46. It was found that overexpression of ZmGRAS46 in Arabidopsis promotes the formation of rosette leaves and flower buds and delays flowering time in Arabidopsis, and experiments have shown that ZmGRAS46 represses the expression of FLOWERING LOCUS T (FT), SUPPRESSOR OF CONSTANS1 (SOC1), CONSTANS (CO), and LEAFY (LFY). Our results indicated the possibility that ZmGRAS46 represses flowering through the CO-FT-SOC1-mediated photoperiodic flowering pathway. The delayed flowering phenotype of overexpressing ZmGRAS46 Arabidopsis could be rescued by applying GA3. The experimental results indicate that ZmGRAS46 depends on the GA3 pathway to regulate flowering in Arabidopsis.

Auxin response factors(ARFs) are a class of transcription factors that regulate the expression of auxin response genes and play a crucial role in plant growth and development. Florigen plays a crucial role in the process of flowering. However, the process by which auxin regulates the accumulation of florigen remains largely unclear. This study found that the expression of ZmARF16 in maize increases during flowering, and the genetic transformation of ZmARF16 accelerates the flowering process in Arabidopsis and maize. Furthermore, ZmARF16 was found to be positively correlated with the transcription of the ZCN12 gene. Similarly, the FT-like gene ZCN12 in maize rescues the late flowering phenotype of the FT mutation in Arabidopsis. Moreover, ZCN12 actively participates in the accumulation of florigen and the flowering process. Further research revealed that ZmARF16 positively responds to the auxin signal, and that the interaction between ZmARF16 and the ZCN12 promoter, as well as the subsequent promotion of ZCN12 gene expression, leads to early flowering. This was confirmed through a yeast one-hybrid and dual-luciferase assay. Therefore, the study provides evidence that the ZmARF16-ZCN12 module plays a crucial role in regulating the flowering process of maize.