Heat stress (HS), caused by extremely high temperatures, is one of the most severe forms of abiotic stress in pepper. In the present study, we studied the transcriptome and metabolome of a heat-tolerant cultivar (17CL30) and a heat-sensitive cultivar (05S180) under HS. Briefly, we identified 5754 and 5756 differentially expressed genes (DEGs) in 17CL30 and 05S180, respectively. Moreover, we also identified 94 and 108 differentially accumulated metabolites (DAMs) in 17CL30 and 05S180, respectively. Interestingly, there were many common HS-responsive genes (approximately 30%) in both pepper cultivars, despite the expression patterns of these HS-responsive genes being different in both cultivars. Notably, the expression changes of the most common HS-responsive genes were typically much more significant in 17CL30, which might explain why 17CL30 was more heat tolerant. Similar results were also obtained from metabolome data, especially amino acids, organic acids, flavonoids, and sugars. The changes in numerous genes and metabolites emphasized the complex response mechanisms involved in HS in pepper. Collectively, our study suggested that the glutathione metabolic pathway played a critical role in pepper response to HS and the higher accumulation ability of related genes and metabolites might be one of the primary reasons contributing to the heat resistance.

Salinity is a significant limiting factor that adversely affects plant growth, distribution and crop yield. Ethylene responsive factors play crucial roles in plant responses to and tolerance of various abiotic stresses. Recently, we revealed that OsERF2 is involved in root growth by transcriptionally regulating hormone and sugar signaling in rice. Here, we report that OsERF2 is a direct target gene of OsEIL1 and negatively regulates salt tolerance in rice. Compared to the wild type, the gain-of-function mutant of OsERF2 (nsf2857) and the knockdown of OsERF2 via an artificial microRNA (Ami-ERF2) exhibited decreased and increased salt tolerance, respectively. The enhanced salt tolerance observed in Ami-OsERF2 lines was associated with lower accumulations of malondialdehyde and reactive oxygen species (ROS) under salt stress, while the opposite was true for nsf2857 plants, which exhibited decreased salt tolerance. At the transcriptional level, several stress-related genes encoding ROS and NAD(P)H-related oxidoreductases were downregulated in nsf2857 plants but upregulated in Ami-ERF2 plants. Furthermore, yeast one-hybrid and ChIP assays revealed that OsEIL1 can bind to the of EBS cis element present in the promoter of OsERF2 (-bp), suggesting that OsEIL1 may directly regulate the expression of OsERF2. Collectively, our findings indicate that OsERF2 is a direct downstream factor involved in the regulation of salt tolerance in rice, highlighting its potential application in the genetic improvement of tolerance to abiotic stresses in this crop.

Limited knowledge is available for phosphorylation modifications in pepper (Capsicum annuum L.), especially in pepper fruit development. In this study, we conducted the first comprehensive phosphoproteomic analysis of pepper fruit at four development stage by Tandem Mass Tag proteomic approaches. A total of 2639 unique phosphopeptides spanning 1566 proteins with 4150 nonredundant sites of phosphorylation were identified, among which 2327 peptides in 1413 proteins were accurately quantified at four different stages. Mature Green (MG) to breaker stage showed the largest number of differentially expressed phosphoproteins and the number of downregulated phosphoproteins was significantly higher than that of upregulated after MG stage. Twenty seven phosphorylation motifs, including 22 pSer motifs and five pThr motifs and 85 kinase including 28 serine/threonine kinases, 14 receptor protein kinases, six mitogen-activated protein kinases, seven calcium-dependent protein kinases, two casein kinases, and some other kinases were quantified. Then the dynamic changes of phosphorylated proteins in ethylene and abscisic acid signaling transduction pathways during fruit development were analyzed. Our results provide a cascade of phosphoproteins and a regulatory network of phosphorylation signals, which help to further understand the mechanism of phosphorylation in pepper fruit development.

As the only harvest organ of pepper, fruit size is an important yield determinant. To elucidate the molecular mechanisms underlying pepper fruit size, we performed histological, physiological, and transcriptomic analyses on the pepper varieties QB6 (large fruit) and CXJ82 (small fruit). High contents of auxin and cytokinin in the early stage of fruit development promoted the rapid division of fruit cells in both varieties, which provided sufficient cells for subsequent fruit enlargement. High gibberellin accumulation induced the elongation and expansion of QB6 pericarp cells. Transcriptome analysis showed that genes related to cell division, cell wall polysaccharide degradation, and photosynthesis were highly expressed in QB6 fruit, likely contributing to its larger size. In the hormone–signal transduction factor–gene interaction network, GID6, GID1, IAA12, MYC30, and SAUR36 exhibited high correlations with numerous genes related to cell division, the cell wall, and photosynthesis, emerging as key signal transduction factors for the hormone-mediated regulation of pepper fruit size. Weighted gene co-expression network analysis identified the transcription factors OFP20, HD-ZIP6, and HD-ZIP13 as fundamental for pepper fruit size regulation. Our results expand the understanding of hormone regulation of pepper fruit size, providing a foundation for the breeding and improvement of excellent pepper varieties.