Lingxia Wang

Lycium barbarum L. possesses great salt tolerance and medicinal values, studying its salt tolerance contribute to variety improvement, as well as the increase in yield and quality. The study integrated the tandem mass tags (TMT) phosphoproteomics and physiological indexes of L. barbarum exposed to different concentrations of NaCl, with the aim of characterizing salt adaptation characteristics of L. barbarum. The findings indicated that a total of 2189 differentially phosphorylated peptides were identified, functional analysis revealed their involvement in glycolysis, plant hormone signal transduction, mitogen-activated protein kinase (MAPK) signal transduction and other pathways, and that the enzyme activities and substances related to glycolysis and signal transduction underwent significant changes under salt stress. Salt stress enhanced the glycolysis pathway through protein phosphorylation, and the changes in related enzymes activity accelerated the conversion of intermediate metabolites and energy supply. Salt stress led to the accumulation of abscisic acid (ABA), jasmonic acid (JA), and salicylic acid (SA) levels, triggering signal transduction events regulated by phosphorylated proteins to improve salt tolerance for L. barbarum in saline environments. The phosphorylation of MAPK signaling pathway-related proteins is triggered by reactive oxygen species (ROS) and ABA as signal molecules to induce the expression of downstream salt stress response factors. This study provides a foundation for further analysis of the molecular regulatory mechanism of protein phosphorylation in L. barbarum for salt stress.

Lysine malonylation (Kmal) is an evolutionarily conserved post-translational modification (PTM) that has been demonstrated to be involved in cellular and organismal metabolism. However, the role that Kmal plays in response to drought stress of the terrestrial cyanobacteria N. flagelliforme is still unknown. In this study, we performed the first proteomic analysis of Kmal in N. flagelliforme under different drought stresses using LC-MS/MS. In total, 421 malonylated lysine residues were found in 236 different proteins. GO and KEGG enrichment analysis indicated that these malonylated proteins were highly enriched in several metabolic pathways, including carbon metabolism and photosynthesis. Decreased malonylation levels were found to hinder the reception and transmission of light energy and CO₂ fixation, which led to a decrease in photosynthetic activity. Kmal was also shown to inhibit the flux of the TCA cycle and activate the gluconeogenesis pathway in response to drought stress. Furthermore, malonylated antioxidant enzymes and antioxidants were synergistically involved in reactive oxygen species (ROS) scavenging. Malonylation was involved in lipid degradation and amino acid biosynthesis as part of drought stress adaptation. This work represents the first comprehensive investigation of the role of malonylation in dehydrated N. flagelliforme, providing an important resource for understanding the drought tolerance mechanism of this organism.

Acetylation represents an extensively occurring protein post-translational modification (PTM) that plays a key role in many cellular physiological and biochemical processes. However, studies on PTMs such as acetylation of lysine (LysAc) in cyanobacteria are still rare. In this study, a quantitative LysAc approach (acetylome) on the strains of Nostoc flagelliforme subjected to different dehydration treatments was conducted. We observed that starch contents were significantly accumulated due to dehydration treatments, and we identified 2474 acetylpeptides and 1060 acetylproteins based on acetylome analysis. Furthermore, an integrative analysis was performed on acetylome and nontargeted metabolism, and the results showed that many KEGG terms were overlapped for both omics analyses, including starch and sucrose metabolism, transporter activity, and carbon metabolism. In addition, time series clustering was analyzed, and some proteins related to carbon metabolism and the ROS scavenging system were significantly enriched in the list of differentially abundant acetylproteins (DAAPs). These protein expression levels were further tested by qPCR. A working model was finally proposed to show the biological roles of protein acetylation from carbon metabolism and the ROS scavenging system in response to dehydration in N. flagelliforme. We highlighted that LysAc was essential for the regulation of key metabolic enzymes in the dehydration stress response.