iTRAQ proteome analysis reveals the underling mechanisms of foliage zinc-spraying to improve photosynthetic capacity and seed yields of Peaonia ostii 'Fengdan'.

Zinc (Zn) deficiency is a significant nutritional limitation to crop yield globally, particularly in calcareous soil environments. Tree peony of Peaonia ostii 'Fengdan' is regarded as an oil crop due to its seeds rich in alpha-linolenic acid, a beneficial compound for health promotion. However, low seed yield remains a primary challenge in attaining sufficient seed oil from tree peony. In this study, Zn fertilization was applied to soil or foliage of P. ostii 'Fengdan' in the growth period before fruit development. Our findings reveal that foliar Zn-spraying, as opposed to soil application, proves to be a more effective method for augmenting seed yield, Zn accumulation and photosynthetic capacity in 'Fengdan'. Comparative analyses of the leaf proteome of 'Fengdan' using iTRAQ profiling under foliar Zn-spraying identified 115 differentially expressed proteins (DEPs), including 36 upregulated proteins, which likely contribute to the observed increase in seed yields of 'Fengdan' caused by foliage Zn-spraying. Specifically, Zn2+ stimulation of phosphatidylinositol signaling initiates a cascade of metabolic regulations. Firstly, ATP synthesis promotes leaf photosynthetic capacity, facilitated by improved sucrose metabolism through upregulated pullulanase and 1,4-alpha-glucan-branching enzyme. Furthermore, lipid synthesis and transport are facilitated by upregulated lipoyl synthase and plastid lipid-associated proteins. Additionally, DEPs involved in secondary metabolism are upregulated in the production of various metabolites conducive to 'Fengdan' growth. Overall, our results demonstrate that foliage Zn-spraying enhances seed yield in P. ostii 'Fengdan' by elevating Zn content and secondary metabolite synthesis in leaves, thereby augmenting leaf photosynthetic capacity and lipid synthesis. This study provides an effective way to increase seed yield of tree peony by exogenous Zn application.

Effect of polysaccharide extract SPSS1 from Apostichopus japonicas spermary on HepG2 cells via iTRAQ-based proteome analysis.

In this study, polysaccharide extract was prepared from Apostichopus japonicus spermary and purified by ion-exchange chromatography and gel filtration chromatography. Two main fractions named SPSS1 and SPSS2 were obtained and analyzed by ultraviolet spectroscopy and mixed with KBr, respectively. Chemical components analysis proved that SPSS1 and SPSS2 were rich in sulfate. Monosaccharide analysis indicated that in addition to the high content of lactose in both kinds of polysaccharides, the highest content of monosaccharide in SPSS1 was galactose, while in SPSS2 it was fucose. Further, the antitumor study of SPSS1 was carried and the results showed that SPSS1 treatment inhibited the proliferation of HepG2 cells. Through the iTRAQ-based proteome analysis, there were 208 differential proteins between control tumor cells and SPSS1 treatment of tumor cells. Compared to control tumor cells, 135 proteins were upregulated and 73 proteins were downregulated in treatment tumor cells. PRACTICAL APPLICATIONS: Our study suggested that polysaccharide from sea cucumbers had the potential to be further developed as antitumor drugs.

Transcriptomics and iTRAQ-Proteomics Analyses of Bovine Mammary Tissue with Streptococcus agalactiae-Induced Mastitis.

Mastitis is a highly prevalent disease in dairy cows that causes large economic losses. Streptococcus agalactiae is a common contagious pathogen and a major cause of bovine mastitis. The immune response to intramammary infection with S. agalactiae in dairy cows is a very complex biological process. To understand the host immune response to S. agalactiae-induced mastitis, mammary gland of lactating Chinese Holstein cows was challenged with S. agalactiae via nipple tube perfusion. Visual inspection, analysis of milk somatic cell counts, histopathology, and transmission electron microscopy of mammary tissue were performed to confirm S. agalactiae-induced mastitis. Microarray and isobaric tags for relative and absolute quantitation (iTRAQ) were used to compare the transcriptomes and proteomes of healthy and mastitic mammary tissue. Compared with healthy tissue, a total of 129 differentially expressed genes (DEGs, fold change >2, p < 0.05) and 144 differentially expressed proteins (DEPs, fold change >1.2, p < 0.05) were identified in mammary tissue from S. agalactiae-challenged cows. Among the concordant 18 DEGs/DEPs, immunoglobulin M precursor, cathelicidin-7 precursor, integrin alpha-5, and complement C4-A-like isoform X1 were associated with mastitis. Intramammary infection with S. agalactiae triggered a complex host innate immune response that involved complement and coagulation cascades, ECM-receptor interaction, focal adhesion, and phagosome and bacterial invasion of epithelial cells pathways. These results provide candidate genes or proteins for further studies in the context of prevention and targeted treatment of bovine mastitis.

Isobaric Tags for Relative and Absolute Quantification (iTRAQ)-Based Untargeted Quantitative Proteomic Approach To Identify Change of the Plasma Proteins by Salbutamol Abuse in Beef Cattle.

Salbutamol, a selective ß2-agonist, endangers the safety of animal products as a result of illegal use in food animals. In this study, an iTRAQ-based untargeted quantitative proteomic approach was applied to screen potential protein biomarkers in plasma of cattle before and after treatment with salbutamol for 21 days. A total of 62 plasma proteins were significantly affected by salbutamol treatment, which can be used as potential biomarkers to screen for the illegal use of salbutamol in beef cattle. Enzyme-linked immunosorbent assay measurements of five selected proteins demonstrated the reliability of iTRAQ-based proteomics in screening of candidate biomarkers among the plasma proteins. The plasma samples collected before and after salbutamol treatment were well-separated by principal component analysis (PCA) using the differentially expressed proteins. These results suggested that an iTRAQ-based untargeted quantitative proteomic strategy combined with PCA pattern recognition methods can discriminate differences in plasma protein profiles collected before and after salbutamol treatment.

Complex and extensive post-transcriptional regulation revealed by integrative proteomic and transcriptomic analysis of metabolite stress response in Clostridium acetobutylicum.

BACKGROUND: Clostridium acetobutylicum is a model organism for both clostridial biology and solvent production. The organism is exposed to its own toxic metabolites butyrate and butanol, which trigger an adaptive stress response. Integrative analysis of proteomic and RNAseq data may provide novel insights into post-transcriptional regulation. RESULTS: The identified iTRAQ-based quantitative stress proteome is made up of 616 proteins with a 15 % genome coverage. The differentially expressed proteome correlated poorly with the corresponding differential RNAseq transcriptome. Up to 31 % of the differentially expressed proteins under stress displayed patterns opposite to those of the transcriptome, thus suggesting significant post-transcriptional regulation. The differential proteome of the translation machinery suggests that cells employ a different subset of ribosomal proteins under stress. Several highly upregulated proteins but with low mRNA levels possessed mRNAs with long 5'UTRs and strong RBS scores, thus supporting the argument that regulatory elements on the long 5'UTRs control their translation. For example, the oxidative stress response rubrerythrin was upregulated only at the protein level up to 40-fold without significant mRNA changes. We also identified many leaderless transcripts, several displaying different transcriptional start sites, thus suggesting mRNA-trimming mechanisms under stress. Downregulation of Rho and partner proteins pointed to changes in transcriptional elongation and termination under stress. CONCLUSIONS: The integrative proteomic-transcriptomic analysis demonstrated complex expression patterns of a large fraction of the proteome. Such patterns could not have been detected with one or the other omic analyses. Our analysis proposes the involvement of specific molecular mechanisms of post-transcriptional regulation to explain the observed complex stress response.