Label-Free Quantitative Proteomics Analysis of Cotton Leaf Response to Nitric Oxide

Abstract

To better understand nitric oxide (NO) responsive proteins, we investigated the proteomic differences between untreated (control), sodium nitroprusside (SNP) treated, and carboxy-PTIO potassium salt (cPTIO, NO scavenger) followed by SNP treated cotton plants. This is the first study to examine the effect of different concentrations of NO on the leaf proteome in cotton using a label-free approach based on nanoscale ultraperformance liquid chromatography-electrospray ionization (ESI)-low/high-collision energy MS analysis (MS(E)). One-hundred and sixty-six differentially expressed proteins were identified. Forty-seven of these proteins were upregulated, 82 were downregulated, and 37 were expressed specifically under different conditions. The 166 proteins were functionally divided into 17 groups and localized to chloroplast, Golgi apparatus, cytoplasm, and so forth. The pathway analysis demonstrated that NO is involved in various physiological activities and has a distinct influence on carbon fixation in photosynthetic organisms and photosynthesis. In addition, this is the first time proteins involved in ethylene synthesis were identified to be regulated by NO. The characterization of these protein networks provides a better understanding of the possible regulation mechanisms of cellular activities occurring in the NO-treated cotton leaves and offers new insights into NO responses in plants.

NO to drought-multifunctional role of nitric oxide in plant drought: Do we have all the answers?

“…Comparative proteome profiles of leaf samples of Gossypium hirsutum treated with SNP and NO scavenger followed by NO donor treatment revealed 166 differentially expressed proteins belonging to different cellular compartments and involved in diverse pathways. Of these, 47 were upregulated, 82 were downregulated and 37 were condition-specific [84]. The emerging experimental evidence demonstrates that NO possibly operates through post-translational modification of proteins, mainly via S-nitrosylation, metal nitrosylation, carbonylation and tyrosine nitration [54,67,70,85].…”

Two-dimensional gel electrophoresis-based analysis provides global insights into the cotton ovule and fiber proteomes

“…However, scarce information was available about the largest portion of 2DE-detectable protein spots, usually more than 1,000 protein spots, beyond differentially expressed proteins. Gel-free high-throughput MS approaches have been rapidly developed in recent years and have identified 1,000-5,000 or more proteins in cotton tissues (Hu et al, 2013;Meng et al, 2011;Wang et al, 2015). However, these data were still lacking global proteome information in some aspects that could be provided by 2DE maps, such as protein spot visualization, reliable evidence for existing protein isoforms and the abundances of individual protein spots.…”

“…Furthermore, the complex allotetraploid genome sequences of Gh TM-1 were reported in 2015 (Cao, 2015;Li et al, 2015;Zhang et al, 2015). Based on the cotton genome sequences, significant advances have been made in the last few years in the development of gel-free and high-throughput MS protein identification technologies (He, 2013;Hu et al, 2013;Meng et al, 2011). Comparative proteomics research of fiber initiation based on isobaric tags for relative and absolute quantification (iTRAQ) has been reported, and 5,884 proteins from 0 day post-anthesis (DPA) wild-type and fuzzless-lintless (fl) ovules, of which 2,927 proteins preferentially accumulated in wild-type ovules, have been identified .…”

Comparative Proteomic Analysis of Differentially Expressed Proteins Induced by Hydrogen Sulfide in Spinacia oleracea Leaves

“…Carbon dioxide fixation is an essential process of photosynthesis, and this pathway involves in many enzymes that catalyze and regulate energy generation [40]. In this study, we identified several photosynthesis-associated proteins and their expression levels were significantly up-regulated after NaHS treatment, e.g., the ribulose-1,5-bisphosphate carboxylase/oxygenase large subunit (rubisco LSU, spots 3, 7, 8 and 53) and rubisco activase precursor (spots 1 and 34).…”