Phosphoproteomic Study Of Upland Cotton Ovule Based On Labeling Quantification

Cotton is one of the most important crops in the world,and its fiber provides a good material for the textile industry.Gossypium hirsutum L.is most widely cultivated species in the world’s four largest cotton cultivated species(Gossypium hirsutum L.,Gossypium barbadense L.,Gossypium arboretum L.,Gossypium herbaceum L.).Gossypium hirsutum L.has a fiberless-lintless mutant(fl)screened from nature.Its fiberless phenotype can be inherited steadily.The difference between the mutant and wild type(WT)has always been a hot topic for cotton researchers.The protein is the main bearer of life activities,so this paper selected the wild type and mutant after flowering 0 days,3 days and 5 days of ovule material as the research object,then using the stable isotope dimethyl labeling to labeling the enriched phosphorylated protein,at the same time the application of TMT(tandem mass tags)kit to mark protein.Finally,by means of mass spectrometry identification and bioinformatics analysis,we try to analyze the difference between wild type and mutant from the protein.The main results of this paper include the following 7 aspects:(1)It was identified a large number of non-redundant non-phosphorylated proteins(13394),phosphoproteins(4702),phosphopeptides(6814),phosphosites(9102),and the relative quantitative ratio obtained from various proteins in wild type and mutant ovules in each sample.(2)Characterize all phosphosites.(3)Through the functional analysis of the identified phosphoproteins,cotton specific phosphoproteins,phosphorylation modified transcription factors and kinases,etc.were screened.(4)According to the t-test and differential multiple screening of the two technical duplicate data,the phosphoproteins that were differentially expressed between the wild type and the mutants at 0,3 and 5 days after flowering were obtained respectively.Overexpression analysis of differentially expressed proteins excavates the biological processes with enrichment effects.(5)Analysis of the BRs signal pathway and the endoreduplication signal pathway which RBRI involved in.(6)Cluster analysis of quantitative protein data and joint analysis of transcriptome quantitative data.(7)Combined analysis of multiples of differential expression,protein function annotation and sequence analysis etc.to obtain candidate proteins that can be studied in depth,such as:EPS 15,RBR1.