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Identification of key proteinIdentification of key proteins related to cashmere fibre diameter by integrated proteomics and bioinformatic analysiss related to cashmere fibre diameter by integrated proteomics and bioinformatic analysis

ABSTRACT: Goat (Capra hircus) has always been a source of fibre for human use and holds an important place in international high-end textiles. Fibre diameter is the most concerned economic indicator for producers. Understanding the formation mechanism of fibre diameter and related key proteins can help optimize and control the production of cashmere. Although mass spectrometry (MS)-based workflows have made great progress in achieving near-genome-wide coverage, the equivalent complete map of cashmere proteome is still elusive. Here, we performed label-free liquid chromatography coupled with tandem MS to profile the cashmere proteome. Firstly, by measuring the fibre diameter, it was found that the average fibre diameter of Alxa cashmere was significantly smaller than that of Alpas. With the help of proteomics technology,170 proteins were detected with high confidence. A total of 68 differentially expressed proteins were identified in the two cashmere, 131 proteins were specifically expressed in Alpas, and 40 proteins were specifically expressed in Alxa. Through Gene Ontology enrichment analysis, the Kyoto Encyclopedia of Genes and Genomes, Protein-Protein Interaction network construction, and the CytoHubba plugin of the Cytoscape software were used to interpret the proteomic data. Finally, KRT10, KRT14, KRT17, and KRT82 played a key role in leading to the difference in the diameter of the two fibres. Combined with bioinformatics, the structure and function of these proteins were further studied to provide a more com-prehensive perspective for understanding the regulation mechanism of cashmere diameter. At the same time, combined with the information on genetics and gene expression, the genetic factors and epigenetic regulation mechanisms affecting cashmere fibre diameter can be further studied.

INSTRUMENT(S): TripleTOF 5600

ORGANISM(S): Capra Hircus (goat)

TISSUE(S): Skin

SUBMITTER: Chongyan Zhang

LAB HEAD: Chongyan Zhang

PROVIDER: PXD050828 | Pride | 2024-03-21

REPOSITORIES: Pride

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Project description:Comparative analysis of transcriptome profiles of G. arboreum L. cv. and its fuzzy-lintless mutant (ANOI 1960) at 0 and 10 dpa. Cotton is one of the most commercially important fibre crops in the world and used as a source for natural textile fibre and cottonseed oil. The fuzzy-lintless ovules of cotton mutants are ideal source for identifying genes involved in fibre development by comparing with fibre bearing ovules of wild-type. To decipher molecular mechanisms involved in fibre cell development, transcriptome analysis has been carried out by comparing G. arboreum cv. (wild-type) with its fuzzy-lintless mutant (ANOI 1960). Fuzzed-lintless mutant line was generated by back cross breeding between FL and Fl (recurrent parent) lines (personal communication by Dr. I. S. Katageri). Basically Fibre less type was a RIL, first recovered from cross between G.arboreum (linted) and G. anomalum (lint less). This RIL was used as donor parent and crossed with normal arboreum (as recurrent parent) to develop G. arboreum FL and G. arboreum Fl isogenic lines. This G. arboreum Fl line is named as ANOI 1960. Cotton bolls were collected at fibre initiation (0 dpa/days post anthesis) and elongation (10 dpa) and gene expression profiles were analyzed in wild-type and ANOI 1960 mutant using Affymetrix cotton GeneChip Genome array. Cotton plants were grown under field condition. Flowers were tagged and cotton bolls were collected during fibre development stages. Total RNA was isolated from fibre bearing ovules of wild-type (WT) and fuzzy-lintless ovules of mutant (ANOI 1960) collected at various (0 and 10 dpa) fibre development stages using SpectrumTM Plant Total RNA kit (Sigma, USA) according to the manufacturerM-bM-^@M-^Ys protocol. Affymetrix cotton GeneChip Genome array (Affymetrix, USA) having 23,977 probe sets representing 21,854 cotton transcripts was used for transcriptome analysis. Three biological replicates were maintained to test the reproducibility and quality of the chip hybridization. cDNA labeling, array hybridization, staining and washing procedures were carried out as described in the Affymetrix protocols. CEL files having estimated probe intensity values were analyzed with GeneSpring GX-11.5 software (Agilent Technologies, USA) to get differentially expressed transcripts. The Robust Multiarray Average (RMA) algorithm was used for the back ground correction, quantile normalization and median polished probe set summarization to generate single expression value for each probe set. Normalized expression values were log2-transformed and differential expression analysis was performed using unpaired t-test. The p-values were corrected by applying the false discovery rate (FDR) correction (Benjamini and Hochberg, 2000).

Dataset Information

Identification of key proteinIdentification of key proteins related to cashmere fibre diameter by integrated proteomics and bioinformatic analysiss related to cashmere fibre diameter by integrated proteomics and bioinformatic analysis

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