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

引用

Science China(Life Sciences) 2016年第2期59卷 154-163页

作者： Xiang Jin Limin Wang Liping He Weiqiang Feng Xuchu Wang Institute of Tropical Biosciences and Biotechnology Chinese Academy of Tropical Agricultural Sciences The State Key Laboratory of Protein and Plant Gene Research College of Life Sciences Peking University College of Horticulture and Landscape Hainan University

Proteomic analysis of upland cotton was performed to profile the global detectable proteomes of ovules and fibers using two-dimensional electrophoresis(2DE).A total of 1,203 independent protein spots were collected from representative 2DE gels,which were digested with trypsin and identified by matrix-assisted laser desorption and ionization-time-offlight/time-of-flight(MALDI-TOF/TOF)mass *** mass spectrometry or tandem mass spectrometry(MS or MS/MS)data were then searched against a local database constructed from gossypium hirsutum genome sequences,resulting in successful identification of 975 protein spots(411 for ovules and 564 for fibers).Functional annotation analysis of the 975identified proteins revealed that ovule-specific proteins were mainly enriched in functions related to fatty acid elongation,sulfur amino acid metabolism and post-replication repair,while fiber-specific proteins were enriched in functions related to root hair elongation,galactose metabolism and D-xylose metabolic *** annotation analysis of the most abundant protein spots showed that 28.96%of the total proteins in the ovule were mainly located in the golgi apparatus,endoplasmic reticulum,mitochondrion and ribosome,whereas in fibers,27.02%of the total proteins were located in the cytoskeleton,nuclear envelope and cell *** real-time polymerase chain reaction(q RT-PCR)analyses of the ovule-specific protein spots P61,P93 and P198 and fiber-specific protein spots 230,477 and 511 were performed to validate the proteomics ***-protein interaction network analyses revealed very different network cluster patterns between ovules and *** work provides the largest protein identification dataset of 2DE-detectable proteins in cotton ovules and fibers and indicates potentially important roles of tissue-specific proteins,thus providing insights into the cotton ovule and fiber proteomes on a global scale.

关键词： cotton 2DE MALDI-TOF/TOF go enrichment protein-protein interaction networks

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