Abstract
The distinct stages of cotton fiber development and maturation serve as a single-celled model for studying the molecular mechanisms of plant cell elongation, cell wall development and cellulose biosynthesis. However, this model system of plant cell development is compromised for proteomic studies due to a lack of an efficient protein extraction method during the later stages of fiber development, because of a recalcitrant cell wall and the presence of abundant phenolic compounds. Here, we compared the quality and quantities of proteins extracted from 25 dpa (days post anthesis) fiber with multiple protein extraction methods and present a comprehensive quantitative proteomic study of fiber development from 10 dpa to 25 dpa. Comparative analysis using a label-free quantification method revealed 287 differentially-expressed proteins in the 10 dpa to 25 dpa fiber developmental period. Proteins involved in cell wall metabolism and regulation, cytoskeleton development and carbohydrate metabolism among other functional categories in four fiber developmental stages were identified. Our studies provide protocols for protein extraction from maturing fiber tissues for mass spectrometry analysis and expand knowledge of the proteomic profile of cotton fiber development.
Highlights
Being the world’s leading natural textile fiber, the economically-valuable cotton fibers of upland cotton (Gossypium hirsutum L.) are unique in the plant kingdom due to their size and chemical composition [1,2]
This study presents the first proteome of the 35 dpa cotton fiber, a highly recalcitrant tissue, and presents a comprehensive proteomic study of fiber development from 10 dpa to 25 dpa
We found that an isoform of profilin (A0A0B0PF49) was downregulated during secondary wall deposition (25 dpa) when compared to an earlier fiber development stage (15 dpa), while actin depolymerizing factor 7 (A1XJ44) was found to be upregulated in 15 dpa when compared to 10 dpa and downregulated in 25 dpa fiber when compared to 15 dpa
Summary
Being the world’s leading natural textile fiber, the economically-valuable cotton fibers of upland cotton (Gossypium hirsutum L.) are unique in the plant kingdom due to their size and chemical composition [1,2]. During primary wall elongation (within 20 days post anthesis (dpa)) and secondary wall deposition and thickening (from 20 to 35 dpa), sizeable amounts of polysaccharide components are synthesized and deposited, producing a cell wall 3 to 4 μm thick, made up of more than 94% cellulose [3,7,8]. These advantageous features make cotton fiber an excellent single-celled model for studying the molecular mechanisms of plant cell elongation, cell wall development and cellulose biosynthesis [2,5,9,10,11]. Cotton fiber proteins were extracted by directly homogenizing cotton fibers in aqueous buffer followed by organic solvent precipitation [7,23]
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