ABSTRACT
Abstract Through exploring potential analogies between cotton seed trichomes (or cotton fiber) and arabidopsis shoot trichomes we discovered that CesAs from either the primary or secondary wall phylogenetic clades can support secondary wall thickening. CesA genes that typically support primary wall synthesis, AtCesA1,2,3,5, and 6, underpin expansion and secondary wall thickening of arabidopsis shoot trichomes. In contrast, apparent orthologs of CesA genes that support secondary wall synthesis in arabidopsis xylem, AtCesA4,7, and 8, are up-regulated for cotton fiber secondary wall deposition. These conclusions arose from: (a) analyzing the expression of CesA genes in arabidopsis shoot trichomes; (b) observing birefringent secondary walls in arabidopsis shoot trichomes with mutations in AtCesA4, 7, or 8; (c) assaying up-regulated genes during different stages of cotton fiber development; and (d) comparing genes that were co-expressed with primary or secondary wall CesAs in arabidopsis with genes up-regulated in arabidopsis trichomes, arabidopsis secondary xylem, or cotton fiber during primary or secondary wall deposition. Cumulatively, the data show that: (a) the xylem of arabidopsis provides the best model for secondary wall cellulose synthesis in cotton fiber; and (b) CesA genes within a "cell wall toolbox" are used in diverse ways for the construction of particular specialized cell walls.
Subject(s)
Arabidopsis/metabolism , Cell Wall/metabolism , Glucosyltransferases/classification , Gossypium/metabolism , Phylogeny , Plant Proteins/classification , Plant Shoots/metabolism , Arabidopsis/genetics , Arabidopsis Proteins/classification , Arabidopsis Proteins/genetics , Arabidopsis Proteins/physiology , Cotton Fiber , Gene Expression Regulation, Plant/genetics , Gene Expression Regulation, Plant/physiology , Glucosyltransferases/genetics , Glucosyltransferases/physiology , Gossypium/genetics , Oligonucleotide Array Sequence Analysis , Plant Proteins/genetics , Plant Proteins/physiology , Plant Shoots/geneticsABSTRACT
A new procedure has been developed for the isolation of wild-type and mutant Arabidopsis trichomes. The isolated trichomes maintained enzymatic activity and were used for DNA, protein, and RNA isolation. The RNA was used to generate probes suitable for Affymetrix analysis. The validity of the Affymetrix results was confirmed by quantitative PCR analysis on a subset of genes that are preferentially expressed in trichomes or leaves. Sufficient quantities of trichomes were isolated to probe the biochemical nature of trichome cell walls. These analyses provide evidence for the presence of lignin in Arabidopsis trichome cell walls. The monosaccharide analysis and positive staining with ruthenium red indicates that the walls also contain a large portion of pectin. The 2.23-fold ratio of pectin-related sugars compared with potential cellulosic glucose suggests that the polysaccharides of the trichome cell walls are more like those of typical primary walls even though the wall becomes quite thick. Overall, these analyses open the door to using the Arabidopsis trichome cell wall as an excellent model to probe various questions concerning plant cell wall biosynthesis.
Subject(s)
Arabidopsis/cytology , Cell Separation/methods , Cell Wall/chemistry , Gene Expression Profiling , Arabidopsis/chemistry , Arabidopsis/genetics , Arabidopsis Proteins/isolation & purification , DNA, Plant/genetics , DNA, Plant/isolation & purification , Genes, Plant , Genome, Plant , Lignin/chemistry , Microscopy, Electron, Transmission , Monosaccharides/chemistry , Plant Epidermis/cytology , Plant Epidermis/genetics , Plant Leaves/cytology , Plant Leaves/genetics , Polymerase Chain Reaction , RNA, Plant/genetics , RNA, Plant/isolation & purificationABSTRACT
Cotton fibers are single-celled extensions of the seed epidermis. They can be isolated in pure form as they undergo staged differentiation including primary cell wall synthesis during elongation and nearly pure cellulose synthesis during secondary wall thickening. This combination of features supports clear interpretation of data about cell walls and cellulose synthesis in the context of high throughput modern experimental technologies. Prior contributions of cotton fiber to building fundamental knowledge about cell walls will be summarized and the dynamic changes in cell wall polymers throughout cotton fiber differentiation will be described. Recent successes in using stable cotton transformation to alter cotton fiber cell wall properties as well as cotton fiber quality will be discussed. Futurec prospects to perform experiments more rapidly through altering cotton fiberwall properties via virus-induced gene silencing will be evaluated.