A new method for isolating large quantities of Arabidopsis trichomes for transcriptome, cell wall and other types of analyses.
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.
Marks
M David
MD
Department of Plant Biology, University of Minnesota, 1445 Gortner Ave., St Paul, MN 55108, USA. marks004@umn.edu
Betancur
Lissete
L
Gilding
Edward
E
Chen
Fang
F
Bauer
Stefan
S
Wenger
Jonathan P
JP
Dixon
Richard A
RA
Haigler
Candace H
CH
D017360:Arabidopsis / Q000737:chemistry / Q000166:cytology* / Q000235:genetics; D029681:Arabidopsis Proteins / Q000302:isolation & purification; D002469:Cell Separation / Q000379:methods*; D002473:Cell Wall / Q000737:chemistry*; D018744:DNA, Plant / Q000235:genetics / Q000302:isolation & purification; D020869:Gene Expression Profiling; D017343:Genes, Plant; D018745:Genome, Plant; D008031:Lignin / Q000737:chemistry; D046529:Microscopy, Electron, Transmission; D009005:Monosaccharides / Q000737:chemistry; D019441:Plant Epidermis / Q000166:cytology / Q000235:genetics; D018515:Plant Leaves / Q000166:cytology / Q000235:genetics; D016133:Polymerase Chain Reaction; D018749:RNA, Plant / Q000235:genetics / Q000302:isolation & purification
D029681:Arabidopsis Proteins; D018744:DNA, Plant; D009005:Monosaccharides; D018749:RNA, Plant; D008031:Lignin