A new method for isolating large quantities of Arabidopsis trichomes for transcriptome, cell wall and other types of analyses
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作者:
Marks, M. David
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Univ Minnesota, Dept Plant Biol, St Paul, MN 55108 USAUniv Minnesota, Dept Plant Biol, St Paul, MN 55108 USA
Marks, M. David
[1
]
Betancur, Lissete
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N Carolina State Univ, Dept Plant Biol, Raleigh, NC 27695 USAUniv Minnesota, Dept Plant Biol, St Paul, MN 55108 USA
Betancur, Lissete
[2
]
Gilding, Edward
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Univ Minnesota, Dept Plant Biol, St Paul, MN 55108 USAUniv Minnesota, Dept Plant Biol, St Paul, MN 55108 USA
Gilding, Edward
[1
]
Chen, Fang
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机构:
Samuel Roberts Noble Fdn Inc, Div Plant Biol, Ardmore, OK 73401 USAUniv Minnesota, Dept Plant Biol, St Paul, MN 55108 USA
Chen, Fang
[3
]
Bauer, Stefan
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Carnegie Inst Washington, Dept Plant Biol, Stanford, CA 94305 USAUniv Minnesota, Dept Plant Biol, St Paul, MN 55108 USA
Bauer, Stefan
[4
]
Wenger, Jonathan P.
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Univ Minnesota, Dept Plant Biol, St Paul, MN 55108 USAUniv Minnesota, Dept Plant Biol, St Paul, MN 55108 USA
Wenger, Jonathan P.
[1
]
Dixon, Richard A.
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机构:
Samuel Roberts Noble Fdn Inc, Div Plant Biol, Ardmore, OK 73401 USAUniv Minnesota, Dept Plant Biol, St Paul, MN 55108 USA
Dixon, Richard A.
[3
]
Haigler, Candace H.
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N Carolina State Univ, Dept Plant Biol, Raleigh, NC 27695 USA
N Carolina State Univ, Dept Crop Sci, Raleigh, NC 27695 USAUniv Minnesota, Dept Plant Biol, St Paul, MN 55108 USA
Haigler, Candace H.
[2
,5
]
机构:
[1] Univ Minnesota, Dept Plant Biol, St Paul, MN 55108 USA
[2] N Carolina State Univ, Dept Plant Biol, Raleigh, NC 27695 USA
[3] Samuel Roberts Noble Fdn Inc, Div Plant Biol, Ardmore, OK 73401 USA
[4] Carnegie Inst Washington, Dept Plant Biol, Stanford, CA 94305 USA
[5] N Carolina State Univ, Dept Crop Sci, Raleigh, NC 27695 USA
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.