A rapid preparation procedure for laser microdissection-mediated harvest of plant tissues for gene expression analysis

Plant Methods
Stian Olsen, Kirsten Krause

Abstract

Gene expression changes that govern essential biological processes can occur at the cell-specific level. To gain insight into such events, laser microdissection is applied to cut out specific cells or tissues from which RNA for gene expression analysis is isolated. However, the preparation of plant tissue sections for laser microdissection and subsequent RNA isolation usually involves fixation and embedding, processes that are often time-consuming and can lower the yield and quality of isolated RNA. Infection sites of the parasitic plant Cuscuta reflexa growing on its compatible host plant Pelargonium zonale were sectioned using a vibratome and dried on glass slides at 4 °C before laser microdissection. High quality RNA (RQI > 7) was isolated from 1 mm2, 3 mm2 and 6 mm2 total surface areas of laser microdissection-harvested C. reflexa tissue, with the yield of RNA correlating to the amount of collected material (on average 7 ng total RNA/mm2). The expression levels of two parasite genes previously found to be highly expressed during host plant infection were shown to differ individually between specific regions of the infection site. By drying plant sections under low pressure to reduce the dehydration time, the induced express...Continue Reading

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Citations

Nov 20, 2021·The Plant Journal : for Cell and Molecular Biology·Joseph SwiftC Robertson McClung

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Methods Mentioned

BETA
Electrophoresis
RNA-Seq
RNA
Assay
PCR

Software Mentioned

Excel
NormFinder
CFX Manager
PALMRobo

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