Identification of Arabidopsis thaliana phloem RNAs provides a search criterion for phloem-based transcripts hidden in complex datasets of microarray experiments.
Phloem-mobile signals play a major role in plant nutrition, development and communication. In the latter context, phloem-mobile RNAs have been associated with signalling between plant tissues. In this study, we focused on the identification of transcripts in the shoot phloem of the model plant Arabidopsis thaliana. To isolate transcripts expressed in phloem parenchyma cells and in companion cell-sieve element complexes, we used laser microdissection coupled to laser pressure catapulting (LMPC). Mobile transcripts in sieve elements were isolated from leaf phloem exudates. After optimization of sampling and fixation, RNA of high quality was isolated from both sources. The modifications to the RNA amplification procedure described here were well suited to production of RNA of sufficient yield and quality for microarray experiments. Microarrays hybridized with LMPC-derived phloem tissue or phloem sap RNA allowed differentiation between phloem-expressed and mobile transcript species. Using this set of phloem transcripts and comparing them with microarrays derived from databases of light, hormone and nutrient treatment experiments, we identified phloem-derived RNAs as mobile, potential long-distance signals. Our dataset thus provides a search criterion for phloem-based signals hidden in the complex datasets of microarray experiments. The availability of these comprehensive phloem transcript profiles will facilitate reverse-genetic studies and forward-genetic screens for phloem and long-distance RNA signalling mutants.
Deeken
Rosalia
R
Julius-von-Sachs-Institute for Biosciences, University of Würzburg, Julius-von-Sachs-Platz 2, D-97082 Würzburg, Germany.
Ache
Peter
P
Kajahn
Inga
I
Klinkenberg
Joern
J
Bringmann
Gerhard
G
Hedrich
Rainer
R
D017360:Arabidopsis / Q000235:genetics* / Q000378:metabolism; D005122:Exudates and Transudates / Q000378:metabolism; D020869:Gene Expression Profiling; D017343:Genes, Plant; D005961:Glucosinolates / Q000096:biosynthesis; D042282:Microdissection; D020411:Oligonucleotide Array Sequence Analysis; D052585:Phloem / Q000235:genetics* / Q000378:metabolism; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D018547:Plant Stems / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics*; D015398:Signal Transduction
D005961:Glucosinolates; D012333:RNA, Messenger; D018749:RNA, Plant