title: A novel procedure for the quantitative analysis of metabolites, storage products and transcripts of laser microdissected seed tissues of Brassica napus.
abstract: BACKGROUND
  The biology of the seed is complicated by the extensive non-homogeneity (spatial gradients) in gene expression, metabolic conversions and storage product accumulation. The detailed understanding of the mechanisms underlying seed growth and storage therefore requires the development of means to obtain tissue-specific analyses. This approach also represents an important priority in the context of seed biotechnology.
  
  
  RESULTS
  We provide a guideline and detailed procedures towards the quantitative analysis of laser micro-dissected (LM) tissues in oilseed rape (Brassica napus). This includes protocols for laser microdissection of the seed, and the subsequent extraction and quantitative analysis of lipids, starch and metabolites (sugars, sugar phosphates, nucleotides, amino acids, intermediates of glycolysis and citric acid cycle). We have also developed a protocol allowing the parallel analysis of the transcriptome using Brassica-specific microarrays. Some data are presented regarding the compartmentation of metabolites within the oilseed rape embryo.
  
  
  CONCLUSIONS
  The described methodology allows for the rapid, combined analysis of metabolic intermediates, major storage products and transcripts in a tissue-specific manner. The protocols are robust for oilseed rape, and should be readily adjustable for other crop species. The suite of methods applied to LM tissues represents an important step in the context of both the systems biology and the biotechnology of oilseeds.
authors: Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr, 3, 06466 Gatersleben, Germany. rollet@ipk-gatersleben.de.
authors: Silke
authors: S
authors: Schiebold
authors: Henning
authors: H
authors: Tschiersch
authors: Ljudmilla
authors: L
authors: Borisjuk
authors: Nicolas
authors: N
authors: Heinzel
authors: Ruslana
authors: R
authors: Radchuk
authors: Hardy
authors: H
authors: Rolletschek