title: Gene-sequence-tag expression analyses of 1,800 genes related to chloroplast functions.
abstract: Quantification of the expression levels of nuclear genes encoding plastid proteins under different genetic or environmental conditions can contribute to the genetic dissection of plastid functions. To facilitate such measurements, a set of 1,827 Arabidopsis thaliana genes coding for plastid proteins was PCR-amplified from genomic DNA and spotted on nylon membranes to generate an array of chloroplast-specific gene-sequence-tags (GSTs). The sensitivity and reliability of the experimental system was evaluated and a procedure was developed for detecting differential gene expression. The GST array was found to serve as a reliable monitor of changes in gene expression induced by environmental and genetic alteration of chloroplast functions. Based on comparisons of dark- versus light-grown seedlings, and wild-type versus prpl11-1 plants, lists of differentially expressed genes are provided which include 193/7 and 25/42 up/down-regulated genes, respectively. The cut-off values for differential expression were 2.5-times (up) and 0.40 (down). Additional up-regulated genes with relatively low expression ratios (from 1.5- to 2.5-times) or down-regulated with relatively high ratios (0.4-0.67) can be accessed at the website: http://www.mpiz-koeln.mpg.de/~richly/GST-array.html. A sample of genes analysed by quantitative reverse transcription PCR confirmed the expression profiles monitored by the GST array. Differential hybridisation experiments with the prpl11-1 mutant revealed the existence of regulatory networks sensing the protein state of the chloroplast and transmitting the signal to the nucleus.
authors.affiliation: Zentrum zur Identifikation von Genfunktionen durch Insertionsmutagenese bei Arabidopsis thaliana (ZIGIA), Max-Planck-Institut für Züchtungsforschung, Carl-von-Linné-Weg 10, 50829 Köln, Germany.
authors.forename: Joachim
authors.initials: J
authors.lastname: Kurth
authors.forename: Claudio
authors.initials: C
authors.lastname: Varotto
authors.forename: Paolo
authors.initials: P
authors.lastname: Pesaresi
authors.forename: Alexander
authors.initials: A
authors.lastname: Biehl
authors.forename: Erik
authors.initials: E
authors.lastname: Richly
authors.forename: Francesco
authors.initials: F
authors.lastname: Salamini
authors.forename: Dario
authors.initials: D
authors.lastname: Leister
mesh_terms: D017360:Arabidopsis / Q000235:genetics* / Q000254:growth & development / Q000528:radiation effects; D002467:Cell Nucleus / Q000235:genetics / Q000502:physiology; D002736:Chloroplasts / Q000235:genetics / Q000502:physiology*; D003624:Darkness; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant / Q000528:radiation effects; D008027:Light; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D018749:RNA, Plant / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction
chemical_list: D018749:RNA, Plant