title: Quantitative RT-PCR Platform to Measure Transcript Levels of C and N Metabolism-Related Genes in Durum Wheat: Transcript Profiles in Elevated [CO2] and High Temperature at Different Levels of N Supply.
abstract: Only limited public transcriptomics resources are available for durum wheat and its responses to environmental changes. We developed a quantitative reverse transcription-PCR (qRT-PCR) platform for analysing the expression of primary C and N metabolism genes in durum wheat in leaves (125 genes) and roots (38 genes), based on available bread wheat genes and the identification of orthologs of known genes in other species. We also assessed the expression stability of seven reference genes for qRT-PCR under varying environments. We therefore present a functional qRT-PCR platform for gene expression analysis in durum wheat, and suggest using the ADP-ribosylation factor as a reference gene for qRT-PCR normalization. We investigated the effects of elevated [CO(2)] and temperature at two levels of N supply on C and N metabolism by combining gene expression analysis, using our qRT-PCR platform, with biochemical and physiological parameters in durum wheat grown in field chambers. Elevated CO(2) down-regulated the photosynthetic capacity and led to the loss of N compounds, including Rubisco; this effect was exacerbated at low N. Mechanistically, the reduction in photosynthesis and N levels could be associated with a decreased transcription of the genes involved in photosynthesis and N assimilation. High temperatures increased stomatal conductance, and thus did not inhibit photosynthesis, even though Rubisco protein and activity, soluble protein, leaf N, and gene expression for C fixation and N assimilation were down-regulated. Under a future scenario of climate change, the extent to which C fixation capacity and N assimilation are down-regulated will depend upon the N supply.
authors.affiliation: Institute of Natural Resources and Agrobiology of Salamanca, CSIC, Cordel de Merinas 40-52, Salamanca, 37008 Spain.
authors.forename: Rubén
authors.initials: R
authors.lastname: Vicente
authors.affiliation: Institute of Natural Resources and Agrobiology of Salamanca, CSIC, Cordel de Merinas 40-52, Salamanca, 37008 Spain.
authors.forename: Pilar
authors.initials: P
authors.lastname: Pérez
authors.affiliation: Institute of Natural Resources and Agrobiology of Salamanca, CSIC, Cordel de Merinas 40-52, Salamanca, 37008 Spain.
authors.forename: Rafael
authors.initials: R
authors.lastname: Martínez-Carrasco
authors.affiliation: Institute for Biology 1, RWTH Aachen University, Aachen, D-52062 Germany IBG-2: Plant Sciences, Institute of Bio- and Geosciences, Forschungszentrum Jülich, Leo-Brandt-Straße, Jülich, D-52425 Germany.
authors.forename: Björn
authors.initials: B
authors.lastname: Usadel
authors.affiliation: Institute of Natural Resources and Agrobiology of Salamanca, CSIC, Cordel de Merinas 40-52, Salamanca, 37008 Spain Present address: Department of Agrochemistry and Soil Science, Agricultural University, 12 Mendeleev Street, Plovdiv, 4004 Bulgaria.
authors.forename: Svetla
authors.initials: S
authors.lastname: Kostadinova
authors.affiliation: Institute of Natural Resources and Agrobiology of Salamanca, CSIC, Cordel de Merinas 40-52, Salamanca, 37008 Spain rosa.morcuende@irnasa.csic.es.
authors.forename: Rosa
authors.initials: R
authors.lastname: Morcuende
mesh_terms: D020727:ADP-Ribosylation Factors / Q000235:genetics; D002244:Carbon / Q000378:metabolism*; D002245:Carbon Dioxide / Q000494:pharmacology*; D018506:Gene Expression Regulation, Plant; D006358:Hot Temperature; D009584:Nitrogen / Q000378:metabolism*; D010788:Photosynthesis / Q000502:physiology*; D018515:Plant Leaves / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D010940:Plant Proteins / Q000235:genetics; D018517:Plant Roots / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012273:Ribulose-Bisphosphate Carboxylase / Q000235:genetics; D014908:Triticum / Q000187:drug effects / Q000235:genetics* / Q000502:physiology
chemical_list: D010940:Plant Proteins; D002245:Carbon Dioxide; D002244:Carbon; D020727:ADP-Ribosylation Factors; D012273:Ribulose-Bisphosphate Carboxylase; D009584:Nitrogen
keywords: Elevated CO2; High temperature; Nitrogen; Primary metabolism; Triticum durum; qRT–PCR