Normalizing gene expression by quantitative PCR during somatic embryogenesis in two representative conifer species: Pinus pinaster and Picea abies

摘要: Suitable internal control genes to normalize qPCR data from different stages of embryo development and germination were identified in two representative conifer species. Clonal propagation by somatic embryogenesis has a great application potentiality conifers. Quantitative PCR (qPCR) is widely used for gene expression analysis during germination. No single reference universal, so systematic characterization endogenous concrete conditions fundamental accuracy. We suitable obtained at steps (embryonal mass proliferation, maturation germination) species, Pinus pinaster Picea abies. Candidate included commonly conifers, previously tested model plants, with lower variation the along according genome-wide transcript profiling studies. Three algorithms evaluate stability. The geometric average values elongation factor-1α, α-tubulin histone 3 P. pinaster, α-tubulin, adenosine kinase CAC abies adequate studies throughout embryogenesis. However, improved accuracy was achieved when using other combinations experiments samples developmental stage. importance selecting use tissues or within one close instability genes, highlighted.