All plant material was derived from two tissues (endosperm and embryo) of coffea arabica seeds at 150 and 200 days after flowering (stage 5 and 6, respectively). Total RNA was also extracted from separated endosperm and zygotic embryos using the RNeasy™ Lipid Tissue Kit (Qiagen). cDNA were tagged independently (Titanium kit, Roche Diagnostics) then mixed for sequencing by GATC Biotech AG (Germany) using GS FLX Titanium (Roche).. Integrative analysis of the late maturation programme and desiccation tolerance mechanisms in intermediate coffee seeds

The intermediate seed category was defined in the early 1990s using coffee (Coffea arabica) as a model. In contrast to orthodox seeds, intermediate seeds cannot survive complete drying, which is a major constraint for seed storage, for both biodiversity conservation and agricultural purposes. However, intermediate seeds are considerably more tolerant to drying than recalcitrant seeds, which are highly sensitive to desiccation. To gain insight into the mechanisms governing such differences, changes in desiccation tolerance (DT), hormone content and the transcriptome were analysed in developing coffee seeds. Acquisition of DT coincided with a dramatic transcriptional switch characterised by the repression of primary metabolism, photosynthesis and respiration, and the upregulation of genes coding for late embryogenesis abundant (LEA) proteins, heat shock proteins (HSP) and antioxidant enzymes. Analysis of heat-stable proteome in the mature coffee seed confirmed the accumulation of LEA proteins identified at the transcript level. Transcriptome analysis also suggests a major role for ABA and for the transcription factors CaHSFA9, CaDREB2G, CaANAC029, CaPLATZ and CaDOG-like in DT acquisition. The ability of CaHSFA9 and CaDREB2G to trigger HSP gene transcription was validated by Agrobacterium-mediated transformation of coffee somatic embryos.