The objective of this action was to extend our knowledge on the molecular basis of apomixis in the Gramineae and generate tools for harnessing this trait for the benefit of agriculture. The role of putative master genes in determining the switch between sexuality and apomixis is being validated in the species available from the PROCROP partners. A website (www.procropproject.eu) meant to contain all the information about the project was created and all the papers produced within the project are published there. Several meetings were carried out, in order to establish and to monitor PROCROP activities and secondments. A kick off conference was conducted in Milan. Then, most project participants met again in Montpellier during the EU mid-term evaluation meeting. Two meetings were organized in Argentina involving some of the beneficiaries (IRD, UMIL and UNIPG) participating together with the CONICET scientists. The meeting included also a 3-day workshops and also a final meeting to take stock of the situation and to define the last deliverables. The group was very satisfied by the number of manuscripts published or under preparation and scientific outcomes reached in the 4 years.
The work conducted in the first term of the project was aimed at: completing the sequencing and bioinformatics analysis of the sexual and apomictic floral transcriptomes from Poa pratensis (Illumina), Paspalum notatum (454) and Eragrostis curvula (454); elaborating a list of candidate genes differentially represented or displaying splice variants in the sexual and apomictic floral transcriptomes of P. pratensis, P. notatum and E. curvula; identifying miRNAs presenting differential activity in flowers of sexual and apomictic plants of P. notatum and E. curvula; detecting mRNAs controlled by silencing mechanisms in sexual and apomictic plants of P. notatum and E. curvula; identifying novel predicted miRNAs in sexual and apomictic P. notatum and E. curvula genotypes; identifying transcripts and miRNAs differentially expressed in flowers of apomictic and sexual E. curvula genotypes under control and water stress conditions; carrying out an MSAP wide genome methylation analysis in apomictic and sexual P. notatum genotypes, which led to the identification of one candidate gene (Pn-SCD1) epigenetically regulated in the nucellus of apomictic plants just before the onset of aposporous initials; conducting an analysis of the RdDM pathway activity in ovules of Eragrostis curvul; performing wide genome sequencing of Paspalum notatum by using Illumina (the raw data generated will be used in the next term to assemble the Paspalum notatum genome sequence); analyzing genes located within the ASR and expressed in the P. notatum libraries; obtaining a mapping population of E. curvula segregrating for the reproductive mode; identifying several developmental routes with different activity in apomictic and sexual plants; starting functional analysis of selected candidate genes.
In the second part of the project we established the list of selected candidate genes to be further examined through functional analysis; selection was based on positional linkage, expression association and relevant reproductive functional annotation criteria; completed a list of candidates showing te same expression pattern alterations in aposporous and diplosporous plants; completed an incomplete assembly of the P. notatum genome and the ASR structural analysis; mapped the sRNA databases onto the ASR and the DSR sequences; hypothesized on the ASR and the DSR functional role; carried out deep P. notatum Illumina floral transcriptome sequencing at specific developmental stages, and assembled it on the 454 reference transcriptome previously constructed, in order to detect additional candidate genes; completed functional analysis of selected candidate genes; and finally, organized a closing meeting
