1a.Objectives (from AD-416):
Fine mapping, metabolomics and transcriptomics of genes for soybean aphid resistance.
1b.Approach (from AD-416):
Recombinant inbred line (RIL) populations and near isogenic lines (NILs) will be used for conducting the fine mapping, metabolomics, trancritomics and proteomics studies with help from collaborators.
This was a one year grant, thus this one year report also represents the life time overview. More than 5000 near-isogenic line progeny were harvested in November 2012 for this project. Nearly 200 kilobases in the Rag5 region of two parental lines is being sequenced after amplifying with long range PCR (10-15kb each). SNP markers will be identified in the region and NILs will be genotyped to identify recombinants to further fine map the Rag5 region, (ii) the study on metabolomics was completed and manuscript was submitted for journal review, (iii) the transcriptomic study on the Rag5 NILs are in progress. RNAs were extracted from leaf tissues collected at five time points (0, 8, 16, 24 and 48 hrs) after infestation of soybean seedlings with the soybean aphid biotype 2. Preparation of cDNA libraries using the RNA samples are in progress for sequencing.
Collaborators at Univ. of Missouri Columbia completed studies on proteomics and transcriptomics of the Rag2 NILs. A manuscript on the proteomics and transcriptomics data is in preparation. This project relates to and expands on objective 2B (Genetic characterization and molecular mapping of aphid resistance genes) of the parent project.