identification of causal tnt1 insertions in Medicago truncatula by RNA-seq analysis

The mutations causing Medicago truncatula TNT1 insertion lines to fail producing functional symbiotic nodules were identified through the analysis of chimeric transcripts using RNA-seq. In each line, individuals with a WT or mutant phenotype were pooled and the transcriptome of their nodules were sequenced. This protocol allowed the rapid identification of causal mutations in several lines.