RNA Extraction and Quantitative Real-Time PCR (qRT-PCR) Analysis

Total RNA from different M. truncatula tissues was extracted using the Trizol method, and RNA was treated with Turbo DNase (Ambion) to remove genomic DNA. Five micrograms of total RNA was used for cDNA synthesis by using the SuperScript^® III First-Strand Synthesis System for RT-PCR kit (Life Technologies). qRT-PCR was performed using the ROCHE LightCycler^® 96 detect system with the TransStart Tip Green qPCR SuperMix (TransGen Biotech). qRT-PCR data were obtained using three biological replicates and transcripts were normalized to MtActin. Primers used were listed in Supplementary Table S1.