RNAseq of leaf tissue of Medicago truncatula R108 WT and mtftb1/2 gene-edited double mutant at two different developmental stages and vernalised conditions
RNAseq of leaf tissue of Medicago truncatula R108 WT and mtftb1/2 gene-edited double mutant at two different developmental stages and vernalised conditions
Fully expanded leaf tissue was sampled of Mtftb1/2 and control plants grown under LD (16h light/8h dark) for 2 weeks after 14 days of vernalization (Vernalized conditions) or plants grown for 60 days under LD (16h light/8h dark) without previous vernalization (non-vernalised conditions).  Scarified seeds (seed surface scratched using fine sandpaper) were water imbibed and germinated at room temperature in a Petri dish in the dark. When radicles extended to a sufficient length (~10 mm), the seedlings were transferred into growth media (3 parts soil (Yates Professional Potting Mix, Auckland, New Zealand) and 1 part vermiculite) in controlled environment rooms. Plants were grown in a growth cabinet supplied with white light (TL5 28W low-pressure mercury discharge lamp, Philips, Auckland, New Zealand) ~90–110 µmol/m−2s−1) in 65% relative humidity at 25 °C or in a growth room, supplied with white light (~80–100 µmol/m−2s−1) in ~60% relative humidity at 18–23 °C. For vernalized conditioning, Medicago seeds were scarified and water imbibed overnight in the dark at room temperature before being placed at 4 °C in the dark for two weeks in a Petri dish. The seedlings were then planted into soil and put into the growth cabinet under the specified light conditions. Total RNA was extracted using the Spectrum Plant Total RNA kit (Sigma-Aldrich, St. Louis, MO) Library construction and sequencing were performed at Otago Genomics Facility (OGF) (https://www.otago.ac.nz/genomics)