design: Barley (Hordeum vulgare cv. RGT Planet) grown in glasshouse for three weeks. Each sample comprised of 24 (2 mm) biopsy punches taken from leaf tissue; 24 punches were made up from 3 punches taken from two regions 10-15 cm from the tip of the second leaf from 4 leaves, each from a different plant. Total RNA was extracted using Norgen Plant/Fungi Total RNA Purification Kit. An equal amount of RNA from the six control samples was pooled into one sample and 1000 ng was taken from the pool for library preparation using the Oxford Nanopore Technologies (ONT) Direct RNA sequencing kit (SQK-RNA004); recommended protocol was kept except for changing RCS volume to 0.1 uL. The libraries were loaded on a FLO-PRO004RA cell and sequenced on an ONT PromethION 2 Solo.