flo.a spike transcriptome project
Bowman and flo.a were grown in a greenhouse conditions (20°C in day / 15°C at night, 16h of light / 8h of dark) in a 24-well plate. Immature spike samples of Bowman and flo.a plants were collected at Waddington stages W2.0 (Double ridge), W3.0 (Lemma primordium), W4.5 (Awn primordium) and W5.5 (White anther). Due to the 'flo' spikelet phenotype only existing in the upper-mid portion of spike, the whole spike was crossed-sectioned into two parts (upper-mid and basal) at stages W3.0, W4.5 and W5.5 under a stereoscopic microscope. A total of 15-48 spikes per sample and three biological replicates were harvested and used for RNA extraction. All sample collections were conducted between 11:00 am – 14:00 pm within a day. After verifying RNA quality, the sequencing libraries were prepared and sequencing was conducted using the NovaSeq 6000 platform.