RNA-Seq of Saccharum officinarum: young leaf
100 ug of purified mRNA from the same sample was used to construct RNA sequencing library using the Ion Total RNA-seq Kit v2 (Thermo Fisher Scientific, USA) according to manufacturers instruction.mRNA samples were fragmented using RNaseIII (Thermo Fisher Scientific, USA) to obtain samples with a size distribution around 200 nt. Subsequently, the adapters were ligated to fragmented mRNA, and the first strand cDNA synthesis was performed using SuperScriptTMIII (Thermo Fisher Scientific, USA). Ion XpressTM RNA-seq Barcodes (Thermo Fisher Scientific, USA) were added the samples during the second strand cDNA synthesis. Amplified DNA was quantified and multiplexed in equimolar concentration into libraries, which were sequenced on the Ion Proton using Ion PITM Chip Kit v3 chips(Thermo Fisher Scientific, USA).