RNA-seq of T. thianschanica seeds
Total RNA was extracted using Trizol reagent (Invitrogen, CA, USA). following the manufacturer's procedure. The total RNA quantity and purity were analysis of Bioanalyzer 2100 and RNA 1000 Nano LabChip Kit (Agilent, CA, USA) with RIN number > 7.0. Poly(A) RNA is purified from total RNA(5 ug) using poly-T oligo-attached magnetic beads using two rounds of purification. Following purification, the mRNA is fragmented into smallpieces using divalent cations under elevated temperature. Then the cleaved RNA fragments werereverse-transcribed to create the final cDNA library in accordance with the protocol for the mRNASeq sample preparation kit (Illumina, San Diego, USA), the average insert size for the paired-endlibraries was 300 bp (50 bp). And then we performed the paired-end sequencing on an Illumina Novaseq 6000 at the (LC Sceiences,USA) following the vendors recommended protocol.