Transcriptome Analysis of Aluminum Stress

Leaf samples from the treatments with Al concentration of 0 (L0), 1 (L1) and 4 (L4) mM were collected for RNA-Seq analysis. The Al concentration at 1 mM, was used as the control (CK) because of good root growth at that concentration. Equal amounts of frozen leaves from three individual plants were mixed as one biological replicate. There were three biological replicates for each treatment. Total RNA was extracted using an RNeasy Plant Mini Kit (TaKaRa, Dalian, China), according to the manufacturer’s protocol. The concentration and quality of total RNA were examined using a NanoDrop 2000c spectrophotometer (NanoDrop, Wilmington, DE, USA) and 1% w/v agarose gel electrophoresis. The cDNA libraries were sequenced by a llumina Hiseq Xten platform by the Bioacme Biotechnology Co., Ltd. (Wuhan, China). The high-quality clean reads from each sample were mapped to the reference genome^{48 (link)} using Hisat2.