Heat stress response of perennial ryegrass (PRJCA021952)
Fourteen-day-old perennial ryegrass leaves treated at 42oC for 0, 2, 24, and 48 hours were used for transcriptome sequencing. Each treatment or control contained three replicate samples. The total RNA was extracted using the RNAprep Pure Plant Kit (Tiangen, Beijing, China). A total amount of 1 µg RNA per sample was used as input material for the RNA sample preparations. Sequencing libraries were generated using Hieff NGS Ultima Dual-mode mRNA Library Prep Kit for Illumina (Yeasen Biotechnology (Shanghai) Co., Ltd.). The libraries were sequenced on an Illumina NovaSeq platform to generate 150 bp paired-end reads. The raw reads were further processed with a bioinformatic pipelinetool, BMKCloud(www.biocloud.net) online platform. Trimmomatic (Bolger et al., 2014) was used to remove adapter and low-quality sequences. HISAT2 (Kim et al., 2015) was used to quickly and precisely compare Clean Reads with perennial ryegrass genomes (Nagy et al., 2022). StringTie (Pertea et al., 2015) was used to assemble reads and calculate their expression levels.