Rnaex solution

RnaExTM solution (Generay) was used for total RNA extraction. First-strand cDNA was synthesized by a reverse transcription kit (Transgen). Real-Time quantitative PCR (RT-qPCR) was performed as previously described [62 (link)]. The primers for gene expression analysis were used in this study can be found in Additional file 10. The relative expression levels were calculated using PpAdePRT [57 (link)] as expression control.

WT and Ppatg3 plants were grown on BCD medium for 28d. Total RNA of the whole plant was extracted by using a RnaExTM solution (Generay). cDNA library construction, sequencing and data analysis were carried out by the BGISEQ-500 platform established by BGI-Tech (Wuhan, China). The reads alignment and expression level calculation were conducted by using Bowtie 2 [59 (link)] and RSEM [60 (link)], respectively. A differential expression transcript/gene with at least two-fold change and p-value ≤0.001 was used as further analysis [61 (link)]. R program “princomp”, “phyper” and “pheatmap” were used to conduct PCA analysis, KEGG pathway enrichment and construction of heat maps, respectively. The RNA-Seq data of the present study had been deposited at the BIG genome sequence archive (GSA) under Bioproject identifier PRJCA001964 with accession number: SAMC116811 to SAMC116814.