Quantitative RT-PCR Analysis of Poplar Genes

cDNA was synthesized from 1.0 µg total RNA and digested with DNase I using a PrimeScript RT reagent kit for RT-PCR (Takara, Dalina, China) according to the manufacturer’s instructions. The specific primers for poplar (Table S1) were synthesized by the Sangon Biotechnology Company (Shanghai, China). Real-time quantitative RT-PCR was performed with a SYBR Premix ExTaqTM II kit (Takara) with a Roche LightCycler 480 (Roche, Penzberg, Germany). To quantify the relative expression levels of the target genes, the reference gene β-TUB from P. massoniana was used as the internal control [51 (link)], and the primer sequences were as follows: 5’-GTCGTGAATCATGGCATGGC-3’ (forward); 5’-GCCTCACTATCGGTTTCCCA-3’ (reverse). Fold changes were calculated by the relative expressions of treatments compared to the control. Three independent biological repeats were performed for each selected gene (at least two technical repeats per biological repeat).