quantitative real-time PCR (qRT-PCR)
To verify the expression levels of the DEGs, qRT-PCR amplifications of
17 randomly selected genes were performed on a LightCycler 480 II
instrument (Roche Diagnostics GmbH, Germany) using the RNAs as for the
library construction and illumina sequencing. Gene PU1 (Gohir.A01G131900 ) was used as internal control. The primers of
these genes for the qRT-PCR analysis were designed and listed in Table
S2. Relative expression of these genes were calculated according to the
2–∆∆CT method (Schmittgen & Livak, 2008). T-test was
applied for significant differences among treatments at P < 0.05 level. The results were shown as the means ± standard
error of three replicates.