RNA extraction and RT-qPCR
Total RNA was isolated using TRIzol reagent (Invitrogen) according to
the manufacturer’s instructions. RNA quantity and purity were assessed
with NanoQuant spectrophotometer. First strand cDNAs were synthesized
from DNase I-treated total RNA using cDNA Synthesis Kit (Takara). Gene
expression was normalized using tubulin (At5g23860) as the
reference gene, and relative gene expression was calculated as the mean
of three biological replicates and three technical replicates. Gene
specific primers used for RT-qPCR are listed in Table S1.