RNA isolation and cDNA synthesis
The infected leaves and root collars from three biological replicates
(one replicate representing a pool of three plants) were flash-frozen
after collection and separately ground with a pestle and mortar. Total
RNA was extracted using AccuZolTM kit (Bioneer, Korea), according to the
manufacturer’s instructions. The genomic DNA in the total RNA was
eliminated by digestion with RNase-free DNase (Thermo Scientific)
following the manufacturer’s protocol. The quantity and purity of total
RNA was evaluated using a Nanodrop NP80 spectrophotometer (Implen
Germany). The cDNA was synthesized from 5 μg of total RNA using the cDNA
Synthesis commercial kit (YTA, Yekta Tajhiz Azma, Iran) according to the
manufacturer’s instructions.