2.8 qRT-PCR
Total RNA samples were isolated with a SuperPure Plantpoly RNA Kit (Gene
Answer) following the manufacturer’s instructions. RNase-free DNAse I
(New England Biolabs) was used to remove any trace genomic DNA.
First-strand complementary DNA (cDNA) was synthesized with Reverse
Transcriptase M-MLV (Takara) and oligo (dT) 12-18 as a primer. The PCR
reactions were performed with SYBR Green kit (Roche) on a
LightCycler® 96 SW 1.1 cycler (Roche). TheNtL25 gene was chosen as the internal reference control
(Schmidt & Delaney, 2010), and the
realtive expression levels of the target genes were calculated as
described previously (Livak & Schmittgen,
2001). All the gene-specific primers for qRT-PCR were listed in
Supplemental Table S3.