Analysis of Gene Expression by qRT-PCR

Total RNA was prepared using Trizol methods (Invitrogen, Carlsbad, CA, USA). The first-strand cDNA was synthesis by SuperScript II reverse transcriptase using oligo (dT) as a primer (Invitrogen). Reverse transcriptional products were used as a template for real-time PCR and actin7 of Arabidopsis and GAPDH gene in rice as inner control separately. The RT-PCR amplification step was performed using the SYBR^® Premix Ex Taq^TM II (TaKaRa) and a RT-PCR detector (TaKaRa Smart Cycler II system) by using the SYBR Green I chimeric fluorescence method according to the manufacturer’s instruction. Reactions were performed in triplicate, including the no-template and no-reverse-transcriptase controls, and were monitored using an Applied Biosystems 7500 RT-PCR instrumentation outfitted with SDS software version 1.3.1.