Quantitative RT-PCR Analysis of Salt Tolerance Genes in Rice

Quantitative real time PCR was conducted according to Liu et al. [109 (link)] to measure the gene expression of key transporters related to salt tolerance in rice. Total RNA from rice leaves was extracted in the 4th week after stress application using TRIzol reagent (Invitrogen, Carlsbad, CA, USA), and reverse transcription was performed using a SensiFAST cDNA synthesis kit (Bioline, London, UK) according to the manufacturer’s instructions. The expression of genes (HAK1, NHX1, HKT1;4, SOS1, and VHA-c) known to be involved with salt tolerance was assessed by real-time quantitative RT-PCR using a Quantinova Sybr Green Kit (Qiagen, Valencia, CA, USA) in a Rotor-Gene 3000 quantitative PCR thermocycler (Corbett Research, Mortlake, NSW, Australia). G6PDH and Elfa were used as reference genes.