RNA extraction and quantitative real-time PCR (qRT-PCR)
Total RNA was extracted from 80–100 mg of frozen, homogenizedArabidopsis tissue using the MagMAX Plant RNA Isolation Kit (Applied Biosystems, Foster City, CA, United States) following the manufacturer’s instructions. cDNA was synthesized using the NovoScript Plus All-in-one 1st Stand cDNA Synthesis, SuperMix Kit. Quantitative RT-PCR was performed using the NovoStart SYBR qPCR SuperMix Kit in a QuantStudio TM 12K Flex Real-Time PCR system (Applied Biosystems, Foster City, CA, United States). The thermal cycling conditions were as follows: 95°C for 2 min; 40 cycles of 95°C for 20 sec; and 60°C for 30 sec. The primers were shown in Table S3. Data were analyzed using QuantStudio TM 12K Flex software (Applied Biosystems, Foster City, CA, United States). Significant differences were evaluated using Student’s t -test, and asterisks indicated significant