The total RNA was extracted from samples of fungal inoculated leaves using the optimized extraction procedure described by Zhang et al. (2014) (link).
The SYBR Green Premix Ex Taq™ II quantitative PCR system (Takara, Dalian) was used for qPCR analysis. All experiments involving q-PCR were performed on a Q7 Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). The actin gene (GenBank: aK458277.1) was used as the reference gene. The PCR reaction and program were modified according to the manual. The PCR reaction (a total reaction volume of 10 µL) comprised 5 µL 2 × SYBR Green PCR Master Mix, 3 µL of the cDNA product, 1 µL of primer mix, and 1 µL of DNase/RNase-free water. The quantitative PCR thermal cycler program included 95 °C for 10 s, followed by 40 cycles at 95 °C for 5 s and 60 °C for 31 s. All primers for q-PCR were synthesized by the same company (AoKe, Yangling) (Table S4 ).
The SYBR Green Premix Ex Taq™ II quantitative PCR system (Takara, Dalian) was used for qPCR analysis. All experiments involving q-PCR were performed on a Q7 Real-Time PCR System (Applied Biosystems, Foster City, CA, USA). The actin gene (GenBank: aK458277.1) was used as the reference gene. The PCR reaction and program were modified according to the manual. The PCR reaction (a total reaction volume of 10 µL) comprised 5 µL 2 × SYBR Green PCR Master Mix, 3 µL of the cDNA product, 1 µL of primer mix, and 1 µL of DNase/RNase-free water. The quantitative PCR thermal cycler program included 95 °C for 10 s, followed by 40 cycles at 95 °C for 5 s and 60 °C for 31 s. All primers for q-PCR were synthesized by the same company (AoKe, Yangling) (
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