*2.6. RNA Extraction, Quantification, and cDNA Synthesis*

The total RNA from the leaves was extracted by using a ThermoFisher scientific® Gene JET plant RNA purification kit, according to the manufacturer's protocol. The RNA concentration was calculated by Nanodrop, and was utilized for cDNA synthesis, using a ThermoFisher scientific® cDNA synthesis kit.

#### *2.7. Primer Designing and RT-PCR*

RT-PCR (BIO-RAD) was performed, to examine the expressions of thaumatin-like protein, β-1,3-glucanase and chitinase genes. Total cDNA was used as a template. The primers used in this experiment are given in Table 1. PCR was carried out in a 25 μL reaction mixture comprising 16 μL of water, 2.5 μL of buffer, 1.5 μL of MgCl2, 1.5 μL of dNTPs, 0.5 μL of Taq, 1 μL of template, and 1 μL of both forward and reverse primers. The thermal profile was as follows: 5 min at 94 ◦C, 25 cycles of 40 s at 94 ◦C, 1 min at 49 ◦C, 1 min at 72 ◦C, and a one-step final extension of 5 min at 72 ◦C.


**Table 1.** Primers used in this experiment.
