Total RNA isolation and first-strand cDNA synthesis of all samples were performed as described by Qian et al. (2018) [49]. For qRT-PCR analysis, 20 μL reaction volumes, including 10 μL SYBR Premix Ex Taq, 1.6 μL forward/reverse primers, 2 μL cDNA and 6.4 μL distilled water, were analyzed on a Roche 384 real-time PCR machine (Roche). The qRT-PCR program began with 95 °C for 10 min, followed by 45 cycles of 94 °C for 10 s, 60 °C for 15 s and 72 °C for 12 s; thus, a melting curve was added. CsPTB [90], as an actin gene, was used to quantify the relative expression levels of each CsARG according to the 2-ΔCt or 2−ΔΔCt methods [91]. Three replications were generated for each RNA sample for quantitative analysis, and the qRT-PCR primers are listed in Additional file 1.

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