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To verify the reliability and reproducibility of the results obtained in the RNA-Seq, some key genes involved in metabolic pathways were selected and further verified by RT-qPCR. The RT-qPCR primers are listed in Table S5. RNA samples were treated with DNase I and further reverse transcribed to cDNA using SYBR Premix Ex Taq II (Tli RNaseH Plus) kit. The RT-qPCR was run on a CFX96 RT-qPCR Detection System. The housekeeping gene ZmActin1 was set as the endogenous control [66]. The relative expression of each gene was calculated with the 2-ΔΔCt method [67, 68].
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