Total RNA was extracted using TaKaRa MiniBEST Plant RNA Extraction Kit (Takara, Dalian, China) from roots according to the manufacturer’s instruction. Four samples (Yanjiang, Yanjiang NT, 9901, 9901 NT) were collected following the same samples treatments procedure as that in RNA sequencing. For each sample, 3 µg of total RNA was used to synthesize first-strand cDNA with SuperScriptII reverse transcriptase (Takara, Dalian, China). For qRT-PCR, the reaction preparation, application parameter settings and quantitative analysis were performed as previously described (Chen et al., 2018). The reactions were performed using the ABI Prism 7000 Real-time PCR system (Applied Biosystems, USA). The Salix purpurea Actin1 gene (SapurV1A.0655s0050.1) were used as reference genes. The gene-specific primers for the 15 selected genes are listed in Table S1.

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