Total RNA of WFTs was extracted with the RNAiso Plus reagent (Takara) following the manufacturer’s protocol. After digestion with a gDNA digester (Yeasen), cDNA was synthesized using the RevertAid First-Strand cDNA Synthesis Kit (Yeasen). The reaction mixture for PCR amplification consisted of 2 μL of cDNA, 0.25 μL of forward and reverse primers, and 5 μL TB Green Premix Ex Taq II (Takara) in a final 10-μL reaction volume. The relative expression levels of target genes were calculated by the 2-ΔΔCT method (54 (link), 55 (link)). Gene expression levels in WFTs were measured by qRT-PCR between day 2 and day 5 of the bioassay. The elongation factor 1α (EF1α) was chosen as a reference gene (56 (link)).
Protocol detail
Gene Expression Analysis of WFTs
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Corresponding Organization :
Other organizations : Hubei University, Max Planck Institute of Molecular Plant Physiology
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Variable analysis
independent variables
- Day of the bioassay (between day 2 and day 5)
- Relative expression levels of target genes
- Elongation factor 1α (EF1α) as a reference gene
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
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