Quantitative Expression Analysis of Maize Genes

For expression analysis of Zm00001d023265 and Zm00001d023267, total RNA was extracted using an RNAprep Pure Plant kit (Tiangen) from ear leaves of Jing2416K and Jing2416 at three time points: 48 Days After Sowing (DAS) (before the appearance of visible lesions in ear leaves of Jing2416), 55 DAS (lesions just visible) and 66 DAS (lesions clearly visible). First-strand cDNA was synthesized using a SuperScript II kit (Takara) following the manufacturer’s recommendations. Primer pairs were designed using GenScript (https://www.genscript.com/tools/real-time-pcr-taqman-primer-design-tool) and are listed in Supplementary Table S3. Quantitative real-time PCR amplifications were carried out using a SYBR Premix Ex Taq kit (Takara) as described previously (Wang et al., 2017 (link)), with the maize Actin gene (Zm00001d012277) used as an internal reference. Relative expression levels of the two genes were determined by the 2^−ΔΔCT method (Livak and Schmittgen, 2001 (link)).

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Wang S., Zhang R., Shi Z., Zhao Y., Su A., Wang Y., Xing J., Ge J., Li C., Wang X., Wang J., Sun X., Liu Q., Chen Y., Zhang Y., Wang S., Song W, & Zhao J. (2020). Identification and Fine Mapping of RppM, a Southern Corn Rust Resistance Gene in Maize. Frontiers in Plant Science, 11, 1057.

Publication 2020

RNAprep Pure Plant kit SuperScript II kit SYBR Premix Ex Taq kit

Actin Cdna Expression genes Gene Maize Plant Primer Quantitative real time pcr

Corresponding Organization :

Other organizations : Beijing Academy of Agricultural and Forestry Sciences

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Variable analysis

independent variables

Genotype (Jing2416K and Jing2416)
Time point (48 DAS, 55 DAS, 66 DAS)

dependent variables

Expression levels of Zm00001d023265 and Zm00001d023267

control variables

Maize Actin gene (Zm00001d012277) used as internal reference

controls

No positive or negative controls were explicitly mentioned.

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