Transcriptional Analysis of Wheat Biotic Stress

The qPCR analyses were performed to see the changes in the expression profile of key genes of phenylpropanoid cascade and MAPK and WRKY transcription factors along with the heteromeric G protein, Gα, and Gβ in the wheat genotypes treated with B. sorokiniana, crude toxins, and different doses of Bipolaroxin at 7 days of inoculation under glasshouse conditions. Total RNA was extracted using RNA isolation kit (Agilent, Santa Clara, CA, USA) and cDNA was synthesized using cDNA synthesis kit (BioRAD, Hongkong, China) following the manufacturers’ instructions. The quality and quantity of cDNA were analyzed using a nanodrop spectrophotometer (Thermo Scientific, Waltham, MA, USA). Expression analyses were performed using gene-specific primers (Supplementary Tables S2–S4) using sybr green master mix (Agilent, India) on the BioRAD Real Time PCR System (Model: MJ MiniOpticon, BioRAD, India) according to Singh et al. [42 (link)]. Actin was taken as the internal control. The relative transcript levels were calculated using the 2^−ΔΔC_T method [43 (link)].