Quantification of Foc colonization through qPCR in resistant and susceptible cultivars grown under elevated CO2
The chickpea seedlings of susceptible cv. JG 62 and resistant cv. WR 315 were grown in open top chambers at different CO2 concentrations [ambient (~380 ppm) and elevated ( 550 ppm and 700 ppm). Seven day old seedlings were inoculated with Foc and samples were collected 7, 14 and 21 days of post inoculation. The sequence specific primers for quantification of Foc were designed using 18S sequences of FOC. qPCR was carried out using pure gDNA and a standard curve was generated by plotting the cycle threshold value (Ct) versus the logarithm of the concentration of each serial dilution of DNA in a 10 fold over a 7-log range from 10 to 1 × 10−4 ng/μL. The quantification of Foc in chickpea was quantified using DNA of infected plant samples through real-time PCR.