Centre : Central Potato Research Institute(CPRI)
Name of the Protocol
Detection and quantification of P. infestans in host tissues using PCR protocols
Nature : Modification of existing protocol
Crop / Pathogen : Potato/Phytophthora infestans
The rapid NaOH tissue assay was used with primer set INF FW2 and INF REV for detection of P. infestans from artificially infected potato tubers, seed tubers (storage), infected potato leaf, pure culture of P. infestans. A single PCR product approximately 600 bp in size was detected. DNA from healthy tubers did not amplify with these primers and could not be detected by ethidium staining on agarose gel. The standardised PCR protocol could detect up to a level of 10 fg of genomic DNA. P. infestans was detected in artificially infected tubers (Cv Kufri Jyoti) by the PCR assay and ethidium bromide staining of agarose gels at 4 & 6 days post-inoculation but not at 2 days post-inoculation, before the occurrence of visible symptoms. Moreover, amplified products of expected size were obtained from tuber lesions and symptomless areas at various distances from the tuber lesions (5, 10 & 20 mm away from the lesion).