Centre : Central Tuber Crops Research Institute(CTCRI),TVM
Name of the technology :
PCR based diagnosis of P. colocasiae
Early, rapid and reliable identification and quantification of pathogen is an integral part of any disease management initiative. Stepping away the conventional approach, here we have developed and standardized a conventional and real time PCR exploiting three target regions viz. RAS-related protein (Ypt1), G protein alpha-subunit (GPA1) and Phospho-ribosylanthranilate isomerase (TRP1) for the detection and quantification of P. colocasiae in infected propagules, field and seed material. The PCRs were conducted in a volume of 25 µL, which included 100 ng of template DNA, 100 µM each deoxynucleotide triphosphate, 10 ng of each primer, 1.5 mM MgCl2, 19 Taq buffer (10 mM Tris-HCl pH 9.0, 50 mM KCl, 0.01% gelatin), 1 U of Taq DNA polymerase (Merck GeNei, India). The following optimized PCR regime was used: denaturation at 95 °C for 2 min, followed by 35 cycles of 95 °C for 30 s, 55 °C for 45 s, 72 °C for 1 min and a final extension at 72 °C for 8 min.
The developed PCR assay proved to be a robust and reliable technique to detect P. colocasiae in taro planting material and for assessing the distribution of pathogen within fields, thus aid in mitigating taro leaf blight.
Highly specific, sensitive and robust than the available technology
Status of Commercialization : Not applicable