1Central Potato Research Institute, Shimla-171 001, Himachal Pradesh, India
2Present address: Central Tuber Crops Research Institute, Thiruvananthapuram – 695 017, Kerala, India
*Corresponding author: jeevalatha_a@yahoo.co.in
Online published on 4 December, 2019.
Primer pairs specific to Potato leafroll virus (PLRV), Potato virus X (PVX), Potato spindle tuber viroid (PSTVd) and elongation factor 1-α (ef1α) were designed and used in RT-PCR analysis. Based on specificity, amplicon size and annealing temperature one pair of primers for each target were selected for standardization of duplex and multiplex RT-PCR protocols. An expected amplicons of 267 bp, 492 bp, 566 bp and 703 bp sizes specific to PSTVd, PLRV, PVX and ef1α of potato, respectively were observed from infected leaves and stored potato tubers both in single and mixed infections. The sensitivity of the protocol was tested with randomly collected field samples, glass house, germplasm collections and tissue culture raised microplants and the protocol was found better than DAS-ELISA. Amplification of the internal control, ef1α gene was found throughout the experiments and also in potato tubers stored for more than one month which clearly indicated its use in routine detection of potato viruses in seed production system.
Detection, PLRV, PVX, PSTVd, Multiplex RT-PCR, elongation factor 1-α, internal control
