PJB-2010-342
MOLECULAR DETECTION OF POTATO LEAF ROLL POLEROVIRUS THROUGH REVERSE TRANSCRIPTION POLYMERASE CHAIN REACTION IN DORMANT POTATO TUBERS
ALI RAZA AWAN1, 3*, IKRAM UL HAQ2, MASROOR ELLAHI BABAR1 AND IDREES AHMAD NASIR3
Abstract
A Reverse Transcription-Polymerase Chain Reaction (RT-PCR) technique was applied for the detection of Potato leaf roll polerovirus (PLRV) in dormant potato tubers. A primer pair was designed from the coat protein-encoding fragment of the PLRV genome that amplified a 336-bp product. The amplified product was detected in nucleic acid preparations from leaves and tubers of 5 cultivars and from purified virions. The specificity of the RT-PCR product was confirmed through southern blot analysis. The primer pair used in the RT-PCR did not produce any non-specific product from 7 other potato viruses. Sensitivity of RT-PCR was confirmed by detecting PLRV from known mixture of PLRV and randomly selected potato virus. Dilution of 1:1000–1:4000 and 1:200–1:1000 were used to detect viral load from foliage and tuber, respectively. RT-PCR efficiently detected PLRV in sprouting tubers as well as dormant tubers stored at 20– 25°C for 4 months.
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