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The feasibility of tetraplex RT-PCR in the determination of
PVS, PLRV, PVX and PVY from dormant potato tubers
Hidayet Bostan* and Pinar Kubra Peker
Atatürk University, Faculty of
Agriculture, Department of Plant Protection, Campus,
TR-25240, Erzurum, Turkey.
*Corresponding author. E-mail:
hibostan@yahoo.com.
Accepted 13 August, 2009 |
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Dormant potato tubers belonging to cvs., namely, Agria,
Granola and Marfona known to be infected with potato viruses
(Potato leafroll virus, PLRV; Potato virus S,
PVS; Potato virus X, PVX and Potato virus Y,
PVY) were tested with uniplex RT-PCR and strong bands
specific to each virus were obtained from cultivars. When
cDNA synthesized for uniplex RT-PCR was used for tetraplex
RT-PCR, the bands obtained from PVS, PLRV and PVX were too
faint to be photographed and there is no any observed band
for PVY. To improve the band density, the concentration of
oligo dT primer in RT was increased from 20 to 40 ng in the
subsequent experiments. The increasing of oligo dT primer
concentration in RT increased the band density for PVS and
PVX, but not PVY. Upon this, different amount of total RNA
were tested in RT stage. The best result was obtained from 5
µl of total RNA and followed by 3.5 and 2.5 µl applications.
In order to determine the effect of cDNA amount in PCR, 2 µl
cDNA + 23 µl PCR, 5 µl cDNA + 20 µl PCR and 5 µl cDNA + 25
µl PCR mixture were compared. However, no distinct
differences were observed among various cDNA amounts. As a
result, instead of tetraplex RT-PCR, it is suggested the use
of triplex RT-PCR for reliable detection of PLRV, PVS and
PVX. However, uniplex PCR could be suggested for reliable
detection of PVY from this study by using the same cDNA.
Key words:
Potato viruses, detection, dormant tubers, tetraplex RT-PCR. |
