Simultaneous Detection Of Two Viroids Infecting Grapevines In Taiwan By Multiplex Rt-Pcr

Multiplex RT-PCR was used to detect Hop stunt viroid (HSVd) and Grapevine yellow speckle viroid-1 (GYSVd-1). Twenty five samples were collected from grapevine farms located at Changhua county of Taiwan. Specific primer pairs used for the detection of HSVd and GYSVd-1 were designed according to the conserved sequences of the viroid genomes. Multiplex one-step RT-PCR followed gel electrophoresis analysis showed that two DNA fragments with the sizes of 367 bp and 300 bp indicated the infection of GYSVd-1 and HSVd, respectively. The preliminary data suggests that the frequency of GYSVd-1 and HSVd infection is 80% and 90%, respectively.