RT-PCR detection and identification of three species of cucumoviruses witha genus-specific single pair of primers

Reverse transcription and polymerase chain reaction (RT-PCR) was used for d etection and identification of three cucumoviruses (cucumber mosaic virus, CMV; peanut stunt virus, PSV; tomato aspermy virus, TAV) in various plants sources with a single pair of primers, designed as CPTALL-3 and CPTALL-5. T he pair of cucumovirus genus-specific primers that flank the coat protein g ene were designed and used to amplify a DNA fragment of approximately rangi ng from 938 to 966 bp. The RT-PCR with the set of primers specifically ampl ified the target size of DNA fragment in all the tested cucumoviruses (CMV S-TA, S-IB and S-II, PSV and TAV). No DNA product of any length was produce d when brome mosaic virus or tobacco mosaic virus RNA was used as templates . The cucumoviruses examined were differentiated by PCR-restriction fragmen t length polymorphism with different enzymes. This indicates that the desig ned primers are only specific for the cucumoviruses and useful for reliable information of identification of members of the Cucumovirus genus, (C) 199 9 Elsevier Science B.V. All rights reserved.