Abstract
Reverse transcription polymerase chain reaction (RT‐PCR) was used for the detection of Citrus exocortis viroid (CEVd) in total nucleic acids from infected citrus indicator hosts or directly from field trees. Grapevine yellow speckle viroid‐1 (GYSVd‐1) and Australian grapevine viroid (AGVd) were also detected by this technique in total nucleic acids from infected grapevine plants. Three protocols for nucleic acid extraction were tested: a phenol‐based method, a protocol using a commercial reagent (Trizol) and a method in which extraction with a high‐salt buffer was combined with partial purification using non‐ionic cellulose. The third protocol, coupled to RT‐PCR, seems to be the most suitable, allowing detection of the three viroids in Tunisian citrus and grapevine samples. This method should be used in a testing programme to produce certified viroid‐free planting material.
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