Routine Detection of Chrysanthemum Stunt Viroid in Chrysanthemum Plants by Reverse Transcription - Polymerase Chain Reaction

Abstract

For detection of chrysanthemum stunt viroid (CSVd) in chrysanthemum plants, the reverse transcription-polymerase chain reaction (RT-PCR) method described by Hooftman et al. (1996) has been adjusted for practical use. This led to minor changes in the homogenization of leaf material, extraction method and conditions during PCR. The results obtained by the RT-PCR method were similar to those obtained by the current method, i.e. return-polyacrylamide gel electrophoresis (R-PAGE). With healthy chrysanthemum plants, also a slight reaction was obtained in the RT-PCR method. In this case however, the PCR product was some nucleotides smaller than in the case of infected plants. It appeared that the present RT-PCR method can be used for the detection of CSVd in chrysanthemum at practical scale. The reliability of the method still has to be proved in large-scale application, during which the results of this method are compared with those of the current method.