Detection of avocado sunblotch viroid (ASBV) by dot-spot self-hybridization with a [(32)P]-labelled ASBV-RNA

RNA was extracted from plants infected with avocado sunblotch viroid (ASBV) and was analyzed by electrophoresis in polyacrylamide gel. The ASBV related fraction was eluted from the gel, labelled with [(32)P] using polynucleotide kinase and used as a probe for hybridization with a purified ASBV-RNA preparation dot spotted on nitrocellulose paper. Positive self-hybridization indicated a high degree of internal complementarity. Dot spots of whole cell RNA and of leaf sap from ASBV infected plants were shown to hybridize with the labelled probe. This hybridization procedure proved to be 16-64 times more sensitive in diagnosing ASBV when compared with polyacrylamide gel analysis.