Rapid Detection of Potato Spindle Tuber Viroid (PSTV) by Dot Blot Hybridization

A PSTV probe of 350 bases, prepared in our laboratory, was used to detect picogram quantities of viroids by a simplified, improved, dot blot, hybridization technique in crude potato and tomato extracts. Optimum conditions for certification of PSTV-free potato plants were established, involving formamide concentration, washing stringencies, exposure time during autoradiography and method of probe radiolabelling. Ten pg of purified PSTV in water and 50 pg of purified PSTV added to healthy plant extract were detected. Also hybridization signals could be detected from as little as 0.075 mg of infected plant tissue.

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