Characterisation of synthetic DNA probe detecting potato spindle tuber viroid

Abstract

Chemically synthesized DNA fragments complementary to selected regions of the potato spindle tuber viroid (PSTV) genome were cloned into Escherichia coli plasmid pUC9. One of the recombinant plasmids (pIBB4) with a 87 bp insert representing the central region of the PSTV genome (nucleotides 88 to 174) was used after labelling by nick translation for detecting PSTV by dot-blot hybridization. The molecular probe was almost as sensitive as the one carrying the full genomic PSTV copy (pAV401), detecting down to 20 pg of PSTV RNA in 8-15 micrograms of infected tissue. The specificity of the test was high; no signals were created by extracts from healthy plants or plants infected with a variety of common potato viruses. The probe is potentially useful in studies on mixed infections of potatoes with different viruses, and in selection of certified seed material.