Rapid detection and discrimination of fabaviruses by flow-through hybridisation with genus- and species-specific riboprobes

Abstract

Viruses cause significant damage in agricultural crops worldwide. Disease management requires sensitive and specific tools for virus detection and identification. Also, detection techniques need to be rapid to keep pace with the continuous emergence of new viral diseases. The genus Fabavirus is composed of five viruses infecting many economically important crops worldwide. This research describes the development of a procedure based on flow-through hybridisation (FTH), which is faster than and as sensitive as conventional hybridisation for virus detection in tissue-prints from infected plants. Six digoxigenin-labelled RNA probes were synthesised with two levels of specificity: (a) five specific for each viral species within this genus, and (b) a genus-specific probe that hybridises with a nucleotide sequence signature only found in the 5-untranslated region of the genus Fabavirus, which is the first of this type reported for plant viruses. The new procedure developed is useful for rapid detection and discrimination of the five fabaviruses identified so far and opens the possibility of discovering new species of this genus.