Detection of Ralstonia solanacearum (biovar 2A) in stems of symptomless plants before harvest of the potato crop using post-enrichment DAS-ELISA

Abstract

The detection of Ralstonia solanacearum (biovar 2A) in stems of symptomless plants before harvest of the potato crop, instead of tubers, would not only save highly valued planting material but would be less time-consuming and would also enhance farmers' market decisions. Although pathogen detection in stems has been proven efficient for ring rot, this has never been investigated for bacterial wilt (BW). Therefore the possibility of detecting BW latent infection in stem pieces about three weeks before harvest was assessed in 57 fields of the Andean highlands of Peru. Two sensitive, specific and user-friendly serological methods were used to detect the pathogen in latently infected tubers and stems: double-antibody sandwich (DAS)-ELISA and indirect ELISA on nitrocellulose membrane (NCM) after enrichment of the plant extracts in a semi-specific broth. Optimum sample sizes of stems and tubers were evaluated for 37 potato crops showing between 0 and 0·1% BW incidence using a binomial distribution model to calculate the detection probabilities. Although results of detection using the two serological techniques had 100% concordance, detection probabilities were higher using DAS-ELISA, whatever the plant part tested. BW detection probabilities were higher for tubers than for stems; a 99% detection probability was obtained by analysing 400 stems sections or 250 tubers using DAS-ELISA. Detection of BW infection in symptomless plants 20 days before harvest using post-enrichment DAS-ELISA is a reliable and user-friendly technique that can easily be used by national plant protection services and seed programmes in developing countries. © 2009 BSPP.