A method to detect and quantify Eutypa lata and Diplodia seriata-complex DNA in grapevine pruning wounds

Jérôme Pouzoulet, Philippe E. Rolshausen, Marco Schiavon, Sebastiaan Bol, Renaud Travadon, Daniel P. Lawrence, Kendra Baumgartner, Vanessa E. Ashworth, Gwénaëlle Comont, Marie France Corio-Costet, Romain J.G. Pierron, Xavier Besson, Alban Jacques

Producción científica: Articlerevisión exhaustiva

21 Citas (Scopus)

Resumen

Trunk diseases are factors that limit sustainability of vineyards worldwide. Botryosphaeria and Eutypa diebacks are caused by several fungi belonging to the Botryosphaeriaceae and Diatrypaceae, respectively, with Diplodia seriata and Eutypa lata being two of the most common species. Previous information indicated that the traditional isolation method used to detect these pathogens from plant samples could underestimate their incidence levels. In the present study, we designed two sets of primers that target the β-tubulin gene and that are amenable for quantitative real-time PCR (qPCR) Sybr-Green assays for the detection and quantification of D. seriata-complex (DseCQF/R) and E. lata (ElQF/R) DNA. The design of a species-specific assay was achieved for E. lata. For D. seriata, a species-specific assay could not be designed. The low interspecific diversity across b-tubulin genes resulted in an assay that could not discriminate D. seriata from some closely related species either not yet reported or presenting a low prevalence on grapevine, such as D. intermedia. We validated our technique on grapevine spur samples naturally and artificially infected withD. seriata and E. lata during the dormant season. Experimental grapevines were located in two counties of northern California where the incidence of both pathogens was previously reported. The qPCR assays revealed that a high frequency of pruning wound infections (65%) was achieved naturally by E. lata, while low infection frequency (less than 5%) was observed using the reisolation method. For D. seriata-complex, low (5%) to no natural infection frequencies were observed by the qPCR and the reisolation method, respectively. These results also provided evidence that our qPCR detection methods were more sensitive to assess the incidence of E. lata and D. seriata-complex in plant samples, than traditional isolation techniques. Benefits of molecular methods for the detection of canker pathogens in the field under natural conditions are discussed.

Idioma originalEnglish (US)
Páginas (desde-hasta)1470-1480
Número de páginas11
PublicaciónPlant Disease
Volumen101
N.º8
DOI
EstadoPublished - ago 2017
Publicado de forma externa

ASJC Scopus subject areas

  • Agronomy and Crop Science
  • Plant Science

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