LANGRELL, Stephen R.H.; MOREL, Olivier; ROBIN, Cécile

doi:10.1016/j.funbio.2011.04.009

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LANGRELL, Stephen R.H.
JRC Institute for Prospective Technological Studies [IPTS]

MOREL, Olivier
Biodiversité, Gènes & Communautés [BioGeCo]

ROBIN, Cécile
Biodiversité, Gènes & Communautés [BioGeCo]

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Fungal Biology. 2011, vol. 115, n° 7, p. 672-682

Elsevier

Résumé en anglais

Soil borne Phytophthora cinnamomi and Phytophthora cambivora are considered the most pathogenic species associated with chestnut (Castanea sativa) decline in Europe. Mapping their incidence and distribution from nursery and plantation soils may offer valuable information for limiting spread. As conventional biological baiting and taxonomic confirmation is generally time consuming, labour, logistically and space intensive, we have focused on the development of a specific touchdown nested multiplex Polymerase Chain Reaction (PCR) approach for the simultaneous detection of both species direct from soil. Pre-existing and novel primers, based on Internal Transcribed Spacer (ITS) sequences, have been evaluated for their specificity and use in a multiplex capacity in various combinations. Coupled to this we have modified a mechanical lysis procedure for DNA extraction from up to 10 g of chestnut under storey soils (ranging from 0.5 to 25 μg DNA g−1 fresh soil). Using serial dilutions and/or polyvinylpolypyrrolidone chromatography purification, both species have been successfully detected, in artificially and naturally infected soils. Levels of assay detection are comparable to other Phytophthora species where PCR based diagnostic systems have been reported. A qualitative evaluation of this approach against conventional baiting is presented.< Réduire

Mots clés

CHESTNUT

SOIL

RELATION PLANTE-SOL

GESTION DE LA MALADIE

Mots clés en anglais

PHYTHOPHTHORA CINNAMOMI

PHYTHOPHTHORA CAMBIVORA

DETECTION

DISEASE MANAGEMENT

INK DISEASE

NESTED-POLYMERASE CHAIN REACTION (PCR)

MULTIPLEX

TOUCHDOWN

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Touchdown nested multiplex PCR detection of Phytophthora cinnamomi and P. cambivora from French and English chestnut grove soils

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