Touchdown nested multiplex PCR detection of Phytophthora cinnamomi and P. cambivora from French and English chestnut grove soils
Language
en
Article de revue
This item was published in
Fungal Biology. 2011, vol. 115, n° 7, p. 672-682
Elsevier
English Abstract
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 ...Read more >
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.Read less <
Keywords
CHESTNUT
SOIL
RELATION PLANTE-SOL
GESTION DE LA MALADIE
English Keywords
PHYTHOPHTHORA CINNAMOMI
PHYTHOPHTHORA CAMBIVORA
DETECTION
DISEASE MANAGEMENT
INK DISEASE
NESTED-POLYMERASE CHAIN REACTION (PCR)
MULTIPLEX
TOUCHDOWN
Origin
Hal imported