Identification of <Emphasis Type="Italic">Ditylenchus</Emphasis> species associated with Fabaceae seeds based on a specific polymerase chain reaction of ribosomal DNA-ITS regions |
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Authors: | Mohamed Kerkoud Magali Esquibet Olivier Plantard Myriam Avrillon Corinne Guimier Martine Franck Joël Léchappé Rene Mathis |
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Institution: | 1.GEVES-SNES,Beaucouze Cedex,France;2.UMR ??BiO3P??, INRA-Agrocampus Rennes,Le Rheu Cedex,France;3.GEVES,Guyancourt Cedex,France |
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Abstract: | A technique based on the use of specific primers for polymerase chain reaction (PCR) was developed for the identification
of the stem and bulb nematode belonging to the Ditylenchus dipsaci species complex. The internal transcribed spacer region ITS1 and ITS2, the gene 5.8 S and part of genes 18 S and 26 S of
twenty populations of the D. dipsaci species complex belonging to both D. dipsaci sensu stricto and Ditylenchus sp. B (corresponding to populations of giant individuals associated to Vicia faba) and three congeneric species were amplified with two universal ribosomal primers. PCR-amplified DNA samples were digested
with five restriction enzymes in order to reveal some polymorphism allowing the identification of D. dipsaci populations associated with Fabaceae seeds. The polymorphism among species was confirmed by the sequencing of the PCR products.
A primer (DdpS2) was designed in a region conserved in all populations of both D. dipsaci
sensu stricto and D. sp. B studied in the present work. The other Anguinidae species (except a few species from Central Asia associated to Astereaceae
and D. sp. G associated to Plantago maritima) differ in two to four nucleotides at the 3′ extremity of this region. This sequence portion coincides with a TspEI restriction site. In combination with a primer located in the ribosomal region, this first primer is a good candidate for
identification by PCR of populations of the D. dipsaci species complex found in Fabaceae seeds. A second primer (DdpS1) was designed in a similar way and was specific to D. dipsaci sensu stricto. The utility of these two sets of primers is discussed against the background of quarantine regulation. |
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Keywords: | Diagnostics Ditylenchus ITS Pathogen PCR Seed |
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