Berry S., Fargette Mireille, Morand Serge, Cadet Patrice. (2007). Reliability of PCR-based techniques for detection and discrimination of plant-parasitic nematodes of sugarcane. Nematology, 9 (4), p. 499-514. ISSN 1388-5545.
Titre du document
Reliability of PCR-based techniques for detection and discrimination of plant-parasitic nematodes of sugarcane
Nematology, 2007,
9 (4), p. 499-514 ISSN 1388-5545
In order to develop diagnostic tools for plant-parasitic nematodes of sugarcane in South Africa, this work investigated the potential biodiversity of these nematodes in the amount of nucleic acid available, the discrimination of species by a PCR-based method (targeting the ITS I region) and the effect of some experimental conditions on their detection or discrimination. The ITS I amplified fragment varied in size, ranging from 400 bp (Meloidogyne javanica) to 1200 bp (Longidorus pisi, Paratrichodorus minor, Xiphinema elongatum, Xiphinema Zulu). Some species discrimination was possible by discernible differences in ITS I amplified fragment sizes. The potential. as well as shortcomings (inhibitions. competition) of this approach are assessed and further steps/outcomes discussed. Part of the work also focused on DNA extraction. Nematode genomic nucleic acid was extracted front five nematode species(Prati,lenchus zeae, Helicotylenchus dihystera, Meloidogyne javanica. Xiphinema elongatum and Paratrichodorus lobatus) and we discuss the significant differences obtained between species in the amount of nucleic acid extracted. There were also marked differences in the quantity and quality of nucleic acid extracted from nematode Suspensions originating from soils, depending on the extraction buffer used.
