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XII
Congreso de la Sociedad Española de Fitopatología (SEF),
2004
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3962 |
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Aplicación de un método de extracción de ADN
para la identificación de nematodos fitopatógenos por
técnicas moleculares
Abelleira, A.; Aguín,
O.; Balado, M.; Mansilla, J.P.
Estación Fitopatolóxica do
Areeiro, Excma. Diputación Provincial de Pontevedra, Subida a la Robleda
s/n, 36153 Pontevedra, España.
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ABSTRACT: Morphological and
morphometric determination of phytopathogenic nematodes in diagnosis
laboratories, , is difficult, given the wide range of existing species. In the
last years, molecular techniques were developed, mainly for the identification
of the most important species. Nowadays these methods are being implemented
since are strongly helpful in the diagnosis of these species. Before
identifying a particular nematode using an appropriate molecular marker, it is
necessary to carry out DNA extraction, being this the most difficult task in
molecular control methods. Depending on the genus, several methods were
reported. For a diagnosis laboratory it is a time-consuming and difficult task
to use extraction specific protocols for each nematode, so in the present work
the efficiency of a single DNA extraction protocol for the main genus of plant
pathogenic nematodes was studied. The protocol studied was the one proposed by
Bulman and Marshall (1997) for Globodera DNA extraction. The method was
applied to nematodes belonging to the genus Globodera, Heterodera
and Meloidogyne, using different nematode stages: cysts, adults,
juveniles and eggs. Results proved that this was a fast, accurate and
reproducible method for obtaining enough genetic material of nematodes
belonging to these genus, that will be subsequently used in identification
molecular techniques.
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