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Genetic differentiation of charcoal rot pathogen, Macrophomina phaseolina, into specific groups using URP-PCR

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Abstract:

Forty isolates of Macrophomina phaseolina, a pathogen causing charcoal dry root rot of soybean, cotton, and chickpea, were genetically characterized with universal rice primers (URP; primers derived from DNA repeat sequences in the rice genome) using polymerase chain reaction (URP-PCR). Out of 12 URPs used in this study, 5 primers were effective in producing polymorphic fingerprint patterns from the DNA of M. phaseolina isolates. Three primers (URP-2F, URP-6R, and URP-30F) were quite informative and produced high levels of polymorphism among the isolates of M. phaseolina. Analysis of the entire fingerprint profiles using unweighted pair-group method with arithmetic averages (UPGMA) clearly differentiated M. phaseolina isolates obtained from soybean, cotton, and chickpea hosts into specific groups. In this study, we found for the first time transferability and use of PCR primers derived from plant genomes to generate host-specific fingerprint profiles of M. phaseolina, a broad host range plant pathogenic fungus. These results demonstrate that URPs are sensitive and technically simple to use for assaying genetic variability in M. phaseolina populations.Key words: Macrophomina phaseolina, molecular variability, soybean, cotton, chickpea.

Quarante isolats de Macrophomina phaseolina, un pathogène de la pourriture charbonneuse sèche de racines de soya, de coton et de pois chiches ont été caractérisés génétiquement à l'aide d'amorces universelles du riz (URP – des amorces dérivées de séquence de répétition d'ADN du génome du riz) et de la réaction de la polymérase en chaîne (URP-PCR). Des 12 URP utilisés dans cette étude, 5 amorces ont pu générer efficacement des empreintes polymorphes à partir de l'ADN d'isolats de M. phaseolina. Trois amorces, URP-2F, URP-6R et URP-30F étaient particulièrement informatives et ont mis en évidence un degré élevé de polymorphisme chez les isolats de M. phaseolina. Une analyse de UPGMA (« unweighted pair-group method with arithmetic averages ») des empreintes génétiques ont permis de séparer clairement les isolats de M. phaseolina issues de soya, de coton et de pois chiches en groupes spécifiques. Dans cette étude, nous avons découvert pour la première fois la possibilité de transférer et d'utiliser des amorces de PCR dérivées d'un génome de plante afin de générer des empreintes génétiques spécifiques à l'hôte pour le champignon pathogène à large spectre infectieux M. phaseolina. Ces résultats démontrent que les URP constituent une méthode simple et sensible pour l'analyse de la variabilité génétique des populations de M. phaseolina.Mots clés : Macrophomina phaseolina, variabilité moléculaire, soya, coton, poids chiches.[Traduit par la Rédaction]

Document Type: Research Article

Publication date: February 1, 2005

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  • Published since 1954, this monthly journal contains new research in the field of microbiology including applied microbiology and biotechnology; microbial structure and function; fungi and other eucaryotic protists; infection and immunity; microbial ecology; physiology, metabolism and enzymology; and virology, genetics, and molecular biology. It also publishes review articles and notes on an occasional basis, contributed by recognized scientists worldwide.
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