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Molecular characterisation of 18 Pratylenchus species using rDNA Restriction Fragment Length Polymorphism

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For more content, see Nematologica.

The RFLP technique was used to establish a reliable diagnostic method for 18 Pratylenchus species: Pratylenchus agilis, P. bolivianus, P. brachyurus, P. coffeae, P. crenatus, P. fallax, P. goodeyi, P. loosi, P. mediterraneus, P. neglectus, P. penetrans, P. pratensis, P. pseudocoffeae, P. scribneri, P. subranjani, P. thornei, P. vulnus and P. zeae. The polymerase chain reaction (PCR) amplified the ITS regions from all species and populations examined and revealed large differences in length, ranging in size from approximately 900 to 1250 bp. The rDNA fragments were digested with five restriction enzymes (CfoI, DdeI, HindIII, HpaII, and PstI). All Pratylenchus species can be differentiated from each other by a combination of at least two enzymes. CfoI differentiated all nematode species with the exception of P. fallax, P. penetrans and P. pseudocoffeae. P. fallax was further separated by a DdeI restriction, and P. pseudocoffeae by a PstI digestion. Intraspecific RFLP were observed. Upon CfoI, DdeI, HindIII, or HpaII digestion, it was possible to separate the three P. coffeae populations studied from each other.

La technique RFLP a été utilisée pour créer une méthode fiable de diagnostic pour 18 espèces de Pratylenchus: Pratylenchus agilis, P. bolivianus, P. brachyurus, P. coffeae, P. crenatus, P. fallax, P. goodeyi, P. loosi, P. mediterraneus, P. neglectus, P. penetrans, P. pratensis, P. pseudocoffeae, P. scribneri, P. subranjani, P. thornei, P. vulnus et P. zeae. La réaction de polymérisation en chaîne (PCR) a amplifié les régions de l’ITS pour toutes les espèces et populations étudiées et a mis en évidence de grandes différences dans la taille des gammes de longueur, de 900 à 1250 bp approximativement. Les fragments de rDNA ont été digérés à l’aide de cinq enzymes de restriction (CfoI, DdeI, HindIII, HpaII, and PstI). Toutes les espèces de Pratylenchus ont pu être différenciées les unes des autres par une combinaison d’au moins deux enzymes. CfoI a différencié toutes les espèces à l’exception de P. fallax, P. penetrans et P. pseudocoffeae. P. fallax a été ultérieurement séparé par une restriction DdeI, et P. pseudocoffeae par une digestion PstI. Des RFLP intraspécifiques ont été observés. Par les digestions CfoI, DdeI, HindIII, ou HpaII, il s’est révélé possible de séparer les unes des autres les trois populations étudiées de P. coffeae.

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/content/journals/10.1163/156854100509024
2000-05-01
2016-12-06

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