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

The RFLP technique was used to establish a reliable diagnostic method for 1 8 Pratylenchus species: Pratylenchus agilis, P. bolivianus, P. brachyurus, P. coffeae, P. crenatus, P.fallax, P. goodeyi, P. loosi, P. mediterraneus, P. neglectus, P. penstrans P. pratensis, P. pseudocoffeae, P. scribneri, P. subranjani, P. thornei, P. vulnus and P. zene. The polymerase chain reacti on (PCR) amplified the ITS regions from all species and populations examine d and revealed large differences in length, ranging in size from approximat ely 900 to 1250 bp. The rDNA fragments were digested with five restriction enzymes (CfoI, DdeI, HindIII, HpaII, and PstI). All Pratylenchus species ca n be differentiated from each other by a combination of at least two enzyme s. CfoI differentiated all nematode species with the exception of P. fallax , P. penetrans and P. pseudocoffeae. P. fallax was further separated by a D deI restriction, and P. pseudocoffeae by a PstI digestion. Intraspecific RF LP were observed. Upon CfoI, DdeI, HindIII, or HpaII digestion, it was poss ible to separate the three P. coffeae populations studied from each other.