Specificity of different PCR primers for Verticillium dahliae isolated from olive trees in Jordan

Abstract EN

Three pairs of polymerase chain reaction (PCR) primers were compared to amplify the characteristic DNA fingerprints of Verticillium dahliae Kleb, (the causal agent of olive vascular wilt disease).

The primers include several combinations of the internal transcribed spacer (ITS) regions of nuclear ribosomal RNA (rRNA) genes and one pair from repetitive nuclear DNA sequences.

Deoxyoligonucleotide primers specific to V.

dahliae were synthesized based on the identified variable nucleotide sequences of the nuclear ITS regions of different Verticillium species that cause vascular wilt diseases.

The PCR primers of nuclear repetitive DNA showed variable band intensities for the different isolates of V.

dahliae isolated from olive trees in Jordan.

This suggests genetic variabilities of the local isolates collected from olive.

Primers based on the ITS sequences produced more consistently homogeneous and characteristic fingerprints.

The detection limit of these ITS primers was further improved using nested PCR and to show the high specificity of the ITS primers.

No nested PCR amplification was produced using the DNA isolated from different fungi in the same taxonomic class or other classes.

These ITS-specific primer pairs may be useful in developing diagnostic procedures of Verticillium wilt disease using single or nested PCR.