Detection of Meloidogyne incognita and Pochonia chlamydosporia by fluorogenic molecular probes.

Fluorescent molecular probes were applied for detection of the plant parasitic nematode Meloidogyne incognita and the nematode-egg parasitic fungus Pochonia chlamydosporia var. chlamydosporia. A region in the M. incognitar DNA including ITS2 was selected for amplification and recognition with a real-time PCR assay, based on a combination of three specific motifs, each recognized by a specific fluorescent probe. Similarly, a Scorpion probe was designed for the RT-PCR quantification of P. c. chlamydosporia. For this purpose, the ITS-2 rDNA gene of the fungus was sequenced from a number of Italian isolates. A conserved region unique for P. c. chlamydosporia found in the ITS-2 rDNA gene was used. The probes allowed recognition of single juveniles of M. incognita and of the mycelium- or soil-extracted fungal DNA. The potentialities of the detection procedures are discussed.