A real-time quantitative PCR assay for the detection of Sphaeropsis sapinea from inoculated Pinus nigra shoots

Pinus nigra (Austrian pine) is a tree widely planted in central and southern Europe in the reforestation of areas with poor soils. One of the major constrains of P. nigra is a fungal disease caused by Sphaeropsis sapinea which causes shoot tip dieback and tree mortality. This paper describes a real-time quantitative PCR assay based on the partial sequence of the small subunit ribosomal RNA of S. sapinea to detect and quantify DNA of the fungal pathogen in S. sapinea inoculated shoots. The study found that real-time quantitative PCR was a reliable technique to diagnose this disease and suggests that it can also be used to study fungal behaviour in host tissue and particularly to quantify fungal growth in the latent or endophytic phase.