Real-time PCR quantification and spatio-temporal distribution of airborne inoculum of Mycosphaerella graminicola in Belgium

Two kinds of propagules play a role in Mycosphaerella graminicola dissemination: splash-dispersed pycnidiospores and airborne sexual ascospores. A method based on real-time polymerase chain reaction (PCR) assay and using Burkard spore traps was developed to quantify M. graminicola airborne inoculum. The method was tested for its reliability and applied in a spore trap network over a 2-year period in order to investigate the spatio-temporal distribution of airborne inoculum in Belgium. At four experimental sites, airborne inoculum was detected in both years. A seasonal distribution was observed, with the highest mean daily quantities (up to 351.0 cDNA) trapped in July and with clusters detected from September to April. The first year of trapping, a mean daily quantity of 15.7 cDNA of M. graminicola airborne inoculum was also detected in the air above a building in a city where the spatio-temporal distribution showed a similar pattern to that in the field. Mean daily quantities of up to 60.7 cDNA of airborne inoculum were measured during the cereal stem elongation and flowering stages, suggesting that it contributes to the infection of upper leaves later in the season. Most detection, however, tended to occur between flowering and harvest, suggesting significant production of pseudothecia during that period. Variations in mean daily quantities from 1.0 to 48.2 cDNA were observed between sites and between years in the patterns of airborne inoculum. After stem elongation, the quantities detected at a site were positively correlated with the disease pressure in the field. Quantities trapped at beginning of the growing season were also well correlated with the disease level the previous year. Multiple regressions revealed that some factors partly explain the daily variations of airborne inoculum.