Partial MAT-2 gene structure and the influence of temperature on mating success in Gibberella circinata

Genetic analysis of Gibberella circinata, the causative agent of pitch canker disease of pines, historically has been thwarted by a low frequency of mating success in the laboratory. We describe two findings that should facilitate genetic analysis of this fungus and related species. First, we determined that previously described degenerate primers could be used to amplify a portion of the MAT-2 mating type gene from G. circinata. This led to the cloning and sequencing of a fragment of the MAT-2 gene, which in turn made it possible to distinguish between G. circinata isolates of opposite mating types. Second, we discovered that of the 18 G. circinata field isolates in our collection, the 1 female fertile isolate expressed its fertility at 15 and 20 degrees C but not at 25 degrees C, the temperature used for crossing many Gibberella species. It is evident, therefore, that when sexual reproduction in other closely related species is initially being investigated, the crosses should be established at a variety of temperatures. Once we learned that female fertility in this G. circinata isolate was expressed at 20 degrees C, a high frequency of mating success was achieved.