An effective medium for isolating Sphaeropsis sapinea from asymptomatic pines

The pathogen Sphaeropsis sapinea can persist in stems and branches of asymptomatic pines and can later induce disease when triggered by host stress. Several experiments were conducted to test if:(i) medium amended with tannic acid (TA) can increase the frequency of cultural detection of this shoot blight and canker pathogen from asymptomatic red pine (Pinus resinosa) stems, and (ii) S. sapinea can persist in asymptomatic red pine in the field following artificial inoculation. TA (0.5% w/v) in 2% (w/v) water agar proved to be the best medium for isolation of S. sapinea among a larger number of tested media. The addition of TA had little or no effect on the growth of two group A and two group B isolates of S. sapinea. However, when TA was added, 11 other fast-growing fungal isolates from stems/branches of red or jack pines (P. banksiana) were inhibited and grew more slowly (p < 0.05) than both S. sapinea groups. The TA-amended medium improved cultural detection of S. sapinea from 2-year-old, asymptomatic red pine nursery seedlings compared with two other methods used for the cultural detection of S. sapinea (32%vs. 8.5% and 18% recovery; p < 0.001 and p = 0.031, respectively). A field test using the TA-amended media established that S. sapinea can persist asymptomatically in red pine trees for at least 1 year. This medium significantly reduces the frequency of false-negatives from asymptomatic field material.