Needle fungi in young Tasmanian Pinus radiata plantations in relation to elevation and rainfall

Background: Needle fungi in conifers have been extensively studied to explore their diversity, but environmental factors influencing the composition of fungal communities in Pinus radiata D. Don needles have received little attention. This study was conducted to examine the influence of the environment as defined by rainfall, elevation and temperature on the composition of fungal communities in pine needles at an age prior to that at which spring needle cast (SNC) is generally observed. Elucidating the entire fungal community in the needles is a first step towards understanding the cause of the disease. Methods: Needle samples were collected from 5-year-old P. radiata trees, their age predating the onset of SNC, from 12 plantations in Tasmania. Interpolated data for the climate variables, including seasonal components for rainfall and temperature, were obtained from an enhanced climate data bank. Pooled needle samples were examined for the fungi they contained using DNA sequencing of cloned polymerase chain reaction (PCR) products. Clones were grouped into operational taxonomic units (OTUs) and identified to their lowest possible taxonomic level by comparison with reference isolates and public DNA databases. Results: DNA sequencing revealed that needle fungal communities differed greatly, depending upon the total annual rainfall and needle age. Needle fungi that have been previously associated with pathogenic behaviour, such as Cyclaneusma minus, Dothistroma septosporum, Lophodermium pinastri, Strasseria geniculata and Sydowia polyspora, were all found in the needles in this study. Also, of 70 ascomycetous OTUs that were discriminated by their DNA sequences, 21 were identified as belonging to the Teratosphaeriaceae. Conclusions: An overview of the endophytic fungi present in the needles of P. radiata has been obtained from trees whose age precedes the onset of SNC. All of the previously known pathogenic species for needle cast diseases were present, in addition to a range of unidentified OTUs belonging to the Teratosphaeriaceae.