Gene expression variation can be partitioned into different components (regulatory, genetic and acclimatory effects) but for lichen-forming fungi, the relative importance of each of these effects is unclear. Here, we studied gene expression in the lichen-forming fungus Lobaria pulmonaria in response to thermal stress and parasitism by the lichenicolous fungus Plectocarpon lichenum. Our experimental procedure was to acclimate lichen thalli to 4 °C over three weeks and then expose them to 15 °C and 25 °C for 2 hours each, sampling infected and visually asymptomatic thalli at each temperature. Quantitative Real-Time PCR was utilized to quantify gene expression of six candidate genes, normalizing expression values with two reference genes. We found that two genes encoding heat shock proteins (hsp88 and hsp98), two polyketide synthase genes (rPKS1, nrPKS3) and elongation factor 1-1-α (efa) were upregulated at higher temperatures. Moreover, we observed higher expression of hsp98 at 25 °C in samples infected by P. lichenum than in uninfected samples. Finally, in partial redundancy analyses, most of the explained variation in gene expression was related to temperature treatment; genetic variation and long- term acclimatization to sites contributed far less. Hence, regulatory effects (i.e. direct adjustments of gene expression in response to the temperature change) dominated over genetic and acclimatory effects in the gene expression variability of L. pulmonaria. This study suggests that L. pulmonaria could become a valuable lichen model for studying heat shock protein responses in vivo. acclimation, heat shock genes, lichenicolous fungi, polyketide synthase genes (PKS), quantitative Real-Time PCR (qPCR), stress response, thermal stress, transcriptome