Optimising in vitro culture conditions for the truffle Tuber brumale

The vegetative propagation of ectomycorrhizal truffle fungi is limited by their slow mycelial growth. Many factors including media, isolate genotypes and environmental conditions can alter fungal mycelial growth rates. This study aimed to improve the in vitro growth rate of Tuber brumale by determining the optimal carbohydrate and nitrogen sources, temperature and pH. After 8 weeks, the highest level of growth and densest hyphal branching were recorded in the medium containing glucose as the main carbohydrate. For nitrogen, glutamine (200 mg N l-1) provided the greatest hyphal growth and density compared to the other amino acid treatments. Regarding temperature, 16°C proved to be optimal for T. brumale growth and branching. Media of pH 6 and pH 7 were most favourable for the growth of T. brumale. The results from this research provide baseline data on the vegetative nutrition of T. brumale and have implications for the in vitro culture of winter truffle hyphae