Tuber melanosporum can be found in hillside copses, where the trees are fairly far apart and are surrounded by vegetation-free areas called "pianelli" (also used in Italian to denote the strip of land between two rows of vines).

It thrives on stony-calcareous (permeable) ground with a compact subsoil, which helps the development of the roots of the trees on the surface. This soil is generally packed with calcium carbonate and lacking in organic residues.

Above-all it must contain no more than 40% of clay, as poor porosity would lead to asphyxia. Tree coverage must be no higher than 30%, and there must be little or no shrubland, and practically no grass at all. Light must penetrate well, providing good heating of the soil.

Tuber melanosporum symbionts plants

• Pubescent oak (Quercus pubescens)
  
• Evergreen oak (Quercus ilex)
  
• Turkey oak (Quercus cerris)
  
• Large Leaved Lime (Tilia playphyllos)
  
• Hazel tree (Corylus avellana)
  
• European Hop-hornbeam (Ostrya carpinifolia)
  
• Rockrose (Cistus spp.)