Fig.S1.tiff

The Asian black truffle <i>Tuber himalayense</i> is harvested from natural habitats for sale. However, its populations have declined since the 1990s owing to habitat degradation and overharvesting. To ensure sustainable production and to conserve wild genetic resources, cultivation and a better understanding of reproductive dynamics are essential. We inoculated <i>Quercus serrata</i> seedlings with <i>T. himalayense</i> ascospores sourced from two localities and planted into a nursery in Japan. After 7½ years, two ascocarps appeared, followed a year later by 12 more. To confirm their identity, we performed molecular phylogenetic analysis by using the rDNA internal transcribed spacer region and multilocus genotyping with 13 simple sequence repeats (SSR) markers. All ascocarps matched one inoculated strain. Further analysis of SSR and mating-type markers from spore and glebal DNA showed that all ascocarps were homozygotes. This study marks the first successful cultivation of <i>T. himalayense</i> in Japan. It also provides rare data on mating dynamics from the first year of ascocarp formation—an aspect rarely captured in<i> Tuber </i>studies.