Truffle Typing
Democratek
is working on a PCR based test for determining mating type in truffles. Truffle fungi live in symbiosis with a number
of species of trees, drawing carbohydrates from the roots in turn for supplying essential nutrients from the soil.
Truffles are the fruiting body and reproductive spore dispersal mechanism for the truffle fungus. These subterranean
fruits are relished by a number of mammals and birds, drawn by the seductive scent of the truffle. This prized fungus
brings a high price in the market, and has generated great interest in cultivation for more reliable harvests. Early
cultivation efforts are rudimentary. Most commonly, the spore slurry method is employed on young seedlings,
with a truffle smoothie dripped or injected into the root system of the host plant. More modern mycelial cultivation methods employed
broadly in the mushroom cultivation industry have been limited by the fastidious growth of truffle fungus in pure
culture. Furthur, efforts to uncover details of the truffle reproductive cycle have revealed that truffle mycelia are
heterothallic, or of two types, and must combine to form the reproductive structure of the truffle. Thus, in order for
successful orchards to be established, two mycelial types must be present in the same orchard. For mycelial pure
cultures to be used to generate productive truffle orchards, cultures of both mating types must be employed in the
inoculation of truffle trees. Mating type genes have been identified in a few truffle trees, but tools to test the
genotype of truffles, truffle trees, and truffle cultures have remained elusive. Democratek aims to develop a rapid
Real-Time PCR assay, akin to the COVID PCR test, to analyze truffles, truffle cultures, and truffle tree roots for
truffle mating type.
PCR is like Xerox for DNA, utilizing enzymes to make
thousands and thousands of copies of DNA. Other biotechnologies such as fluorescent dyes and synthetic DNA allow us
to target DNA sequences unique to each mating type and cause the reaction tubes to glow when the specific DNA sequences
accumulate. Genome analysis allows us to determine the specific sequences that we need to target
and so we are able to develop this test for those truffle species with sequenced genomes. The first truffle that we
are focusing our attention on is Tuber canaliculatum, or the Appalachian truffle. This truffle is a native American
truffle, is highly regarded, and is extremely rare. Efforts to cultivate it are ongoing and early attempts suggest
that it may be easier to grow native American truffles in the United States than it is to grow European truffles.
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