US 12,365,867 B2
Rapid isolation, cultivation methods and utilities of truffle fungi
Gary A. Strobel, Bozeman, MT (US); and Kelly W. Gaisford, Bozeman, MT (US)
Assigned to BLACK BOAR TRUFFLE, LLC, Bozeman, MT (US)
Filed by Black Boar Truffle, LLC, Bozeman, MT (US)
Filed on Mar. 29, 2024, as Appl. No. 18/621,769.
Claims priority of provisional application 63/455,304, filed on Mar. 29, 2023.
Prior Publication US 2024/0327779 A1, Oct. 3, 2024
Int. Cl. C12N 1/14 (2006.01)
CPC C12N 1/145 (2021.05) [C12N 2500/34 (2013.01); C12N 2500/76 (2013.01)] 19 Claims
 
1. A method of growing truffle fungi comprising the steps of:
obtaining a spore-producing fruiting body of an ascomycetous fungus from soil, wherein the ascomycetous fungus is a Tuber spp. or an Imaia spp.;
scraping the inside of the spore-producing fruiting body of the ascomycetous fungus to collect a fruiting body sample;
placing the fruiting body sample on a first surface comprising a first nutrient substrate, wherein the first nutrient substrate comprises:
a simple sugar;
a botanical isolate; and
agar;
incubating the fruiting body sample on the first nutrient substrate with an antimicrobial volatile mixture until mycelium formation, wherein the antimicrobial volatile mixture is derived from the volatile organic compounds produced by Muscodor crispans, and wherein the presence of mycelium indicates new fungal growth; and
transferring the new fungal growth into a second surface comprising a second nutrient substrate to obtain a pure culture of the ascomycetous fungus.