US-12618043-B2 - Rapid isolation, cultivation methods and utilities of truffle fungi

US12618043B2US 12618043 B2US12618043 B2US 12618043B2US-12618043-B2

Abstract

Methods of farming, domesticating, or cultivating truffle fungi, including isolation and cultivation methods for producing pure truffle fungi, or mini-truffles, typically a Tuber spp. or an Iamai spp. Examples of truffle include Tuber melanosporum, Tuber magnatum, Tuber aestivum, Tuber uncinatum, Tuber borchii, Imaia spp., Tuber macrosporum, Tuber gibbosum, Tuber oregonense , and Tuber lyonii . The isolated pure truffle may then be cultivated on a nutrient substrate, which may be, for example, a fruit, nut, grain, or a portion thereof, resulting in a truffle-flavored food product. Non-limiting examples of such substrates include rye, barley, lentil, wheat, rice, soybeans, pecan, hazelnut, pine nut, English walnut, coffee beans, mustard, cacao, sesame, sunflower, grapes, blackberries, blueberries, cherries, kiwi, mango, raspberries, and huckleberries.

Inventors

Gary A. Strobel
Kelly W. GAISFORD

Assignees

Black Boar Truffle, LLC

Dates

Publication Date: 20260505
Application Date: 20250613

Claims (9)

1 . A method of making a truffle-flavored food product comprising: 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; incubating a portion of the pure culture of ascomycetous fungus with fruit, nut, grain, or a portion thereof for at least 2 weeks to produce an inoculated botanical sample; and processing the inoculated botanical sample into a food product.
2 . The method of claim 1 , wherein the Tuber spp. is selected from the group consisting of: Tuber melanosporum, Tuber magnatum, Tuber aestivum, Tuber uncinatum, Tuber borchii, Tuber macrosporum, Tuber gibbosum, Tuber oregonense , and Tuber lyonii syn. Texense.
3 . The method of claim 1 , wherein the fruit, nut, grain, or a portion thereof is selected from the group consisting of rye, barley, lentil, wheat, rice, soybeans, pecan, hazelnut, pine nut, English walnut, coffee beans, mustard, cacao, sesame, sunflower, grapes, blackberries, blueberries, cherries, kiwi, mango, raspberries, huckleberries, and combinations thereof.
4 . The method of claim 2 , wherein the fruit, nut, grain, or a portion thereof is selected from the group consisting of rye, barley, lentil, wheat, rice, soybeans, pecan, hazelnut, pine nut, English walnut, coffee beans, mustard, cacao, sesame, sunflower, grapes, blackberries, blueberries, cherries, kiwi, mango, raspberries, huckleberries, and combinations thereof.
5 . The method of claim 1 , wherein the processing step comprises at least one of: using the inoculated botanical sample wet in combination with a food product; drying the inoculated botanical sample; grinding the inoculated botanical sample into a powder; and creating a mash using the inoculated botanical sample; and mixing the powder or mash with the food product.
6 . The method of claim 2 , wherein the processing step comprises at least one of: using the inoculated botanical sample wet in combination with a food product; drying the inoculated botanical sample; grinding the inoculated botanical sample into a powder; and creating a mash using the inoculated botanical sample; and mixing the powder or mash with the food product.
7 . The method of claim 3 , wherein the processing step comprises at least one of: using the inoculated botanical sample wet in combination with a food product; drying the inoculated botanical sample; grinding the inoculated botanical sample into a powder; and creating a mash using the inoculated botanical sample; and mixing the powder or mash with the food product.
8 . The method of claim 4 , wherein the processing step comprises at least one of: using the inoculated botanical sample wet in combination with a food product; drying the inoculated botanical sample; grinding the inoculated botanical sample into a powder; and creating a mash using the inoculated botanical sample; and mixing the powder or mash with the food product.
9 . The method of claim 1 , wherein before the incubation of a portion of the pure culture of ascomycetous fungus with fruit, nut, grain, or a portion thereof, the method further comprises a. Starting with about 1 liter volume of the fruit, nut, grain, or portion thereof; b. Adding about 0.6 to 3.0 liters of water; c. Adding about 5 g glucose, 1 g peptone, and 2 g yeast extract; and d. Stirring thoroughly.

Description

RELATED APPLICATIONS This application is a divisional of U.S. patent application Ser. No. 18/621,769 filed Mar. 29, 2024, which claims the benefit of U.S. provisional patent application 63/455,304, filed Mar. 29, 2023, the entirety of the disclosures of which are hereby incorporated by this reference. INCORPORATION-BY-REFERENCE OF MATERIAL ELECTRONICALLY FILED Incorporated by reference in its entirety herein is a computer-readable nucleotide/amino acid sequence listing submitted concurrently herewith and identified as follows: One 7,511 byte XML file named “22630.002US-PAT” created on Mar. 28, 2024. TECHNICAL FIELD This disclosure relates to methods of farming or domesticating truffle fungi. In some aspects, the disclosure relates to isolation and cultivation methods for producing pure truffle fungi. BACKGROUND The fruiting bodies of the truffle fungi have been sought after for thousands of years and are prized for their flavor making them extremely expensive in all markets. Most commonly, they are collected in the soil surrounding certain tree species and certain dogs and trained pigs are used to locate the prized truffles. Once located, the truffle is then dug from the soil and shipped for commercial distribution. The truffle itself ranges from about 1 to 6-8 or more cm in diameter with a rough and undulating surface and can be used for the enhancement of flavor in all types of foods. Cultivation of the truffle has traditionally been done by dedicating land to certain tree types (usually oak species but others as well including pecan, and hazelnut) and then inoculating the roots of the trees being planted with harvested truffle material that carries spores and mycelial fragments that invade the young roots. Eventually the infected roots begin to support the growth of the fungus, which is attached to them, to produce a root enlargement that contains the truffle fungus, plant tissues and a plethora of soil related microbes. The fungal growth begins as a small ball of enlarged root growth eventually enlarging to the full-sized truffle. Full truffle development takes five to ten years. The long processes involved in truffle production; their short shelf life once dug from the soil as well as the demand for the truffle itself add to the cost of this sought-after culinary delicacy. Given the multitude of costs and time involved in their production prices for truffle can be expensive (for example, $700-$1000/lb. or more for black truffle, and $3,000/lb. or more for white truffle). To satisfy market demand, a method for farming truffles is needed. SUMMARY OF THE DISCLOSURE Growing truffles is a long and involved process that contributes to the high costs of this culinary delicacy. A method to grow and farm truffles would aid in lowering the cost associated with truffle production by increasing market availability without having to rely on the traditional collection methods to obtain the truffles. The present disclosure is directed to a method of isolating and culturing truffle fungi species that provides an alternative for the domestication of major truffle species. The disclosure includes methods for growing commercially valuable truffles without the need for soil, therefore reducing the time needed for truffle production. The disclosure includes a method to isolate pure culture of ascomycetous fungus that can be grown on plates, such method provides an alternative to obtain inoculants free of soil-borne contamination. Some embodiments of the present disclosure comprise a method of growing truffles comprising the steps of obtaining a spore-producing fruiting body of an ascomycetous fungus from soil, 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 nutrient substrate, wherein the nutrient substrate comprises: a simple sugar, a botanical isolate and agar, incubating the fruiting body sample on the nutrient substrate with an antimicrobial volatile mixture until mycelium formation, wherein the presence of mycelium indicates new fungal growth; and transferring the new fungal growth into a second surface comprising the nutrient substrate to obtain a pure culture of the ascomycetous fungus. Some embodiments of the present disclosure comprise soil-less method of growing truffle comprising the steps of obtaining a spore-producing fruiting body of an ascomycetous fungus from soil, 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 nutrient substrate, incubating the fruiting body sample on the nutrient substrate with an antimicrobial volatile mixture until mycelium formation, and transferring the new fungal growth into a second surface comprising the nutrient substrate to obtain a pure culture of the ascomycetous fungus. Presence of mycelium indicates new fung