Tuber planting formula and method for shortening the initial production period of truffles

Abstract

The present application belongs to the technical field of truffle planting, and particularly relates to a truffle planting formula and a planting method for shortening the initial yield period of truffles. In view of the problems of poor growth and development of mycorrhizal roots, poor mycorrhizal root vitality, small number of mycorrhizal roots, high contamination rate of ectomycorrhizal miscellaneous bacteria, long initial yield period of truffles, and low yield, the present application provides a truffle planting formula and a planting method. The formula can effectively promote the growth of truffle seedlings, the development of truffle mycorrhizal roots, reduce the contamination rate of ectomycorrhizal miscellaneous bacteria, accelerate the formation of fire rings, shorten the initial yield period of truffles by 2-5 years, and improve the yield of truffle planting.

Description

Technical Field

[0001] This invention belongs to the field of truffle cultivation technology, specifically relating to truffle cultivation formulas and methods for shortening the initial production period of truffles. Background Technology

[0002] truffles ( Tuber Truffles, also known as truffles, are a rare, large edible fungus. Biologically, they belong to the phylum Ascomycota, class Discotyledonae, order Discotyledales, family Truffleaceae. Tuberaceae ), Truffles ( Tuber Truffles, renowned worldwide for their distinctive aroma, are considered a top-tier ingredient, ranking alongside caviar and foie gras as one of the world's three most prized delicacies, earning the reputation of "kitchen diamonds." Mature black truffles (Indian truffles and Chinese summer truffles) in China sell for 600-1000 yuan / kg, while Panzhihua white truffles sell for 3000-5000 yuan / kg. To effectively protect and sustainably utilize wild truffle resources, the research, development, demonstration, and promotion of truffle cultivation techniques are urgently needed.

[0003] Truffles are mycorrhizal edible fungi that must form a symbiotic relationship with a host plant to complete their life cycle; otherwise, they cannot form fruiting bodies. Truffle cultivation has demanding requirements regarding climate, environment, and soil, needing to satisfy both the growth of the truffle and the host plant. Therefore, artificial cultivation is technically challenging, resulting in a long initial production cycle, low yield, and poor yield stability. While *Ichthyophthirius multifiliis* truffle cultivation has a long history and relatively mature technology abroad, the initial production period is still 5-8 years. In China, truffle cultivation is an emerging industry, still in its early stages, with an initial production period of 5-10 years, low yield, and poor yield stability. Therefore, shortening the initial production period and increasing and stabilizing truffle yield are the technical bottlenecks in truffle cultivation. Summary of the Invention

[0004] To address the problems of poor mycorrhizal growth and development, low mycorrhizal viability, low mycorrhizal quantity, high contamination rate of ectomycorrhizal fungi, long initial production period, low yield, and unstable yield in truffle cultivation, this invention provides a truffle cultivation formula and method. The formula effectively promotes the growth of truffle seedlings and the development of truffle mycorrhizae, reduces the contamination rate of ectomycorrhizal fungi, accelerates the formation of burn rings, and shortens the initial production period of truffles by 2–5 years.

[0005] In a first aspect, the present invention provides a truffle cultivation formula, which includes: a soil conditioner and a truffle mycorrhizal growth regulator;

[0006] The soil conditioner components, by weight, include: 25-28 parts of edible fungus residue, 25-28 parts of cow dung, 10-15 parts of sugarcane bagasse, 8-10 parts of distiller's grains, 5-8 parts of peat moss, 3-5 parts of silkworm excrement, 5-8 parts of fern powder, 3-5 parts of mulberry branch and leaf powder, 3-5 parts of fragrant wood branch and leaf powder, and 1-2 parts of Bacillus subtilis.

[0007] The truffle mycorrhizal growth conditioner comprises, by weight, 25-30 parts vermiculite, 25-30 parts perlite, 24-28 parts peat moss, 8-12 parts limestone powder, 3-5 parts truffle spore powder, 0.005-0.015 parts gibberellin (GA), 0.005-0.015 parts naphthaleneacetic acid (NAA), 1-3 parts potassium humate, and 1-3 parts extract of *Cinnamomum camphora* branches and leaves.

[0008] Preferably, the soil conditioner comprises, by weight, 26 parts of edible fungus residue, 26 parts of cow dung, 12 parts of sugarcane bagasse, 9 parts of distiller's grains, 6 parts of peat moss, 4 parts of silkworm excrement, 7 parts of fern powder, 4 parts of mulberry branch and leaf powder, 4 parts of fragrant wood branch and leaf powder, and 2 parts of Bacillus subtilis.

[0009] The truffle mycorrhizal growth conditioner comprises, by weight, 28 parts vermiculite, 28 parts perlite, 26 parts peat moss, 10 parts limestone powder, 3.98 parts mature truffle spore powder, 0.01 parts GA, 0.01 parts NAA, 2 parts potassium humate, and 2 parts extract of *Cinnamomum camphora* branches and leaves.

[0010] Furthermore, the edible mushroom residue is the waste residue from mushroom bags grown with cooked substrate.

[0011] Furthermore, the cow dung mentioned is from cows raised in captivity.

[0012] Furthermore, the lees are beer lees, which mainly utilize the yeast in beer lees.

[0013] Furthermore, the peat moss has an organic matter content of ≥75wt% and a humic acid content of ≥30wt%.

[0014] Furthermore, the particle size of the pulverized fern, mulberry branches and leaves, and fragrant wood branches and leaves is 0.2–1.0 cm.

[0015] Furthermore, the vermiculite has a particle size of 3–6 mm.

[0016] Furthermore, the perlite has a particle size of 3–6 mm.

[0017] Furthermore, the limestone powder is ≤10mm.

[0018] Furthermore, the peat moss in the truffle mycorrhizal growth conditioner has an organic matter content of ≥75wt% and a humic acid content of ≥30wt%.

[0019] Furthermore, the truffle spore powder has a maturity of ≥90% and a particle size of ≤1mm.

[0020] Furthermore, the selected truffles are mature Indian truffles with round fruit shape, strong aroma, and a diameter ≥ 5 cm.

[0021] Furthermore, the extract of *Corydalis tangutica* branches and leaves is prepared by crushing *Corydalis tangutica* branches and leaves into powder with a diameter ≤0.2mm, soaking them at room temperature for 24-48 hours at a material-to-water ratio of 1-2:20, boiling them at a temperature >95℃ for 1-2 hours, cooling them to below 30℃, adding 5-7wt% honey residue (based on the weight of *Corydalis tangutica* branches and leaves), letting them stand for 48-72 hours, filtering them to obtain the extract, and storing it at 2-3℃.

[0022] Secondly, the present invention also provides a method for preparing the above-mentioned truffle cultivation soil formula, comprising the following steps:

[0023] A. Preparation of soil conditioner, including the following steps:

[0024] a. Crush edible fungus residue, ferns, mulberry branches and leaves, and fragrant wood branches and leaves, mix them with sugarcane residue, pre-moisten with water at room temperature, cover with film and pile for 24-48 hours.

[0025] b. After adding cow dung, peat moss, silkworm excrement, distiller's grains, and Bacillus subtilis and mixing well, adjust the moisture content to 60-65%;

[0026] c. Build a material pile, cover it with a film for fermentation, and turn the pile over 3 times during fermentation to obtain a soil conditioner;

[0027] Furthermore, in step c, the material pile has a width of 1.0 to 1.5 m, a height of 0.8 to 1 m, and an unlimited length.

[0028] Furthermore, in step c, when the temperature inside the material is ≥65℃ each time, fermentation is maintained for 48-72 hours before turning the pile; this is repeated 3 times, and fermentation continues for another 48-72 hours after the third turning.

[0029] Furthermore, turning the pile inside out and top to bottom ensures even fermentation of the material.

[0030] Furthermore, in step c, the soil conditioner has a water content of 20-30 wt% and is stored at a temperature of 20-25°C.

[0031] B. Preparation of truffle mycorrhizal growth conditioner includes the following steps: weigh the components of the truffle mycorrhizal growth conditioner and mix them evenly.

[0032] Thirdly, the present invention also provides a method for cultivating truffles, which includes the following steps:

[0033] Mix limestone powder with soil, add soil conditioner, plant truffle mycorrhizal seedlings, and carry out regular watering, weeding and maintenance; in the second year of planting, inoculate the base of the tree with truffle mycorrhizal growth conditioner, and cover with topsoil after inoculation.

[0034] Furthermore, the method for mixing the limestone powder with the soil is as follows: calculate the volume of the original soil according to the area and depth of the soil to be modified, measure the corresponding volume of limestone powder according to the proportion, and mix it evenly into the soil.

[0035] Further, by volume, the composition comprises 85–95 parts soil and 8–12 parts limestone powder. Preferably, it comprises 90 parts native soil and 10 parts limestone powder.

[0036] Furthermore, the mixing depth of the limestone powder, soil conditioner, and soil is ≥20cm.

[0037] Furthermore, the method for inoculating the truffle mycorrhizal growth conditioner involves drilling a hole in each of the four directions (north, south, east, and west) directly below the drip line of the tree canopy, exposing the tree roots, inoculating the mycorrhizal growth conditioner into the holes, and then covering it with topsoil.

[0038] Furthermore, the amount of soil conditioner added is 20-25 kg / plant.

[0039] Preferably, the hole has a diameter of 10-15 cm and a depth of 20-25 cm.

[0040] Preferably, the amount of the truffle mycorrhizal growth conditioner added is 0.8–1.2 kg / well.

[0041] Preferably, the host plant of the truffle is a chestnut tree.

[0042] Beneficial Effects: This invention provides a truffle cultivation formula and method, comprising a truffle cultivation soil conditioner, a truffle mycorrhizal growth regulator, and a soil improvement method. The soil composition formula improves soil aggregate structure, fertility, and microbial community composition, effectively promoting truffle seedling growth and mycorrhizal development, reducing the contamination rate of ectomycorrhizal fungi, accelerating the formation of burn zones, and shortening the initial production period of truffles by 2-5 years. Examples show that this invention achieves initial production of truffles in 3 years, with the heaviest single truffle reaching 650g in 4 years, the highest yield per plant in 5 years reaching 1054g, and the highest yield per plant in 6 years, setting new domestic records for the shortest initial production period, heaviest single truffle, and highest yield per plant. It establishes a new dual-benefit cultivation model of "fruiting on the tree, producing truffles under the tree." The cultivation soil conditioner and truffle mycorrhizal growth regulator of this invention are simple to formulate, use readily available raw materials, are low in cost, and have significant effects. They can provide technical support for the sustainable development of the entire truffle cultivation industry chain and have broad application prospects. Detailed Implementation

[0043] In one embodiment of the present invention, the truffle cultivation formula includes: a soil conditioner and a mycorrhizal growth regulator; wherein, by weight, the soil conditioner comprises: 25-28 parts of edible fungus residue, 25-28 parts of cow dung, 10-15 parts of sugarcane bagasse, 8-10 parts of distiller's grains, 5-8 parts of peat moss, 3-5 parts of silkworm excrement, 5-8 parts of fern powder, 3-5 parts of mulberry branches and leaves powder, 3-5 parts of fragrant wood branches and leaves powder, and 1-2 parts of Bacillus subtilis;

[0044] The truffle mycorrhizal growth conditioner comprises, by weight, 25-30 parts vermiculite, 25-30 parts perlite, 24-28 parts peat moss, 8-12 parts limestone powder, 3-5 parts truffle spore powder, 0.005-0.015 parts gibberellin (GA), 0.005-0.015 parts naphthaleneacetic acid (NAA), 1-3 parts potassium humate, and 1-3 parts extract of *Cinnamomum camphora* branches and leaves.

[0045] In a preferred embodiment of the present invention, the soil conditioner comprises, by weight, 26 parts of edible fungus residue, 26 parts of cow dung, 12 parts of sugarcane bagasse, 9 parts of distiller's grains, 6 parts of peat moss, 4 parts of silkworm excrement, 7 parts of fern powder, 4 parts of mulberry branch and leaf powder, 4 parts of fragrant wood branch and leaf powder, and 2 parts of Bacillus subtilis.

[0046] The mycorrhizal growth conditioner comprises, by weight, 28 parts vermiculite, 28 parts perlite, 26 parts peat moss, 10 parts limestone powder, 3.98 parts mature truffle spore powder, 0.01 parts GA, 0.01 parts NAA, 2 parts potassium humate, and 2 parts extract of *Cinnamomum camphora* branches and leaves.

[0047] The aforementioned soil conditioner, specifically the edible mushroom residue, mainly contains abundant organic matter, amino acids, and biological enzymes, which can increase the soil's organic matter content, making it easier for truffle seedlings to absorb and utilize nutrients and minerals in the soil, thus promoting mycorrhizal growth and development. Sugarcane bagasse contains carbon sources and organic matter necessary for truffle mycelial growth, which can regulate the carbon-nitrogen ratio in the soil and facilitate the formation of new mycorrhizae. Distillers' grains mainly contain yeast, protein, and sugar, providing nutrients for plant mycorrhizal growth and development. Silkworm excrement is rich in enzymes and vitamins, providing vitamins and trace elements for plant and mycorrhizal growth. Fern shreds provide active calcium for mycorrhizal formation, promoting the growth and development of truffle mycorrhizae. Shredded branches and leaves of *Cinnamomum camphora* and *Aquilaria sinensis* can inhibit the growth and reproduction of pathogenic bacteria, provide abundant organic matter, change soil looseness, and promote mycorrhizal growth.

[0048] Mycorrhizal growth conditioners vermiculite and perlite primarily regulate soil moisture content and improve the water-holding capacity of the soil matrix. Gibberellin (GA) and naphthaleneacetic acid (NAA) promote the germination of new roots and the formation of numerous lateral roots in the host plant. Peat moss, rich in organic matter and humic acid, promotes the germination of truffle spores. Potassium humate is beneficial for the reproduction of truffle mycelia after spore germination. The addition of truffle spore powder promotes the rapid formation of mycorrhizae from new roots, regulates the polarity ratio of mycorrhizae, and shortens the initial production period of truffles. Extracts from mulberry branches and leaves inhibit contaminating bacteria and promote mycorrhizal growth and development.

[0049] In a preferred embodiment of the present invention, the soil conditioner is prepared by the following steps: After weighing the raw materials according to the proportions of the raw material components, the waste residue from the mature substrate mushroom cultivation bag, ferns, *Cinnamomum camphora* branches and leaves, and *Indiana macrophylla* branches and leaves are crushed and mixed with sugarcane bagasse. The mixture is pre-moistened with water at room temperature, covered with a film, and piled for 48 hours. Cow dung, peat moss, silkworm excrement, distiller's grains, and *Bacillus subtilis* are added and mixed thoroughly, and the moisture content is adjusted to 63%. The mixture is then piled into strips 1.0–1.5 m wide, 0.8–1 m high, and of unlimited length, and covered with a plastic film. Fermentation is carried out. When the temperature inside the material reaches above 65℃, fermentation is maintained for 48 hours, and the pile is turned over for the first time. When turning, the pile is turned inside out and top to bottom. When the temperature inside the pile exceeds 65℃ again, fermentation is maintained for 48 hours, and the pile is turned over for the second time. After the second turning, the temperature inside the pile exceeds 65℃ again, and fermentation is maintained for another 48 hours, and the pile is turned over for the third time. Then, fermentation continues for another 48 hours. After three turnings, the soil conditioner is obtained. At this time, the moisture content of the soil conditioner is 28wt%, and it is stored at 25℃ for later use.

[0050] In a preferred embodiment of the present invention, a method for cultivating truffles is provided, comprising the following steps: Calculating the volume of the original soil according to the area and depth of the soil to be modified; measuring the corresponding volume of limestone powder according to the proportion; spreading the limestone powder evenly on the soil surface; mixing the limestone with the soil using a rotary tiller to a depth of 20cm; evenly spreading 20kg / plant soil conditioner within a 0.5m radius centered on the seedling; mixing it into the soil again using a rotary tiller to a depth of 20cm; planting truffle mycorrhizal seedlings; installing a sprinkler irrigation system; watering once every half month from February to June each year; weeding 2-3 times by machine from July to September each year; in the second year after planting, drilling a hole 10cm in diameter and 20cm deep in each of the four directions (east, south, west, and north) directly below the drip line of the tree canopy, exposing the tree roots; inoculating the mycorrhizal growth conditioner into the holes at a rate of 1kg / hole; covering with topsoil after inoculation.

[0051] In the above planting method, the volume of the original soil is calculated according to the area and depth of the soil to be improved. A corresponding volume of limestone powder is then measured and evenly mixed into the soil. The volume percentage is 85-95 parts soil and 8-12 parts limestone powder. Preferably, the ratio is 90 parts original soil and 10 parts limestone powder.

[0052] This invention improves soil quality by adding an appropriate amount of limestone, which can increase and regulate soil pH, provide the calcium needed for mycorrhizal formation, increase soil aeration, and reduce the contamination rate of foreign mycorrhizal bacteria.

[0053] Of the planting methods described above, 20 kg of soil conditioner per plant is optimal. Excessive soil conditioner increases planting costs, reduces rhizosphere soil content, alters soil micronutrients and minerals, and negatively impacts the rhizosphere microbial community, hindering mycorrhizal growth and truffle production. Conversely, insufficient addition results in soil lacking looseness, organic matter, and other nutrients, also negatively impacting mycorrhizal reproduction and truffle production.

[0054] Similarly, adding too much mycorrhizal growth conditioner will increase the cost of truffle cultivation, and enlarged holes will increase root damage, affect the amount of mycorrhizal formation, and delay truffle production.

[0055] The present invention will be explained below with reference to embodiments. Those skilled in the art will understand that the following embodiments are for illustrative purposes only and should not be considered as limiting the scope of the invention. Where specific techniques or conditions are not specified in the embodiments, they are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. Reagents or instruments whose manufacturers are not specified are all conventional products that can be obtained commercially.

[0056] Example 1: Preparation of soil conditioner and mycorrhizal growth regulator.

[0057] (1) Preparation of soil conditioner:

[0058] A. Soil conditioner components, by weight, are: 26 parts waste residue from edible mushroom substrate cultivation bags, 26 parts cow manure from penned livestock, 12 parts sugarcane bagasse, 9 parts brewer's grains, 6 parts peat moss, 4 parts silkworm excrement, 7 parts pulverized fern material with a particle size of 0.2–1.0 cm, 4 parts pulverized branches and leaves of *Cinnamomum camphora* with a particle size of 0.2–1.0 cm, 4 parts pulverized branches and leaves of *Aristolochia debilis* with a particle size of 0.2–1.0 cm, and 2 parts *Bacillus subtilis*.

[0059] B. After weighing the raw materials according to the above proportions of soil conditioner components, crush the waste residue of edible fungi substrate bags, ferns, mulberry branches and leaves, and fragrant wood branches and leaves, and mix them with sugarcane bagasse. Pre-moisten with water at room temperature, cover with film and pile for 48 hours. Add cow dung, peat moss, silkworm excrement, distiller's grains, and Bacillus subtilis and mix well, adjusting the moisture content to 63%. Build a strip-shaped pile with a width of 1.0-1.5m, a height of 0.8-1m, and unlimited length. Cover with plastic film for fermentation. When the temperature inside the pile reaches above 65℃, maintain fermentation for 48 hours and then turn the pile for the first time, turning it inside out and top to bottom. When the temperature inside the pile exceeds 65℃ again, maintain fermentation for 48 hours and then turn it for the second time. After the second turning, if the temperature inside the pile exceeds 65℃ again, maintain fermentation for another 48 hours and then turn it for the third time. Continue fermentation for another 48 hours. After three turnings, the soil conditioner is obtained. At this point, the moisture content of the soil conditioner is 28wt%. Store at 25℃ for later use.

[0060] (2) Preparation of mycorrhizal growth conditioner:

[0061] A. Mycorrhizal growth conditioner composition by weight: 28 parts vermiculite with a particle size of 3-6 mm, 28 parts perlite with a particle size of 3-6 mm, 26 parts peat moss, 10 parts limestone powder with a particle size of ≤10 mm, 3.98 parts mature Indian truffle spore powder with a particle size of ≤0.2 mm and a maturity of ≥90%, 0.01 parts GA, 0.01 parts NAA, 2 parts potassium humate, and 2 parts extract of tamarisk branches and leaves;

[0062] The truffle spore powder is Indian truffle spore powder. The maturity of the truffle spores is detected under an Olympus microscope, and then the spores are subjected to aseptic surface treatment and crushed into truffle spore powder with a particle size ≤0.2mm.

[0063] The extract of *Corydalis tangutica* branches and leaves is prepared by crushing *Corydalis tangutica* branches and leaves into powder with a diameter ≤0.2mm, soaking them at room temperature for 24 hours at a material-to-water ratio of 1:20, boiling them at a temperature >95℃ for 1 hour, cooling them to below 30℃, adding 5% honey residue (based on the weight of *Corydalis tangutica* branches and leaves) and letting them stand for 48 hours, filtering them to obtain the extract, and storing it at 2-3℃ for later use.

[0064] B. Weigh out each component of the truffle mycorrhizal growth conditioner according to the above proportions, and mix them evenly to obtain the mycorrhizal growth conditioner.

[0065] Example 2 uses the truffle cultivation formula of Example 1 to cultivate Indian truffles.

[0066] The first step is to calculate the volume of the original soil (2m x 2m, 20cm deep) according to the ratio of 90 parts original soil and 10 parts limestone powder. Then, calculate the volume of limestone powder according to the formula ratio. Spread the limestone evenly in the 2m x 2m area and use a rotary tiller to mix the limestone with the soil to a depth of 20cm.

[0067] The second step is to evenly spread 20 kg of soil conditioner within a 0.5 m radius of the seedling, then mix it into the soil with a rotary tiller to a depth of 20 cm. Plant chestnut Indian truffle mycorrhizal seedlings, install a sprinkler system, and water once every half month from February to June each year. Weed 2-3 times a year from July to September. In the second year after planting, make a hole with a diameter of 10 cm and a depth of 20 cm in each of the four directions (east, south, west, and north) directly below the drip line of the tree canopy, exposing the tree roots. Inoculate the mycorrhizal growth conditioner into the hole at a rate of 1 kg / hole, and cover with topsoil after inoculation.

[0068] Using this technology, 30 chestnut truffle seedlings were planted. In the third year, 2 seedlings began to produce fruit, with the heaviest single seedling weighing 509g and the heaviest single truffle weighing 154g, with an average yield of 331.5g / seedling. In the fourth year, 10 seedlings began to produce fruit, with the heaviest single seedling weighing 650g and the heaviest single truffle weighing 650g, with an average yield of 276.47g / seedling. In the fifth year, 12 seedlings began to produce fruit, with the heaviest single seedling weighing 1054g and the heaviest single truffle weighing 183g, with an average yield of 637g / seedling. In the sixth year, 14 seedlings began to produce fruit, with the heaviest single seedling weighing 1066g and the heaviest single truffle weighing 375g, with an average yield of 529g / seedling.

[0069] Comparative Example 1: Cultivation of Indian Truffle using Traditional Methods

[0070] Traditional chestnut cultivation: The first step is to dig pits according to the planting spacing design, plant chestnut truffle mycorrhizal seedlings in the center of the pits, install a sprinkler system, water once every half month from February to June each year, and weed by machine 2 to 3 times from July to September each year.

[0071] Thirty chestnut truffle seedlings were planted using traditional methods, but none of them produced any fruit after six years of cultivation.

[0072] Comparative Example 2: Indian Truffles were cultivated using limestone-modified soil.

[0073] The first step is to calculate the volume of the original soil (2m x 2m, 20cm deep) according to the ratio of 90 parts original soil and 10 parts limestone powder. Then, calculate the volume of limestone according to the formula ratio. Spread the limestone evenly in the 2m x 2m area and use a rotary tiller to mix the limestone with the soil to a depth of 20cm.

[0074] The second step involves digging holes according to the planting spacing design, planting chestnut Indian truffle mycorrhizal seedlings, and installing a sprinkler irrigation system. Watering is done every two weeks from February to June each year, and weeding is carried out 2-3 times a year from July to September. The development of the truffle mycorrhizae, the first year of truffle production, and the yield are shown in Tables 1 and 2.

[0075] Comparative example 3 plants were cultivated with soil conditioner to grow Indian truffles

[0076] Spread 20kg of soil conditioner evenly within a 0.5m radius of the seedling, then mix it into the soil with a rotary tiller to a depth of 20cm. Dig holes according to the planting row and plant chestnut Indian truffle mycorrhizal seedlings. Install a sprinkler irrigation system and water once every half month from February to June each year. Weed 2 to 3 times a year from July to September.

[0077] The mycorrhizal development of the truffles, the first production year of the truffles, and the yield are shown in Tables 1 and 2.

[0078] Comparative example: 4 specimens were planted with Indian truffles after adding mycorrhizal growth conditioner.

[0079] According to the planting spacing design, dig pits and plant chestnut truffle mycorrhizal seedlings in the center of the pits. Install a sprinkler irrigation system and water once every half month from February to June each year. Weed with a machine 2 to 3 times each year from July to September. In the second year after planting, make a hole with a diameter of 10cm and a depth of 20cm in each of the four directions (east, south, west, and north) directly below the drip line of the tree canopy, exposing the tree roots. Inoculate the mycorrhizal growth conditioner into the holes at a rate of 1kg / hole, and cover with topsoil after inoculation.

[0080] The mycorrhizal development of the truffles, the first production year of the truffles, and the yield are shown in Tables 1 and 2.

[0081] Table 1. Tuber mycorrhizal development in different improved embodiments.

[0082]

[0083] Table 2. Fruiting effect of truffles in different improvement implementation cases

[0084]

[0085] As shown in Tables 1 and 2 above, the technology of this invention can induce truffle mycorrhizae to grow towards the soil surface, effectively increasing the amount of chestnut Indian truffle mycorrhizae, promoting the formation of burn rings on truffle trees, reducing the contamination rate of mycorrhizal fungi, shortening the initial production period of truffles, and increasing truffle yield. Chestnut Indian truffle seedlings begin to produce fruit 3 years after planting, 2-5 years earlier than the traditional 5-8 years. The highest yield per plant is 1066g, and the heaviest single fruiting body is 650g, setting new domestic records for the earliest initial production period, highest yield per plant, and heaviest single fruiting body of Indian truffles.

Claims

1. A truffle cultivation formula, characterized in that: include: Soil conditioner and truffle mycorrhizal growth regulator; The soil conditioner components, by weight, include: 25-28 parts of edible fungus residue, 25-28 parts of cow dung, 10-15 parts of sugarcane bagasse, 8-10 parts of distiller's grains, 5-8 parts of peat moss, 3-5 parts of silkworm excrement, 5-8 parts of fern powder, 3-5 parts of mulberry branch and leaf powder, 3-5 parts of fragrant wood branch and leaf powder, and 1-2 parts of Bacillus subtilis. The truffle mycorrhizal growth conditioner comprises, by weight, 25-30 parts vermiculite, 25-30 parts perlite, 24-28 parts peat moss, 8-12 parts limestone powder, 3-5 parts truffle spore powder, 0.005-0.015 parts gibberellin, 0.005-0.015 parts naphthaleneacetic acid, 1-3 parts potassium humate, and 1-3 parts extract of *Cinnamomum camphora* branches and leaves.

2. The truffle cultivation formula according to claim 1, characterized in that: The soil conditioner comprises, by weight, 26 parts of edible fungus residue, 26 parts of cow dung, 12 parts of sugarcane bagasse, 9 parts of distiller's grains, 6 parts of peat moss, 4 parts of silkworm excrement, 7 parts of fern powder, 4 parts of mulberry branch and leaf powder, 4 parts of fragrant wood branch and leaf powder, and 2 parts of Bacillus subtilis. The truffle mycorrhizal growth conditioner comprises, by weight, 28 parts vermiculite, 28 parts perlite, 26 parts peat moss, 10 parts limestone powder, 3.98 parts mature truffle spore powder, 0.01 parts gibberellin, 0.01 parts naphthaleneacetic acid, 2 parts potassium humate, and 2 parts extract of *Cinnamomum camphora* branches and leaves.

3. The truffle cultivation formula according to claim 1 or 2, characterized in that: Meet at least one of the following: The edible mushroom residue is the waste residue from the cooked substrate mushroom bags; The cow dung mentioned is from cows raised in enclosures; The lees are beer lees; The peat moss has an organic matter content of ≥75wt% and a humic acid content of ≥30wt%. The particle size of the pulverized fern, mulberry branches and leaves, and fragrant wood branches and leaves is 0.2–1.0 cm; The vermiculite has a particle size of 3-6 mm; The perlite has a particle size of 3-6 mm; The limestone powder is ≤10mm; The truffle spore powder has a maturity of ≥90% and a particle size of ≤1mm; The extract of *Corydalis tangutica* branches and leaves is prepared by crushing *Corydalis tangutica* branches and leaves into powder with a diameter ≤0.2mm, soaking them at room temperature for 24-48 hours at a material-to-water ratio of 1-2:20, boiling them at a temperature >95℃ for 1-2 hours, cooling them to below 30℃, adding 5-7wt% honey residue (based on the weight of *Corydalis tangutica* branches and leaves), and letting them stand for 48-72 hours. After filtration, the extract is stored at 2-3℃.

4. The method for preparing the truffle cultivation formula according to any one of claims 1 to 3, characterized in that: Includes the following steps: A. Preparation of soil conditioner, including the following steps: a. Crush edible fungus residue, ferns, mulberry branches and leaves, and fragrant wood branches and leaves, mix them with sugarcane residue, pre-moisten with water at room temperature, cover with film and pile for 24-48 hours. b. After adding cow dung, peat moss, silkworm excrement, distiller's grains, and Bacillus subtilis and mixing well, adjust the moisture content to 60-65%; c. Build a material pile, cover it with a film for fermentation, and turn the pile over three times during fermentation to obtain a soil conditioner; B. Preparation of truffle mycorrhizal growth conditioner includes the following steps: weigh the components of the truffle mycorrhizal growth conditioner and mix them evenly.

5. The method for preparing the truffle cultivation formula according to claim 4, characterized in that: In preparing soil conditioners, step c must satisfy at least one of the following: The material pile has a width of 1.0 to 1.5 m, a height of 0.8 to 1 m, and a length that is unlimited. When the temperature inside the material is ≥65℃ each time, fermentation is maintained for 48-72 hours and then the pile is turned over. This process is repeated 3 times. After the third turning, fermentation continues for another 48-72 hours. Turn the pile over both inside and outside, top and bottom, to ensure even fermentation of the material. The soil conditioner has a water content of 20-30 wt% and is stored at a temperature of 20-25℃.

6. A method for cultivating truffles containing the truffle cultivation formula according to any one of claims 1 to 5, characterized in that: Includes the following steps: Mix limestone powder with soil, add soil conditioner, plant truffle mycorrhizal seedlings, and carry out regular watering, weeding and maintenance; in the second year of planting, inoculate the base of the tree with truffle mycorrhizal growth conditioner, and cover with topsoil after inoculation.

7. The truffle cultivation method according to the truffle cultivation formula of claim 6, characterized in that: The method for mixing limestone powder with soil is as follows: Calculate the volume of the original soil according to the area and depth of the soil to be improved, measure the corresponding volume of limestone powder according to the proportion, and mix it evenly into the soil; by volume, 85-95 parts soil and 8-12 parts limestone powder.

8. The truffle cultivation method according to the truffle cultivation formula according to claim 7, characterized in that: 90 parts soil, 10 parts limestone powder.

9. The truffle cultivation method according to the truffle cultivation formula according to claim 6, characterized in that: The method for inoculating the truffle mycorrhizal growth conditioner is to make a hole in each of the four directions (north, south, east, and west) directly below the drip line of the tree canopy, exposing the tree roots, inoculating the mycorrhizal growth conditioner into the holes, and then covering it with topsoil. The holes are 10-15 cm in diameter and 20-25 cm deep.

10. The truffle cultivation method according to the truffle cultivation formula of claim 6, characterized in that: The soil conditioner is added at a rate of 20–25 kg per plant.

11. The truffle cultivation method according to the truffle cultivation formula of claim 9, characterized in that: The amount of the truffle mycorrhizal growth conditioner added is 0.8–1.2 kg / well.

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