A method for cultivating and applying *Quercus mongolica* truffle seedlings

Abstract

This invention discloses a method for cultivating and applying *Quercus mongolica* mycorrhizal seedlings. It belongs to the field of truffle cultivation technology. This invention establishes for the first time the symbiotic relationship between *Quercus mongolica* and *Quercus mongolica*. A *Quercus mongolica* inoculum solution is prepared using *Quercus mongolica* inoculation solution, *Pseudomonas*, *Bacillus subtilis*, gibberellin, glucose, and biochar. This solution is then inoculated into the root system of *Quercus mongolica* seedlings at specific growth stages for mycorrhizal seedling cultivation. This method significantly shortens the mycorrhizal synthesis time, increases the infection rate and survival rate of mycorrhizae, and improves the cultivation effect. Furthermore, the symbiosis with *Quercus mongolica* also improves the growth indicators of *Quercus mongolica*, providing technical support for mycorrhizal seedling cultivation.

Description

Technical Field

[0001] This invention relates to the field of truffle mycorrhizal seedling cultivation technology, and more specifically to a method and application for cultivating *Quercus mongolica* truffle mycorrhizal seedlings. Background Technology

[0002] Truffles belong to the phylum Ascomycota, order Tubrales, family Tuberaceae, and genus Tuber. The main edible species are *Tuber nigricans*, *Tuber spp.*, and *Tuber spp.* Truffles possess a unique aroma, texture, and nutritional value. Humans have consumed truffles for thousands of years, making them a delicacy among wild mushrooms. They are considered one of the three great delicacies, alongside caviar and foie gras.

[0003] *Tube aestivum* Vitt. is a type of truffle. Morphological characteristics: The ascocarp is irregularly spherical, often with a concave base, 3-6 cm in diameter, with 5-6 angular tubercles on the surface, the tubercles reaching up to 0.5 cm in diameter. The interior (spore body) is pale white, later turning creamy or dirty yellowish-gray, sometimes with red or purple marbled patterns, containing interconnected and branching veins. The asci are initially club-shaped, later becoming nearly spherical, with an indistinct ascostalk, and generally contain 4 ascospores. The spores are elliptical, with a reticulate surface, 25-35 μm × 25-26 μm. When mature, the fruiting body of *Tube aestivum* is rich in nutrients, has a distinctive aromatic odor, and lives in symbiosis with its host. It is mainly found in Northern Europe, including Britain, and under broad-leaved forests in North America. In recent years, it has been discovered in the mountainous areas of northern and southwestern my country. In Europe, it is mostly cultivated semi-artificially. The commercially available truffles in the domestic market are primarily sourced from wild resources. However, due to the unique growth characteristics of truffles and improper harvesting methods, natural cultivation ponds and their ecological environments have been severely damaged, leading to a sharp decline in truffle production that falls far short of market demand. Therefore, developing artificial cultivation methods for summer truffles has become increasingly important.

[0004] Currently, the artificial cultivation procedure for truffles is as follows: seedling cultivation → inoculation of truffle spores at the roots of seedlings to form infected seedlings → transplanting of infected seedlings → management such as root turning and pruning → fruiting bodies begin to form after 5-7 years, and the fruiting bodies can continue to develop for 20-30 years. Existing technologies also include some artificial cultivation methods for *Tricholoma matsutake* mycorrhizal seedlings, such as using chestnut, *Pinus armandii*, and *Pinus yunnanensis* as hosts and *Tricholoma matsutake* as the parasitic fungus. However, these methods result in long mycorrhizal emergence times, low infection rates, and low seedling survival rates, leading to unsatisfactory cultivation results.

[0005] Therefore, how to provide a more effective method for cultivating summer truffle mycorrhizal seedlings is a technical problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0006] In view of this, the present invention provides a method for cultivating and applying *Quercus mongolica* mycorrhizal seedlings, which shortens the time for mycorrhizal emergence, increases the infection rate and survival rate of mycorrhizae, and greatly improves the cultivation effect.

[0007] To achieve the above objectives, the present invention adopts the following technical solution:

[0008] A method for cultivating *Quercus mongolica* truffle seedlings includes the following steps:

[0009] (1) Preparation of truffle spore suspension: Take mature fruiting bodies of truffle spores, disinfect them, crush them, add sterile water to prepare truffle spore suspension, and store it for later use.

[0010] (2) Preparation of *Tuber spore* inoculum: The *Tuber spore* suspension obtained in step (1) was mixed with *Pseudomonas*, *Bacillus subtilis*, gibberellin, glucose and biochar in a weight ratio of 20-25 parts: 4-6 parts: 0.5-1 parts: 3-4 parts: 10-15 parts: 15-20 parts to prepare *Tuber spore* inoculum.

[0011] (3) Inoculation: The aseptic seedlings of Mongolian oak were dipped in the inoculation solution of summer truffle for root treatment; the root-dipping seedlings were planted in the cultivation substrate, and the remaining inoculation solution was poured into the cultivation substrate;

[0012] (4) Post-inoculation cultivation: After the seedlings are planted in step (3), seedling management is carried out to prevent the occurrence of diseases and pests. After 3-3.5 months of cultivation, summer truffle mycorrhizal seedlings are obtained.

[0013] Preferably, in step (1), the fruiting body is disinfected by soaking in 75% alcohol for 3-5 seconds; the pulverization speed is 8000-19000 rpm and the pulverization time is 10-30 seconds; the storage temperature is 0-4℃.

[0014] Preferably, the mycelium block size of the spore suspension of *Ustilago maydis* in step (2) is 0.2-0.4 mm; and the particle size of the biochar is 0.6-0.7 mm.

[0015] Preferably, the effective spore count in the *Tuber spp.* inoculation solution in step (2) is ≥6 × 10⁻⁶. 6 The concentration of *Tuber spp.* inoculum is 7.5-8.5.

[0016] Preferably, the number of fibrous roots of the aseptic Mongolian oak seedlings in step (3) is 3-4, and the length of the fibrous roots is 5-6cm; the inoculation amount for each seedling is 0.1-0.2mL of summer truffle inoculation solution.

[0017] Preferably, the cultivation substrate in step (3) comprises the following components in parts by weight: 10-20 parts peat moss, 5-8 parts perlite and 10-15 parts vermiculite; wherein the peat moss, perlite and vermiculite are all sterilized at 121°C.

[0018] Preferably, the pH of the cultivation substrate in step (3) is 7.5-8.5 and the moisture content is 25-30%.

[0019] Preferably, the culture temperature in step (4) is 20-25℃, the substrate humidity is 25-28%, and the light conditions are 2000-5000 Lux.

[0020] Another object of the present invention is to provide the application of the above-described cultivation method in the cultivation of *Tuber spp.* mycorrhizal seedlings.

[0021] As can be seen from the above technical solution, compared with the prior art, the present invention has the following beneficial effects:

[0022] This invention utilizes the symbiotic relationship between Mongolian oak and summer truffles. A summer truffle inoculum is prepared using Pseudomonas, Bacillus subtilis, gibberellin, glucose, and biochar. This inoculum is then inoculated into the roots of Mongolian oak seedlings at specific growth stages to cultivate mycorrhizal seedlings. This significantly shortens the time for mycorrhizal emergence, increases the infection rate and survival rate, and improves the cultivation effect. Furthermore, the parasitism of the truffles improves the growth status of Mongolian oak, providing technical support for the cultivation of truffle mycorrhizae. Attached Figure Description

[0023] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0024] Figure 1 For: the morphology of mycorrhizae;

[0025] Figure 2 Image: Magnified image of mycorrhizal morphology. Detailed Implementation

[0026] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0027] Example 1

[0028] A method for cultivating *Quercus mongolica* truffle seedlings includes the following steps:

[0029] (1) Preparation of truffle sporospore suspension: Take mature truffle fruiting bodies at room temperature, wash the surface, disinfect in 75% alcohol for 3-5 seconds, blot the alcohol off the surface with filter paper, peel off the epidermis of the fruiting body with a sterile knife, and cut into small squares of about 1cm×1cm for later use; wipe the mixer with 75% alcohol, rinse with sterile water, repeat 3 times, weigh 65g of the small pieces of fruiting body each time and put them into the prepared mixer, pulverize at 8000rpm for 30s, add sterile water, and prepare truffle sporospore suspension with a particle size of 0.2-0.4mm, and store at 0℃ for later use;

[0030] (2) Preparation of *Tuber spp.* inoculum: The *Tuber spp.* spore suspension obtained in step (1) was mixed with *Pseudomonas aeruginosa* (a strain already disclosed in the prior art, preservation number CGMCC No. 5040), *Bacillus subtilis*, gibberellin, glucose, and biochar at a weight ratio of 22:5:1:3:12:15 to prepare the *Tuber spp.* inoculum. The number of effective spores in the *Tuber spp.* inoculum was statistically analyzed to be ≥6×10⁻⁶. 6 The concentration of *Tuber spp.* was 1 / mL; the pH of the *Tuber spp.* inoculum was 7.5; and the particle size of the biochar was 0.6-0.7 mm.

[0031] (3) Inoculation: Place the aseptic seedlings of Mongolian oak with 3-4 fibrous roots and 5-6 cm in length into the inoculation solution of summer truffle obtained in step (2) and dip the roots (cover the base of the stem and soak for 15 min);

[0032] Mix 15 parts peat moss, 6 parts perlite and 12 parts vermiculite, sterilize at 121℃, adjust the pH of the substrate to 7.5 and the moisture content to 25% with lime or Ca(OH)2, and use it as the cultivation substrate.

[0033] Fill the prepared substrate into a container with a bottom diameter ≥8.5cm, a mouth diameter ≥10cm, and a height ≥18cm. The total amount of substrate should be 1 / 2 of the container. Plant the seedlings with roots dipped in the substrate into the container, cover them with soil, and leave the soil layer 2cm from the mouth of the container. Pour the remaining truffle inoculation solution into the cultivation substrate. The inoculation amount for each seedling is 0.1mL of truffle inoculation solution.

[0034] (4) Post-inoculation cultivation: After the seedlings are planted in step (3), seedling management is carried out. During the cultivation period, water should be irrigated to keep the substrate moist. The substrate humidity should be controlled at 25%, the cultivation temperature should be controlled at 20℃, and the light conditions should be 2000 Lux. After 3-3.5 months of cultivation, summer truffle mycorrhizal seedlings are obtained. When the mycorrhizal seedlings first develop powdery mildew, spray the diseased plants with 0.5mol / L lime water or remove the diseased plants.

[0035] Example 2

[0036] A method for cultivating *Quercus mongolica* truffle seedlings includes the following steps:

[0037] (1) Preparation of truffle spore suspension: Take mature truffle fruiting bodies at room temperature, wash the surface, disinfect in 75% alcohol for 3-5 seconds, blot the alcohol off the surface with filter paper, peel off the epidermis of the fruiting body with a sterile knife, and cut into small squares of about 1cm×1cm for later use; wipe the mixer with 75% alcohol and rinse with sterile water, repeat 3 times, weigh 65g of the fruiting body cut into small pieces each time and put them into the prepared mixer, grind at 19000rpm for 10s, add sterile water to prepare truffle spore suspension, the particle size of the fungal blocks is 0.2-0.4mm, store at 2℃ for later use;

[0038] (2) Preparation of *Tuber spp.* inoculum: The *Tuber spp.* spore suspension obtained in step (1) was mixed with *Pseudomonas aeruginosa* (preservation number CGMCC No. 5040), *Bacillus subtilis*, gibberellin, glucose, and biochar in a weight ratio of 20:4:0.5:4:10:20 to prepare the *Tuber spp.* inoculum. The number of effective spores in the *Tuber spp.* inoculum was statistically analyzed to be ≥6×10⁻⁶. 6 The concentration of *Tuber spp.* was 1 / mL; the pH of the *Tuber spp.* inoculum was 8.0; and the particle size of the biochar was 0.6-0.7 mm.

[0039] (3) Inoculation: Place the sterile seedlings of Mongolian oak with 3-4 fibrous roots and 5-6 cm in length into the inoculation solution of summer truffle obtained in step (2) and dip the roots (covering the base of the stem and soaking for 25 min);

[0040] Mix 10 parts peat moss, 5 parts perlite and 10 parts vermiculite, sterilize at 121℃, adjust the pH of the substrate to 8.0 with lime or Ca(OH)2 and adjust the moisture content to 30% to use as the cultivation substrate.

[0041] Fill the prepared substrate into a container with a bottom diameter ≥8.5cm, a mouth diameter ≥10cm, and a height ≥18cm. The total amount of substrate should be 2 / 3 of the container. Plant the seedlings with roots dipped in the substrate in the container, cover them with soil, and leave the soil layer 3cm from the mouth of the container. Pour the remaining truffle inoculation solution into the cultivation substrate. The inoculation amount for each seedling is 0.2mL of truffle inoculation solution.

[0042] (4) Post-inoculation cultivation: After the seedlings are planted in step (3), seedling management is carried out. During the cultivation period, water should be irrigated to keep the substrate moist. The substrate humidity should be controlled at 28%, the cultivation temperature should be controlled at 25℃, and the light conditions should be 5000 Lux. After 3-3.5 months of cultivation, summer truffle mycorrhizal seedlings are obtained. When the mycorrhizal seedlings first develop powdery mildew, spray the diseased plants with 0.5mol / L lime water or remove the diseased plants.

[0043] Example 3

[0044] A method for cultivating *Quercus mongolica* truffle seedlings includes the following steps:

[0045] (1) Preparation of *Tuber spore suspension*: Take mature fruiting bodies of *Tuber spore* at room temperature, wash the surface, disinfect in 75% alcohol for 3-5 seconds, blot the alcohol off the surface with filter paper, peel off the epidermis of the fruiting body with a sterile knife, and cut into small squares of about 1cm×1cm for later use; wipe the mixer with 75% alcohol, rinse with sterile water, repeat 3 times, weigh 65g of the fruiting body cut into small pieces each time and put them into the prepared mixer, grind at 15000rpm for 20s, add sterile water to prepare *Tuber spore suspension*, the particle size of the fungal blocks is 0.2-0.4mm, and store at 0℃ for later use;

[0046] (2) Preparation of *Tuber spp.* inoculum: The *Tuber spp.* spore suspension obtained in step (1) was mixed with *Pseudomonas aeruginosa* (preservation number CGMCC No. 5040), *Bacillus subtilis*, gibberellin, glucose, and biochar at a weight ratio of 25:6:0.75:3.5:15:17 to prepare the *Tuber spp.* inoculum. The number of effective spores in the *Tuber spp.* inoculum was statistically analyzed to be ≥6×10⁻⁶. 6 The concentration of *Tuber spp.* was 1 / mL; the pH of the *Tuber spp.* inoculum was 8.5; and the particle size of the biochar was 0.6-0.7 mm.

[0047] (3) Inoculation: Place the sterile seedlings of Mongolian oak with 3-4 fibrous roots and 5-6 cm in length into the inoculation solution of summer truffle obtained in step (2) and dip the roots (cover the base of the stem and soak for 20 min);

[0048] Mix 20 parts peat moss, 8 parts perlite and 15 parts vermiculite, sterilize at 121℃, adjust the pH of the substrate to 8.5 and the moisture content to 28% with lime or Ca(OH)2, and use it as the cultivation substrate.

[0049] Fill the prepared substrate into a container with a bottom diameter ≥8.5cm, a mouth diameter ≥10cm, and a height ≥18cm. The total amount of substrate should be 2 / 3 of the container. Plant the seedlings with soaked roots in the container, cover them with soil, and leave the soil layer 2cm from the mouth of the container. Pour the remaining truffle inoculation solution into the cultivation substrate. The inoculation amount for each seedling is 0.15mL of truffle inoculation solution.

[0050] (4) Post-inoculation cultivation: After the seedlings are planted in step (3), seedling management is carried out. During the cultivation period, water should be irrigated to keep the substrate moist. The substrate humidity should be controlled at 26%, the cultivation temperature should be controlled at 25℃, and the light conditions should be 3000 Lux. After 3-3.5 months of cultivation, summer truffle mycorrhizal seedlings are obtained. When powdery mildew first appears on the mycorrhizal seedlings, spray the diseased plants with 0.5mol / L lime water or remove the diseased plants.

[0051] Comparative Example 1

[0052] It is basically the same as Example 3, except that:

[0053] (2) Preparation of *Tuber spp.* inoculum: The *Tuber spp.* spore suspension obtained in step (1) was mixed with *Bacillus subtilis*, gibberellin, glucose, and biochar at a weight ratio of 25:0.75:3.5:15:17 to prepare the *Tuber spp.* inoculum. The number of effective spores in the *Tuber spp.* inoculum was statistically analyzed to be ≥6×10⁻⁶. 6 The concentration of *Tuber spp.* was 1 / mL; the pH of the *Tuber spp.* inoculum was 7.5; and the particle size of the biochar was 0.6-0.7 mm.

[0054] Comparative Example 2

[0055] It is basically the same as Example 3, except that:

[0056] The symbiotic tree species are different; they are both species of the genus *Quercus*, specifically *Quercus suber*.

[0057] Comparative Example 3

[0058] It is basically the same as Example 3, except that:

[0059] (3) Inoculation: Place the sterile seedlings of Mongolian oak with 2 fibrous roots and a length of 4 cm into the inoculation solution of summer truffle obtained in step (2) and dip the roots (covering the base of the stem and soaking for 20 min).

[0060] Comparative Example 4

[0061] It is basically the same as Example 3, except that:

[0062] (3) Inoculation: Place the sterile seedlings of Mongolian oak with 6 fibrous roots and a length of 8 cm into the inoculation solution of summer truffle obtained in step (2) and dip the roots (covering the base of the stem and soaking for 20 min).

[0063] To evaluate the cultivation of *Tuber spp.* mycorrhizal seedlings from different groups, continuous sampling and testing of mycorrhizal infection were conducted. The specific sampling method is as follows:

[0064] Random sampling: Random sampling is conducted at irregular intervals during the seedling stage. When the roots of the seedlings with soil attached grow to the surface of the substrate and mycorrhizae are found, a handheld magnifying glass of ≥20x is used to observe the development of the mycorrhizae. If the mycorrhizae are normal and uncontaminated, they are put back into the container for continued cultivation.

[0065] Microscopic examination: Mycorrhizal seedlings are randomly selected at irregular intervals, and mycorrhizal sections are taken for microscopic examination of mycorrhizae. The sections should have the morphological structure of truffle mycorrhizae.

[0066] Molecular detection: The mycorrhizae of qualified mycorrhizal seedlings are cut and examined under a microscope, and DNA testing and sequence comparison are performed. Contaminated and non-tuber mycorrhizae are removed and discarded.

[0067] The specific seedling sampling rules are shown in Table 1, and the sampling results are shown in Table 2. The specific mycorrhizal morphology is as follows: Figure 1 and Figure 2 As shown.

[0068] Table 1. Number of seedlings sampled for testing

[0069] Number of seedlings Number of plants tested 500～1000 50 1001～10000 100 10001～50000 250 50001～100000 350 100001～500000 500 500001 and above 750

[0070] Table 2. Mycorrhizal infection status of different groups

[0071]

[0072] Results analysis: According to Table 1, the truffle mycorrhizal seedlings of Examples 1-3 of the present invention have a short formation time, relatively high infection rate and survival rate, indicating that the cultivation method of the present invention has achieved significant results.

[0073] Furthermore, a comparison between Example 3 and Comparative Example 1 shows that adding Pseudomonas (CGMCC No. 5040) to the truffle inoculation solution can significantly shorten the mycorrhizal formation time from 50 days to about 24 days, while also increasing the infection rate. The reason for this may be that Pseudomonas can secrete a variety of plant hormones, which can promote root branching and growth, providing more infection sites for truffle mycelia. At the same time, Pseudomonas can improve the soil microenvironment and secrete some antibiotic or signaling molecules, which can not only promote the symbiotic relationship between truffle spores and plant roots, accelerate the mycorrhizal formation time, and increase the infection rate, but also promote the growth of symbiotic plants.

[0074] The comparative experiments of Example 3 and Comparative Example 2 show that the symbiotic relationship between truffles and plants is very subtle. Different truffle species and different tree species have different symbiotic affinity with truffles due to differences in the physiological characteristics and root structure of the tree species. In Comparative Example 2, another tree species in the genus Quercus, Quercus suber, was used as the symbiotic plant and inoculated with truffle inoculum. Its mycorrhizal growth was far inferior to that of summer truffle and Mongolian oak.

[0075] As can be seen from Example 3 and Comparative Examples 3 and 4, different inoculation times also affect the overall inoculation effect. When the number of fibrous roots of Mongolian oak seedlings is 3-4 and the length of the fibrous roots is 5-6 cm, the seedlings have the strongest affinity with the inoculated truffle spores and the inoculation effect is the best.

[0076] To verify the effect of symbiosis between truffle mycorrhizae and Mongolian oak on the growth of Mongolian oak, the changes in the growth status of different groups of Mongolian oak after 2 months of symbiotic culture were measured compared to the initial stage of symbiosis. A blank control group without truffle inoculation was also set up.

[0077] The experimental results are shown in Table 3:

[0078] Table 3. Growth status of different groups of Mongolian oak

[0079]

[0080] Results analysis: As shown in Table 3, this cultivation method can not only improve the infection effect of mycorrhizae, but also, due to the symbiotic relationship between mycorrhizae and tree species, can improve the absorption capacity of Mongolian oak of key nutrients in the soil to a certain extent, thus promoting tree growth and achieving a mutually beneficial symbiotic effect.

[0081] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on the differences from other embodiments. The same or similar parts between the various embodiments can be referred to each other.

[0082] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for cultivating Quercus mongolica summer truffle mycorrhizal seedlings, characterized by, It comprises the following steps: (1) Preparation of Tricholoma populinum spore suspension: Take the mature fruiting bodies of Tricholoma populinum, sterilize them, crush them, add sterile water, and prepare Tricholoma populinum spore suspension for storage; (2) Preparation of Tricholoma populinum inoculum: Mix the Tricholoma populinum spore suspension obtained in step (1) with Pseudomonas, Bacillus subtilis, gibberellin, glucose and biochar according to the weight ratio of 20-25:4-6:0.5-1:3-4:10-15:15-20, to prepare Tricholoma populinum inoculum; (3) Inoculation: Dip the sterile seedlings of Quercus mongolica in the Tricholoma populinum inoculum; plant the dipped seedlings in the cultivation substrate, and pour the remaining inoculum into the cultivation substrate; (4) Post-inoculation cultivation: After planting the seedlings in step (3), carry out seedling management and prevent the occurrence of diseases and pests, and cultivate for 3-3.5 months to obtain Tricholoma populinum mycorrhizal seedlings.

2. The cultivation method of the Quercus mongolica-summer block truffle mycorrhizal seedling according to claim 1, characterized by, In step (1), the fruiting bodies are soaked in 75% alcohol for 3-5 s; the crushing speed is 8000-19000 rpm, and the crushing time is 10-30 s; the storage temperature is 0-4℃.

3. The method of cultivating the Quercus mongolica-summer block truffle mycorrhizal seedling according to claim 1, characterized by, In step (2), the particle size of the Tricholoma populinum spore suspension is 0.2-0.4 mm; the particle size of the biochar is 0.6-0.7 mm.

4. The method of cultivating the Quercus mongolica-summer block truffle mycorrhizal seedling according to claim 1, characterized by, The effective spore number in the Grifola frondosa inoculum solution in step (2) is ≥ 6 x 10 6 The pH of the Grifola frondosa inoculum solution is 7.5-8.

5.

5. The method of cultivating the Quercus mongolica-summer block truffle mycorrhizal seedling according to claim 1, characterized by, In step (3), the number of root hairs of the sterile seedlings of Quercus mongolica is 3-4, and the length of the root hairs is 5-6 cm; the inoculation amount of each seedling is 0.1-0.2 mL of Tricholoma populinum inoculum.

6. The method of cultivating the Quercus mongolica-summer block truffle mycorrhizal seedling according to claim 1, characterized by, In step (3), the cultivation substrate comprises the following components by weight: 10-20 parts of peat soil, 5-8 parts of perlite and 10-15 parts of vermiculite; the peat soil, perlite and vermiculite are all subjected to sterilization treatment at 121℃.

7. The method of cultivating the Quercus mongolica-summer block truffle mycorrhizal seedling according to claim 1, characterized by, In step (3), the pH of the cultivation substrate is 7.5-8.5, and the moisture content is 25-30%.

8. The method of cultivating the Quercus mongolica-summer block truffle mycorrhizal seedling according to claim 1, characterized by, In step (4), the cultivation temperature is 20-25℃, the substrate humidity is 25-28%, and the light condition is 2000-5000 Lux.

9. The use of the cultivation method of any one of claims 1-8 in the cultivation of Tricholoma populinum mycorrhizal seedlings.

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