Multi-tree-species synergistic summer truffle mycorrhiza seedling cultivation method and application

Abstract

The invention discloses a multi-tree-species cooperative summer truffle mycorrhiza seedling cultivation method and application. Belongs to the technical field of truffle cultivation. According to the method, Mongolian oak and pinus sylvestris are adopted as symbiotic hosts of summer truffles for the first time, the mycorrhizal fungi are subjected to seedling raising and cultivation in a two-tree-species synergistic mode, the synergistic effect is achieved, the mycorrhizal seedling mycorrhiza forming time is remarkably shortened, the mycorrhiza infection rate and the mycorrhiza seedling survival rate are increased, and the survival rate of mycorrhiza seedlings is increased. The increase of the yield of the mycorrhiza of the summer truffles is greatly promoted, and a technical support is provided for artificial cultivation of the summer truffles.

Description

Technical Field

[0001] The invention relates to the technical field of truffle mycorrhizal seedling raising and cultivation, and more particularly to a multi-tree species coordinated truffle mycorrhizal seedling raising and cultivation method and application. Background Art

[0002] Summer truffle (Tube aestivum Vitt.) belongs to the genus Tuber, Tubrales, Tuberaceae, Ascomycota. As a kind of truffle, it is mainly produced in Europe and other countries, and is a favorite kitchen ingredient in European countries. When the fruiting body of summer truffle matures, it is rich in nutrition and has a special aromatic smell. It coexists with the host and is mostly produced in the broad-leaved forests of Northern Europe such as the United Kingdom and North America. In recent years, it has been found in the mountainous forests in northern and southwestern my country. At present, the commercial truffles in the domestic market are basically derived from wild resources. Due to the particularity of truffle growth and incorrect excavation methods, natural fungus ponds and their ecological environment have been severely damaged, and the truffle production has dropped sharply, which is far from meeting market demand. Therefore, it is becoming increasingly important to develop artificial cultivation methods for summer truffles.

[0003] At present, the artificial cultivation procedure of truffles is: cultivate seedlings → inoculate truffle spores at the roots of seedlings to form infected seedlings → transplant infected seedlings → root turning and trimming and other management → fruiting bodies begin to form in 5 to 7 years, and the fruiting bodies can continue to form for 20 to 30 years. In the prior art, the artificial cultivation method of summer truffle mycorrhizal seedlings is mostly based on chestnut, red pine, American pecan, etc. as hosts for artificial cultivation of truffles, but the above-mentioned artificial cultivation method is mostly a parasitic relationship of a single tree species, which will significantly reduce the species diversity in forest or woodland ecosystems, leading to the simplification of soil microbial community structure, soil nutrient imbalance, and decreased fertility; at the same time, a single tree species is easy to form a large area of ​​homogeneous environment, providing an ideal habitat for specific types of pathogens or pests, and increasing the possibility of pests and diseases. The occurrence of the above problems has limited the growth of truffles to a certain extent, resulting in a long mycorrhizal appearance time, low infection rate, low survival rate of mycorrhizal seedlings, and unsatisfactory cultivation effect.

[0004] Therefore, how to provide a more effective multi-tree species collaborative summer truffle mycorrhizal seedling cultivation method is a technical problem that technical personnel in this field urgently need to solve. Summary of the invention

[0005] In view of this, the present invention provides a multi-tree species collaborative summer truffle mycorrhizal seedling cultivation method and application, using Mongolian oak and Larix gmelinii collaboratively as symbiotic hosts to carry out summer truffle mycorrhiza seedling cultivation and cultivation, which significantly improves the seedling and cultivation effects.

[0006] In order to achieve the above object, the present invention adopts the following technical solution:

[0007] A multi-tree species cooperative summer truffle mycorrhizal seedling cultivation method comprises the following steps:

[0008] (1) Preparation of summer truffle fruiting body powder: sterilize and grind the mature summer truffle fruiting bodies to obtain summer truffle fruiting body powder, and store it at low temperature for later use;

[0009] (2) Preparation of liquid culture solution: Add the fruiting body powder of Truffle sutchuenensis and Fusarium oxysporum to a liquid culture medium, place at 22-25° C., and culture with shaking for 7-10 days to obtain a liquid culture solution;

[0010] (3) Inoculation and cultivation: placing the sterile seedlings of Mongolian oak and Mongolian pine in the liquid culture solution obtained in step (2), covering the stem base, and soaking for 10-20 minutes; planting the soaked Mongolian oak seedlings and Mongolian pine seedlings in a mixed inter-row pattern of 1-2 rows: 2-3 rows in a cultivation medium, and pouring the remaining liquid culture solution into the cultivation medium along the roots of the seedlings for cultivation, to obtain Mongolian oak and Mongolian pine summer truffle mycorrhizal seedlings;

[0011] (4) Transplanting: The Mongolian oak and Mongolian pine summer truffle mycorrhizal seedlings obtained in step (3) are transplanted in a mixed inter-row manner of 1-2 rows: 4-6 rows. After transplanting, regular management is performed to prevent the occurrence of diseases and insect pests, and summer truffles are harvested after cultivation.

[0012] Preferably, in step (1), the fruiting bodies are immersed in 75% alcohol for disinfection for 3-5 seconds; the rotation speed of the crushing is 8000-19000 rpm, and the crushing time is 10-30 seconds; and the storage temperature is 0-4°C.

[0013] Preferably, the composition of the liquid culture medium in step (2) is as follows: glucose 15-25 g / L, maltose 10-13 g / L, peptone 3-4 g / L, KH2PO4 4-5 g / L, MgSO4·7H2O 2-3 g / L and vitamin B1 0.006-0.007 g / L.

[0014] Preferably, the dry weight of mycelium in the liquid culture solution obtained in step (2) is 1-2 g / L; the content of Fusarium oxysporum in the liquid culture solution obtained is 2×10 6 -8×10 6 CFU / mL; adjust the pH of the liquid culture solution to 7.5-8.0.

[0015] Preferably, the rotation speed of the shaking culture in step (2) is 100-150 rpm, and the culture is carried out in the dark.

[0016] Preferably, the root length of the Mongolian oak seedling in step (3) is 6-8 cm, and the stem length is 5-10 cm; the root length of the Mongolian pine is 5-10 cm, and the stem length is 10-20 cm.

[0017] Preferably, the cultivation medium in step (3) is composed of: 40-50 g humus, 30-35 g perlite, 15-20 g loam and 10-15 g limestone particles; the pH of the cultivation medium is 7.5-8.5.

[0018] Preferably, in step (3), the inoculation amount of the Mongolian oak seedlings is 0.2-0.4 ml of liquid culture solution per seedling; the inoculation amount of the Larix gmelinii seedlings is 0.2-0.3 ml of liquid culture solution per seedling; the culture temperature is 22-25°C, the humidity of the cultivation medium is 35-45%, and the lighting conditions are 1500-4000 Lux.

[0019] Another object of the present invention is to provide: application of the above-mentioned multi-tree species cooperative summer truffle mycorrhizal seedling cultivation method in summer truffle production.

[0020] It can be seen from the above technical solution that compared with the prior art, the present invention has the following beneficial effects:

[0021] The present invention is directed to an improved artificial cultivation method for summer truffles. On the basis of the original single symbiotic host, a symbiotic relationship between Mongolian oak and Scots pine and summer truffles is constructed. Under the symbiotic relationship, Mongolian oak, Scots pine and summer truffles reach a relatively stable symbiotic balance, promote each other, and mutually benefit and symbiosis, forming a nutrient complementary effect, improving the ecological stability under the symbiotic environment, and significantly shortening the time of mycorrhizal formation compared to the symbiosis between a single tree species and summer truffles, improving the infection rate and survival rate, and obtaining a great improvement in yield. At the same time, the summer truffle mycorrhizal seedlings prepared by the method of the present invention have strong stress resistance and can effectively tolerate the influence of adverse temperature and humidity on mycorrhizal growth. DETAILED DESCRIPTION

[0022] The technical solutions in the embodiments of the present invention are described clearly and completely below. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0023] The Fusarium oxysporum used in the present invention is a strain disclosed in the prior art and is deposited in the General Microbiological Center of China Microbiological Culture Collection Committee with a deposit number of CGMCC No. 19031.

[0024] Example 1

[0025] A multi-tree species cooperative summer truffle mycorrhizal seedling cultivation method comprises the following steps:

[0026] (1) Preparation of summer truffle fruiting body powder: Take mature summer truffle fruiting bodies, soak them in 75% alcohol for 3-5 seconds to disinfect them, and then crush them at a speed of 8000 rpm for 30 seconds to prepare summer truffle fruiting body powder, which is stored at a low temperature of 0-4°C for later use;

[0027] (2) Preparation of liquid culture solution: The fruiting body powder of summer tuber and Fusarium oxysporum were added to a liquid culture medium consisting of 15 g / L glucose, 10 g / L maltose, 3 g / L peptone, 4 g / L KH2PO4, 2 g / L MgSO4·7H2O and 0.006 g / L vitamin B1. The culture was carried out at 22-25°C, protected from light, and shaken at 100 rpm for 10 days to obtain a liquid culture solution. The dry weight of mycelium in the liquid culture solution was determined to be 1.24 g / L, and the content of Fusarium oxysporum was 4.32×10 6 CFU / mL; adjust the pH of the liquid culture solution to 7.5-8.0;

[0028] (3) Inoculation and cultivation: A sterile seedling of Mongolian oak with a root length of 7 cm and a stem length of 9 cm and a sterile seedling of Pinus sylvestris with a root length of 5 cm and a stem length of 10 cm are placed in the liquid culture solution obtained in step (2), covering the stem base, and soaked for 10-20 minutes; the soaked Mongolian oak seedlings and the Mongolian pine seedlings are planted in a cultivation medium (40 g humus, 30 g perlite, 15 g loam and 10 g limestone particles, pH 7.5-8.5) in a 1:3 row inter-row mixed manner, and the remaining liquid culture solution is poured into the cultivation medium along the roots of the seedlings, the inoculation amount of the Mongolian oak seedlings is 0.4 ml of liquid culture solution / seedling, and the inoculation amount of the Mongolian pine seedlings is 0.3 ml of liquid culture solution / seedling, and the cultivation is carried out at 22-25° C., 35-45% humidity, and 1500-4000 Lux to obtain Mongolian oak and Mongolian pine summer truffle mycorrhizal seedlings;

[0029] (4) Transplanting: The Mongolian oak and Mongolian pine summer truffle mycorrhizal seedlings obtained in step (3) are transplanted in a 1-row:4-row inter-row mixed planting manner. After transplanting, regular management is performed to prevent the occurrence of diseases and insect pests, and summer truffles are harvested after cultivation.

[0030] Example 2

[0031] A multi-tree species cooperative summer truffle mycorrhizal seedling cultivation method comprises the following steps:

[0032] (1) Preparation of summer truffle fruiting body powder: Take mature summer truffle fruiting bodies, soak them in 75% alcohol for 3-5 seconds to disinfect them, and then crush them at a speed of 19000 rpm for 10 seconds to prepare summer truffle fruiting body powder, which is stored at a low temperature of 0-4°C for later use;

[0033] (2) Preparation of liquid culture solution: The fruiting body powder of summer tuber and Fusarium oxysporum were added to a liquid culture medium consisting of 25 g / L glucose, 13 g / L maltose, 4 g / L peptone, 5 g / L KH2PO4, 3 g / L MgSO4·7H2O and 0.007 g / L vitamin B1. The culture was incubated at 22-25°C, protected from light, and shaken at 150 rpm for 7 days to obtain a liquid culture solution. The dry weight of mycelium in the liquid culture solution was 1.36 g / L, and the content of Fusarium oxysporum was 3.84×10 6 CFU / mL; adjust the pH of the liquid culture solution to 7.5-8.0;

[0034] (3) Inoculation and cultivation: placing sterile seedlings of Mongolian oak with a root length of 8 cm and a stem length of 10 cm and a sterile seedling of Scots pine with a root length of 10 cm and a stem length of 20 cm in the liquid culture solution obtained in step (2), covering the stem base, and soaking for 10-20 minutes; planting the soaked Mongolian oak seedlings and Scots pine seedlings in a 2-row:3-row inter-row mixed planting method in a cultivation medium (50 g humus, 35 g perlite, 20 g loam and 15 g limestone particles, pH 7.5-8.5), and pouring the remaining liquid culture solution into the cultivation medium along the roots of the seedlings, the inoculation amount of the Mongolian oak seedlings is 0.2 ml of liquid culture solution / each, and the inoculation amount of the Scots pine seedlings is 0.2 ml of liquid culture solution / each, and the cultivation is carried out at 22-25° C., 35-45% humidity, and 1500-4000 Lux to obtain Mongolian oak and Scots pine summer truffle mycorrhizal seedlings;

[0035] (4) Transplanting: The Mongolian oak and Mongolian pine summer truffle mycorrhizal seedlings obtained in step (3) are transplanted in a 2-row:6-row interplanting manner. After transplanting, regular management is performed to prevent the occurrence of pests and diseases, and summer truffles are harvested after cultivation.

[0036] Example 3

[0037] A multi-tree species cooperative summer truffle mycorrhizal seedling cultivation method comprises the following steps:

[0038] (1) Preparation of summer truffle fruiting body powder: Take mature summer truffle fruiting bodies, soak them in 75% alcohol for 3-5 seconds to disinfect them, and then crush them at a speed of 10000 rpm for 20 seconds to prepare summer truffle fruiting body powder, which is stored at a low temperature of 0-4°C for later use;

[0039] (2) Preparation of liquid culture solution: The fruiting body powder of summer tuber and Fusarium oxysporum were added to a liquid culture medium consisting of 20 g / L glucose, 12 g / L maltose, 3 / L peptone, 4 g / L KH2PO4, 3 g / L MgSO4·7H2O and 0.007 g / L vitamin B1. The culture was incubated at 22-25°C, protected from light, and shaken at 150 rpm for 10 days to obtain a liquid culture solution. The dry weight of mycelium in the liquid culture solution was 1.64 g / L, and the content of Fusarium oxysporum was 4.52 × 10 6 CFU / mL; adjust the pH of the liquid culture solution to 7.5-8.0;

[0040] (3) Inoculation and cultivation: A sterile seedling of Mongolian oak with a root length of 6 cm and a stem length of 8 cm and a sterile seedling of Scots pine with a root length of 8 cm and a stem length of 16 cm is placed in the liquid culture solution obtained in step (2), covering the stem base, and soaked for 10-20 minutes; the soaked Mongolian oak seedlings and Scots pine seedlings are planted in a cultivation medium (45g humus, 30g perlite, 15g loam and 12g limestone particles, pH 7.5-8.5) in a 2-row:3-row inter-row mixed manner, and the remaining liquid culture solution is poured into the cultivation medium along the roots of the seedlings, the inoculation amount of the Mongolian oak seedlings is 0.3ml of liquid culture solution / seedling, and the inoculation amount of the Scots pine seedlings is 0.25ml of liquid culture solution / seedling, and the cultivation is carried out at 22-25°C, 35-45% humidity, and 1500-4000Lux to obtain Mongolian oak and Scots pine summer truffle mycorrhizal seedlings;

[0041] (4) Transplanting: The Mongolian oak and Mongolian pine summer truffle mycorrhizal seedlings obtained in step (3) are transplanted in a 2-row:5-row inter-row mixed planting manner. After transplanting, regular management is performed to prevent the occurrence of diseases and insect pests, and summer truffles are harvested after cultivation.

[0042] Comparative Example 1

[0043] It is basically the same as Example 3, except that:

[0044] In the step (2) of preparing the liquid culture solution, no Fusarium oxysporum is added.

[0045] Comparative Example 2

[0046] It is basically the same as Example 3, except that:

[0047] Step (3) The root length of Quercus mongolica is 9 cm and the stem length is 12 cm; the root length of Pinus sylvestris var. mongolica is 4 cm and the stem length is 10 cm.

[0048] Comparative Example 3

[0049] It is basically the same as Example 3, except that:

[0050] Step (3) The root length of Quercus mongolica is 5 cm, and the stem length is 5 cm; the root length of Pinus sylvestris var. mongolica is 11 cm, and the stem length is 22 cm.

[0051] Comparative Example 4

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

[0053] Step (3) The Quercus mongolica seedlings and the Pinus sylvestris var. mongolica seedlings are planted in a cultivation medium in a 3-row:2-row inter-row mixed planting pattern.

[0054] Comparative Example 5

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

[0056] Step (4) The Mongolian oak and Mongolian pine summer tuber mycorrhizal seedlings are transplanted in a 3-row:3-row interplanting manner.

[0057] Comparative Example 6

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

[0059] In step (3), only the Pinus sylvestris var. mongolica seedlings and the liquid culture solution are provided for inoculation and cultivation.

[0060] Comparative Example 7

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

[0062] Step (3) provides a Mongolian pine seedling and a Korean pine seedling with a liquid culture solution for inoculation and cultivation, wherein the Mongolian pine seedling has a root length of 8 cm and a stem length of 16 cm, and the Korean pine seedling has a root length of 8 cm and a stem length of 16 cm.

[0063] In order to evaluate the cultivation of mycorrhizal seedlings of different groups of summer tuber, the infection of mycorrhiza was continuously sampled and tested. The specific results are as follows:

[0064] Table 1 Infection of mycorrhizae in different groups

[0065]

[0066] Result analysis: It can be seen from the content of Table 1 that the formation time of the truffle mycorrhizal seedlings of Examples 1-3 of the present invention is short, the infection rate and the survival rate are at a high level, and the yield is generally higher than the yield of the prior art; at the same time, through the comparison results of Example 3 with Comparative Examples 6 and 7, it can be seen that Comparative Example 6 uses a single tree species, Pinus sylvestris var. mongolica, for mycorrhizal seedling cultivation and cultivation, and the mycorrhizal formation time, infection rate, survival rate and yield are all inferior to the experimental results of Comparative Example 3, and Comparative Example 7 replaces the symbiotic tree species, and replaces the compatibility of Quercus mongolica and Pinus sylvestris var. mongolica with the compatibility of Pinus koraiensis and Pinus sylvestris var. mongolica, but its mycorrhizal formation time, infection rate, survival rate and yield are also inferior to those of Example 3. It can be seen that the multiplicity of tree species and the specific compatibility types have an impact on the mycorrhizal seedling cultivation effect. The present invention selects Quercus mongolica and Pinus sylvestris var. mongolica for compatibility, which can significantly improve the cultivation effect of truffles.

[0067] In addition, by comparing Example 3 with Comparative Example 1, it can be seen that the addition of Fusarium oxysporum can affect the mycorrhizal formation time, infection rate, survival rate and yield. The reason may be that Fusarium oxysporum can secrete some enzymes, such as cellulase and ligninase, and release some antibiotic antibacterial substances as well as some phenolic and sugar substances, which promote the colonization of truffles and the growth of mycelium to a certain extent.

[0068] Furthermore, by comparing Example 3 with Comparative Examples 2 and 3, it can be seen that in the process of cooperative summer truffle mycorrhizal seedling cultivation and cultivation of Mongolian Oak and Larix gmelinii, the root length and stem length of Mongolian Oak seedlings and Larix gmelinii seedlings must be within a specific range to achieve the technical effect of the present invention. When it is not within the scope of the present invention, the growth competition and growth promotion of Mongolian Oak seedlings and Larix gmelinii seedlings cannot reach an effective balance, resulting in the inability to maximize the synergistic effect and even produce antagonistic effects.

[0069] By comparing Example 3 with Comparative Examples 4 and 5, it can be seen that different mixed planting methods have an impact on the final mycorrhizal seedling and cultivation effects. When the growth and competition of Mongolian Oak and Scots pine reach a relatively stable equilibrium, the root distribution depths and nutrient absorption methods of different tree species are different, forming a nutrient complementary effect and improving ecological stability. On the contrary, if the mixed planting method cannot achieve a stable balance between the growth and competition of Mongolian Oak and Scots pine, it will affect the truffle cultivation effect.

[0070] At the same time, in order to verify the symbiotic effect of truffle mycorrhiza with Mongolian oak and Pinus sylvestris, the present invention conducted stress resistance experiments on mycorrhizal seedlings cultivated from different groups of Example 3 and Comparative Examples 6 and 7. The experimental process is as follows:

[0071] The mycorrhizal seedlings of different groups were placed at 15°C, 25°C and 35°C for transplanting experiments. Specifically, the growth rate, fruiting body formation and yield of truffle mycelium in the mature stage after transplanting (5-7 years after transplanting) were measured. The experimental results are shown in Table 2:

[0072] Table 2 Temperature tolerance of mycorrhizal seedlings in different groups

[0073]

[0074]

[0075] Analysis of results: It can be seen from the data in Table 2 that when different groups of mycorrhizal seedlings are transplanted in different environments, temperature changes have different effects on the mycorrhizal growth conditions of different groups of mycorrhizal seedlings. By comparison, it can be seen that the mycorrhizal seedlings obtained in Example 3 of the present invention have better stress resistance than those of the mycorrhizal seedlings in Comparative Examples 6 and 7. Affected by temperature, the fluctuation range of the mycorrhizal growth rate and the number of fruiting bodies is smaller, indicating that the mycorrhizal seedlings of summer truffles co-cultivated with Mongolian oak and mongolica pine in the present invention can significantly improve the stress resistance of mycorrhizal seedlings compared with single mongolica pine or the combination of mongolica pine and red pine.

[0076] At the same time, the symbiotic effect of truffle mycorrhiza with Mongolian oak and Pinus sylvestris was verified. The present invention conducted stress resistance experiments on mycorrhizal seedlings cultivated from different groups of Example 3 and Comparative Examples 6 and 7. The experimental process is as follows:

[0077] The mycorrhizal seedlings of different groups were placed in soil moisture of 20%, 50% and 80% for transplanting experiments. Specifically, the growth rate, fruiting body formation and yield of truffle mycelium in the mature stage after transplanting (5-7 years after transplanting) were measured. The experimental results are shown in Table 3:

[0078] Table 3 Humidity tolerance of mycorrhizal seedlings in different groups

[0079]

[0080]

[0081] Analysis of results: It can be seen from the data in Table 3 that when different groups of mycorrhizal seedlings are transplanted in different environments, changes in substrate humidity have different effects on the mycorrhizal growth conditions of different groups of mycorrhizal seedlings. By comparison, it can be seen that the mycorrhizal seedlings obtained in Example 3 of the present invention have better stress resistance than those of the mycorrhizal seedlings in Comparative Examples 6 and 7. Affected by humidity, the fluctuation range of the mycorrhizal growth rate and the number of fruiting bodies is smaller, indicating that the mycorrhizal seedlings of summer truffles co-cultivated with Mongolian oak and mongolica pine in the present invention can significantly improve the stress resistance of mycorrhizal seedlings compared with single mongolica pine or the combination of mongolica pine and red pine.

[0082] The various embodiments in this specification are described in a progressive manner, and each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referenced to each other.

[0083] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one 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 present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A multi-species cooperative summer tuber mycorrhizal seedling cultivation method, characterized in that: The steps include: (1) Preparation of summer truffle fruiting body powder: sterilize and grind the mature summer truffle fruiting bodies to obtain summer truffle fruiting body powder, and store it at low temperature for later use; (2) Preparation of liquid culture solution: Add the fruiting body powder of Truffle sutchuenensis and Fusarium oxysporum to a liquid culture medium, place at 22-25° C., and culture with shaking for 7-10 days to obtain a liquid culture solution; (3) Inoculation and cultivation: placing the sterile seedlings of Mongolian oak and Mongolian pine in the liquid culture solution obtained in step (2), covering the stem base, and soaking for 10-20 minutes; planting the soaked Mongolian oak seedlings and Mongolian pine seedlings in a mixed inter-row pattern of 1-2 rows: 2-3 rows in a cultivation medium, and pouring the remaining liquid culture solution into the cultivation medium along the roots of the seedlings for cultivation, to obtain Mongolian oak and Mongolian pine summer truffle mycorrhizal seedlings; (4) Transplanting: The Mongolian oak and Mongolian pine summer truffle mycorrhizal seedlings obtained in step (3) are transplanted in a mixed inter-row manner of 1-2 rows: 4-6 rows. After transplanting, regular management is performed to prevent the occurrence of diseases and insect pests, and summer truffles are harvested after cultivation.

2. The multi-species cooperative summer tuber mycorrhizal seedling cultivation method according to claim 1, characterized in that: In step (1), the fruiting bodies are immersed in 75% alcohol for disinfection for 3-5 seconds; the rotation speed of the crushing is 8000-19000 rpm, and the crushing time is 10-30 seconds; and the storage temperature is 0-4°C.

3. The multi-species cooperative summer tuber mycorrhizal seedling cultivation method according to claim 1, characterized in that: The composition of the liquid culture medium in step (2) is as follows: glucose 15-25 g / L, maltose 10-13 g / L, peptone 3-4 g / L, KH2PO4 4-5 g / L, MgSO4·7H2O 2-3 g / L and vitamin B1 0.006-0.007g / L.

4. The multi-species cooperative summer tuber mycorrhizal seedling cultivation method according to claim 1, characterized in that: The dry weight of mycelium in the liquid culture solution obtained in step (2) is 1-2 g / L; the content of Fusarium oxysporum in the liquid culture solution obtained is 2×10 6 -8×10 6 CFU / mL; adjust the pH of the liquid culture solution to 7.5-8.

0.

5. The multi-species cooperative summer tuber mycorrhizal seedling cultivation method according to claim 1, characterized in that: The rotation speed of the shaking culture in step (2) is 100-150 rpm, and the culture is carried out in the dark.

6. The multi-species cooperative summer tuber mycorrhizal seedling cultivation method according to claim 1, characterized in that: The root length of the Mongolian oak seedling in step (3) is 6-8 cm, and the stem length is 5-10 cm; the root length of the Mongolian pine is 5-10 cm, and the stem length is 10-20 cm.

7. The multi-species cooperative summer tuber mycorrhizal seedling cultivation method according to claim 1, characterized in that: The cultivation medium in step (3) is composed of: 40-50g humus, 30-35g perlite, 15-20g loam and 10-15g limestone particles; the pH of the cultivation medium is 7.5-8.

5.

8. The multi-species cooperative summer tuber mycorrhizal seedling cultivation method according to claim 1, characterized in that: In step (3), the inoculation amount of the Mongolian oak seedlings is 0.2-0.4 ml of liquid culture solution per seedling; the inoculation amount of the Mongolian pine seedlings is 0.2-0.3 ml of liquid culture solution per seedling; the culture temperature is 22-25° C., the humidity of the cultivation medium is 35-45%, and the lighting condition is 1500-4000 Lux.

9. Application of the multi-species cooperative summer truffle mycorrhizal seedling cultivation method according to any one of claims 1 to 8 in summer truffle production.

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