A method for cultivating truffle mycorrhizal seedlings

Abstract

The present invention provides a truffle mycorrhizal seedling cultivation method, and relates to the technical field of plant cultivation. The truffle mycorrhizal seedling cultivation method mainly comprises the following steps: culturing host seedlings, slurrying together truffles with anthocyanins and folic acid-mixed nutrient agents to prepare a suspension, mixing the suspension with indoleacetic acid and a growth agent with tea saponin to prepare a bacterial agent, and inoculating the host seedlings with the bacterial agent to perform seedling hardening and cultivation. The present invention overcomes the shortcomings of the prior art, effectively shortens the formation time of mycorrhizae, promotes the growth of mycorrhizal seedlings, ensures the survival rate of subsequent commercial seedling transplanting, comprehensively reduces the loss of cultivation, and improves economic benefits.

Description

Technical Field

[0001] The invention relates to the technical field of plant cultivation, and in particular to a method for cultivating truffle mycorrhiza seedlings. Background Art

[0002] Truffles refer to true truffle-like fruiting bodies that grow underground, also known as truffles. Truffles have important economic, edible and health values. Mature truffles have marble patterns, rich aromas and unique tastes, and are favored by the international edible fungi market. Truffles contain a variety of active substances, such as truffle polysaccharides, ceramides and α-androstanol, which can resist viruses, enhance immunity, regulate endocrine, improve sexual function, etc., and have high economic benefits.

[0003] Truffles are typical ectomycorrhizal edible fungi. They must form mycorrhizae with plants and obtain the nutrients necessary for growth through symbiotic exchange to complete their life cycle and form fruiting bodies (truffles). Therefore, the cultivation method of truffles is essentially different from that of saprophytic fungi such as oyster mushrooms. At present, truffles come from the collection of wild resources and artificially cultivated resources. In the natural environment, it usually takes 7-10 years for truffles to complete their life cycle and produce truffles. Through artificial cultivation, the production cycle of truffles can be shortened to 3-5 years. Therefore, artificial cultivation of truffles has a high planting economic benefit. The most important step in the artificial cultivation of truffles is to cultivate truffle mycorrhizal seedlings. The existing mycorrhizal infection method mainly uses truffle suspension to directly infect and cultivate mycorrhizae. On this basis, the infection rate of mycorrhizae is mostly around 80%, and the mycorrhizal formation time is even as long as 2-3 months, which increases the cultivation time of mycorrhizal seedlings and causes certain losses in actual cultivation. Therefore, how to quickly achieve truffle infection mycorrhizae to cultivate qualified commercial seedlings is a major research direction at this stage. Summary of the invention

[0004] In view of the deficiencies in the prior art, the present invention provides a truffle mycorrhizal seedling cultivation method, which can effectively shorten the formation time of mycorrhizae, promote the growth of mycorrhizal seedlings, ensure the survival rate of subsequent commercial seedling transplanting, comprehensively reduce the loss of cultivation, and improve economic benefits.

[0005] To achieve the above objectives, the technical solution of the present invention is implemented through the following technical solutions:

[0006] A method for cultivating truffle mycorrhizal seedlings, the method comprising the following steps:

[0007] S1. Cultivation of host plant seedlings: Selecting seeds of host plants, sterilizing the seeds, sowing them in a sterilized medium, and then germinating and cultivating them into host seedlings for later use;

[0008] S2. Preparation of truffle suspension: summer truffle fruiting bodies are selected for sterilization, and then the sterilized summer truffle fruiting bodies are mixed with nutrients and crushed by a beater to prepare a suspension for standby use, and the formula of the nutrient is: distilled water + anthocyanin 20-30 mg / L + folic acid 30-40 mg / L;

[0009] S3, preparation of bacterial agent: the above suspension and highly absorbent acrylic resin are mixed evenly at a volume ratio of 400:1 to obtain a premix, and then the premix is ​​mixed with a growth agent at equal volumes to obtain a bacterial agent for standby use; and the formula of the growth agent is: distilled water + indoleacetic acid 120-150 mg / L + tea saponin 30-45 mg / L;

[0010] S4, inoculation: fill the inoculation cup with 1 / 3 of the total volume of sterilized matrix, then place the host seedlings with excess roots and leaves cut off in the center of the inoculation cup, spray the roots of the host seedlings with fungicides, and then cover with a layer of sterilized matrix to complete the inoculation;

[0011] S5. Seedling hardening and cultivation: After hardening the inoculated cups for 15-30 days, place them in a greenhouse for cultivation for 6-24 months to obtain commercial seedlings ready for transplanting.

[0012] Preferably, the host plant is any one of Pinus yunnanensis, Pinus armandii, and Castanea mollissima.

[0013] Preferably, the seeds are disinfected in step S1 by soaking them in 30% hydrogen peroxide for 15-20 minutes, and then rinsing them with distilled water for 1-3 minutes.

[0014] Preferably, the sterilized matrix is ​​prepared by mixing vermiculite and peat soil of equal mass and sterilizing them at 121-126° C. for 2 hours.

[0015] Preferably, the summer truffle fruiting bodies in step S2 are sterilized by first cleaning the surface soil and then soaking them in 75% alcohol for 3-5 minutes, and then placing them on a flame for rapid burning.

[0016] Preferably, the volume ratio of the sterilized truffle fruiting bodies mixed with the nutrient agent in step S2 is 1:5, and the particle size of the suspension prepared by crushing with a pulper is less than 1 mm.

[0017] Preferably, the dosage of bacteria sprayed on the root of each host seedling in step S4 is 3-5 mL.

[0018] Preferably, during the hardening of the seedlings in step S5, the temperature is controlled at 15-25° C., and the humidity of the sterilized substrate is 50%-80%.

[0019] Preferably, the temperature of the greenhouse in step S5 is 20-30°C, and the humidity of the sterilized substrate is 20%-30%.

[0020] The present invention provides a method for cultivating truffle mycorrhizal seedlings, which has the advantages of:

[0021] The invention adds anthocyanidins, folic acid and other substances as auxiliary materials in the preparation of the truffle suspension, and subsequently mixes the suspension with a growth agent prepared by substances such as indoleacetic acid and tea saponin to co-inoculate host seedlings, so that rapid infection and formation of mycorrhizae can be achieved, and the mycorrhizal seedlings after infection can be ensured to grow effectively, and have a high seedling rate and survival rate during the entire seedling raising period, and also have a high activity after transplanting. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is a schematic diagram of the growth of Yunnan pine seedlings in experimental group 1 of the present invention 40 days after inoculation with bacterial agent 1;

[0023] Figure 2 This is a schematic diagram of the growth of Pinus chinensis seedlings in experimental group 8 of the present invention 40 days after inoculation with bacterial agent 1;

[0024] Figure 3 This is a schematic diagram of the growth of chestnut seedlings in experimental group 15 of the present invention 40 days after inoculation with bacterial agent 1;

[0025] Figure 4 Schematic diagram of mycorrhizal formation of Pinus yunnanensis seedlings in experimental group 1 of the present invention;

[0026] Figure 5 This is a schematic diagram of the chestnut seedlings in the experimental group 15 of the present invention growing for 2 months;

[0027] Figure 6 This is a schematic diagram of the growth of Yunnan pine seedlings in experimental group 1 of the present invention for 3 months;

[0028] Figure 7 This is a schematic diagram of the growth of Pinus chinensis seedlings of experimental group 8 of the present invention for 3 months;

[0029] Figure 8 This is a schematic diagram of the growth of Yunnan pine seedlings in experimental group 1 of the present invention after 7 days of transplantation;

[0030] Fig. 9 This is a schematic diagram of the growth of Pinus chinensis seedlings in experimental group 8 of the present invention after 30 days of transplantation. DETAILED DESCRIPTION

[0031] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention is clearly and completely described below in combination with the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. 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.

[0032] The sterilized matrix used in the following examples was prepared by mixing vermiculite and peat soil in equal proportions and then sterilizing at 121-126° C. for 2 h. Example

[0033] Cultivation of truffle mycorrhizal seedlings:

[0034] 1. Cultivation of host seedlings:

[0035] (1) Select Yunnan pine, Armand pine, and chestnut seeds respectively, remove the shells of each seed, soak them in 30% hydrogen peroxide for 18 minutes, and then take them out and rinse them with distilled water for 2 minutes to sterilize the seeds;

[0036] (2) Add sterilized substrate to the seedling frame, then sow the sterilized seeds, water them and germinate them at room temperature. When the germination rate reaches 80%, transfer them to the greenhouse and control the humidity of the sterilized substrate to 50%-80% at a temperature of 15-30℃. After 2 months of seedling cultivation, Yunnan pine seedlings, Armand pine seedlings, and chestnut seedlings are obtained as host seedlings.

[0037] 2. Preparation of microbial agent:

[0038] (1) Select the fruiting bodies of summer truffles, first scrub the dirt off the surface of the fruiting bodies, then soak them in 75% alcohol for 3 minutes, take them out and quickly burn them on the flame of an alcohol lamp to complete the sterilization;

[0039] (2) The sterilized fruiting bodies are mixed with the nutrient agent at a volume ratio of 1:5 and crushed in a beater to form a suspension with a particle size of less than 1 mm for later use;

[0040] (3) the suspension is mixed with highly absorbent acrylic resin in a volume ratio of 400:1 to form a premix, and then the premix is ​​mixed with a growth agent in equal volumes to obtain a bacterial agent;

[0041] And different bacterial agents were prepared according to the nutrients and nutrients with different formulas in Table 1 below:

[0042] Table 1

[0043]

[0044] 3. Inoculation, cultivation and treatment:

[0045] (1) Fill the inoculation cup with 1 / 3 of the total volume of sterilized matrix, select the host seedlings, cut off the excess roots and leaves and place them in the inoculation cup, then spray the roots of the host seedlings with the fungicide at 4 mL / plant, and then cover with sterilized matrix to complete the inoculation;

[0046] (2) After inoculation, place the inoculation cup on a culture rack at 15-25°C, control the humidity of the sterilized substrate to 50%-80%, and cultivate the seedlings for 30 days;

[0047] (3) After the seedling hardening is completed, place the inoculation cup in a greenhouse at a temperature of 20-30°C and control the humidity of the sterilized substrate to 20%-30%.

[0048] Set different host seedlings and inoculants according to the selections in Table 2 below:

[0049] Table 2

[0050]

[0051] Detection:

[0052] The above experimental groups were tested every 10 days to see whether mycorrhiza was formed (mycelium was observed to spread on the root surface to form a mycelium sheath, and under a microscope, mycelium could be seen to spread on the root surface to form a Hatti network, which was mycorrhiza formation). The time of mycorrhiza formation in each group was recorded, and the survival rate and mycorrhiza infection rate were calculated after 6 months of cultivation in the greenhouse. The specific results are shown in Table 3 below:

[0053] Table 3

[0054]

[0055] It can be seen from the above table that the host seedlings treated with bacterial agent 1 and bacterial agent 2 can form mycorrhiza in about 40 days, and have a high survival rate and infection rate, realizing the efficient cultivation of mycorrhizal seedlings.

[0056] The mycorrhizal seedlings of experimental groups 1-14, which had been cultivated for 8 months, were transplanted outdoors (transplanting temperature was 15-25°C), and the survival rates of each group were tested 7d, 15d, and 30d after transplanting. The specific results are shown in Table 4 below:

[0057] Table 4

[0058]

[0059] It can be seen from the above table that the mycorrhizal seedlings of experimental groups 1-2 and 8-9 have a higher transplant survival rate.

[0060] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for cultivating truffle mycorrhizal seedlings, characterized in that: The cultivation method comprises the following steps: S1. Cultivation of host plant seedlings: Selecting seeds of host plants, sterilizing the seeds, sowing them in a sterilized medium, and then germinating and cultivating them into host seedlings for later use; S2, preparation of truffle suspension: summer truffle fruiting bodies are selected for sterilization, and then the sterilized summer truffle fruiting bodies are mixed with nutrients and crushed by a pulper to prepare a suspension for standby use, and the formula of the nutrient is: distilled water + anthocyanin 20-30 mg / L + folic acid 30-40 mg / L; the volume ratio of the sterilized summer truffle fruiting bodies mixed with nutrients is 1:5, and the particle size of the suspension crushed by the pulper is less than 1 mm; S3, preparation of bacterial agent: the above suspension and highly absorbent acrylic resin are mixed evenly at a volume ratio of 400:1 to obtain a premix, and then the premix is ​​mixed with a growth agent at equal volumes to obtain a bacterial agent for standby use; and the formula of the growth agent is: distilled water + indoleacetic acid 120-150 mg / L + tea saponin 30-45 mg / L; S4, inoculation: fill the inoculation cup with 1 / 3 of the total volume of the sterilized matrix, then place the host seedlings with excess roots and leaves cut off in the center of the inoculation cup, spray the roots of the host seedlings with a fungicide, and then cover with a layer of sterilized matrix to complete the inoculation; and the sterilized matrix is ​​prepared by mixing vermiculite and peat soil of equal quality and sterilizing at 121-126°C for 2h; S5. Seedling hardening and cultivation: After hardening the inoculated cups for 15-30 days, place them in a greenhouse for cultivation for 6-24 months to obtain commercial seedlings ready for transplanting.

2. The cultivation method according to claim 1, characterized in that: The host plant is any one of Pinus yunnanensis, Pinus armandii and Castanea mollissima.

3. The cultivation method according to claim 1, characterized in that: The method of seed disinfection in step S1 is to soak the seeds in 30% hydrogen peroxide for 15-20 minutes, and then rinse them with distilled water for 1-3 minutes.

4. The cultivation method according to claim 1, characterized in that: In step S2, the summer truffle fruiting bodies are sterilized by first cleaning the surface soil and then soaking them in 75% alcohol for 3-5 minutes, and then placing them on a flame for rapid burning.

5. The cultivation method according to claim 1, characterized in that: The dosage of bacteria sprayed on the root of each host seedling in step S4 is 3-5 mL.

6. The cultivation method according to claim 1, characterized in that: During the hardening of the seedlings in step S5, the temperature is controlled to be 15-25° C., and the humidity of the sterilized substrate is 50%-80%.

7. The cultivation method according to claim 1, characterized in that: In step S5, the temperature of the greenhouse is 20-30° C., and the humidity of the sterilized substrate is 20%-30%.

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