Method for collecting hyphae of arbuscular mycorrhizal fungi

Abstract

The invention discloses an arbuscular mycorrhizal fungus hypha collection method, and particularly relates to the technical field of hypha collection, and the arbuscular mycorrhizal fungus hypha collection method comprises the following steps: S01, soaking quartz sand in 0.1 mol / L diluted hydrochloric acid, cleaning with tap water until the quartz sand is neutral, and drying with a drying oven; s02, cutting a nylon net with the aperture of 10-20 microns, and making the nylon net into a nylon net bag by using a sealing machine; s03, 70g of quartz sand and a proper amount of slow-release fertilizer are put into a nylon mesh bag, and then the nylon mesh bag is subjected to opening binding through a sealing machine to be made into a hypha collecting bag. The device is suitable for potting and field experiments, a large number of mycelia can be efficiently collected, and the root system of a plant can be prevented from entering the mycelia bag.

Description

Technical Field

[0001] The invention relates to the technical field of hyphae collection, and in particular to a method for collecting hyphae of arbuscular mycorrhizal fungi. Background Art

[0002] Arbuscular mycorrhizal fungi (AMF) can coexist with more than 80% of plants on land, infect the root system of host plants to form arbuscular and vesicle structures, and the hyphae outside the root system form a dense hyphal network in the soil. Through the hyphal network, AMF can deliver 90% of the phosphorus and 20% of the nitrogen required by plants to plants (Smith et al., 2003), and plants can supply up to 30% of photosynthetic products to fungi (Nehls & Hammpp, 2000; Nehls et al., 2001). Since AMF is a symbiotic fungus that depends on host plants for carbon sources, it is difficult to achieve pure culture. Although it has been reported that sodium myristate can be used as a carbon source to enable AMF to grow and produce spores under non-symbiotic conditions (Sugiura et al., 2019), this discovery has not been widely used. It is difficult to collect and purify hyphae, so there are certain limitations in studying the functional traits of AMF fungal hyphae.

[0003] Hattingh et al. first developed the mycelium compartment method (Hattinghetal, 1973), which can purify mycelium. Such as glass bead split chamber culture system, double chamber culture system, etc. (Wang et al., 2014), but these methods are only suitable for greenhouse pot experiments, and cannot be applied to field experiments. An improved method is to use a mesh bag filled with glass beads or sand to collect ectomycorrhizal (EM) fungal mycelium (Wallander et al., 2001), but the collected mycelium is not enriched and purified. However, the growth bag with only quartz sand lacks the nutrients contained in the soil, especially nitrogen and phosphorus, which is not conducive to the collection of a large number of mycelium. In addition, the mesh diameter of the mycelium bag mesh used in previous studies is about 40μm, and root hairs with smaller diameters may enter the mycelium bag and affect the detection results of mycelium. Summary of the invention

[0004] The invention aims to provide a method for collecting arbuscular mycorrhizal fungi hyphae, so as to solve the existing situation that it is difficult to collect arbuscular mycorrhizal fungi hyphae.

[0005] In order to achieve the above object, the present invention provides the following technical solution: a method for collecting mycelium of arbuscular mycorrhizal fungi, comprising the following steps:

[0006] S01, quartz sand is soaked in 0.1mol / L dilute hydrochloric acid, washed with tap water until neutral, and then placed in an oven for drying;

[0007] S02, cutting a nylon mesh with a pore size of 10-20 μm, and using a sealing machine to make a nylon mesh bag from the nylon mesh;

[0008] S03. Put 70g of quartz sand and an appropriate amount of slow-release fertilizer into a nylon mesh bag, and then use a sealing machine to seal the bag to make a mycelium collection bag.

[0009] S04. The contents of the harvested mycelium bag are placed in a larger container and washed with tap water to enrich and purify the mycelium.

[0010] Preferably, the soaking time in step S01 is 50-70 minutes.

[0011] Preferably, the temperature of the oven in step S01 is 120°C.

[0012] Preferably, in step S03, before putting 70 g of quartz sand into the nylon mesh bag, 70 g of quartz sand needs to be mixed with 0.5 g of slow-release fertilizer.

[0013] Preferably, the nylon mesh in step S02 has a pore size of 10 μm.

[0014] In the above technical scheme, the present invention provides a method for collecting arbuscular mycorrhizal fungal mycelium, which has the following beneficial effects: relatively pure arbuscular mycorrhizal fungal mycelium with high biomass can be obtained, and the actual operation is relatively simple, and it can be used in potted plants and in the wild, and has strong applicability. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the embodiments of the present application or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments recorded in the present invention. For ordinary technicians in this field, other drawings can also be obtained based on these drawings.

[0016] Figure 1 A process diagram provided for an embodiment of the present invention;

[0017] Figure 2 A flow chart of mycelium bag cleaning provided in an embodiment of the present invention.

[0018] Figure 3 The diameter of mycelium extracted from the experimental group provided in the embodiment of the present invention. DETAILED DESCRIPTION

[0019] In order to enable those skilled in the art to better understand the technical solution of the present invention, the present invention will be further described in detail below with reference to the accompanying drawings.

[0020] like Figure 1 As shown, a method for collecting arbuscular mycorrhizal fungal hyphae comprises:

[0021] Example 1

[0022] The collection method includes the following steps:

[0023] S01, quartz sand was soaked in 0.1mol / L dilute hydrochloric acid (50-70min), then washed with tap water until neutral, and dried in an oven (120℃);

[0024] S02, cutting a nylon mesh with a pore size of 10-20 μm (the specifications of the cut nylon mesh are 15 cm in length and 4 cm in width), and using a sealing machine to make the nylon mesh into a nylon mesh bag;

[0025] S03, 70g of quartz sand and an appropriate amount of slow-release fertilizer were put into a nylon mesh bag, and then the bag was sealed by a sealing machine to make a mycelium collection bag;

[0026] S04. Bury the mycelium collection bag and harvest it after one month.

[0027] Example 2

[0028] The collection method includes the following steps:

[0029] S01, quartz sand was soaked in 0.1mol / L dilute hydrochloric acid (60min), then washed with tap water until neutral, and finally dried in an oven (120℃);

[0030] S02, cutting a nylon mesh with a pore size of 10 μm (the specifications of the cut nylon mesh are 15 cm in length and 4 cm in width), and using a sealing machine to make a nylon mesh bag from the nylon mesh;

[0031] S03, 70g of quartz sand and an appropriate amount of slow-release fertilizer were put into a nylon mesh bag, and then the bag was sealed by a sealing machine to make a mycelium collection bag;

[0032] S04. Bury the mycelium collection bag and harvest it after two months.

[0033] Example 3

[0034] The collection method includes the following steps:

[0035] S01, quartz sand was soaked in 0.1 mol / L dilute hydrochloric acid (60 min), then washed with tap water until neutral, and dried in an oven (120 °C);

[0036] S02, cutting a nylon mesh with a pore size of 10 μm (the specifications of the cut nylon mesh are 15 cm in length and 4 cm in width), and using a sealing machine to make a nylon mesh bag from the nylon mesh;

[0037] S03, putting 70g of quartz sand into a nylon mesh bag, and then sealing it with a sealing machine to make a mycelium collection bag;

[0038] S04. Bury the mycelium collection bag and harvest it after one month.

[0039] Example 4

[0040] The collection method includes the following steps:

[0041] S01, quartz sand was soaked in 0.1 mol / L dilute hydrochloric acid (60 min), then washed with tap water until neutral, and dried in an oven (120 °C);

[0042] S02, cutting a nylon mesh with a pore size of 10 μm (the specifications of the cut nylon mesh are 15 cm in length and 4 cm in width), and using a sealing machine to make a nylon mesh bag from the nylon mesh;

[0043] S03, first mix 70g of quartz sand and 0.5g of slow-release fertilizer, then put them into a nylon mesh bag, and then use a sealing machine to tie the mouth to make a mycelium collection bag;

[0044] S04. Bury the mycelium collection bag and harvest it after one month.

[0045] Comparative Example 1

[0046] The mycelium collection bags prepared by the processes of Example 4 and Example 3 were extracted and experiments were carried out respectively, wherein Example 4 was an experimental group and Example 3 was a control group.

[0047] Experimental environment: During the growing season, a plot with homogeneous plants and no fertilizer was added was selected at the Institute of Applied Ecology, Shenyang, Chinese Academy of Sciences, Ergun Forest-Grassland Transitional Zone Ecosystem Research Station (119°30′E, 50°12′N). A 15 cm deep hole was drilled with a 3 cm diameter earth drill, and the mycelium collection bag was buried in the hole. The mycelium was harvested one month later.

[0048] Subjects:

[0049] Three samples were prepared using the process of Example 4, namely Experimental Group 1, Experimental Group 2 and Experimental Group 3;

[0050] Three samples were prepared using the process of Example 3, namely control group 1, control group 2 and control group 3.

[0051] The experimental results are shown in the following table:

[0052]

[0053]

[0054] In summary, combined with Figure 2 As shown, the dry weight of mycelium of experimental group 1, experimental group 2 and experimental group 3, and comparison group 1, comparison group 2 and comparison group 3 were sampled one month later. The experimental results show that the mycelium collection bag mixed with 0.5g slow-release fertilizer can collect more mycelium than the mycelium collection bag without mixed slow-release fertilizer.

[0055] The above description is only by way of illustration of certain exemplary embodiments of the present invention. It is undoubted that those skilled in the art can modify the described embodiments in various ways without departing from the spirit and scope of the present invention. Therefore, the above drawings and descriptions are illustrative in nature and should not be construed as limiting the scope of protection of the claims of the present invention.

Claims

1. A method for collecting mycelium of arbuscular mycorrhizal fungi, characterized in that A large amount of purified mycelium can be efficiently collected, comprising the following steps: S01, quartz sand is soaked in 0.1mol / L dilute hydrochloric acid, then washed with tap water until neutral, and dried in an oven; S02, cutting a nylon mesh with a pore size of 10-20 μm, and using a sealing machine to make a nylon mesh bag from the nylon mesh; S03. Put 70g of quartz sand and an appropriate amount of slow-release fertilizer into a nylon mesh bag, and then use a sealing machine to seal the bag to make a mycelium collection bag. S04. When harvesting, place the contents of the mycelium collection bag into a 1L container, add an appropriate amount of tap water or purified water and stir. This can separate the mycelium and quartz sand and extract the purified mycelium.

2. The method for collecting arbuscular mycorrhizal fungi hyphae according to claim 1, characterized in that: The soaking time in step S01 is 50-70 minutes.

3. The method for collecting arbuscular mycorrhizal fungi hyphae according to claim 1, characterized in that: The temperature of the oven in step S01 is 120°C.

4. The method for collecting arbuscular mycorrhizal fungi hyphae according to claim 1, characterized in that: The specifications of the nylon mesh bag cut in step S02 are 15 cm in length and 4 cm in width.

5. The method for collecting arbuscular mycorrhizal fungi hyphae according to claim 1, characterized in that: The nylon mesh in step S02 has a pore size of 10 μm.

6. The method for collecting arbuscular mycorrhizal fungi hyphae according to claim 1, characterized in that: Before putting 70 g of quartz sand into the nylon mesh bag in step S03, 70 g of quartz sand needs to be mixed with 0.5 g of slow-release fertilizer.

7. The method for collecting arbuscular mycorrhizal fungi hyphae according to claim 1, characterized in that: In step S04, tap water or purified water is used for washing to extract and purify mycelium.

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