A propagation matrix for promoting spore production of arbuscular mycorrhizal fungi and a propagation method thereof
Through the synergistic effect of ingredients such as strigolactones and naphthaleneacetic acid, combined with specific matrices and methods, the problem of unstable spore count of AM fungi was solved, efficient propagation of arbuscular mycorrhizal fungi was achieved, and its industrial application was promoted.
Patent Information
- Application Number
- CN202211345584.7
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2022-10-31
- Publication Date
- 2025-09-30
- Estimated Expiration
- 2042-10-31
AI Technical Summary
The lack of professional AM fungus propagation matrix on the market has resulted in unstable spore counts and low concentrations in AM fungus inoculants, limiting their industrial application.
Using strigolactone and naphthaleneacetic acid as the main ingredients, combined with vermiculite, earthworm castings, peat, indoleacetic acid and calcium magnesium phosphate, a propagation matrix that promotes spore production of arbuscular mycorrhizal fungi was prepared. The spore number and root infection rate were increased through a specific propagation method.
The root infection rate and spore count of arbuscular mycorrhizal fungi were significantly improved, and large-scale production and application of AM fungal agents were realized with low cost and simple operation.
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Abstract
Description
Technical Field
[0001] The invention relates to the technical field of bacterial preparations, and in particular to a propagation matrix for promoting spore production of arbuscular mycorrhizal fungi and a propagation method thereof. Background Art
[0002] Arbuscular mycorrhizal fungi (AM fungi) are a very important class of beneficial microorganisms in the soil, forming stable symbiotic relationships with over 80% of terrestrial plants. AM fungi can form vast hyphal networks in the soil, reaching areas inaccessible to roots. This effectively expands the root area and rhizosphere, promoting plant nutrient absorption. Furthermore, AM fungi can help plants defend against infection by nematodes, fungi, bacteria, and viruses, enhancing their resistance to soil-borne diseases by strengthening the structure of plant root cell walls, inducing the production of disease-resistant metabolites, regulating microbial community structure, and competing with pathogens for ecological niches.
[0003] The commercialization of AM fungi requires the availability of AM fungal inoculants with high spore concentrations. However, the market still lacks specialized AM fungal propagation substrates, resulting in unstable spore counts and low spore concentrations in AM fungal inoculants, limiting the industrial application of AM fungi. Summary of the Invention
[0004] In view of this, the purpose of the present invention is to provide a propagation matrix and a propagation method thereof for promoting spore production of arbuscular mycorrhizal fungi, so as to solve the above problems.
[0005] The technical solution of the present invention is achieved as follows:
[0006] A propagation substrate for promoting sporulation of arbuscular mycorrhizal fungi, comprising strigolactones and naphthylacetic acid.
[0007] Preferably, 1 to 40 mg of strigolactone and 1 to 40 mg of naphthaleneacetic acid are added per liter of propagation medium.
[0008] Preferably, 1 to 20 mg of strigolactone and 20 to 39 mg of naphthaleneacetic acid are added per liter of propagation medium.
[0009] More preferably, 2-15 mg of strigolactone and 25-38 mg of naphthaleneacetic acid are added per liter of propagation medium.
[0010] Preferably, vermiculite, earthworm castings, peat, indoleacetic acid, humic acid and calcium magnesium phosphate fertilizer are also included.
[0011] More preferably, based on weight, it also includes 50-60 parts of vermiculite, 20-30 parts of earthworm castings, 15-18 parts of peat, 0.006-0.012 parts of indoleacetic acid, 0.4-0.5 parts of humic acid and 1-2 parts of calcium magnesium phosphate.
[0012] The present invention provides a propagation method of a propagation substrate for promoting spore production of arbuscular mycorrhizal fungi, comprising the following steps:
[0013] S1: Using the propagation matrix as the base material, dissolve the strigolactone and naphthaleneacetic acid and spray them onto the propagation matrix, stir, sprinkle the arbuscular mycorrhizal fungus agent raw materials on the propagation matrix, and then lay a 3-5 cm thick propagation matrix;
[0014] S2: Using corn as the host plant, corn seeds were sown in a propagation medium 2.5 to 3.5 cm from the surface. After emergence, the seeds were watered with Hoagland nutrient solution.
[0015] S3: From seedling emergence to the 21st week, remove the above-ground part of the corn, air-dry the roots together with the propagation medium, and then break the roots and mix them with the propagation medium to obtain the target arbuscular mycorrhizal fungal agent.
[0016] Preferably, in step S2, one week after seedling emergence, water with Hoagland nutrient solution 1 to 2 times every other week for 7 consecutive weeks; the amount of Hoagland nutrient solution for watering is doubled in the 8th and 9th weeks; the amount of Hoagland nutrient solution for watering is reduced to 48 to 52% in the 10th week; the amount of Hoagland nutrient solution for watering is reduced to 18 to 22% from the 11th week to the 16th week; and no Hoagland nutrient solution is watered from the 17th week to the 20th week.
[0017] The invention provides an application of a propagation matrix for promoting spore production of arbuscular mycorrhizal fungi in increasing the spore production quantity of arbuscular mycorrhizal fungi and improving the infection rate of arbuscular mycorrhizal fungi on corn root systems.
[0018] Preferably, the arbuscular mycorrhizal fungi are Funneliformis mosseae, Rhizophagus irregularis and Glomus etunicatum.
[0019] Compared with the prior art, the present invention has the following beneficial effects:
[0020] 1. The present invention utilizes the synergistic effect of strigolactone and naphthaleneacetic acid to increase the root infection rate and spore count of arbuscular mycorrhizal fungi.
[0021] 2. The present invention selects the synergistic effect of strigolactone and naphthaleneacetic acid, uses earthworm castings and peat as main raw materials, and combines vermiculite, indoleacetic acid, humic acid and calcium magnesium phosphate to prepare a propagation matrix for arbuscular mycorrhizal fungi, which effectively improves the root infection rate of arbuscular mycorrhizal fungi and the number of arbuscular mycorrhizal fungal spores; the arbuscular mycorrhizal fungi propagation matrix provided by the present invention has low cost and simple operation, and can realize the large-scale production and application of arbuscular mycorrhizal fungal agents.
[0022] 3. The propagation matrix for promoting spore production of arbuscular mycorrhizal fungi of the present invention effectively promotes the root infection rate and spore production of Funneliformis mosseae, Rhizophagus irregularis and Glomus etunicatum. BRIEF DESCRIPTION OF THE DRAWINGS
[0023] Figure 1 This is the effect of different addition amounts of strigolactone and naphthaleneacetic acid on the number of F. mosseae spores in Example 1 of the present invention.
[0024] Figure 2 This is the effect of different addition amounts of strigolactone and naphthylacetic acid on the root infection rate of corn in Example 1 of the present invention.
[0025] Figure 3 This is the effect of different addition amounts of strigolactone and naphthylacetic acid on the number of spores of different arbuscular mycorrhizal fungi in Example 2 of the present invention.
[0026] Figure 4 This is the effect of different addition amounts of strigolactone and naphthaleneacetic acid on the infection rate of different arbuscular mycorrhizal fungi in Example 2 of the present invention.
[0027] Figure 5 This is the effect of different propagation substrates on the number of arbuscular mycorrhizal fungal spores in Example 3 of the present invention.
[0028] Figure 6 This is the effect of different propagation substrates on the infection rate of arbuscular mycorrhizal fungi in Example 3 of the present invention. DETAILED DESCRIPTION
[0029] In order to better understand the technical content of the present invention, specific examples are provided below to further illustrate the present invention.
[0030] Unless otherwise specified, the experimental methods used in the examples of the present invention are all conventional methods.
[0031] Unless otherwise specified, the materials, reagents, etc. used in the examples of the present invention can be obtained from commercial sources.
[0032] The vermiculite particle size used in the embodiment of the present invention is less than 2 mm, the peat particle size is less than 1 mm, and the vermicompost is obtained by the following method: cow dung is high-temperature composted for 20 to 30 days, and the compost product is further vermicomposted for 30 to 60 days to obtain vermicompost.
[0033] Example 1
[0034] Using vermiculite as the base material, weigh the monochalcone and naphthaleneacetic acid according to the design in Table 1, dilute and dissolve them with an organic solvent to a uniform volume, then evenly spray them into the vermiculite and stir them evenly. Sprinkle the AM fungal agent - Funneliformis mosseae (F.mosseae) evenly on the vermiculite, and then lay 4 cm thick vermiculite. The amount of AM fungal agent added is 8% of the mass of the vermiculite. Corn (Xianyu 335) is used as the host plant, and corn seeds are sown in vermiculite 3 cm from the surface, with a sowing density of 30 plants / m 3 , one week after germination, retain 20 plants.
[0035] During the propagation phase, corn plants were watered with Hoagland nutrient solution containing 50% of the standard phosphorus content one week after seedling emergence. Watering was continued every other week for seven consecutive weeks. The nutrient solution dosage was doubled in weeks 8 and 9. In week 10, the nutrient solution dosage was reduced to 50%. From weeks 11 to 16, the nutrient solution dosage was reduced to 20%. No fertilizer was applied in week 17, but normal watering was used. From weeks 18 to 19, no fertilizer was applied, and moderate drought was maintained. In week 20, no watering was applied, allowing the plants to dry out normally. In week 21, the aboveground parts of the corn were removed, and the roots, along with the vermiculite, were air-dried. The roots were then crushed and mixed with the vermiculite to produce the AM fungal inoculant. Sampling was then conducted to analyze the effects of the different treatments on spore count and corn root infection rates.
[0036] Table 1 Design table of different addition amounts of strigolactone and naphthaleneacetic acid (mg / L matrix)
[0037]
[0038] from Figure 1 It can be seen that when the addition amounts of strigolactone and naphthaleneacetic acid were 5 and 35 mg / L respectively (treatment T8), the number of F. mosseae spores was significantly higher than that of other treatments; Figure 2 It can be seen that when the addition amounts of strigolactone and naphthacetic acid were 2-15 mg / L and 25-38 mg / L (treatments T5-T10), respectively, the infection rate of F. mosseae was significantly higher than that of other treatments.
[0039] Example 2
[0040] Based on the experimental results of Example 1, the propagation effects of different arbuscular mycorrhizal fungi inoculants were specifically evaluated when strigolactone addition levels were 2-15 mg / L and naphthaleneacetic acid addition levels were 25-38 mg / L. The specific addition levels of strigolactone and naphthaleneacetic acid are shown in Table 2. The arbuscular mycorrhizal fungi inoculants selected were F. mosseae, Rhizophagus irregularis (R. irregularis), and Glomus etunicatum (C. etunicatum). The experimental methods and propagation management were the same as in Example 1.
[0041] Table 2 Design table of different addition amounts of strigolactone and naphthaleneacetic acid (mg / L matrix)
[0042]
[0043] from Figure 3 、 Figure 4 As can be seen from the results, both strigolactones and naphthaleneacetic acid significantly increased the spore count and mycorrhizal infection rate of the arbuscular mycorrhizal fungus F. mosseae. While strigolactones and naphthaleneacetic acid significantly increased the infection rates of R. irregularis and C. etunicatum, they had no significant effect on the spore count of either species. Therefore, strigolactones and naphthaleneacetic acid can significantly promote the spore count and mycorrhizal infection rate of F. mosseae to a certain extent. Further comprehensive analysis ultimately determined that the recommended addition levels for strigolactones should be 2-10 mg / L and for naphthaleneacetic acid 30-38 mg / L.
[0044] Example 3
[0045] To further enhance the propagation of arbuscular mycorrhizal fungi, peat, earthworm castings, humic acid, and calcium magnesium phosphate fertilizer were added to the initial materials to create a propagation matrix for arbuscular mycorrhizal fungi. This matrix, by weight, consists of 50-60 parts vermiculite, 20-30 parts earthworm castings, 15-18 parts peat, 0.006-0.012 parts indoleacetic acid, 0.4-0.5 parts humic acid, and 1-2 parts calcium magnesium phosphate. Furthermore, 2-10 mg of strigolactone and 30-38 mg of naphthaleneacetic acid were added per liter of matrix.
[0046] To further promote the application of this propagation medium, the prepared propagation medium was tested for its effectiveness. Maize (Xianyu 335) and arbuscular mycorrhizal fungi (F. mosseae) were used as test materials. The experimental design included 18 treatments, each with three replicates and one pot per replicate. The specific experimental design is as follows.
[0047] Treatment 1: A propagation matrix prepared from the following materials in parts by weight: 50 parts of vermiculite, 30 parts of earthworm castings, 18 parts of peat, 0.012 parts of indoleacetic acid, 0.5 parts of humic acid, and 1.488 parts of calcium magnesium phosphate, with 2 mg of strigolactone and 30 mg of naphthaleneacetic acid added per liter of matrix.
[0048] Treatment 2: A propagation matrix prepared from the following materials in parts by weight: 55 parts of vermiculite, 26.541 parts of earthworm castings, 16 parts of peat, 0.009 parts of indoleacetic acid, 0.45 parts of humic acid, and 2 parts of calcium magnesium phosphate, with 6 mg of strigolactone and 34 mg of naphthylacetic acid added per liter of matrix.
[0049] Treatment 3: A propagation matrix prepared from the following materials in parts by weight: 60 parts of vermiculite, 23.594 parts of earthworm castings, 15 parts of peat, 0.006 parts of indoleacetic acid, 0.4 parts of humic acid, and 1 part of calcium magnesium phosphate, with 10 mg of strigolactone and 38 mg of naphthylacetic acid added per liter of matrix.
[0050] Treatment 4: A propagation matrix prepared from the following materials in parts by weight: 60 parts of vermiculite, 20 parts of earthworm castings, 17.988 parts of peat, 0.012 parts of indoleacetic acid, 0.5 parts of humic acid, and 1.5 parts of calcium magnesium phosphate, with 8 mg of strigolactone and 35 mg of naphthaleneacetic acid added per liter of matrix.
[0051] Control 1-1: The propagation medium prepared in Treatment 1 was not added with strigolactone.
[0052] Control 1-2: Treatment 2 without the addition of strigolactone to the propagation medium.
[0053] Controls 1-3: Treatment 3 without the addition of strigolactone to the propagation medium.
[0054] Controls 1-4: Treatment 4 without the addition of strigolactone to the enrichment medium.
[0055] Control 2-1: The enrichment medium prepared in Treatment 1 was not added with naphthaleneacetic acid.
[0056] Control 2-2: The enrichment medium prepared in Treatment 2 was not added with naphthaleneacetic acid.
[0057] Control 2-3: The enrichment medium prepared in treatment 3 was not added with naphthaleneacetic acid.
[0058] Controls 2-4: enrichment media prepared without adding naphthaleneacetic acid in treatment 4.
[0059] Control 3-1: The enrichment medium prepared in Treatment 1 was not added with strigolactone and naphthaleneacetic acid.
[0060] Control 3-2: The enrichment medium prepared in Treatment 2 was not added with strigolactone and naphthaleneacetic acid.
[0061] Control 3-3: The enrichment medium prepared in Treatment 3 was not added with strigolactone and naphthaleneacetic acid.
[0062] Control 3-4: The enrichment medium prepared in treatment 4 was not added with strigolactone and naphthaleneacetic acid.
[0063] Absolute control 1: vermiculite.
[0064] Absolute control 2: peat.
[0065] According to the above experimental design, the propagation matrix was prepared and a potted test was carried out. The experimental method and propagation management were the same as in Example 1. After 20 weeks of propagation, the aboveground part of the corn was removed, the roots and matrix were collected, air-dried, the roots and matrix were crushed, and mixed to obtain the arbuscular mycorrhizal fungal agent. At the same time, the number of arbuscular mycorrhizal fungal spores and the corn root infection rate in each treatment were measured. The results are shown in Table 1. Figure 5 、 Figure 6 .
[0066] from Figure 5 、 Figure 6 It can be seen that compared with other treatments, the use of the enrichment matrix of the present invention (Treatments 1-4) can significantly increase the number of arbuscular mycorrhizal fungal spores and root infection rate. At the same time, the number of arbuscular mycorrhizal fungal spores and root infection rate in the treatments with only naphthaleneacetic acid added (Controls 1-1 to 1-4) and the treatments with only strigolactone added (Treatments 2-1 to 2-4) were significantly lower than those in the treatments with both strigolactone and naphthaleneacetic acid added (Treatments 1-4). This shows that strigolactone and naphthaleneacetic acid have a certain synergistic effect in increasing the number of arbuscular mycorrhizal fungal spores and root infection rate.
[0067] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.
Claims
1. A propagation matrix for promoting spore production of arbuscular mycorrhizal fungi, characterized in that: Including strigolactone and naphthaleneacetic acid, add 1-40 mg of strigolactone and 1-40 mg of naphthaleneacetic acid per liter of propagation medium; The arbuscular mycorrhizal fungus is Pseudomonas mosseae ( Funneliformis mosseae ).
2. A propagation substrate for promoting spore production of arbuscular mycorrhizal fungi according to claim 1, characterized in that: Add 1-20 mg of strigolactone and 20-39 mg of naphthaleneacetic acid per liter of propagation medium.
3. A propagation matrix for promoting spore production of arbuscular mycorrhizal fungi according to claim 2, characterized in that: Add 2-15 mg of strigolactone and 25-38 mg of naphthaleneacetic acid per liter of propagation medium.
4. A propagation substrate for promoting spore production of arbuscular mycorrhizal fungi according to claim 1, characterized in that: It also includes vermiculite, earthworm castings, peat, indoleacetic acid, humic acid and calcium magnesium phosphate fertilizer.
5. A propagation substrate for promoting spore production of arbuscular mycorrhizal fungi according to claim 4, characterized in that: Calculated by weight, it also includes 50-60 parts of vermiculite, 20-30 parts of earthworm castings, 15-18 parts of peat, 0.006-0.012 parts of indoleacetic acid, 0.4-0.5 parts of humic acid and 1-2 parts of calcium magnesium phosphate.
6. The method for propagating a propagation substrate for promoting spore production of arbuscular mycorrhizal fungi according to any one of claims 1 to 5, characterized in that: The steps include: S1: Sprinkle the arbuscular mycorrhizal fungal agent on the propagation substrate, and then lay a 3-5 cm thick propagation substrate; S2: Using corn as the host plant, corn seeds were sown in the propagation medium 2.5-3.5 cm from the surface. After emergence, the seeds were watered with Hoagland nutrient solution. S3: From seedling emergence to the 21st week, remove the above-ground part of the corn, air-dry the roots together with the propagation medium, and then break the roots and mix them with the propagation medium to obtain the target arbuscular mycorrhizal fungal agent.
7. A method for propagating a propagation substrate for promoting spore production of arbuscular mycorrhizal fungi according to claim 6, characterized in that: In step S2, the seeds are watered with Hoagland nutrient solution one week after emergence; thereafter, the seeds are watered with Hoagland nutrient solution 1 to 2 times every other week for 7 consecutive weeks; the amount of Hoagland nutrient solution used for watering is doubled in the 8th and 9th weeks; the amount of Hoagland nutrient solution used for watering is reduced to 48 to 52% in the 10th week; the amount of Hoagland nutrient solution used for watering is reduced to 18 to 22% from the 11th to the 16th week; and no Hoagland nutrient solution is used for watering from the 17th to the 20th week.
8. A use of a propagation matrix for promoting spore production of arbuscular mycorrhizal fungi as claimed in claim 1 in increasing the number of spores produced by arbuscular mycorrhizal fungi and increasing the infection rate of arbuscular mycorrhizal fungi on corn roots, wherein the arbuscular mycorrhizal fungi is Pseudomonas mosseae ( Funneliformis mosseae ).