Three-river-source moderately-degraded alpine meadow microbial complex inoculant synergistic reseeding rapid recovery method and application

Abstract

The invention provides a three-river-source moderately-degraded alpine meadow microbial complex microbial inoculant synergistic reseeding rapid recovery method and application, and belongs to the technical field of soil remediation. The invention provides a microbial complex microbial inoculant and application of synergistic reseeding of the microbial complex microbial inoculant in degraded meadow repair. The microbial complex microbial inoculants comprise rhizopus intro-rhizosporium and streptomyces microflavus; the reseeded plant seeds comprise seeds of gramineous plants, leguminous plants and miscellaneous grass plants. By introducing two dominant bacteria, the utilization rate of plants to soil resources is improved, the plants have higher environmental stress coping capacity, and by means of the synergistic reseeding technology, the vegetation community composition and the species important value are improved. According to the method disclosed by the invention, the microbial complex microbial inoculants are applied to the restoration of the moderately degraded alpine meadow in the Yangtze-Yangtze-Yangtze-Yangtze-Yangtze-Yangtze-Yangtze-Yangtze-Yangtze-Yangtze-Yangtze-Yangtze-Yangtze-Yangtze-Yangtze-

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of soil remediation, and particularly relates to a method for rapidly restoring degraded alpine meadow in the Three Rivers Source region through microbial compound inoculant and complementary seeding and application thereof. BACKGROUND

[0002] Under the driving of global climate change and human activities, the structure, species diversity and function of grassland ecosystems have undergone tremendous changes. Alpine grassland vegetation is influenced by soil microorganisms, nutrients and environmental factors such as temperature and humidity. Among them, microorganisms participate in the cycling of soil fertility and nutrients, are an important part of the soil ecosystem, and are an important indicator of soil quality and evaluation of soil restoration performance. Soil microbial community and diversity are closely related to soil quality and the health of the grassland ecosystem. Studies have shown that microbial inoculant addition can significantly affect soil microbial community composition, increase bacterial and fungal abundance and microbial diversity index under different degradation levels, and improve soil microbial community structure. Seed source addition can significantly affect alpine grassland vegetation coverage and community composition, and can affect soil microbial community structure and soil nutrient composition through the vegetation-soil feedback mechanism. Microbial inoculants have achieved good results in many crops, but there are deficiencies in restoring degraded grasslands and maintaining regional ecosystem stability. SUMMARY

[0003] In view of the defects in the prior art, the purpose of the present application is to provide a microbial compound inoculant which can rapidly restore degraded meadow in cooperation with complementary seeding and maintain the stability of the meadow ecosystem.

[0004] The purpose of the present application is achieved by the following technical solutions:

[0005] The present application provides a microbial compound inoculant, which comprises Endogone intraradices and Streptomyces microflavus; the Endogone intraradices comprises Endogone intraradices BGC BJ09; and the Streptomyces microflavus comprises Streptomyces microflavus with strain number CGMCC No.4.6556.

[0006] The present application provides a preparation method of the microbial compound inoculant according to the above technical solutions, which comprises:

[0007] Mixing Endogone intraradices with degraded meadow soil to obtain mixed inoculant soil;

[0008] Mixing Streptomyces microflavus with the mixed inoculant soil to obtain a microbial compound inoculant.

[0009] The present application provides application of the microbial compound inoculant according to the above technical solutions or the microbial compound inoculant prepared by the preparation method according to the above technical solutions in the restoration of degraded meadow.

[0010] The application provides application of a microbial compound inoculant in degenerated meadow restoration, the microbial compound inoculant comprises the microbial compound inoculant in the technical solution or the microbial compound inoculant prepared by the preparation method in the technical solution; the plant seeds for the supplementary sowing comprise seeds of grass family plants, legume plants and miscellaneous grass plants.

[0011] Preferably, the restoration comprises any one or two or more of the following (1)-(7):

[0012] (1) improving soil enzyme activity;

[0013] (2) increasing soil organic carbon content;

[0014] (3) improving soil microbial diversity;

[0015] (4) increasing soil available phosphorus content;

[0016] (5) increasing soil ammonium nitrogen content;

[0017] (6) increasing plant aboveground biomass and / or underground biomass;

[0018] (7) increasing the vegetation coverage of the degenerated patch.

[0019] Preferably, the grass family plants comprise Poa palustris; the legume plants comprise Oxytropis ochrocephala; and the miscellaneous grass plants comprise Potentilla tibetica.

[0020] Preferably, the method for degenerated meadow restoration based on the microbial compound inoculant and supplementary sowing comprises the following steps:

[0021] Before greening, seeds of grass family plants, legume plants and miscellaneous grass plants are sown in the degenerated meadow, and then the microbial compound inoculant is covered above the seeds.

[0022] Preferably, when sowing, the seeds of grass family plants, legume plants and miscellaneous grass plants are mixed in a ratio of (0.7-1):(0.7-1):(0.7-1) and then sown.

[0023] Preferably, when sowing, the sowing amount is 600-1000 seeds / m 2 .

[0024] Preferably, when covering the microbial compound inoculant, the thickness of the covering is 1-1.5 cm.

[0025] The application has the following beneficial effects:

[0026] The present application provides a microbial compound microbial inoculant, comprising endorhiza and streptomyces microflavus; the endorhiza comprises endorhiza BGC BJ09; the streptomyces microflavus comprises the strain numbered as CGMCC No.4.6556 streptomyces microflavus. In the present application, the endorhiza can invade the plant root cortex cells through mycelium, form vesicle-branch structure, promote the plant to absorb the available nitrogen and phosphorus in soil, and can secrete saccarase to decompose soil organic matter, and enhance the resistance of plant to environmental stress; the streptomyces microflavus can reduce the soil pH value by secreting organic acid and the like, increase the solubility of the insoluble phosphorus in soil, make it transform into available phosphorus which can be absorbed by plant, and can secrete various enzymes (such as cellulase, protease, amylase, etc.), not only promote the decomposition of organic matter such as plant residues and animal manure, but also improve the activity of original enzymes in soil, thereby promote the soil nutrient transformation and improve the availability of nutrients for vegetation. The present application introduces two dominant bacteria in the form of microbial compound microbial inoculant, improves the utilization rate of soil resources for plant, and makes the plant have stronger environmental stress response ability. On this basis, the synergistic interplanting can quickly restore the degraded meadow, and is also beneficial to maintaining the stability of meadow ecosystem. The experimental results show that the application of microbial compound microbial inoculant and the interplanting of seeds can significantly improve the soil enzyme activity, increase the available nutrients in soil, and improve the vegetation coverage and underground biomass in the degraded patches.

[0027] The present application provides the application of microbial compound microbial inoculant and synergistic interplanting in the restoration of degraded meadow, the microbial compound microbial inoculant comprises endorhiza and streptomyces microflavus; the interplanted plant seeds comprise seeds of gramineous plants, leguminous plants and forbs. The microbial compound microbial inoculant and synergistic interplanting of the present application mainly introduce two dominant bacteria, improve the utilization rate of soil resources for plant, make the plant have stronger environmental stress response ability, and improve the vegetation community composition and species importance value through synergistic interplanting technology. The results of the examples show that the application of microbial compound microbial inoculant and synergistic interplanting technology in the moderately degraded alpine meadow in the Yangtze River source area of Sanjiangyuan National Park can quickly improve the soil enzyme activity, soil available nutrients, vegetation coverage and biomass in the moderately degraded meadow, and maintain the stability of meadow ecosystem. In summary, the combined application of compound microbial inoculant and interplanting can quickly restore the degraded alpine meadow. BRIEF DESCRIPTION OF DRAWINGS

[0028] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed in the embodiments. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without creative labor.

[0029] Figure 1 Schematic diagram of the method for rapid restoration of moderately degraded alpine meadow in Example 1;

[0030] Figure 2 Layout diagram of the test in the moderately degraded alpine meadow in Example 1;

[0031] Figure 3 Result graph of the influence of the restoration treatment on the characteristics of the vegetation community in the moderately degraded alpine meadow in Example 1;

[0032] Figure 4 Result graph of the influence of the restoration treatment on the vegetation diversity in the moderately degraded alpine meadow in Example 1;

[0033] Figure 5 Result graph of the influence of the restoration treatment on the underground biomass in the moderately degraded alpine meadow in Example 1;

[0034] Figure 6 Result graph of the influence of the restoration treatment on the surface soil enzyme activity in the moderately degraded alpine meadow in Example 1;

[0035] Figure 7 Result graph of the influence of the restoration treatment on the surface soil nutrients in the moderately degraded alpine meadow in Example 1. DETAILED DESCRIPTION

[0036] The application provides a microbial compound microbial agent, which comprises Endogone intraradices and Streptomyces microflavus; the Endogone intraradices comprises Endogone intraradices BGC BJ09; and the Streptomyces microflavus comprises Streptomyces microflavus with a strain number of CGMCC No.4.6556.

[0037] In the application, the microbial compound microbial agent comprises Endogone intraradices and Streptomyces microflavus. In the application, the Endogone intraradices comprises Endogone intraradices with a strain number of BGC BJ09. As an optional embodiment of the application, the product form of the Endogone intraradices can be Endogone intraradices with a spore number of 15 / g. In the application, the Streptomyces microflavus comprises Streptomyces microflavus with a strain number of CGMCC No.4.6556. As an optional embodiment of the application, the product form of the Streptomyces microflavus can be Streptomyces microflavus with a microbial activity of 2x10 9 CFU / g.

[0038] The application provides a preparation method of the microbial compound microbial agent.

[0039] The Endogone intraradices is mixed with degraded meadow soil to obtain mixed microbial agent soil.

[0040] The Streptomyces microflavus is mixed with the mixed microbial agent soil to obtain the microbial compound microbial agent.

[0041] In the present application, the degraded meadow soil can be the soil of the degraded meadow to be repaired, or the soil of the meadow with the same degradation degree as the degraded meadow to be repaired. In the present application, the product of the above technical solution of the root-endoparasitic Glomus and the product of the above technical solution of the Streptomyces microflavus are preferably directly used for the preparation of the microbial compound microbial agent. As an optional embodiment of the present application, the mass ratio of the root-endoparasitic Glomus to the degraded meadow soil can be 1:(40-50), or 1:40, 1:41, 1:42, 1:43, 1:44, 1:45, 1:46, 1:47, 1:48, 1:49 or 1:50; the mass ratio of the Streptomyces microflavus to the mixed microbial agent soil can be 1:(800-1000), or 1:800, 1:810, 1:820, 1:830, 1:840, 1:850, 1:860, 1:870, 1:880, 1:890, 1:900, 1:910, 1:920, 1:930, 1:940, 1:950, 1:960, 1:970, 1:980, 1:990 or 1:1000. The mixing method is not particularly limited in the present application, and the conventional mixing method in the art can be used. In the present application, the microbial compound microbial agent can also be referred to as the degraded meadow soil containing the microbial compound microbial agent. As an optional embodiment of the present application, the microbial compound microbial agent can be directly applied to the top of the supplemental planting seeds.

[0042] The present application provides the application of the microbial compound microbial agent in the above technical solution or the microbial compound microbial agent prepared by the preparation method in the above technical solution in the repair of the degraded meadow. In the present application, the degraded meadow includes any one or two or more of the light degraded meadow, the moderate degraded meadow and the severe degraded meadow.

[0043] The present application provides the application of the microbial compound microbial agent in the repair of the degraded meadow, wherein the microbial compound microbial agent includes the microbial compound microbial agent in the above technical solution; and the supplemental planting seeds include the seeds of the grass family plants, the legume plants and the miscellaneous grass plants.

[0044] In the present application, the meadow includes alpine meadow; the alpine meadow includes the alpine meadow in the Changjiangyuan Garden of the Three-River-Source National Park; the alpine meadow in the Changjiangyuan Garden of the Three-River-Source National Park includes the alpine meadow in the permafrost region of the Changjiangyuan Garden of the Three-River-Source National Park. In the present application, the degraded meadow includes any one or two or more of the following: light degraded meadow, medium degraded meadow and heavy degraded meadow. In the present application, the degradation degree of the degraded meadow is preferably classified according to the reduction rate of the relative percentage of total vegetation coverage. In the present application, when the vegetation coverage is reduced by 0-10%, the degradation degree of the meadow is non-degradation; when the vegetation coverage is reduced by 11%-20%, the degradation degree of the meadow to be repaired is light degradation; when the vegetation coverage is reduced by 21%-30%, the degradation degree of the meadow to be repaired is medium degradation; and when the vegetation coverage is reduced by >30%, the degradation degree of the meadow to be repaired is heavy degradation. In the present application, the medium degraded alpine meadow in the Changjiangyuan Garden of the Three-River-Source National Park in the permafrost region is taken as an example to specifically describe the microbial complex microbial agent and the technical effect thereof.

[0045] In the present application, the microbial complex microbial agent is described in the above technical solution, and will not be described here.

[0046] In the present application, the Gramineae plants, Leguminosae plants and forbs include the main Gramineae plants, Leguminosae plants and forbs in the non-degraded meadow near the degraded meadow. In the selection of the Gramineae plants, Leguminosae plants and forbs in the present application, the non-degraded vegetation community composition near the degraded meadow is investigated, the vegetation community building species and dominant species are investigated by using a 50cm*50cm sample frame, the investigation is carried out for 5 times, and the main vegetation of the Gramineae, Leguminosae and forbs in the meadow is determined. As an optional embodiment of the present application, the Gramineae plants include Poa crymophila; the Leguminosae plants include Oxytropis ochrocephala; and the forbs include Potentilla tibetica. Specifically, the Poa crymophila, Oxytropis ochrocephala and Potentilla tibetica in the present application are obtained by investigating the vegetation community composition of the non-degraded meadow in the permafrost region of the Changjiangyuan Garden of the Three-River-Source National Park.

[0047] As an optional embodiment of the present application, the repairing comprises any one or two or more of the following (1) to (7): (1) improving soil enzyme activity; (2) increasing soil organic carbon content; (3) improving soil microbial diversity; (4) increasing soil available phosphorus content; (5) increasing soil ammonium nitrogen content; (6) increasing plant aboveground biomass and / or underground biomass; and (7) increasing vegetation coverage of degraded patches. As an optional embodiment of the present application, the soil enzymes include any one or two or more of soil sucrose, soil alkaline phosphatase, urease and peroxidase. The present application shows through the example results that the use of the microbial complex microbial agent in combination with supplemental seeding for repairing degraded meadow can significantly increase vegetation coverage, aboveground plant species richness, aboveground vegetation Shannon-Wiener index and dominance index, surface underground biomass, soil sucrose, soil alkaline phosphatase, urease and peroxidase activities, and soil surface organic carbon, ammonia nitrogen and available phosphorus.

[0048] As an optional embodiment of the present application, the method for repairing degraded meadow based on microbial complex microbial agent in combination with supplemental seeding comprises:

[0049] Before greening, the seeds of gramineous plants, leguminous plants and forbs are sown in the degraded meadow, and then the degraded meadow soil containing the microbial complex microbial agent is uniformly covered above the seeds.

[0050] When sowing, the present application can use the quantity ratio of the seeds of gramineous plants, leguminous plants and forbs as (0.7-1):(0.7-1):(0.7-1), or 1:1:1; and the sowing mode is mixed sowing. The sowing amount of the present application can be 600-1000 seeds / m 2 , or 600, 700, 800, 900 or 1000 seeds / m 2 In the present application, the sowing amount refers to the sowing amount in terms of the number of active three kinds of seeds.

[0051] After sowing, the present application uniformly covers the degraded meadow soil containing the microbial complex microbial agent above the seeds. As an optional embodiment of the present application, when covering with the degraded meadow soil containing the microbial complex microbial agent, the thickness of the covering is 1-1.5 cm, or 1, 1.1, 1.2, 1.3, 1.4 or 1.5 cm.

[0052] The Three-River Headwaters National Park is an important ecological security barrier, known as the "Chinese Water Tower", which has far-reaching and important significance. The Three-River Headwaters has a unique alpine wetland ecosystem in the world, which shelters more than 30 endangered and unique species such as snow leopards and Tibetan antelopes, and is the gene pool of biodiversity of the Qinghai-Tibet Plateau. At the same time, its vast glaciers, meadows and peatlands store about 15 billion tons of carbon, which plays a strategic role in mitigating global climate change. Therefore, it is of great significance to maintain the stability of the regional ecosystem to deeply study the restoration method of the degraded grassland in the region. The present application uses a microbial complex inoculant composed of Rhizophagus intraradices and Streptomyces microflavus and a mixed seeding of multiple grasses as a practical example. The constructed microbial complex inoculant and the complementary seeding technology are successfully applied to the restoration of the moderately degraded alpine meadow in the Changjiangyuan Park of the Three-River Headwaters National Park in the permafrost region. The test results show that the application of the microbial complex inoculant and the complementary seeding in the moderately degraded alpine meadow can rapidly improve the soil enzyme activity, increase the plant and microbial diversity, and increase the vegetation coverage and underground biomass in the degraded patches. Therefore, the microbial complex inoculant and the complementary seeding technology of the present application can realize the rapid restoration of the moderately degraded alpine meadow.

[0053] In order to further illustrate the present application, the technical solutions provided by the present application are described in detail below in combination with the drawings and examples, but they should not be understood as limiting the scope of protection of the present application.

[0054] The strain information used in the following technical solutions

[0055] Strain name: Rhizophagus intraradices; Latin name: Rhizophagus intraradices; strain from Chinese arbuscular mycorrhizal fungi germplasm resource bank; institution abbreviation: BGC; address: No. 9, Shuguanghuayuan Middle Road, Haidian District, Beijing; preservation center registration number: BGC BJ09. The product is Rhizophagus intraradices with a spore number of about 15 / g mixed soil. The product can be directly referred to as Rhizophagus intraradices with a spore number of 15 / g.

[0056] Strain name: Streptomyces microflavus; Latin name: Streptomyces microflavus; strain from the preservation agency: China General Microbiological Culture Collection Center; preservation agency abbreviation: CGMCC; address: No. 3, Beichen West Road, Beijing City, Chaoyang District; preservation center registration number: CGMCC No. 4.6556. The product is Streptomyces microflavus with a bacterial activity of 2x10 9 CFU / g.

[0057] The moderately degraded alpine meadow in the following examples and comparative examples is in the same land (an area of 1 km 2)randomly divided into 30 plots (control group, complex microbial agent group, pairwise double broadcasting group (3 groups) and triple broadcasting group, a total of 6 groups, 5 replicates, i.e. 5x6=30 plots). The distribution of test plots is the unrepaired test area, which is used as the control group; the test plot of Example 1 is the complex microbial agent + supplementary planting (triple broadcasting) test plot; the test plot of Comparative Example 1 is the complex microbial agent use group; the test plot of Comparative Example 2 is the complex microbial agent + double broadcasting 1 test plot, the complex microbial agent + double broadcasting 2 test plot and the complex microbial agent + double broadcasting 3 test plot, respectively.

[0058] Example 1

[0059] A schematic diagram of a method for rapid recovery of moderately degraded alpine meadow is shown in Figure 1 , and the specific steps are as follows:

[0060] Experimental site: about 10 kilometers northwest of Yegexiang, Qumalai County, Yushu Tibetan Autonomous Prefecture, Qinghai Province.

[0061] Sample plot information: elevation 4632 m; latitude and longitude: N 34°40'23", E 95°12'5".

[0062] Experimental time: about 5 days before the local plants turn green, and this example started on May 28, 2024.

[0063] S1, degradation assessment: according to the national standard "Classification Index of Natural Grassland Degradation, Desertification and Salinization" (GB 19377-2003), the degradation degree is divided into light, moderate and severe.

[0064] The degradation degree of the grassland to be repaired is divided according to the reduction rate (%) of the total coverage percentage. When the vegetation coverage decreases by 0-10%, the degradation degree of the grassland is not degraded, when the vegetation coverage decreases by 11%-20%, the degradation degree of the grassland to be repaired is light, when the vegetation coverage decreases by 21%-30%, the degradation degree of the grassland to be repaired is moderate, and when the vegetation coverage decreases by >30%, the degradation degree of the grassland to be repaired is severe.

[0065] In this example, moderately degraded alpine meadow is selected as the experimental object, and the test layout of moderately degraded alpine meadow in the example is shown in Figure 2 .

[0066] S2, degradation patch area assessment, estimate the area of each degradation patch by 50cmx50cm quadrat, small patch area is about 0.125m 2 , medium patch area is about 0.25m 2 , and large patch area is about 0.75m 2 .

[0067] S3, investigate the vegetation community composition of the non-degraded alpine meadow near the restoration area, investigate the vegetation community building species and dominant species using a 50cm x 50cm sample frame, investigate 5 times, respectively determine the most important vegetation of the grass family, legume family and forbs of the grassland, finally determine that the most important vegetation of the grass family is Poa crymophila Keng, the most important vegetation of the legume family is Oxytropis ochrocephala, and the most important vegetation of the forbs is Potentilla saundersiana. According to the three most important vegetation determined, the grass family Poa crymophila Keng, the legume family Oxytropis ochrocephala and the forbs Potentilla saundersiana are selected for the restoration of the moderately degraded alpine meadow in this embodiment.

[0068] S4, collect the seeds of Poa crymophila Keng, Oxytropis ochrocephala and Potentilla saundersiana respectively, and screen the seeds to achieve 100% purity.

[0069] S5, use TTC staining method to test seed activity, specifically use triphenyltetrazolium chloride (TTC) staining, observe the staining of the seed embryo to determine whether the seed has activity, and through the activity test, the activity of Poa crymophila Keng, Oxytropis ochrocephala and Potentilla saundersiana is 72%, 64% and 58% respectively.

[0070] S6, test the seed hundred-grain weight respectively, calculate the weight of 200 germinated seeds according to the activity and pack in envelopes, and store in a dry and cool place.

[0071] S7, preparation of microbial compound inoculant soil:

[0072] 1) Soil collection and treatment for microbial compound inoculant preparation: collect soil in the moderately degraded patch outside the experimental area, remove dead branches, leaves and gravel through a 1cm sieve, then manually crush larger soil particles through a 40 mesh (0.4mm) sieve to obtain the experimental soil to be used.

[0073] 2) Mix the endogonales intraradices with a spore number of 15 / g with the above-mentioned treated experimental soil to be used according to a mass ratio of 1:50 to form a mixed inoculant soil.

[0074] 3) Mix the streptomyces microflavus with a strain activity of 2x10 9 CFU / g with the mixed inoculant soil according to a mass ratio of 1:1000 to form a degraded meadow soil containing a microbial compound inoculant.

[0075] S8, supplemental planting method, mix Poa crymophila Keng, Oxytropis ochrocephala and Potentilla saundersiana according to the active seed quantity ratio of 1:1:1, evenly place them in the degraded patch by sowing, determine the seed planting quantity according to the patch area of step S2, and according to 800 grains / m 2The calculation of the amount of the supplementary seeding.

[0076] S9, Specific application of microbial inoculants and supplementary seeding in the degraded patch: The seeds are evenly sown in the patch by S8, and the degraded meadow soil containing microbial complex inoculants prepared in S7 is paved and covered on the sowing area with a thickness of 1.5 cm.

[0077] Management: The experimental area is enclosed, grazing is prohibited, wild animals are prohibited from trampling, mouse holes in the experimental area are checked and the hole is blocked with stones, and every half month, the safety survey of the sample plot is carried out, including the stability of the experimental sample plot fence, the animal footprint in the experimental area, etc., and the overall recovery of the test area is observed to prepare the vegetation survey and soil sample collection before the plant withering period.

[0078] Application Example 1

[0079] The vegetation community survey and soil sample collection after the experiment are carried out 8-15 days before the withering period, and the relevant parameters of the moderately degraded alpine meadow repaired in Example 1 and the moderately degraded alpine meadow without repair are detected and compared.

[0080] The vegetation community characteristics of the moderately degraded alpine meadow repaired in Example 1 (also referred to as complex inoculants + supplementary seeding (three seeding)) and the moderately degraded alpine meadow without repair are counted, the total coverage, abundance and species richness of the grassland in each treatment group are calculated, and the results are shown in Figure 3 . Figure 3 The results of the influence of the repair treatment of Example 1 on the vegetation community characteristics of the moderately degraded alpine meadow are shown in the figure. Among them, A is the influence of the repair treatment on the total coverage; B is the influence of the repair treatment on the vegetation abundance; C is the influence of the repair treatment on the species richness.

[0081] The vegetation diversity of the moderately degraded alpine meadow repaired in Example 1 (also referred to as complex inoculants + supplementary seeding (three seeding)) and the moderately degraded alpine meadow without repair is measured, and the results are shown in Figure 4 . Figure 4 The results of the influence of the repair treatment of Example 1 on the vegetation diversity of the moderately degraded alpine meadow are shown in the figure. Among them, A is the influence of the repair treatment on the Simpson dominance index; B is the influence of the repair treatment on the evenness index; C is the influence of the repair treatment on the Shannon-Wiener index.

[0082] The underground biomass of the moderately degraded alpine meadow repaired in Example 1 (also referred to as complex inoculants + supplementary seeding (three seeding)) and the moderately degraded alpine meadow without repair is measured, and the results are shown in Figure 5 . Figure 5 The results of the influence of the repair treatment of Example 1 on the underground biomass of the moderately degraded alpine meadow are shown in the figure.

[0083] The surface soil enzyme activity of the moderately degraded meadow after the repair of Example 1 (which can also be referred to as compound microbial agent + reseeding (three reseeding)) and the moderately degraded alpine meadow without repair was detected, and the results are shown in Table 2. Figure 6 Figure 6 The influence of the repair treatment of Example 1 on the surface soil enzyme activity of the moderately degraded alpine meadow is shown in the graph of FIG. 2. Among them, A is the influence of the repair treatment on soil sucrase; B is the influence of the repair treatment on soil cellulase; C is the influence of the repair treatment on soil alkaline phosphatase; D is the influence of the repair treatment on soil urease; and E is the influence of the repair treatment on soil peroxidase.

[0084] The surface soil nutrients of the moderately degraded meadow after the repair of Example 1 (which can also be referred to as compound microbial agent + reseeding (three reseeding)) and the moderately degraded alpine meadow without repair were detected, and the results are shown in Table 3. Figure 7 Figure 7 The influence of the repair treatment of Example 1 on the surface soil nutrients of the moderately degraded alpine meadow is shown in the graph of FIG. 3. Among them, A is the influence of the repair treatment on soil total carbon; B is the influence of the repair treatment on soil total nitrogen; C is the influence of the repair treatment on soil total phosphorus; D is the influence of the repair treatment on soil organic carbon; E is the influence of the repair treatment on soil ammonium nitrogen; F is the influence of the repair treatment on soil nitrate nitrogen; and G is the influence of the repair treatment on soil available phosphorus.

[0085] ​​The determination methods of the five soil enzyme activities (urease, phosphatase, sucrase, cellulase and catalase) are as follows. Urease activity determination: collect soil samples, dry, grind and sieve; prepare urea solution and add urease solution; place the reaction system in a constant temperature condition and record the reaction time; take samples at regular intervals during the reaction and determine the content of ammonium ions in the reaction system by ammonium ion determination method (such as Nessler's reagent colorimetry); record the data and calculate the urease activity. Phosphatase activity determination: according to different soil pH values, provide the corresponding pH buffer; add soil samples to the buffer and stir uniformly; incubate under constant temperature conditions for a certain period of time; determine the amount of generated organic groups or inorganic phosphorus by appropriate chemical methods, and calculate the phosphatase activity. Sucrase activity determination: use the polyphenol inhibition method, mix the soil sample with the polyphenol solution, stand for a period of time, then add buffer and phenolphthalein indicator, titrate, record the volume of phenolphthalein titrant required for titration, and calculate the sucrase activity. Cellulase activity determination: prepare reagents and equipment such as acetic acid buffer, CMC solution, hydrated glucose solution, etc.; take an appropriate amount of fresh soil sample, add acetic acid buffer and CMC solution, incubate under constant temperature conditions for a certain period of time; filter the sample, add hydrated glucose solution, heat in a boiling water bath for a certain period of time, cool, then add anhydrous glucose solution, and perform colorimetric determination at a wavelength of 690 nm to calculate the cellulase activity. Catalase determination: prepare reagents such as 0.3% hydrogen peroxide solution, 3N sulfuric acid and 0.1N potassium permanganate solution; accurately weigh a certain amount of reference sodium oxalate and titrate with potassium permanganate solution to calibrate, mix the soil sample with hydrogen peroxide solution, react for a certain period of time, then titrate the remaining hydrogen peroxide with potassium permanganate solution, and calculate the catalase activity.

[0086] Spectrophotometry and elemental analysis are used for soil nutrient testing. Soil total nutrient determination uses a 0.25mm sieve to pass the soil sample, and the determination indicators include soil total carbon, soil total nitrogen (elemental analyzer), and inorganic carbon (soil carbonate determination instrument). Spectrophotometry is used to determine soil available nutrients, and a 2mm sieve is used to pass the soil sample. The determination indicators include soil ammonium nitrogen (potassium chloride extraction-indigo colorimetry), soil available phosphorus (sodium bicarbonate extraction-molybdenum antimony anti-colorimetry), and soil available potassium (acid-soluble flame photometry) [1,2] .

[0087] References:

[0088] [1] Baosidan. Soil agrochemical analysis [M]. 3rd edition, Beijing: China Agricultural Press, 2007: 268-270.

[0089] [2] Pan Xianzhang, Guo Zhiying, Pan Kai. Land ecosystem soil observation indicators and specifications [S]. Beijing: China Environmental Publishing Group, 2019: 185-211.

[0090] The detection results of the related indexes of the moderately degraded meadow after repair in Example 1 and the moderately degraded alpine meadow without repair are shown in Table 1 and Figures 3 to 7 .

[0091] Table 1 Detection results of related indexes of moderately degraded meadow after repair in Example 1 and moderately degraded alpine meadow without repair

[0092]

[0093]

[0094] Note: The control group is the detection results of related indexes of the moderately degraded alpine meadow without repair, and the same below.

[0095] From Table 1 and Figures 3 to 7 , it can be seen that by using the microbial compound inoculant and complementary seeding method in the moderately degraded alpine meadow, the vegetation coverage in the degraded patch is significantly improved by 31%, the number of plant stems before repair is less than 500 stems / m 2 , the number of plant stems above ground after repair is more than 800 stems / m 2 , and the above-ground plant species richness is increased by more than 5 species. The above-ground vegetation Shannon-Wiener index (diversity index) and dominance index are also significantly increased. By taking the soil surface 0-10 cm underground biomass with a root drill, it is found that the surface underground biomass is significantly increased by 800 g / m 2 . In addition, the activities of five enzymes such as soil sucrose, cellulase, soil alkaline phosphatase, urease and peroxidase are tested by using the kit, and except for cellulase, the activities of other enzymes are significantly increased. Through soil nutrient analysis, it is found that the soil surface organic carbon is significantly increased by 5 g / kg, the surface ammonia nitrogen is significantly increased by 25 mg / kg, and the available phosphorus is significantly increased by 2 mg / kg. The experiment shows that the microbial compound inoculant and complementary seeding method used in Example 1 has a significant and rapid repair effect on the vegetation and soil properties of the moderately degraded alpine meadow.

[0096] Comparative Example 1

[0097] Only the compound inoculant in Example 1 is used to repair the moderately degraded alpine meadow degraded patch without complementary seeding.

[0098] Except for no complementary seeding, the other experimental processes are the same as Example 1. Among them, the experimental group is to apply the microbial compound inoculant, and the control group is not to apply the inoculant. The related sample collection and test indexes are also consistent with Example 1.

[0099] Table 2 Detection results of related indexes of moderately degraded meadow after repair in Comparative Example 1 and moderately degraded alpine meadow without repair

[0100]

[0101]

[0102] As shown in Table 2, by using the microbial compound microbial agent method in the moderately degraded alpine meadow, the vegetation abundance in the degraded patch and the soil surface ammonia nitrogen are significantly improved. Compared with Example 1, in the present comparative example, the influence of three sowing on soil and vegetation related indicators is more significant when the microbial compound microbial agent is applied.

[0103] Comparative Example 2

[0104] The compound microbial agent in Example 1 is used in combination with double sowing, and the difference between Example 1 and Comparative Example 2 is that the double sowing of Comparative Example 2 uses any two of the seeds supplemented in Example 1 for double sowing. Except that the supplementing method is double sowing, the other experimental processes are the same as those of Example 1. The related sample collection and test indicators are also consistent with those of Example 1. When double sowing is performed, the two kinds of seeds are mixed according to the ratio of the number of active seeds 1:1, and the supplementing amount is the same as that of Example 1.

[0105] In the present comparative example, the double sowing conditions of Poa crymophila and Oxytropis ochrocephala are defined as compound microbial agent + double sowing 1; the double sowing conditions of Poa crymophila and Potentilla tianschanica are defined as compound microbial agent + double sowing 2; and the double sowing conditions of Oxytropis ochrocephala and Potentilla tianschanica are defined as compound microbial agent + double sowing 3.

[0106] Table 3 Detection results of related indicators of moderately degraded meadow after repair in Comparative Example 2 and moderately degraded alpine meadow without repair

[0107]

[0108]

[0109] As shown in Table 3, from the variance analysis results of Comparative Example 2, compared with the control group, the microbial compound microbial agent and different double sowing combinations can significantly improve the vegetation abundance, BGB, Shannon-Wiener index, urease, soil alkaline phosphatase and peroxidase, but the differences of other soil and vegetation indicators are not significant. Compared with Example 1, in the present comparative example, the influence of three sowing on soil and vegetation related indicators is more significant when the microbial compound microbial agent is applied.

[0110] Although the above examples have made a detailed description of the present application, it is only a part of the embodiments of the present application, not all the embodiments, and other embodiments can be obtained according to the present embodiments without creativity, which all belong to the protection scope of the present application.

Claims

1. A microbial complex inoculant, characterized in that, The root endophytic root rot fungus comprises a root endophytic root rot fungus BGC BJ09; and the fine yellow Streptomyces comprises a fine yellow Streptomyces with a strain number of CGMCC No. 4.6556.

2. The preparation method of the microbial complex microbial agent of claim 1, characterized in that, The application further discloses a preparation method of the microbial composite microbial agent. The root endophytic root rot fungus is mixed with degraded meadow soil to obtain mixed microbial agent soil; The fine yellow Streptomyces is mixed with the mixed microbial agent soil to obtain the microbial composite microbial agent.

3. The microbial composite microbial agent of claim 1 or the microbial composite microbial agent prepared by the preparation method of claim 2 is applied in degraded meadow restoration.

4. Application of a microbial composite microbial agent in degraded meadow restoration in cooperation with supplemental sowing, wherein the microbial composite microbial agent comprises the microbial composite microbial agent of claim 1 or the microbial composite microbial agent prepared by the preparation method of claim 2; and the seeds of the plants to be sown supplementally comprise seeds of grasses, legumes and forbs.

5. Use according to claim 4, characterized in that, The restoration comprises any one or two or more of the following (1)-(7): (1) improving soil enzyme activity; (2) increasing soil organic carbon content; (3) improving soil microbial diversity; (4) increasing soil available phosphorus content; (5) increasing soil ammonium nitrogen content; (6) increasing plant aboveground biomass and / or underground biomass; (7) increasing vegetation coverage of degraded patches.

6. Use according to claim 4, characterized in that, The grasses comprise Poa palustris; the legumes comprise Oxytropis ochrocephala; and the forbs comprise Potentilla tibetica.

7. Use according to claim 4, characterized in that, A method for degraded meadow restoration based on a microbial composite microbial agent in cooperation with supplemental sowing comprises the following steps: Before greening, seeds of grasses, legumes and forbs are sown in the degraded meadow, and then the microbial composite microbial agent is covered above the seeds.

8. Use according to claim 7, characterized in that, When sowing, the seeds of the grasses, the seeds of the legumes and the seeds of the forbs are mixed in a ratio of (0.7-1):(0.7-1):(0.7-1) and then sown.

9. Use according to claim 7, characterized in that, The seeding rate at the time of sowing is 600 to 1000 grains / m 2 .

10. Use according to claim 7, characterized in that, When covering the microbial composite microbial agent, the thickness of the covering is 1-1.5 cm.

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