A physical-biological coupling closed loop method for radically preventing and controlling long-spine burrweed

Abstract

The present application belongs to the technical field of ecological prevention and control of poisonous grass in grassland and ecological restoration of sandy land, and more particularly relates to a method for physically-biologically coupled closed-loop radical control of Amaranthus palmeri S. Wats. The present application is directed to the biological characteristics of Amaranthus palmeri S. Wats. in terms of stratified seed storage and deep dormancy, and through stratified physical operation of surface seed source purification, shallow rotary tillage dormancy breaking, and deep subsoiling pore forming, the soil seed bank in 0-40 cm is disturbed layer by layer, and all the collected Amaranthus palmeri S. Wats. seed sources are concentrated and high-temperature composted during the operation process; the process of surface shallow harrow mixing bacteria and deep subsoiling synchronous bacteria injection is matched, and each layer of dormant seeds is inactivated, and the gradient planting of native grasses is combined with annual closed-loop management and protection, so that the radical control of Amaranthus palmeri S. Wats. and the ecological restoration of sandy land are simultaneously completed, and the needs of large-scale operation, zero ecological pollution, and long-term stable control are met. The present method is free of chemical pesticides throughout the process, is suitable for large-scale operation in sandy soil in the north, and the total inactivation rate of the soil seed bank of Amaranthus palmeri S. Wats. is more than 90%.

Description

Technical Field

[0001] This invention belongs to the field of ecological control of poisonous grasses in grasslands and ecological restoration of sandy land. More specifically, it relates to a method for the radical control of Tribulus terrestris through a physical-biological coupled closed-loop system. Background Technology

[0002] Tribulus terrestris is a malignant invasive annual herbaceous weed with biological characteristics including double seed dormancy in spiny bracts, stratified seed storage in the soil, staged germination, and explosive propagation through tillering. Its seed dormancy period can last for more than 5 years, and its seed bank in the 20-40cm deep soil layer is highly latent. It easily spreads over large areas in sandy soils in northern regions, damaging grassland vegetation communities, reducing land utilization, and impacting livestock production safety. Currently, the control technologies for Tribulus terrestris in China are mainly divided into four categories, all of which have significant technical shortcomings and cannot achieve radical and green control.

[0003] Invasive herbaceous plant substitution and control technologies: Some existing technologies use hybrid grasses such as Napier grass and Jerusalem artichoke, or single native herbs, for broadcasting and planting, relying on plant density to suppress the growth of mature Tribulus terrestris. However, these technologies only inhibit the growth of mature plants and cannot affect the dormant seed bank in the soil, resulting in a recurrence rate of over 60% the following year. The core substitute grass species are introduced species, which have poor adaptability to northern sandy areas, posing a risk of secondary ecological invasion. Single herbaceous plant planting cannot form a stable community, resulting in extremely poor long-term control capabilities.

[0004] Chemical spraying control technology: Some existing technologies use chemical herbicides or plant-derived extracts for whole-area spraying control. In this technology, the agents can only kill weeds that have already emerged on the ground surface and cannot penetrate the soil to act on dormant seeds. Chemical agents leave soil residues, which can damage vegetation and disrupt the soil micro-ecology. Plant-derived agents lack targeting and can easily damage beneficial herbs, which does not meet the requirements of green ecological control.

[0005] Single physical and mechanical control techniques: Some existing technologies control the plant by shallow tilling (18-22cm deep), manual removal, and collection and burning of the spiny husks. However, this approach does not consider the stratified seed storage characteristics of Tribulus terrestris; dormant seeds in the 20-40cm deep soil layer cannot be removed and can germinate upwards year by year; manual removal is extremely inefficient and unsuitable for large-scale operations; burning the spiny husks can easily cause secondary seed dispersal, resulting in poor purification effects.

[0006] Single-strain biological agent control technology: This technology often uses single-function strains such as Alternaria alternata to infect weed plants through spraying or soil mixing. This approach has three significant disadvantages: First, the strains are single-function, only infecting mature plants that have already emerged, and lack the ability to break seed dormancy, lyse seed coats, or inactivate embryos, thus failing to reach dormant seed banks in the soil. Second, without physical disturbance, the agent struggles to penetrate deeper into the soil, offering virtually no control over latent seeds in the 20-40cm soil layer. Third, single strains have a narrow ecological niche in the soil, resulting in poor microbial community stability, short duration of effectiveness, and significant fluctuations in control efficacy in arid and barren sandy environments.

[0007] It is evident that the various control measures in the existing technologies are used independently and without forming a coupled and coordinated system; they have not formed a coordinated system of "breaking dormancy - inactivation - site occupation - management" targeting the core characteristics of Tribulus terrestris's stratified seed storage and deep dormancy, thus failing to eradicate the soil dormant seed bank; most technologies have problems such as ecological risks, residual pollution, high recurrence rate, and poor adaptability, and there is no closed-loop control system that can achieve one-time rectification and long-term stability.

[0008] Therefore, in view of the technical shortcomings of existing control technologies for Tribulus terrestris, such as single control methods, poor eradication, high ecological risk, weak adaptability, and insufficient long-term effectiveness, it is an urgent problem to be solved by those skilled in the art to provide a completely green, chemical-free, physical-biological coupled closed-loop eradication control method. Summary of the Invention

[0009] The purpose of this invention is to provide a physical-biological coupled closed-loop method for the radical control of Tribulus terrestris. Specifically, it relates to a closed-loop control method that combines purely physical and mechanical control with biological control for Tribulus terrestris in arid sandy areas and natural grasslands in northern China. This method is suitable for large-scale green control operations in areas where Tribulus terrestris is heavily invaded.

[0010] To achieve the above objectives, the present invention provides the following solution: One of the technical solutions of this invention is to provide a method for the radical control of Tribulus terrestris using a physical-biological coupled closed-loop system, comprising the following steps: Grazing ban and enclosure management will be implemented in the project area; Before the spring germination of Tribulus terrestris, the soil in the project area was subjected to stratified physical dormancy breaking and seed clearing operations. Subsequently, compound biological agents were applied to the soil in layers to target and eliminate disturbed dormant seeds through the treatment of the agents (inhibiting seed germination and gradually weakening embryo viability through the colonization and metabolism of the strains, thus providing long-term control of dormant seeds in each layer). After the microbial agent control treatment, a gradient replacement of native herbaceous plants was carried out. After establishing native herbaceous plants, an annual closed-loop management system is set up to achieve radical control and long-term prevention and control of Tribulus terrestris.

[0011] This invention implements grazing ban and enclosure management in the project area, which can prevent livestock from entering and damaging the control results, and at the same time block the long-distance spread of Tribulus terrestris thorny husks through livestock carrying and trampling.

[0012] Furthermore, the stratified physical dormancy-breaking and seed-clearing operation is carried out 15-20 days before the spring germination of Tribulus terrestris.

[0013] Layered physical dormancy-breaking and seed-clearing operations are generally carried out from late March to early April, that is, 15 to 20 days after the soil thaws and before the germination of Tribulus terrestris, to ensure that soil disturbance and seed source clearing are completed before the large-scale germination of seeds.

[0014] Furthermore, the steps of the layered physical dormancy-breaking and seed-clearing operation include: surface seed source purification, shallow rotary tillage to break dormancy, and deep loosening and hole creation.

[0015] Optionally, the surface seed source purification step is as follows: the surface burr seeds are collected manually or mechanically and then subjected to high-temperature composting treatment.

[0016] The seeds of Tribulus terrestris seeds are harvested from the surface bracts using a mechanical scraper. The collected bracts are then subjected to high-temperature composting. In areas inaccessible by machinery, the bracts are collected manually. Burning the bracts is strictly prohibited to prevent secondary seed dispersal. The composted organic matter can be used as a natural substrate for subsequent cultivation of native herbs.

[0017] Optionally, the shallow rotary tillage and dormancy-breaking operation steps are as follows: use a rotary tiller to shallowly till 0-20cm of soil, break up the compacted layer of sandy soil, turn over the seeds of Tribulus terrestris buried in the 0-20cm shallow soil layer to the surface, collect the turned-over seeds, remove the mature plants and seedlings of Tribulus terrestris that have already emerged, and centrally treat them at high temperature for decomposition.

[0018] Optionally, the deep tillage and pore-forming operation steps are as follows: A deep tillage machine is used for staggered deep tillage, with the tillage depth controlled at 20-40 cm (preferably 35-40 cm). During the operation, the soil layer is not turned over; only the deep, compacted soil structure is loosened, forming longitudinally connected pores. This breaks the closed microenvironment of deep dormant seeds, creating penetration channels for simultaneous inoculum injection. The applied inoculum can penetrate along the pores to the seed layer, targeting deep dormant seeds and eliminating the long-term germination risk of deep seeds.

[0019] Furthermore, by weight, the compound biological agent is composed of the following components: 20-25 parts of Bacillus subtilis, 15-20 parts of Bacillus megaterium, 10-15 parts of Trichoderma, 10 parts of nitrogen-fixing rhizobium, and 30-45 parts of decomposed grassland substrate.

[0020] The functional divisions of each strain are as follows: Trichoderma can infect the seed coat of the bursa and gradually destroy the seed coat structure through parasitic decomposition, thereby reducing the stability of seed dormancy; Bacillus subtilis secretes metabolites after colonization, inhibiting enzyme activity during seed germination and blocking the seed germination process; Bacillus megaterium activates insoluble phosphorus in the soil and improves the rhizosphere nutrient environment; nitrogen-fixing rhizobia fix nitrogen in symbiosis with leguminous herbs, increasing soil nitrogen levels and strengthening the ecological occupation capacity of herbaceous communities.

[0021] The decomposed grassland substrate used in this invention can be the substrate obtained by high-temperature decomposition in the above-mentioned surface seed source purification step, or it can be a commercially available substrate.

[0022] The composite biological agent of this invention is prepared into granular agent by mixing the components evenly.

[0023] Furthermore, the application of the compound biological agent to the soil in layers includes: The process of spreading and then shallowly harrowing the fungus is adopted. 25-30 kg of compound biological fungicide is evenly applied per mu. The fungicide is thoroughly mixed with the soil and the turned-out thorny seeds by shallow harrowing with a depth of 3-5 cm. The deep loosening and inoculation process is adopted. Inoculation points are set up at horizontal intervals of 20-40cm along the horizontal strip of loosening strip formed by the deep loosening shovel. 30-50g of compound biological agent is applied to each point. The agent penetrates and diffuses downward along the pores of the deep loosening, covering the 20-40cm soil layer of the entire loose strip, and targets the deep dormant seeds. After the compound biological agent is applied, let it stand for 10 to 12 days to complete the activation of the strains and start the soil colonization process.

[0024] This invention combines physical stratification and disturbance with a stratified inoculum application process. After the compound biological agent is applied, the strains are activated and colonized in the soil by relying on the local natural temperature and humidity environment, continuously breaking down the seed dormancy structure and completely eliminating the remaining Tribulus terrestris seeds in the soil.

[0025] Furthermore, the steps for establishing a gradient replacement of native herbaceous plants include: By sowing herbaceous plant seeds according to the root depth gradient, a three-layered vegetation competition barrier of shallow, medium and deep roots is constructed to occupy the ecological niche.

[0026] The herbaceous plant seeds selected in this invention should be based on the local original vegetation and natural conditions, and should be selected as drought-resistant, easy to survive, fast-growing and sustainable perennial native grass species, that is, species contained in the local vegetation. Generally, a combination of legumes and grasses, and a combination of perennial and annual forage grasses are used.

[0027] Optionally, the native herbaceous plant seeds include shallow-rooted plant seeds, medium-rooted plant seeds, and deep-rooted plant seeds.

[0028] Preferably, the shallow-rooted plant seeds include annual foxtail grass; the medium-rooted plant seeds include Mongolian ice grass and / or sheep grass; and the deep-rooted plant seeds include alfalfa No. 3 and / or alfalfa.

[0029] Furthermore, the annual closed-loop management system includes monitoring and early warning, three-stage zeroing, pathway blocking, and biological rejuvenation.

[0030] This invention establishes a closed-loop management system covering the entire lifecycle of "monitoring and early warning → three-stage clearing → pathway blocking → biological rejuvenation" to completely block the accumulation and spread of Tribulus terrestris seed sources, prevent its resurgence, and ensure long-term and stable control effects.

[0031] Optionally, the monitoring and early warning (full-area grid-based monitoring and early warning) includes: dividing the project area into 100m×100m grid units, setting up fixed monitoring points in each grid, and implementing a zoned responsibility system by dedicated maintenance personnel, focusing on monitoring high-risk areas where Tribulus terrestris is prone to invasion and recurrence, such as sparse vegetation areas, soil disturbance areas, roadside areas and fence edges, to achieve early detection and early treatment.

[0032] Optionally, the three-stage zero-COVID policy (three-stage precise zero-COVID prevention and control throughout the year) includes: Once the seeds have entered the germination period, a three-stage eradication operation will be initiated (early to mid-April to early May, i.e., after the seeds have germinated and the seedlings have generally emerged): a comprehensive, grid-like inspection will be conducted once a week. Any scattered Tribulus terrestris seedlings found will be completely removed by manual uprooting and taken out of the project area for centralized high-temperature decomposition. On-site disposal is strictly prohibited (this stage is the critical period with the lowest cost and best results in prevention and control, ensuring that every single seedling is removed). Population interruption during peak growing season (June-July): Conduct a survey every two weeks, focusing on removing any adult plants that have slipped through the net. At the same time, investigate areas with less than 80% vegetation cover and promptly re-sow herbaceous seeds according to the original ratio to quickly form vegetation cover and squeeze the living space of Tribulus terrestris. Seed source elimination before fruiting (before early August): Conduct a comprehensive and in-depth investigation of the entire area to completely remove all Tribulus terrestris plants that have already flowered but have not yet fruited, and absolutely prohibit any plants from entering the fruiting period, thus cutting off the seed source supply for the next year from the source.

[0033] Optionally, the path blocking (transmission path blocking management) includes: regularly checking the integrity of the project area fence to prevent livestock from entering and spreading the disease by carrying thorny buds; promptly cleaning the thorny buds attached to the tires of vehicles and machinery entering the project area; and simultaneously clearing the long-thorny thorny grass within a 500-meter radius of the project area to establish an isolation buffer zone.

[0034] Optionally, the biological rejuvenation (annual soil microbiota and vegetation rejuvenation) includes: every spring (early April), for areas with sparse vegetation or bare patches, lightly harrowing the shallow soil to a depth of ≤3cm, and then applying 5-10kg of compound biological agent per acre to maintain the activity of beneficial microorganisms in the soil, continuously inhibit the germination of residual seeds, and top-dressing and reseeding the herbaceous communities with weakened growth to maintain community structure stability and competitive advantage.

[0035] This invention does not use any chemical herbicides or plant-derived herbicides. It achieves radical control of Tribulus terrestris through the coupled and coordinated use of five steps: grazing ban management, stratified physical and mechanical dormancy breaking and seed clearing, compound biological agent control, herbaceous gradient biological replacement, and annual closed-loop management.

[0036] The present invention discloses the following technical effects: This invention relies on physical and mechanical technology to break seed dormancy and clear the soil seed bank in layers. It combines compound biological agents to target and eliminate dormant seeds, and uses native gradient herbs to build an ecological competitive barrier. With annual dynamic closed-loop maintenance, it completely solves the problem of repeated germination and yearly outbreaks of Tribulus terrestris. It achieves simultaneous eradication of Tribulus terrestris and ecological restoration of sandy land, taking into account the needs of large-scale operation, zero ecological pollution, and long-term stable control.

[0037] This invention significantly improves the control effect, overcoming the core bottleneck of high recurrence rates in existing technologies. Targeting the biological characteristics of Tribulus terrestris's layered seed storage and dual seed dormancy, this invention achieves a technological leap from simply killing adult plants to eradicating the seed bank through a coupled process of physical dormancy disruption and biological control. The total inactivation rate of the soil seed bank reaches 92.7%, which is 2.97 times that of single physical control and 3.25 times that of single biological agent control. It achieves highly efficient control of seeds in soil layers 20-40cm deep, with an inactivation rate of 89.1%, while existing technologies have inactivation rates of less than 20% for deep seeds, completely solving the problem of recurring outbreaks caused by the annual germination of deep seeds. The annual recurrence rate of Tribulus terrestris is only 4.2%, a reduction of more than 93% compared to existing mainstream technologies. This breakthrough overcomes the industry dilemma of annual control and annual outbreaks.

[0038] This invention is entirely green and ecological, combining weed control and soil remediation functions. It does not use any chemical agents, fundamentally avoiding chemical residues. The alternative grass species are native dominant species found locally, with 100% native herbs, avoiding the risk of invasive alien species. In contrast, the hybrid Napier grass used in single ecological substitution technologies is an alien species, posing serious secondary ecological risks. The compound microbial agent and leguminous herbs work synergistically to significantly improve the fertility of sandy soil. At the end of the experiment, the soil organic matter content increased by up to 42.6% compared to the initial value, while the soil fertility of the control group showed no significant improvement.

[0039] The method of this invention has outstanding long-term effectiveness and can significantly reduce long-term control costs. The closed-loop control system constructed by this invention can achieve long-term stability with a single operation. After a complete process operation, the long-term control cycle is ≥5 years, without the need for large-scale tilling, pesticide application, or replanting every year. The cumulative control cost over 5 years is only 326 yuan / mu, which is 55.7% of the cost of single physical control, 50.8% of the cost of single biological agent control, and 68.2% of the cost of single ecological substitution, demonstrating significant long-term economic benefits.

[0040] This invention offers a highly adaptable method suitable for large-scale operations. The entire process can be completed using conventional agricultural machinery, eliminating the need for customized specialized equipment. Mechanized daily operations can cover 50-80 mu (approximately 3.3-4 hectares), significantly exceeding the 0.5-1 mu (approximately 0.06-0.067 hectares) / person / day achieved through manual control. It is well-suited for arid, infertile sandy soils in northern regions and can be widely applied to various invasive species such as natural grasslands, sandy wastelands, field edges, and roadside slopes. Attached Figure Description

[0041] The accompanying drawings, which form part of this invention, are used to provide a further understanding of the invention. The illustrative embodiments of the invention and their descriptions are used to explain the invention and do not constitute an undue limitation of the invention. In the drawings: Figure 1 This is a simplified flowchart illustrating the physical-biological coupled closed-loop method for the eradication and control of Tribulus terrestris according to the present invention.

[0042] Figure 2 This is a schematic diagram illustrating the application and action of the compound biological agent of the present invention. Detailed Implementation

[0043] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0044] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0045] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0046] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0047] The terms “include,” “including,” “have,” and “contain” used in this article are all open-ended terms, meaning that they include but are not limited to.

[0048] The Bacillus subtilis, Bacillus megaterium, Trichoderma, and nitrogen-fixing rhizobium used in the specific embodiments of this invention are all commercially available products, with a bacterial content of not less than 10%. 9 CFU / g.

[0049] Unless otherwise specified, all raw materials and reagents involved in the specific embodiments of this invention are commercially available products.

[0050] Unless otherwise specified, room temperature and normal temperature in the specific embodiments of this invention refer to 20-30°C.

[0051] It should be noted that any aspects not described in detail in this invention are conventional practices in the field and are not the focus of this invention.

[0052] Figure 1 This is a simplified flowchart illustrating the physical-biological coupled closed-loop radical control method for Tribulus terrestris of the present invention.

[0053] Figure 2 This is a schematic diagram illustrating the application and action of the compound biological agent of the present invention.

[0054] Example 1 A method for the radical control of Tribulus terrestris using a physical-biological coupled closed-loop system, comprising the following steps: S1. Grazing ban and enclosure management: Grazing bans are implemented in the project area to prevent livestock from entering and damaging the results of the remediation efforts, while also preventing the long-distance spread of Tribulus terrestris thorny husks through livestock carrying and trampling.

[0055] S2, Layered Physical Sleep Breaking and Seed Clearing Operation: The operation was carried out 20 days before the spring germination of *Tribulus terrestris* to ensure that soil disturbance and seed source clearing were completed before mass seed germination. A combined process of two-layer gradient disturbance and surface burr collection was used to specifically treat the seed bank in the 0-40cm layered soil. S2.1 Surface Seed Source Purification: The seeds of *Tribulus terrestris* are harvested from the surface using a mechanical scraper. The collected seeds are then subjected to high-temperature aerobic composting, with the pile temperature controlled at 55-65℃ for 25-30 days, during which the pile is turned over once a week. In areas inaccessible by machinery, the seeds are collected manually (burning is strictly prohibited to avoid secondary seed dispersal). The composted organic matter can be directly used as a carrier substrate for the compound biological agent (composted grassland soil substrate), or it can be retained as a natural soil amendment substrate for subsequent native herb cultivation.

[0056] S2.2 Shallow Rotary Tillage to Break Dormancy: Use a rotary tiller for shallow rotary tillage, controlling the tillage depth to 15-20cm, to break up the compacted layer of sandy soil, turn over the seeds of Tribulus terrestris buried in the soil at a depth of 0-20cm to the surface, breaking the physical dormancy barrier of the seed shells, collect the turned-up seeds after the operation, and remove the mature plants and seedlings of Tribulus terrestris that have already emerged, and collect them for high-temperature composting treatment.

[0057] S2.3 Deep Tillage and Hole Creation Operation: A deep tillage machine is used for staggered deep tillage, with the deep tillage depth controlled between 20 and 40 cm. During the operation, the soil layer is not turned over, but only the deep compacted soil structure is loosened to form continuous and interconnected pore channels. This breaks the closed microenvironment of deep dormant seeds, creates a penetration channel for simultaneous inoculation, and targets the deep dormant seeds to eliminate the long-term dormant germination risk of deep seeds.

[0058] S3. Application of compound biological agents: S3.1 Preparation of compound biological agent: According to the mass parts, 25 parts of Bacillus subtilis, 20 parts of Bacillus megaterium, 15 parts of Trichoderma, 10 parts of nitrogen-fixing rhizobium, and 30 parts of decomposed grassland substrate are mixed evenly to prepare granular agent.

[0059] S3.2. Based on the physical layered disturbance principle, a layered inoculum application process is adopted: For the shallow 0-20cm soil layer: the microbial agent is mixed by spreading and then shallow harrowing. 30kg of microbial agent is applied evenly per acre. The microbial agent is thoroughly mixed with the soil and the turned-out thorny seeds through shallow harrowing. The depth of shallow harrowing is 3-5cm.

[0060] For the deep 20-40cm soil layer: a deep loosening and inoculation process is adopted. In the soil pores formed by deep loosening, inoculation points are arranged at a horizontal interval of 20-30cm. 50g of granular inoculant is applied to each point to target and control deep dormant seeds.

[0061] After the compound biological agent is applied, it is left to stand for 12 days. Relying on the local natural temperature and humidity environment, the strains are activated and the soil colonization process is initiated. This process continuously breaks down the dormant structure of the seeds, effectively controlling the remaining Tribulus terrestris seeds in the soil for a long time.

[0062] S4, Gradient replacement of native herbaceous plants: S4.1 Selection of Native Grass Species: Based on the local original vegetation and natural conditions, drought-resistant, easy-to-grow, fast-growing, and sustainable perennial native grass species are selected, using a combination of legumes and grasses, and a mix of perennial and annual forage grasses. The herbaceous plants (seeds) used in this embodiment are: Alfalfa No. 3, Safflower, Mongolian Ice Grass, Sheepgrass, and annual foxtail grass.

[0063] Seed quality meets or exceeds the Class II standard stipulated by relevant national standards; when purchasing seeds, an inspection and quarantine certificate, a certificate of conformity, a business license, and a seed quality inspection report must be attached, along with the two certificates and a label; the random inspection pass rate is 100%, and the seeds comply with the provisions of GB6141 and GB6142, and the inspection methods comply with the requirements of GB / T2930.1-2017.

[0064] S4.2 Sowing method: Sowing is carried out using a no-till machine, following the requirements of NY / T1354-2007, completing the four processes of furrowing, sowing, covering with soil and compacting in one go.

[0065] Sowing method and gradient planting: Row sowing is used, with a sowing spacing of 30-40 cm. Gradual stratification planting is implemented based on the root distribution characteristics of different grass species. Shallow root system (annual foxtail grass): Sow at a depth of 0.5-1cm to quickly cover the ground as a pioneer grass.

[0066] Mid-layer root system (sheepgrass, Mongolian ice grass): Sowing depth 1-2cm, forming the main body of mid-layer vegetation.

[0067] Deep root system (Zhongcao No. 3 alfalfa, Sadawang): Sowing depth 2-3cm, long-term stable community structure.

[0068] Seeding rate: The total seeding rate is 4.5 kg / mu. See Table 1 for specific grass seeding rates.

[0069] Table 1 Sowing time: Choose the rainy season when the soil moisture is sufficient, generally from mid-May to early June, and complete all sowing tasks before mid-July.

[0070] Choose a slow-release compound fertilizer with a total nitrogen, phosphorus, and potassium content of ≥45% (N-P2O5-K2O ratio of 15-15-15), refer to GB / T15063-2020 standard, and apply it with no-till sowing of seeds at a rate of 20 kg / mu, with the fertilization time being the same as the sowing time.

[0071] Post-sowing management: Grazing is prohibited for 2 years after sowing, and no livestock are allowed to enter the project area to graze on pasture.

[0072] S5. Establish an annual closed-loop management system: Establish a full-cycle closed-loop management system of "monitoring and early warning → three-stage clearing → path blocking → biological rejuvenation" to completely block the accumulation and spread of Tribulus terrestris seed sources, prevent its resurgence, and ensure long-term and stable control effects.

[0073] S5.1 Comprehensive Grid-Based Monitoring and Early Warning: The project area is divided into 100m×100m grid units, with fixed monitoring points set up in each grid. Dedicated maintenance personnel are responsible for each area. Emphasis is placed on monitoring high-risk areas prone to invasion and recurrence of Tribulus terrestris, such as sparsely vegetated areas, areas with soil disturbance, and areas along roadsides and fence edges, to ensure early detection and timely intervention.

[0074] S5.2. Three-stage precise zero-COVID prevention and control throughout the year: S5.2.1. Spring Sprouting and Elimination (after seeds germinate and seedlings have generally emerged, early to mid-April to early May): Conduct a comprehensive, grid-like inspection once a week. Any scattered *Tribulus terrestris* seedlings found must be completely removed manually by uprooting them and taken out of the project area for centralized high-temperature decomposition. On-site disposal is strictly prohibited. This stage is the critical period with the lowest cost and best results in prevention and control; ensure that every single seedling is removed upon discovery.

[0075] S5.2.2. Population Interception During Peak Growing Season (June-July): Conduct a survey every two weeks, focusing on removing any remaining mature plants. At the same time, investigate areas with vegetation cover below 80% and promptly reseed native herbs according to the original ratio to quickly form vegetation cover and squeeze the living space of Tribulus terrestris.

[0076] S5.2.3. Zero seed source before fruiting (before early August): Conduct a thorough investigation of the entire area to completely remove all Tribulus terrestris plants that have already flowered but have not yet fruited, and absolutely prohibit any plant from entering the fruiting period, thus cutting off the seed source supply for the next year from the source.

[0077] S5.3. Transmission Path Blocking Management: Regularly inspect the integrity of the project area fence to prevent livestock from entering and spreading the thorny buds; promptly remove thorny buds attached to vehicle tires and machinery entering the project area; simultaneously clear long-thorny thorny grass within a 500-meter radius of the project area to establish an isolation buffer zone.

[0078] S5.4. Annual Soil Microbiota and Vegetation Rejuvenation: Every spring (early April), in bare-spot areas or sparsely vegetated areas, perform light harrowing of the shallow soil (harrowing depth ≤3cm), and apply 10kg of compound biological agent per acre to maintain the activity of beneficial microorganisms in the soil and continuously inhibit the germination of residual seeds. For herbaceous communities with weakened growth, apply appropriate topdressing and reseeding to maintain community structure stability and competitive advantage.

[0079] Comparative Example 1 Single physical prevention: S1. Grazing ban and enclosure management: Grazing bans are implemented in the project area to prevent livestock from entering and damaging the results of the remediation efforts, while also preventing the long-distance spread of Tribulus terrestris thorny husks through livestock carrying and trampling.

[0080] S2, the same as step S2 in Example 1.

[0081] Comparative Example 2 Control with a single biological agent: S1. Grazing ban and enclosure management: Grazing bans are implemented in the project area to prevent livestock from entering and damaging the results of the remediation efforts, while also preventing the long-distance spread of Tribulus terrestris thorny husks through livestock carrying and trampling.

[0082] S2, the same as step S3 in Example 1.

[0083] Comparative Example 3 Single-ecosystem alternatives: S1. Grazing ban and enclosure management: Grazing bans are implemented in the project area to prevent livestock from entering and damaging the results of the remediation efforts, while also preventing the long-distance spread of Tribulus terrestris thorny husks through livestock carrying and trampling.

[0084] S2. Use hybrid Napier grass for single broadcasting and planting, with a seeding rate of 2.0 kg / mu and a seeding depth of 1-2 cm. Other field management requirements are the same as step S4 in Example 1.

[0085] Comparative Example 4 Methods to prevent the absence of physical sleep-breaking and seed-clearing steps: S1. Grazing ban and enclosure management: Same as step S1 in Example 1.

[0086] S2. Application of compound biological agent: Same as step S3 in Example 1.

[0087] S3, Herbaceous gradient replacement planting: Same as step S4 in Example 1.

[0088] S4. Establish an annual closed-loop management system: Same as step S5 in Example 1.

[0089] Comparative Example 5 Prevention and control methods that lack the steps of compound biological agent control: S1. Grazing ban and enclosure management: Same as step S1 in Example 1.

[0090] S2, Layered physical sleep-breaking and seed clearing operation: Same as step S2 in Example 1.

[0091] S3, Herbaceous gradient replacement planting: Same as step S4 in Example 1.

[0092] S4. Establish an annual closed-loop management system: Same as step S5 in Example 1.

[0093] Comparative Example 6 Control methods for plants lacking native herbaceous gradient alternatives to establishment steps: S1. Grazing ban and enclosure management: Same as step S1 in Example 1.

[0094] S2, Layered physical sleep-breaking and seed clearing operation: Same as step S2 in Example 1.

[0095] S3. Application of compound biological agent: Same as step S3 in Example 1.

[0096] S4. Establish an annual closed-loop management system: Same as step S5 in Example 1.

[0097] Test case The methods of examples and comparative examples were used to study the severe invasion of Tribulus terrestris in the Horqin Sandy Land of Tongliao City, Inner Mongolia (invasion history ≥ 5 years, initial soil seed bank density ≥ 1200 seeds / m²). 2 A two-year comparative trial was conducted, with five parallel treatments, each covering an area of ​​10 mu (approximately 1.65 acres), and all initial conditions were identical. This included a blank control group (CK4).

[0098] By comparing the effects of single prevention and control technologies with the coupled scheme of this invention in Comparative Examples 1-3, and verifying the synergistic effect of each core step through the default test in Comparative Examples 4-6, the core indicators of different prevention and control technologies are compared in Table 2.

[0099] Table 2 As shown in Table 2, the core indicators of the single control schemes in Comparative Examples 1-3 were far worse than those of the complete coupled scheme of this invention. After Comparative Examples 4-6 were missing any core step, key indicators such as soil seed inactivation rate, weed recurrence rate, and vegetation coverage rate all showed a significant decline. This proves that the three core links of this invention—layered physical dormancy breaking and seed clearing, targeted control with compound biological agents, and gradient replacement of native herbs—have a significant synergistic effect. None of them can be omitted, and their joint coupling achieves the dual effect of controlling Tribulus terrestris and restoring sandy land ecology.

[0100] The various embodiments in this specification are described in a progressive manner. Each embodiment focuses on the differences from other embodiments. The same or similar parts between the various embodiments can be referred to each other.

[0101] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A method for the physical-biological coupled closed-loop radical control of Tribulus terrestris, characterized in that the steps include... include: Grazing ban and enclosure management will be implemented in the project area; Before the spring germination of Tribulus terrestris, a layered physical dormancy-breaking and seed-clearing operation was carried out on the soil seed bank in the project area; Subsequently, a compound biological agent was applied to the soil in layers to target and eliminate disturbed dormant seeds. After the microbial agent control treatment, a gradient replacement of native herbaceous plants was carried out. After establishing native herbaceous plants, an annual closed-loop management system is set up to achieve radical control and long-term prevention and control of Tribulus terrestris.

2. The method as described in claim 1, characterized in that, The stratified physical dormancy-breaking and seed-clearing operation is carried out 15-20 days before the spring germination of Tribulus terrestris. And / or, the steps of the layered physical dormancy-breaking and seed-clearing operation include: surface seed source purification, shallow rotary tillage to break dormancy, and deep loosening and hole creation.

3. The method as described in claim 2, characterized in that, The surface seed source purification step is as follows: the surface prickly bud seeds are collected by manual or mechanical scraping and then subjected to high-temperature composting treatment. And / or, the operation steps of shallow rotary tillage to break dormancy are as follows: use a rotary tiller to shallowly till 0-20cm of soil, break up the compacted layer of sandy soil, turn over the seeds of Tribulus terrestris buried in the 0-20cm shallow soil layer to the surface, break the physical dormancy barrier of the seed shell, pick up and collect the turned-out burrs after the operation, and at the same time remove the mature plants and seedlings of Tribulus terrestris that have emerged, and carry out centralized high-temperature decomposition treatment; And / or, the deep tillage and hole-making operation steps are as follows: a deep tillage machine is used for staggered deep tillage, the deep tillage depth is controlled at 20-40cm, the soil layer is not turned over during the operation, only the deep compacted soil structure is loosened to form longitudinally connected pores, breaking the closed microenvironment of deep dormant seeds, and creating a permeation channel for synchronous inoculation.

4. The method as described in claim 1, characterized in that, The compound biological agent, by weight, consists of the following components: 20-25 parts of Bacillus subtilis, 15-20 parts of Bacillus megaterium, 10-15 parts of Trichoderma, 10 parts of nitrogen-fixing rhizobium, and 30-45 parts of decomposed grassland substrate.

5. The method as described in claim 1, characterized in that, The application of compound biological agents to soil stratification includes: The process of spreading and then shallowly harrowing to mix the bacteria is adopted. 25-30 kg of compound biological agent is evenly applied per mu. The agent is thoroughly mixed with the soil and the turned-out thorny seeds by shallow harrowing with a depth of 3-5 cm. The deep loosening and inoculation process is adopted. Along the horizontal strip of loosening strip formed by the deep loosening shovel, inoculation points are set at a horizontal interval of 20-40cm, and 30-50g of compound biological agent is applied to each point. After the compound biological agent is applied, let it stand for 10-12 days to complete the activation of the strains and start the soil colonization process.

6. The method as described in claim 1, characterized in that, The steps for establishing a gradient replacement of native herbaceous plants include: By sowing herbaceous plant seeds according to the root depth gradient, a three-layered vegetation competition barrier of shallow, medium and deep roots is constructed to occupy the ecological niche.

7. The method as described in claim 6, characterized in that, The native herbaceous plant seeds include shallow-rooted plant seeds, medium-rooted plant seeds, and deep-rooted plant seeds.

8. The method as described in claim 7, characterized in that, The seeds of the shallow-rooted plants include annual foxtail grass; And / or, the seeds of the mid-rooted plants include Mongolian ice grass and / or sheep grass; And / or, the deep-rooted plant seeds include Alfalfa No. 3 and / or Safflower.

9. The method as described in claim 1, characterized in that, The annual closed-loop management system includes monitoring and early warning, three-stage zeroing, pathway blocking, and biological rejuvenation.

10. The method as described in claim 9, characterized in that, The monitoring and early warning system includes: dividing the project area into 100m×100m grid units, setting up fixed monitoring points in each grid, focusing on monitoring high-risk areas where Tribulus terrestris is prone to invasion and recurrence, such as sparse vegetation areas, soil disturbance areas, roadside areas and fence edges, so as to achieve early detection and early treatment. And / or, the path blocking includes: regularly checking the integrity of the project area fence to prevent livestock from entering and spreading the thorny buds; promptly cleaning the thorny buds attached to the tires of vehicles and machinery entering the project area; and simultaneously clearing the long-spined thorny grass within a 500-meter radius of the project area to establish an isolation buffer zone. And / or, the biological rejuvenation includes: every spring, for sparsely vegetated or bare areas, lightly harrowing the shallow soil to a depth of ≤3cm, and after harrowing, applying 5-10kg of compound biological agent per acre to maintain the activity of beneficial bacteria in the soil, continuously inhibit the germination of residual seeds, and topdressing and reseeding herbaceous communities with weakened growth to maintain community structure stability and competitive advantage; And / or, the three-stage zeroing includes: After the seeds enter the germination period, a three-stage clearing operation will be launched: from early to mid-April to early May, a comprehensive screening will be carried out once a week. Any scattered Tribulus terrestris seedlings found will be completely removed by manual uprooting and taken out of the project area for centralized high-temperature decomposition. Population interruption during peak growing season: From June to July, conduct a survey every two weeks, focusing on removing any adult plants that have slipped through the net. At the same time, investigate areas with less than 80% vegetation cover and resow herbaceous seeds to form vegetation cover and squeeze the living space of Tribulus terrestris. Zero seed source before fruiting: Before early August, conduct a comprehensive and in-depth investigation of the entire area to completely remove all Tribulus terrestris plants that have already flowered but have not yet fruited, thus cutting off the seed source supply for the next year from the source.

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