Toxin-control nitrogen-fixing coupling microbial agent and use thereof

Abstract

The present application relates to the field of microorganisms, and in particular to a toxin-control nitrogen-fixation coupling microbial agent and a use thereof. The toxin-control nitrogen-fixation coupling microbial agent is a microbial composition, has a toxin-control nitrogen-fixation coupling function, has the functions of regulating and improving the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the nodule number of leguminous crops, and contains all gene sequences or four or more gene sequences among nucleotide sequences as shown in SEQ ID NOs. 1-12. The toxin-control nitrogen-fixation coupling microbial agent has the toxin-control nitrogen-fixation coupling function, is applied to crop production, realizes the coupling of toxin control and nitrogen fixation, quality improvement and yield increase, involves simple use, has significant social and ecological benefits, and is easy to popularize and apply.

Description

Poison control and nitrogen fixation coupled microbial agent and its use Technical Field

[0001] The present invention belongs to the field of microorganisms, and in particular relates to a toxic-controlling and nitrogen-fixing coupled microbial agent and its use. Background Art

[0002] Oilseeds are important sources of fat and plant protein, two of the three major nutrients for the human body. They play a crucial role in ensuring national grain and oil security, maintaining people's nutritional health, and promoting rural revitalization. Soybeans and peanuts are important grain crops, oil crops, and feed crops in my country, and are fundamental, critical, and strategic industries that are crucial to the national economy and people's livelihood.

[0003] The soybean and peanut industries face two common challenges: First, peanuts and soybeans are susceptible to contamination with highly toxic and carcinogenic aflatoxins, which not only reduce quality and reduce production, but also pose a serious threat to people's health and lives. Aflatoxin B1, for example, is 10 times more toxic than potassium cyanide and is classified as a Class I carcinogen by the WHO's International Agency for Research on Cancer, causing 28.2% of liver cancer worldwide. In recent years, aflatoxin contamination in peanuts has generally increased and has become a major risk factor for the peanut and other industries. Existing methods for controlling aflatoxin in peanuts and soybeans primarily rely on temperature and humidity control during storage, transportation, and processing, resulting in high energy consumption and difficult control. Aflatoxin contamination prevention and control remains a global challenge.

[0004] Secondly, legume crops like soybeans and peanuts, while nodulating and fixing nitrogen symbiotically with soil rhizobia, naturally have few nodules, a short nitrogen fixation period (it's generally believed that no nodules or nitrogen fixation occur in the first month after sowing, and no new nodules form during the pod-filling or fruit-full stages, while existing nodules begin to wither), resulting in low efficiency. Research on biological nitrogen fixation using rhizobia has a history of over 100 years, establishing the classically recognized AON theory—that plants self-regulate nodule number and growth while maintaining total energy conservation. Excessive nodulation inevitably comes at the expense of plant growth. Current approaches primarily rely on selecting and applying optimized rhizobia adapted to specific production environments. This approach is geographically limited and constrained by the AON theory, resulting in limited improvements in nodulation and nitrogen fixation efficiency, typically around 30%. Achieving a doubling of nodulation and nitrogen fixation efficiency while also significantly increasing growth (a challenge to the AON theory) is difficult. Improving the nodulation and nitrogen fixation efficiency of legumes like peanuts and soybeans remains a hotly debated and challenging issue internationally.

[0005] To address these challenges, the inventors' team has conducted over 20 years of continuous research, uncovering the spatiotemporal correlation between the abundance and toxicity of aflatoxin-producing fungi, geography, climate, and 53 other factors, and the occurrence of aflatoxin. They have developed highly sensitive detection and early warning technologies, identified the source of aflatoxin contamination in the soil, constructed a library of aflatoxin-producing strains, and for the first time proposed the scientific concept of coupling soil-source control of aflatoxin with nodulation and nitrogen fixation. By analyzing the microbial population structure within the peanut rhizosphere, isolating and identifying the strains and constructing a library of probiotic strains, and assembling a large number of combinations, they completed laboratory and field screening and identification, thus opening up research and exploration into the coupling of soil-source control of aflatoxin and induced nodulation and nitrogen fixation. Over the past five years, a microbial agent for toxicity control and nitrogen fixation has been successfully developed. This agent effectively controls aflatoxin at the source while simultaneously inducing efficient nodulation and nitrogen fixation in soybeans and peanuts, significantly increasing yields. This agent is easy to use, low-cost, and highly effective. It boasts significant advantages: two fixes (nitrogen and carbon fixation), three increases (increased yield, efficiency, and safety), and five reductions (reduced toxicity, damage, weight, cost, and carbon emissions). This agent has enormous potential for application and has been demonstrated and validated in field trials across major soybean, peanut, and pea production areas across China. It holds significant significance for boosting soybean oilseed production capacity and promoting green, low-carbon, and efficient production in my country. Summary of the Invention

[0006] In response to the deficiencies in the prior art, the present invention provides a coupled microbial agent for controlling toxicity and fixing nitrogen and its use. The agent has a coupled effect of controlling toxicity and fixing nitrogen, and is applied to crop production to achieve the coupling of controlling toxicity and fixing nitrogen, improving quality and increasing yield. The agent is simple to use, has significant social and ecological benefits, and is easy to promote and apply.

[0007] In order to solve the above technical problems, the technical solution adopted by the present invention is:

[0008] The first aspect of the present invention provides a microbial agent coupled with toxicity control and nitrogen fixation, which is a microbial composition having a coupled effect of toxicity control and nitrogen fixation, and has the function of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules in leguminous crops, and contains all the gene sequences in the nucleotide sequences shown in SEQ ID NO.1~12 or 4 or more of them.

[0009] According to the above scheme, the toxic control and nitrogen fixation coupled microbial agent is preferably a composition of more than three kinds of microorganisms, containing 4 or 5 or 6 or 7 or 8 or 9 or more gene sequences of the nucleotide sequences shown in SEQ ID NO.1~12.

[0010] According to the above scheme, the microbial source includes but is not limited to Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus licheniformis, Brevibacillus laterosporus, Bacillus mucilaginosus, Bacillus velezensis, Bacillus siamensis, Paenibacillus polymyxa, Paenibacillus timonensis, Pseudomonas fluorescens, Pseudomonas mendocina, Enterobacter ludwigii, Microbacterium proteolyticum, Leclercia adcarboxglata, Serratia marcescens. marcescens, Empedobacters sp., Priestia priestiamegaterium, Stenotrophomonas maltophilia and other bacteria.

[0011] According to the above scheme, the toxic control and nitrogen-fixing coupled microbial agent has an inhibitory effect on Aspergillus flavus and / or toxins. Furthermore, the inhibition rate of Aspergillus flavus reaches more than 60%, and the inhibition rate of aflatoxin reaches more than 80%. The inhibition of Aspergillus flavus and / or toxins can be detected using other conventional detection methods in the prior art, or the inhibition analysis of Aspergillus flavus and / or its toxins can be performed by inhibition analysis of the marker molecules of Aspergillus flavus toxin-producing fungi. Specifically, the inhibition analysis of Aspergillus flavus and / or its toxins by the toxic control and nitrogen-fixing coupled microbial agent can be performed by inhibition analysis of the marker molecules of Aspergillus flavus toxin-producing fungi.

[0012] According to the above scheme, the toxin-control and nitrogen-fixing coupled microbial agent of the present invention significantly inhibits the expression of the Aspergillus flavus PAB-01 protein. The amino acid sequence of the PAB-01 protein is shown in SEQ ID No. 12. The inhibition rate is greater than 90%, preferably greater than 95%, reflecting the excellent bacteriostatic and toxicity-reducing effects of the toxin-control and nitrogen-fixing coupled microbial agent of the present invention. The method for determining the inhibition rate of the expression of the Aspergillus flavus PAB-01 protein can refer to the method of the following document, Protein biomarker for early diagnosis of microbial toxin contamination: Using Aspergillus flavus as an example, Food Frontiers. 2023, 4, 2013-2023, DOI: 10.1002 / fft2.295.

[0013] According to the above scheme, the toxicity control and nitrogen fixation coupled microbial agent has an inhibitory effect on one or more pathogenic factors / toxins of soil-borne plant pathogens such as Penicillium and Aspergillus other than Aflatoxin, Fusarium, Sclerotium sclerotiorum, Pseudomonas solanacearum, Phytophthora, Sclerotinia sclerotiorum, Pythium, and Rhizoctonia solani. The inhibition rate is greater than 30%.

[0014] According to the above scheme, the gene sequences shown in SEQ ID NO. 1 to 12 may vary to a certain extent in different strains. When the degree of variation is small, such as no more than 10% base variation, preferably no more than 5% base variation, and more preferably no more than 1% base variation, that is, the identity is more than 90%, preferably more than 95%, and more preferably more than 99%, and when they have corresponding biological activity functions, these are called functional equivalents of the DNA sequences shown in SEQ ID NO. 1 to 12, and containing these sequences is equivalent to containing the corresponding sequences of DNA sequences 1 to 12. Microbial compositions containing all the gene sequences in the nucleotide sequences shown in SEQ ID NO. 1 to 12, or 4 or more of them, or functional equivalents of these gene sequences, and having the coupled effects of controlling and fixing toxicity and nitrogen as described above by the microbial agents for controlling and fixing toxicity, regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules in leguminous crops are all coupled microbial agents for controlling and fixing toxicity of the present invention.

[0015] According to the above scheme, the toxicity control and nitrogen fixation coupled microbial agent can promote early nodulation of leguminous crops such as peanuts and soybeans and prolong the nodulation and nitrogen fixation time.

[0016] The above-mentioned DNA sequences 1 to 12 are specific sequences obtained after comparison with Genbank genome big data. They have a direct or indirect correlation with the function of the microbial agent of this patent. When containing 4 or more of the above-mentioned DNA sequences 1 to 12, they have the effects of controlling toxicity and fixing nitrogen, improving quality and increasing yield. They are the toxicity-controlling and nitrogen-fixing coupled microbial agent of the present invention. Furthermore, they can also promote early nodulation of leguminous crops and prolong the nodulation and nitrogen fixation time. These genes may vary to a certain extent in different strains. When the degree of difference is small, such as no more than 10% base variation, and the corresponding biological activity function is maintained, containing them is equivalent to containing all the gene sequences in the DNA sequences 1 to 12 shown in SEQ ID NO. 1-12 or 4 or more of them.

[0017] The toxic control and nitrogen fixation coupled microbial agent provided by the present invention can be, but is not limited to, a combination of three or more of the following microorganisms: Bacillus laterosporus with a deposit number of CCTCC NO: M 20231807, Bacillus timonellia with a deposit number of CCTCC NO: M 20231809, Bacillus subtilis with a deposit number of CCTCC NO: M 20231811, Stenotrophomonas maltophilia with a deposit number of CCTCC NO: M 20231813, Bacillus amyloliquefaciens with a deposit number of CCTCC NO: M 20231816, Bacillus mucilaginosus with a deposit number of CCTCC NO: M 20231817, and Enterobacter ludwigii BG10-1 with a deposit number of CCTCC NO: M 2016014. Specifically, it can be a combination of three, four, five, six, or seven of the above strains.

[0018] Or it can be a combination of one or more of the above-mentioned microorganisms and other microorganisms, so that the combined microbial agent meets the requirements of containing all the gene sequences in the nucleotide sequences shown in SEQ ID NO.1-12 or 4 or more of the gene sequences or their functional equivalents, has a coupled effect of toxicity control and nitrogen fixation, has the effect of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules of leguminous crops, and constitutes the toxicity control and nitrogen fixation coupled microbial agent of the present invention.

[0019] The toxic control and nitrogen fixation coupled microbial agent provided by the present invention can also be a combination of one or more of Bacillus laterosporus of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, Enterobacter ludwigii BG10-1 of CCTCC NO: M 2016014 and the following four strains: Bacillus timundi of CCTCC NO: M 20231809, Bacillus subtilis of CCTCC NO: M 20231811, Stenotrophomonas maltophilia of CCTCC NO: M 20231813, and Bacillus mucilaginosus of CCTCC NO: M 20231817, such as Bacillus laterosporus CB2013 of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, and Bacillus ludwigii BG10-1 of CCTCC NO: M 2016014. a combination of Enterobacter Ludwigii BG10-1 of CCTCC NO: M 20231809 and Bacillus timundi of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, Enterobacter Ludwigii BG10-1 of CCTCC NO: M 2016014 and Bacillus subtilis of CCTCC NO: M 20231811, a combination of Bacillus laterosporus of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, Enterobacter Ludwigii BG10-1 of CCTCC NO: M 2016014 and Stenotrophomonas maltophilia of CCTCC NO: M 20231813, or a combination of Bacillus laterosporus of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, Enterobacter Ludwigii BG10-1 of CCTCC NO: M 2016014 and Stenotrophomonas maltophilia of CCTCC NO: M 20231813, or a combination of Bacillus laterosporus of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816 The combination of Bacillus laterosporus of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, Enterobacter Ludwigii BG10-1 of CCTCC NO: M 2016014, and Bacillus mucilaginosus of CCTCC NO: M 20231817, so that the combined microbial agent satisfies the requirements of containing all gene sequences in the nucleotide sequences shown in SEQ ID NOs. 1 to 12 or four or more gene sequences therein or their functional equivalents, has a coupled effect of toxicity control and nitrogen fixation, has the effect of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops, and increasing the number of nodules in leguminous crops, thereby constituting the toxicity control and nitrogen fixation coupled microbial agent of the present invention.

[0020] The toxic control and nitrogen fixation coupled microbial agent provided by the present invention can also be a combination of Bacillus laterosporus of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, Enterobacter Ludwigii BG10-1 of CCTCC NO: M 2016014 and other bacteria, so that the combined microbial agent meets the requirements of containing all gene sequences in the nucleotide sequences shown in SEQ ID NOs. 1 to 12 or 4 or more gene sequences therein or their functional equivalents, has a toxic control and nitrogen fixation coupled effect, has the effect of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules of leguminous crops, and constitutes the toxic control and nitrogen fixation coupled microbial agent of the present invention.

[0021] Bacillus laterosporus, deposited on September 27, 2023, with a deposit number of CCTCC NO: M 20231807, and a classification name of Brevibacillus laterosporus strain CB2013. The depository is China Center for Type Culture Collection, located at Wuhan University, Wuhan, China.

[0022] Bacillus timonersis, deposited on September 27, 2023, with a deposit number of CCTCC NO: M20231809, a classification name of Paenibacillus timonersisstrain D-DMYB0815, and a depository institution of China Center for Type Culture Collection, located at Wuhan University, Wuhan, China.

[0023] Bacillus subtilis, deposit date is September 27, 2023, deposit number is CCTCC NO: M20231811, classification name is: Bacillus subtilisstrain D-KCY19201, depository name is China Center for Type Culture Collection, address is Wuhan University, Wuhan, China.

[0024] Stenotrophomonas maltophilia, deposit date is September 27, 2023, deposit number is CCTCC NO: M20231813, classification name is: Stenotrophomonas maltophiliastrain D-SMYDB0805, deposit unit name is China Center for Type Culture Collection, address is Wuhan University, Wuhan, China.

[0025] Bacillus amyloliquefaciens, deposited on September 27, 2023, with a deposit number of CCTCC NO: M20231816, a classification name of Bacillus amyloliquefaciens strain JDF2013, and a depository unit named China Center for Type Culture Collection, located at Wuhan University, Wuhan, China.

[0026] Bacillus mucilaginosus, deposited on September 27, 2023, with the deposit number CCTCC NO: M 20231817, the classification name: Bacillus mucilaginosus strain JZ2013, the depository is China Center for Type Culture Collection, and the address is Wuhan University, Wuhan, China.

[0027] Enterobacter Ludwigii BG10-1, with the deposit number CCTCC NO: M2016014, was deposited with the China Center for Type Culture Collection (CCTCC) on January 7, 2016, with the deposit number CCTCC NO: M 2016014, and the classification name: Enterobacter LudwiggiBG10-1. For details, see patent CN 105586300 A.

[0028] According to the above scheme, the proportion of the number of live bacteria of any one strain of the above-mentioned poison-control and nitrogen-fixing coupled microbial agent in the microbial agent, that is, the mixed microbial composition, is greater than or equal to 1%.

[0029] The toxicity-controlling and nitrogen-fixing coupled microbial agent provided by the present invention is a microbial composition. The toxicity-controlling and nitrogen-fixing coupled effect is generated through the synergistic action of the various microorganisms in the microbial composition, exerting the coupled effect of toxicity control and nitrogen fixation. When used in crop production, it has a toxicity-controlling and nitrogen-fixing coupled effect, and has the functions of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules in leguminous crops. Although it is not a rhizobium, it can simultaneously regulate and increase the abundance of rhizobia in the rhizosphere of leguminous crops, increase the number of nodules in leguminous crops, and improve the nitrogenase activity of a single plant. The toxicity-controlling and nitrogen-fixing microbial agent of the present invention can increase the abundance of rhizobia in the rhizosphere of leguminous crops by more than 10% and increase the number of nodules per leguminous crop plant by more than 2 times. It can promote early nodulation of leguminous crops such as peanuts and soybeans and prolong the nodulation and nitrogen fixation time.

[0030] The above-mentioned toxicity-controlling and nitrogen-fixing coupled microbial agent can be prepared by the following method: the microorganisms in the above-mentioned microbial agent are combined and fermented. The above-mentioned fermentation route can adopt conventional fermentation routes of bacteria or fungi disclosed in existing technologies, including existing literature.

[0031] The second aspect of the present invention provides any of the following uses of the above-mentioned toxic control and nitrogen fixation coupled microbial agent in crop production: for improving the quality and safety level of leguminous crop products; for promoting nodulation and nitrogen fixation in leguminous crops; for improving the yield per unit area of ​​leguminous crops; for recruiting indigenous rhizobia and increasing the abundance of rhizobia in the rhizosphere soil of leguminous crops; for promoting early nodulation of leguminous crops and prolonging the nodulation and nitrogen fixation time; for preventing leguminous crops from losing fertilizer and aging prematurely during maturity; for increasing the number of leguminous crop pods; for increasing the fullness of leguminous crop pods and reducing the rate of shrunken pods; for promoting early flowering and early pod setting of leguminous crops; for reducing the occurrence of peanut fruit rot; for reducing the occurrence of bacterial wilt in leguminous crops; for reducing the occurrence of powdery mildew in leguminous crops; for reducing the occurrence of leaf spot in leguminous crops; for reducing It is used to reduce the occurrence of root nematode diseases in leguminous crops; to reduce the occurrence of root rot in leguminous crops; to reduce the occurrence of blight in leguminous crops; to reduce the occurrence of sclerotinia in leguminous crops; to reduce the occurrence of downy mildew in leguminous crops; to reduce the occurrence of wilt in leguminous crops; to reduce the occurrence of white rot in leguminous crops; to reduce the incidence of soybean green spondylosis; to reduce the incidence of corn ear rot and Fusarium toxin; to reduce the abundance of wheat fusarium and reduce the incidence of fusarium toxin; to promote carbon emission reduction in leguminous crops, which is beneficial to soil improvement; to promote the increase of total biomass of leguminous crops; to reduce the abundance of pests such as Aspergillus terreus and Fusarium in the rhizosphere of leguminous crops, which is beneficial to improving the soil microbial population structure; to reduce the surface spots of peanuts and increase commercial value; to promote soybean production in saline-alkali land.

[0032] The third aspect of the present invention provides the use of the above-mentioned poison-controlling and nitrogen-fixing coupled microbial agent in the preparation of poison-controlling and nitrogen-fixing coupled microbial compound fertilizer, which is used to play the role of the quality-improving and nitrogen-fixing coupled microbial agent in production; or the use of the above-mentioned poison-controlling and nitrogen-fixing coupled microbial organic fertilizer, which is used to play the role of the quality-improving and nitrogen-fixing coupled microbial agent in production; or the use of the above-mentioned poison-controlling and nitrogen-fixing coupled microbial fertilizer, which is used to play the role of the quality-improving and nitrogen-fixing coupled microbial agent in production; or the use of the above-mentioned poison-controlling and nitrogen-fixing coupled microbial moisturizing fertilizer, which is used to play the role of the quality-improving and nitrogen-fixing coupled microbial agent in production; or the use of the above-mentioned poison-controlling and nitrogen-fixing coupled microbial inorganic-organic compound fertilizer, which is used to play the role of the quality-improving and nitrogen-fixing coupled microbial agent in production; or the use of the above-mentioned poison-controlling and nitrogen-fixing coupled microbial-rhizobium fertilizer, which is used to play the role of the quality-improving and nitrogen-fixing coupled microbial agent in production; or the use of the above-mentioned poison-controlling and nitrogen-fixing coupled microbial-seed dressing agent / seed dressing agent / seed soaking agent, which is used to play the role of the quality-improving and nitrogen-fixing coupled microbial agent in production.

[0033] The present invention further provides a poison-controlling nitrogen-fixing coupled microbial compound fertilizer, a poison-controlling nitrogen-fixing coupled microbial organic fertilizer, a poison-controlling nitrogen-fixing coupled microbial micro-fertilizer, a poison-controlling nitrogen-fixing coupled microbial moisturizing fertilizer, a poison-controlling nitrogen-fixing coupled microbial inorganic-organic compound fertilizer, a poison-controlling nitrogen-fixing coupled microbial-rhizobium fertilizer, or a poison-controlling nitrogen-fixing coupled microbial-seed dressing agent / seed dressing agent / seed soaking agent obtained by configuring the above-mentioned poison-controlling nitrogen-fixing coupled microbial agent.

[0034] A fourth aspect of the present invention provides a method for preparing a poison-controlling nitrogen-fixing coupled microbial compound fertilizer, a poison-controlling nitrogen-fixing coupled microbial organic fertilizer, a poison-controlling nitrogen-fixing coupled microbial micro-fertilizer, a poison-controlling nitrogen-fixing coupled microbial moisturizing fertilizer, a poison-controlling nitrogen-fixing coupled microbial inorganic-organic compound fertilizer, a poison-controlling nitrogen-fixing coupled microbial-rhizobium fertilizer, or a poison-controlling nitrogen-fixing coupled microbial-seed dressing agent / seed dressing agent / seed soaking agent using the above-mentioned poison-controlling nitrogen-fixing coupled microbial agent:

[0035] Method for preparing a toxic-control and nitrogen-fixing coupled microbial compound fertilizer: according to the conventional dosage of compound fertilizer and the dosage of the toxic-control and nitrogen-fixing coupled microbial agent with a viable count of not less than 80 billion per mu of land, the compound fertilizer and the toxic-control and nitrogen-fixing coupled microbial agent are mixed in proportion, and then the toxic-control and nitrogen-fixing coupled microorganisms are adsorbed and fixed onto the compound fertilizer particles by physical methods to prepare the toxic-control and nitrogen-fixing coupled microbial compound fertilizer;

[0036] Method for preparing toxic-control and nitrogen-fixing coupled microbial organic fertilizer: according to the conventional amount of organic fertilizer per mu of land and the amount of toxic-control and nitrogen-fixing coupled microbial agent with a viable count of not less than 80 billion, the organic fertilizer and the toxic-control and nitrogen-fixing coupled microbial agent are mixed in proportion, and then the toxic-control and nitrogen-fixing coupled microorganisms are adsorbed and fixed on the organic fertilizer particles by physical methods to prepare the toxic-control and nitrogen-fixing coupled microbial compound fertilizer;

[0037] Method for preparing microbial fertilizers for toxin-control and nitrogen-fixing coupling: according to the conventional dosage of trace element fertilizers per mu of land and the dosage of microbial agent for toxin-control and nitrogen-fixing coupling with no less than 80 billion viable bacteria, the trace element fertilizers and the microbial agent for toxin-control and nitrogen-fixing coupling are proportioned, and then mixed by physical methods to adsorb and fix the microorganisms for toxin-control and nitrogen-fixing coupling onto the trace element fertilizer particles to prepare the microbial fertilizer for toxin-control and nitrogen-fixing coupling;

[0038] Method for preparing a poison-controlling and nitrogen-fixing coupled microbial moisturizing fertilizer: according to the conventional dosage of water-retaining agent and the dosage of the poison-controlling and nitrogen-fixing coupled microbial agent with a viable count of not less than 80 billion per mu of land, the trace element fertilizer and the poison-controlling and nitrogen-fixing coupled microbial agent are mixed in proportion, and then the poison-controlling and nitrogen-fixing coupled microbial agent is mixed by conventional physical methods / or the poison-controlling and nitrogen-fixing coupled microbial agent is adsorbed and fixed on the water-retaining agent to prepare the poison-controlling and nitrogen-fixing coupled microbial moisturizing fertilizer;

[0039] A method for preparing a toxic-control and nitrogen-fixing coupled microbial rhizobium fertilizer: according to the conventional amount of rhizobium fertilizer and the amount of the toxic-control and nitrogen-fixing coupled microbial agent with a viable count of not less than 80 billion per mu of land, the rhizobium fertilizer and the toxic-control and nitrogen-fixing coupled microbial agent are proportioned, and then mixed by conventional physical methods, or the two agents are mixed and then fixed on carrier particles to prepare the toxic-control and nitrogen-fixing coupled microbial rhizobium fertilizer;

[0040] Method for preparing toxic-control and nitrogen-fixing coupled microbial organic-inorganic compound fertilizer: according to the conventional dosage of compound fertilizer per mu of land and the dosage of toxic-control and nitrogen-fixing coupled microbial agent with a viable count of not less than 80 billion, the organic-inorganic compound fertilizer and the toxic-control and nitrogen-fixing coupled microbial agent are mixed, and then the toxic-control and nitrogen-fixing coupled microorganisms are adsorbed and fixed on the organic and inorganic particles by physical methods to prepare the toxic-control and nitrogen-fixing coupled microbial organic-inorganic compound fertilizer;

[0041] The method for preparing quality-improving and nitrogen-fixing coupled microorganisms-seed dressing agent / seed mixing agent / seed soaking agent: according to the amount of seed dressing agent or seed mixing agent or seed soaking agent per mu of land and the amount of quality-improving and nitrogen-fixing coupled microbial agent of not less than 80 billion viable bacteria, the seed dressing agent or seed mixing agent or seed soaking agent and the toxicity-controlling and nitrogen-fixing coupled microbial agent are proportioned, and then the toxicity-controlling and nitrogen-fixing coupled microbial agent and the seed dressing agent or seed mixing agent or seed soaking agent are evenly mixed through conventional physical mixing to prepare the toxicity-controlling and nitrogen-fixing coupled microorganisms-seed dressing agent / seed mixing agent / seed soaking agent.

[0042] The fifth aspect of the present invention provides any of the following applications in crop production of a poison-controlling nitrogen-fixing coupled microbial compound fertilizer, a poison-controlling nitrogen-fixing coupled microbial organic fertilizer, a poison-controlling nitrogen-fixing coupled microbial micro-fertilizer, a poison-controlling nitrogen-fixing coupled microbial moisturizing fertilizer, a poison-controlling nitrogen-fixing coupled microbial inorganic-organic compound fertilizer, a poison-controlling nitrogen-fixing coupled microbial-rhizobium fertilizer, or a poison-controlling nitrogen-fixing coupled microbial-seed dressing agent / seed dressing agent / seed soaking agent configured using the above-mentioned poison-controlling nitrogen-fixing coupled microbial agent: for improving the quality and safety level of leguminous crop products; for promoting nodulation and nitrogen fixation of leguminous crops; for improving the yield per unit area of ​​leguminous crops; for recruiting indigenous rhizobia and increasing the abundance of rhizobia in the rhizosphere soil of leguminous crops; for promoting early nodulation of leguminous crops and prolonging the time of nodulation and nitrogen fixation; for preventing leguminous crops from depleting nutrients and aging prematurely during maturity; for increasing the number of leguminous crop pods; for increasing the fullness of leguminous crop pods and reducing the rate of shrunken pods; for promoting early flowering and early pod formation of leguminous crops; for alleviating flowering =The occurrence of fruit rot; used to reduce the occurrence of bacterial wilt in leguminous crops; used to reduce the occurrence of powdery mildew in leguminous crops; used to reduce the occurrence of leaf spot in leguminous crops; used to reduce the occurrence of root nematode disease in leguminous crops; used to reduce the occurrence of root rot in leguminous crops; used to reduce the occurrence of blight in leguminous crops; used to reduce the occurrence of sclerotinia in leguminous crops; used to reduce the occurrence of downy mildew in leguminous crops; used to reduce the occurrence of wilt in leguminous crops; used to reduce the occurrence of white rot in leguminous crops; used to reduce the incidence of soybean green rot; used to reduce the incidence of corn ear rot and Fusarium toxin; used to reduce the abundance of wheat fusarium and reduce the incidence of fusarium toxin; used to promote carbon emission reduction in leguminous crops, which is beneficial to soil improvement; used to promote the increase of total biomass of leguminous crops; used to reduce the abundance of pests such as Aspergillus terreus and Fusarium in the rhizosphere of leguminous crops, which is beneficial to improving the soil microbial population structure; used to reduce the surface spots of peanuts and increase commercial value; used to promote soybean production in saline-alkali land.

[0043] According to the above scheme, the crops include but are not limited to leguminous crops, corn, and wheat. The leguminous crops include soybeans, peanuts, red beans, mung beans, peas, broad beans, cowpeas, green beans, kidney beans, alfalfa, and astragalus.

[0044] In a sixth aspect, the present invention provides a method for crop production, wherein a microbial agent is selected so that the microbial agent contains all or four or more gene sequences in the nucleotide sequences shown in SEQ ID NOs. 1 to 12 after analysis and determination, and has a coupled effect of controlling toxicity and fixing nitrogen, and has the effect of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules in leguminous crops; the microbial agent is applied to crops to control toxicity and fix nitrogen, improve quality and increase yield.

[0045] According to the above scheme, the gene sequences shown in SEQ ID NO.1-12 may have no more than 10% base variation in different strains. When the identity between them and the corresponding DNA sequences is more than 90% and they have the corresponding biological activity functions, they are equivalent to the corresponding gene sequences in the DNA sequences 1-12 shown in SEQ ID No.1-12, which are called functional equivalents of the DNA sequences shown in SEQ ID No.1-12. Containing all the gene sequences in the nucleotide sequences shown in SEQ ID NO.1-12 or 4 or more gene sequences therein or functional equivalents of these gene sequences, and having the coupled effects of controlling and fixing toxicity and nitrogen as described above by the microbial agents for controlling and fixing toxicity and nitrogen, regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules in leguminous crops, all constitute the coupled microbial agents for controlling and fixing toxicity and nitrogen of the present invention, which can be used for controlling toxicity and fixing nitrogen in crops, improving quality and increasing yield.

[0046] In the above scheme, the toxic control and nitrogen fixation coupled microbial agent has an inhibitory effect on aflatoxin and / or toxins. Furthermore, the inhibition rate of aflatoxin reaches more than 60%, and the inhibition rate of aflatoxin reaches more than 80%.

[0047] In the above scheme, the toxicity-control and nitrogen-fixing coupled microbial agent significantly inhibited the expression of the Aspergillus flavus PAB-01 protein, the amino acid sequence of which is shown in SEQ ID No. 12. The inhibition rate was greater than 90%, preferably greater than 95%, reflecting the excellent bacterial control and toxicity reduction effects of the toxicity-control and nitrogen-fixing coupled microbial agent of the present invention.

[0048] In the above scheme, the toxicity control and nitrogen fixation coupled microbial agent has an inhibitory effect on one or more pathogenic factors / toxins of soil-borne plant pathogens such as Penicillium, Aspergillus other than Aspergillus flavus, Fusarium, Sclerotium sclerotiorum, Pseudomonas solanacearum, Phytophthora, Sclerotinia sclerotiorum, Pythium, and Rhizoctonia solani, with an inhibition rate of greater than 30%.

[0049] In the above scheme, the toxic control and nitrogen fixation coupled microbial agent can be, but is not limited to, a combination of three or more of the following microorganisms: Bacillus laterosporus with a deposit number of CCTCC NO: M 20231807, Bacillus timonii with a deposit number of CCTCC NO: M 20231809, Bacillus subtilis with a deposit number of CCTCC NO: M 20231811, Stenotrophomonas maltophilia with a deposit number of CCTCC NO: M 20231813, Bacillus amyloliquefaciens with a deposit number of CCTCC NO: M 20231816, Bacillus mucilaginosus with a deposit number of CCTCC NO: M 20231817, and Enterobacter ludwigii BG10-1 with a deposit number of CCTCC NO: M 2016014. Specifically, it can be a combination of three, four, five, six, or seven of the above strains.

[0050] Or it can be a combination of one or more of the above-mentioned microorganisms and other microorganisms, so that the combined microbial agent meets the requirements of containing all the gene sequences in the nucleotide sequences shown in SEQ ID NO.1-12 or 4 or more of the gene sequences or their functional equivalents, has a coupled effect of toxicity control and nitrogen fixation, has the effect of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules of leguminous crops, and constitutes the toxicity control and nitrogen fixation coupled microbial agent of the present invention.

[0051] In the above scheme, the toxic control and nitrogen fixation coupled microbial agent can also be a combination of one or more of Bacillus laterosporus of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, Enterobacter Ludwigii BG10-1 of CCTCC NO: M 2016014 and the following four strains: Bacillus timundi of CCTCC NO: M 20231809, Bacillus subtilis of CCTCC NO: M 20231811, Stenotrophomonas maltophilia of CCTCC NO: M 20231813, and Bacillus mucilaginosus of CCTCC NO: M 20231817, such as Bacillus laterosporus CB2013 of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, and Bacillus ludwigii BG10-1 of CCTCC NO: M 2016014. a combination of Enterobacter Ludwigii BG10-1 of CCTCC NO: M 20231809 and Bacillus timundi of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, Enterobacter Ludwigii BG10-1 of CCTCC NO: M 2016014 and Bacillus subtilis of CCTCC NO: M 20231811, a combination of Bacillus laterosporus of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, Enterobacter Ludwigii BG10-1 of CCTCC NO: M 2016014 and Stenotrophomonas maltophilia of CCTCC NO: M 20231813, or a combination of Bacillus laterosporus of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, Enterobacter Ludwigii BG10-1 of CCTCC NO: M 2016014 and Stenotrophomonas maltophilia of CCTCC NO: M 20231813, or a combination of Bacillus laterosporus of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816 The combination of Bacillus laterosporus of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, Enterobacter Ludwigii BG10-1 of CCTCC NO: M 2016014, and Bacillus mucilaginosus of CCTCC NO: M 20231817, so that the combined microbial agent satisfies the requirements of containing all gene sequences in the nucleotide sequences shown in SEQ ID NOs. 1 to 12 or four or more gene sequences therein or their functional equivalents, has a coupled effect of toxicity control and nitrogen fixation, has the effect of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops, and increasing the number of nodules in leguminous crops, thereby constituting the toxicity control and nitrogen fixation coupled microbial agent of the present invention.

[0052] In the above scheme, the toxic control and nitrogen fixation coupled microbial agent can also be a combination of Bacillus laterosporus of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, Enterobacter Ludwigii BG10-1 of CCTCC NO: M 2016014 and other bacteria, so that the combined microbial agent meets the requirements of containing all gene sequences in the nucleotide sequences shown in SEQ ID NOs. 1 to 12 or 4 or more gene sequences therein or their functional equivalents, has a toxic control and nitrogen fixation coupled effect, has the effect of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules of leguminous crops, and constitutes the toxic control and nitrogen fixation coupled microbial agent of the present invention.

[0053] In the above scheme, the proportion of the number of live bacteria of any one strain in the poison-controlling and nitrogen-fixing coupled microbial agent in the microbial agent, that is, the mixed microbial composition, is greater than or equal to 1%.

[0054] The beneficial effects of the present invention are:

[0055] 1. The toxin control and nitrogen fixation coupled microbial agent of the present invention can be used to increase the production of legume crops such as peanuts and soybeans. Through a single technology, it achieves green control of the source of aflatoxin while inducing and promoting efficient nodulation and nitrogen fixation in soybeans and peanuts and significantly increasing yields. 2. It is easy to use, low cost, and high benefit. 3. Bacteria control and toxicity reduction are conducive to improving quality and safety levels, which is of great significance to ensuring food safety; 4. Promoting nodulation and nitrogen fixation is conducive to reducing the application of nitrogen fertilizers, which is of great significance to promoting the increase of soybean oil production capacity and green, low-carbon and efficient production in my country. Modes for Carrying Out the Invention

[0056] Part I: Coupling of toxic and nitrogen-fixing microbial agents

[0057] Example 1 Isolation and Identification of Microbial Strains (Series of Microbial Strains, Used in Combinations)

[0058] A series of microbial strains were obtained through the following isolation and identification steps.

[0059] (1) Take whole plants and rhizosphere soil samples of peanuts, soybeans, peas, broad beans, cowpeas, and alfalfa, grind them and mix them evenly, isolate the strains using conventional bacterial isolation methods, and then identify them using conventional 16s rDNA methods. Through the above operations, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus licheniformis, Brevibacillus laterosporus, Bacillus mucilaginosus, Bacillus velezensis, Bacillus siamensis, Paenibacillus polymyxa, Paenibacillus timonensis, Pseudomonas fluorescens, Pseudomonas mendocina, Enterobacter ludwigii, Microbacterium proteolyticum, Leclercia adcarboxglata, and Serratia serratia were obtained. marcescens, Empedobacters sp., Priestia megaterium, Stenotrophomonas maltophilia, and other bacteria. For detailed information, please see Table 1.

[0060] Table 1. Some of the strains isolated and identified from a mixture of major legume crops such as peanuts and soybeans are shown below:

[0061] Strain Code or Deposit Number Strain Name Strain Code or Deposit Number Strain Name CCTCC NO: M 20231807 Brevibacillus laterosporus CCTCC NO: M 20231809 Bacillus paenibacillus timonensis CCTCC NO: M 20231816 Bacillus amyloliquefaciens CCTCC NO: M 20231811 Bacillus subtilis CCTCC NO: M 20231817 Bacillus mucilaginosus CCTCC NO: M 20231813 Stenotrophomonas maltophilia CCTCC NO: M 2016014 Enterobacter ludwigii Strain 8 Bacillus amyloliquefaciens amyloliquefaciens strain 9, Bacillus subtilis strain 10, Bacillus licheniformis strain 11, Brevibacillus laterosporus strain 12, Bacillus mucilaginosus strain 13, Bacillus velezensis strain 14, Bacillus siamensis strain 15, Paenibacillus polymyxa strain 16, Paenibacillus timonensis strain 17, Pseudomonas fluorescens strain 18, Pseudomonas mendocina strain 19, Enterobacter ludwigii strain 20, Microbacterium proteolyticum strain 21, Leclercia nondecarboxylans adcarboxglata strain 22 Serratia marcescens strain 23 Empedobacterium brevis Empedobacters sp.Strain 24 Priestia megaterium, strain 25 Stenotrophomonas maltophilia, strain 26 Enterobacter ludwigii, strain 27 Bacillus velezensis, strain 28 Brevibacillus laterosporus, strain 29 Bacillus amyloliquefaciens, strain 30 Microbacterium proteolyticum, strain 31 is not a rhizobium, the genus and species are not identified, strain 32 is not a rhizobium, the genus and species are not identified, strain 33 is not a rhizobium, the genus and species are not identified, strain 34 is not a rhizobium, the genus and species are not identified, strain 35 is not a rhizobium, the genus and species are not identified, strain 36 is not a rhizobium, the genus and species are not identified, strain 37 is not a rhizobium, the genus and species are not identified, strain 38 is not a rhizobium, the genus and species are not identified.

[0062] Example 2 Preparation of microorganisms and their compositions

[0063] The series of bacterial strains obtained above are amplified and cultured by conventional bacterial culture medium amplification culture method to prepare batches of the above strain fermentation liquid or bacterial powder.

[0064] The fermentation broths or bacterial powders of the above strains were mixed one by one or more than two to form a series of microbial compositions. The information of these microbial compositions is shown in Table 2. The proportion of the number of viable bacteria of any one strain in each microbial combination was greater than or equal to 1%.

[0065] Table 2. Information on microorganisms or microbial compositions

[0066] Composition number Composition strains and proportions (%) Composition number Composition strains and proportions (%) Bacterial agent 1 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / CCTCC NO: M 20231817 Ratio: 30 / 30 / 30 / 10 Bacterial agent 2 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / CCTCC NO: M 20231817 Ratio: 30 / 30 / 10 / 30 Bacterial agent 3 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / CCTCC NO: M 20231817 ratio: 30 / 10 / 30 / 30 Bacterial agent 4 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / CCTCC NO: M 20231817 ratio: 10 / 30 / 30 / 30 Bacterial agent 5 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / CCTCC NO: M 20231817 ratio: 33 / 33 / 33 / 1 Bacterial agent 6 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / CCTCC NO: M 20231817 ratio: 33 / 33 / 1 / 33 Bacterial agent 7 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / CCTCC NO: M 20231817 ratio: 33 / 1 / 33 / 33 Bacterial agent 8 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / CCTCC NO: M 20231817 ratio: 1 / 33 / 33 / 33 Bacterial agent 9 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / CCTCC NO: M 20231809 Ratio: 30 / 30 / 30 / 10 Bacteria 10 Composition: CCTCC NO: M: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / CCTCC NO: M 20231811 Ratio: 30 / 30 / 30 / 10 Bacterial agent 11 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / CCTCC NO: M 20231813 Ratio: 30 / 30 / 30 / 10 Bacterial agent 12 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / strain 10 Ratio: 30 / 30 / 30 / 10 Bacterial agent 13 composition: CCTCC NO: M 20231807 / : CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / CCTCC NO: M 20231817 / Strain 13 Ratio: 30 / 1 / 30 / 10 / 29 Bacterial agent 14 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / Strain 14 Ratio: 30 / 30 / 30 / 10 Bacterial agent 15 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / Strain 15 Ratio: 30 / 30 / 30 / 10 Bacterial agent 16 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / strain 17 Ratio: 30 / 30 / 30 / 10 Bacterial agent 17 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / strain 18 Ratio: 30 / 30 / 30 / 10 Bacterial agent 18 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / strain 20 Ratio: 30 / 30 / 30 / 10 Bacterial agent 19 composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / strain 21 Ratio: 30 / 30 / 30 / 10 Bacterial agent 20 Composition: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCCNO: M 2016014 / strain 22 ratio: 30 / 30 / 30 / 10 composition of microbial agent 21: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / strain 23 ratio: 30 / 30 / 30 / 10 composition of microbial agent 22: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / strain 24 ratio: 30 / 30 / 30 / 10 composition of microbial agent 23: CCTCC NO: M 20231807 / CCTCC NO: M 20231816 / CCTCC NO: M 2016014 / strain 25 Ratio: 30 / 30 / 30 / 10 Bacterial agent 24 composition: strain 12 / strain 26 / strain 27 / strain 28 Ratio: 30 / 30 / 30 / 10 Bacterial agent 25 composition: strain 9 / strain 11 / strain 12 / strain 13 Ratio: 30 / 30 / 30 / 10 Bacterial agent 26 composition: strain 26 / strain 28 / strain 29 / strain 30 Ratio: 30 / 30 / 30 / 10 Bacterial agent 27 composition: CCTCC NO: M 2016014 / strain 8 / strain 12 Ratio: 45 / 45 / 10 Bacterial agent 28 composition: strain 17 / strain 18 / strain 21 / strain 22 Ratio: 30 / 30 / 30 / 10 Bacterial agent 29 composition: strain 23 / strain 24 / strain 25 / strain 30 30 / 30 / 30 / 10 Bacterial agent 30 composition: strain 12 / strain 18 / strain 20 / strain 24 30 / 30 / 30 / 10 Bacterial agent 31 composition: strain 8 Ratio: 100 Bacterial agent 32 composition: strain 11 Ratio: 100 Bacterial agent 33 composition: strain 12 Ratio: 100 Bacterial agent 34 composition: strain 19 Ratio: 100 Bacterial agent 35 composition: strain 21 Ratio: 100 Bacterial agent 36 composition: strain 22 Ratio: 100 Bacterial agent 37 composition: strain 23 Ratio: 100 Bacterial agent 38 composition: strain 24 Ratio: 100 Bacterial agent 39 composition: strain 25 Ratio: 100 Bacterial agent 40 composition: strain 30 Ratio: 100 Bacterial agent 41 composition: strain 31 / strain 32 / strain 33 / strain 34 Ratio: 25 / 25 / 25 / 25 Bacterial agent 42 composition: strain 35 / strain 36 / strain 37 / strain 38 Ratio: 25 / 25 / 25 / 25 Bacterial Agent 43 Composition: Strain 17 / Strain 31 / Strain 32 / Strain 33 Ratio: 30 / 30 / 30 / 10 Bacterial Agent 44 Composition: Strain 30 / Strain 32 / Strain 34 Ratio: 45 / 45 / 10

[0067] Example 3: Sequencing of microbial agents

[0068] A sufficient number of samples were taken from the microorganisms or microbial compositions in Table 2 of Example 2, and total DNA was extracted from these samples in sequence using conventional DNA extraction methods. The DNA sequences of these samples were then determined using conventional DNA sequencing methods. Finally, conventional analysis methods were used to compare the homology of the DNA sequences determined above with the gene sequences provided in this patent text.

[0069] The results of the above homology analysis are shown in Table 3.

[0070] Table 3. Information on microorganisms or microbial compositions

[0071] The number of microorganisms or their compositions contains the 12 characteristic genes, DNA sequence homologies and proportions provided by this patent. The number of microorganisms or their compositions contains the 12 characteristic genes, DNA sequence homologies and proportions provided by this patent. 19 bacterial agents: sequences 1 to 9, all with 100% homology, accounting for 75%. 29 bacterial agents: sequences 1 to 9, all with 100% homology, accounting for 75%. 39 bacterial agents: sequences 1 to 9, all with 100% homology, accounting for 75%. 49 bacterial agents: sequences 1 to 9, all with 100% homology, accounting for 75%. 59 bacterial agents: sequences 1 to 9, all with 100% homology, accounting for 75%. 69 bacterial agents: sequences 1 to 9, all with 100% homology, accounting for 75%. 79 bacterial agents: sequences 1 to 9, all with 100% homology, accounting for 75%. 89 bacterial agents: sequences 1 to 9, all with 100% homology. 910 bacterial agents, accounting for 75% of the total: sequences 1 to 9, 11, with 97% to 100% homology;84% of the bacterial agents: 109 sequences 1 to 9, all with 100% homology; 75% of the bacterial agents: 119 sequences 1 to 9, all with 100% homology; 75% of the bacterial agents: 129 sequences 1 to 9, all with 100% homology; 75% of the bacterial agents: 139 sequences 1 to 9, all with 100% homology; 75% of the bacterial agents: 149 sequences 1 to 9, all with 100% homology; 75% of the bacterial agents: 159 sequences 1 to 9, all with 100% homology; 75% of the bacterial agents: 169 sequences 1 to 9, all with 100% homology; 75% of the bacterial agents: 179 sequences 1 to 9, all with 100% homology; 75% of the bacterial agents: 189 sequences 1 to 9, all with 100% homology 75% of the bacterial agents: 199 sequences 1 to 9, all with 100% homology; 75% of the bacterial agents: 209 sequences 1 to 9, all with 100% homology; 75% of the bacterial agents: 219 sequences 1 to 9, all with 100% homology; 75% of the bacterial agents: 229 sequences 1 to 9, all with 100% homology; 75% of the bacterial agents: 239 sequences 1 to 9, all with 100% homology; 75% of the bacterial agents: 247 sequences 1 to 3, 6 to 8, 10, with 93% to 100% homology; 58% of the bacterial agents: 256 sequences 3, 4, 7, 8, 11, 12, with 90% to 100% homology; 50% of the bacterial agents: 266 sequences 1, 5 to 7, 9, 10, with 90% to 100% homology 50 bacterial agents 278: sequences 1-4, 6-8, 12, 91%-100% homology, accounting for 67% bacterial agent 280, accounting for 0% bacterial agent 290, accounting for 0% bacterial agent 301: sequence 4, 91% homology, accounting for 8% bacterial agent 312: sequences 3, 10, 90% and 91% homology, accounting for 17% bacterial agent 322: sequences 9, 12, 91% and 97% homology, accounting for 17% bacterial agent 331, sequence 5, 96% homology, accounting for 8% bacterial agent 343: sequences 4, 8, 12, 90%-97% homology Accounting for 25% of bacterial agent 350, accounting for 0% of bacterial agent 360, accounting for 0% of bacterial agent 370, accounting for 0% of bacterial agent 380, accounting for 0% of bacterial agent 390, accounting for 0% of bacterial agent 400, accounting for 0% of bacterial agent 415: sequence 1, 2, 4, 6, 11, homology 93%~100%, accounting for 42% of bacterial agent 424: sequence 1, 4, 7, 10, homology 92%~100%, accounting for 33% of bacterial agent 434: sequence 3, 5, 7, 10, homology 90%~99%, accounting for 33% of bacterial agent 444: sequence 2, 8, 9, 12, homology 90%~99%, accounting for 33%;

[0072] Example 4: Determination of the Toxic Control and Nitrogen Fixation Effect of Peanut Microbial Agents

[0073] Taking peanuts as an example, the steps for determining the toxicity control and nitrogen fixation effects of the above-mentioned microbial agents are as follows.

[0074] On the one hand, the bacterial agent in Table 3 of Example 3 above was co-cultured with the toxigenic Aspergillus flavus strain under the same conditions, the expression level of PAB-01 was determined using the literature method, and the inhibition rate of the bacterial agent on PAB-01 expression was calculated. The results are shown in Table 4.

[0075] On the other hand, the microbial agents described in Table 3 of Example 3 were applied to the field along with the peanut sowing base fertilizer, or during the peanut growing season, at a cumulative application rate of 80 billion viable bacteria per mu. A plot without any of the microbial agents was set up as a control, while all other plots were managed using conventional field management. Root nodules were investigated in the seedling stage of peanuts 7 to 12 days after emergence; root nodules were investigated in the mature stage of peanuts 1 to 3 days before harvest; and the effectiveness of toxicity control was investigated from the flowering stage to around harvest. Specifically, the effectiveness of field disease control and the abundance of aflatoxin in peanuts after harvest were investigated. The results of the investigation are shown in Table 4.

[0076] Based on the results in Tables 2-4, agents 1-27 and 41-44 simultaneously possess the following characteristics: 1) They contain all or part of at least four of the DNA sequences 1-12, and these genes may vary by no more than 10% between strains; 2) they significantly inhibit the expression of the protein PAB-01 from Aspergillus flavus, as reported in the literature, or they inhibit one or more pathogenic factors / toxins from soil-borne plant pathogens such as Penicillium, Aspergillus, Fusarium, Sclerotinia, Pseudomonas solanacearum, Phytophthora, Sclerotinia, Pythium, and Rhizoctonia; 3) although not rhizobia themselves, these microbial agents, coupled with toxin control and nitrogen fixation, can simultaneously regulate and increase the abundance of rhizobia in the peanut rhizosphere and the number of peanut root nodules; and 4) they promote early nodulation and prolong the duration of nodulation and nitrogen fixation. Agents 28-40, however, do not meet all four of these characteristics.

[0077] The soil-borne plant pathogens listed above are the pathogenic microorganisms of the corresponding field diseases. Therefore, the prevention and control effect of field diseases is equivalent to the inhibitory effect on the pathogenic factors / toxins of those harmful bacteria.

[0078] Table 4. Results of the microbial agent test on the control of toxicity and nitrogen fixation in peanuts

[0079] Microorganism or its composition number Inhibition rate of PAB-01 (%) Toxicity control effect on Aspergillus flavus pest control rate (%) Toxicity control rate on Penicillium, Aspergillus other than Aspergillus flavus, Fusarium, Sclerotium sclerotiorum, Pseudomonas solanacearum, Phytophthora, Sclerotinia sclerotiorum, Pythium, Rhizoctonia solani, etc. (%) Promotes nodulation and nitrogen fixation. Increase in the abundance of rhizobia / nodule number increase (%) Whether it promotes early nodulation and prolongs the nodulation and nitrogen fixation time of peanut Microorganism or its composition number Inhibition rate of PAB-01 (%) Toxicity control effect on Aspergillus flavus pest control rate (%) Toxicity control rate on Penicillium, Aspergillus other than Aspergillus flavus, Fusarium, Sclerotium sclerotiorum, Pseudomonas solanacearum, Phytophthora, Sclerotinia sclerotiorum, Pythium, Rhizoctonia solani, etc. (%) Promotes nodulation and nitrogen fixation. Increase in the abundance of rhizobia / nodule number increase (%) Whether it promotes early nodulation and prolongs the nodulation and nitrogen fixation time of peanut Microbial agent 199≥67≥4018 / 220 Yes: Bacterial agent 298 ≥ 62 ≥ 4019 / 243 Yes: Bacterial agent 399 ≥ 71 ≥ 4021 / 272 Yes: Bacterial agent 496 ≥ 60 ≥ 4019 / 266 Yes: Bacterial agent 596 ≥ 83 ≥ 4020 / 298 Yes: Bacterial agent 693 ≥ 66 ≥ 4026 / 333 Yes: Bacterial agent 798 ≥ 69 ≥ 4019 / 218 Yes: Bacterial agent 895 ≥ 63 ≥ 4024 / 306 Yes: Bacterial agent 992 ≥ 79 ≥ 4018 / 205 Yes: Bacterial agent 1096 ≥ 69 ≥ 4019 / 265 Yes: Bacterial agent 1195 ≥ 62 ≥ 4019 / 220 Yes: Bacterial agent 1296 ≥ 77 ≥ 4017 / 192 Yes: Bacterial agent 1397 ≥ 71 ≥ 4023 / 320 Yes: Bacterial agent 1499 ≥64 ≥4015 / 178 Yes: Bacterial agent 1593 ≥60 ≥4020 / 256 Yes: Bacterial agent 1692 ≥63 ≥4022 / 306 Yes: Bacterial agent 1791 ≥62 ≥4015 / 185 Yes: Bacterial agent 1894 ≥67 ≥4019 / 235 Yes: Bacterial agent 1993 ≥77 ≥4021 / 312 Yes: Bacterial agent 2097 ≥76 ≥4020 / 261 Yes: Bacterial agent 2191 ≥63 ≥4013 / 168 Yes: Bacterial agent 2293 ≥70 ≥4016 / 195 Yes: Bacterial agent 2394 ≥66 ≥4019 / 226 Yes: Bacterial agent 2496 ≥64 ≥4018 / 202 Yes: Bacterial agent 2597 ≥61 ≥4022 / 322 Yes Bacterial agent 2691 ≥67 ≥4012 / 176 Yes Bacterial agent 2793 ≥65 ≥4020 / 243 Yes Bacterial agent 2886 ≥58 ≤40 No Bacterial agent 290 ≥19 ≤40 No Bacterial agent 3091 ≥55 ≤40 No Bacterial agent 3164 ≥32 ≤40 No Bacterial agent 3262 ≥41 ≤40 No Bacterial agent 3372 ≥32 ≤40 No Bacterial agent 3483 ≥53 ≤40 No Bacterial agent 3558 ≥29 ≤40 No Bacterial agent 3670 ≤40 No Bacterial agent 37160 ≤40 No Bacterial agent 3843 ≥19 ≤40 No Bacterial agent 3947 ≥23 ≤40 No Bacterial agent 4059 ≥27 ≤40No Bacterial agent 4196 ≥ 63 ≥ 4016 / 190 Yes Bacterial agent 4293 ≥ 73 ≥ 4019 / 255 Yes Bacterial agent 4392 ≥ 67 ≥ 4019 / 263 Yes Bacterial agent 4491 ≥ 60 ≥ 4013 / 163 Yes

[0080] Example 5: Determination of the Toxic Control and Nitrogen Fixation Effect of Soybean Microbial Agents

[0081] Taking soybean as an example, the steps for determining the toxicity control and nitrogen fixation effects of the above-mentioned microbial agents are described as follows.

[0082] On the one hand, the bacterial agent in Table 3 of Example 3 above was co-cultured with the toxigenic Aspergillus flavus strain under the same conditions, the expression level of PAB-01 was determined using the literature method, and the inhibition rate of the bacterial agent on PAB-01 expression was calculated. The results are shown in Table 4.

[0083] On the other hand, the microbial agents listed in Table 3 of Example 3 were applied to the field along with soybean sowing base fertilizer, or during the soybean growing season, at a cumulative application rate of 80 billion viable bacteria per mu (approximately 1.5 acre). A control plot was established without any of the microbial agents. Conventional field management was used for all other plots. Root nodulation of soybean seedlings was investigated 7 to 12 days after emergence; root nodulation of peanuts at maturity was investigated 1 to 3 days before harvest; and toxicity control efficacy, i.e., effectiveness in preventing and controlling pest toxin-induced diseases, was investigated from the flowering stage to harvest. The results are shown in Table 5.

[0084] Based on the results in Tables 2, 3, and 5, agents 1–27 and 41–44 simultaneously possess the following characteristics: 1) The toxicity-control and nitrogen-fixing coupled microbial agents contain all or part of DNA sequences 1–12, with at least four of these genes likely to vary by no more than 10% between strains; 2) they significantly inhibit the expression of the Aspergillus flavus PAB-01 protein, or inhibit one or more pathogenic factors / toxins of soil-borne plant pathogens such as Penicillium, Aspergillus, Fusarium, Sclerotinia, Pseudomonas solanacearum, Phytophthora, Sclerotinia, Pythium, and Rhizoctonia; 3) although not rhizobia themselves, the agents simultaneously regulate and increase rhizobium abundance in the soybean rhizosphere and the number of soybean root nodules; and 4) they promote early soybean nodulation and prolong the duration of nodulation and nitrogen fixation. Agents 28–40, however, do not meet all four of these characteristics.

[0085] The soil-borne plant pathogens listed above are the pathogenic microorganisms of the corresponding field diseases. Therefore, the prevention and control effect of field diseases is equivalent to the inhibitory effect on the pathogenic factors / toxins of those harmful bacteria.

[0086] Table 5. Results of the test on the effect of microbial agents on soybean toxicity control and nitrogen fixation

[0087] Microorganism or its composition No. Inhibition rate of PAB-01 (%) Toxicity control effect on Aspergillus flavus pest control rate (%) Toxicity control rate on Penicillium, Aspergillus other than Aspergillus flavus, Fusarium, Sclerotium sclerotiorum, Pseudomonas solanacearum, Phytophthora, Sclerotinia sclerotiorum, Pythium, Rhizoctonia solani, etc. (%) Promotes nodulation and nitrogen fixation. Increase in the abundance of rhizobia / nodule number increase (%) Whether it promotes early nodulation and prolongs the time of nodulation and nitrogen fixation Microorganism or its composition No. Inhibition rate of PAB-01 (%) Toxicity control effect on Aspergillus flavus pest control rate (%) Toxicity control rate on Penicillium, Aspergillus other than Aspergillus flavus, Fusarium, Sclerotium sclerotiorum, Pseudomonas solanacearum, Phytophthora, Sclerotinia sclerotiorum, Pythium, Rhizoctonia solani, etc. (%) Promotes nodulation and nitrogen fixation. Increase in the abundance of rhizobia / nodule number increase (%) Whether it promotes early nodulation and prolongs the time of nodulation and nitrogen fixation Bacterial agent 1 ≥90% ≥63 ≥40% 19 / 218 Yes Bacterial agent 2 ≥90% ≥63 ≥40% 19 / 243 Yes Bacterial agent 3 ≥90% ≥72 ≥40% 20 / 263 Yes Bacterial agent 4 ≥90% ≥65 ≥40% 19 / 266 Yes Bacterial agent 5 ≥90% ≥82 ≥40% 19 / 217 Yes Bacterial agent 6 ≥90% ≥60 ≥40% 26 / 333 Yes Bacterial agent 7 ≥90% ≥67 ≥40% 19 / 202 Yes Bacterial agent 8 ≥90% ≥61 ≥40% 24 / 306 Yes Bacterial agent 9 ≥90% ≥75 ≥40% 17 / 195 Yes Bacterial agent 10 ≥90% ≥63 ≥40% 20 / 302 Yes Bacterial agent 11 ≥90% ≥64 ≥40% 19 / 251 Yes Bacterial agent 12 ≥90% ≥72 ≥40% 18 / 210 Yes Bacterial agent 13 ≥ 90% ≥ 70 ≥ 40% 22 / 198 Yes Bacterial agent 14 ≥ 90% ≥ 60 ≥ 40% 16 / 188 Yes Bacterial agent 15 ≥ 90% ≥ 62 ≥ 40% 20 / 240 Yes Bacterial agent 16 ≥ 90% ≥ 64 ≥ 40% 24 / 336 Yes Bacterial agent 17 ≥ 90% ≥ 63 ≥ 40% 15 / 183 Yes Bacterial agent 18 ≥ 90% ≥ 67 ≥ 40% 20 / 266 Yes Bacterial agent 19 ≥ 90% ≥ 73 ≥ 40% 21 / 314 Yes Bacterial agent 20 ≥ 90% ≥ 77 ≥ 40% 19 / 217 Yes Bacterial agent 21 ≥ 90% ≥ 62 ≥ 40% 14 / 157 Yes Bacterial agent 22 ≥ 90% ≥ 71 ≥ 40% 18 / 197 Yes Bacterial agent 23 ≥90% ≥66 ≥40% 20 / 277 Yes Bacterial agent 24 ≥90% ≥63 ≥40% 18 / 202 Yes Bacterial agent 25 ≥90% ≥64 ≥40% 21 / 312 Yes Bacterial agent 26 ≥90% ≥69 ≥40% 13 / 166 Yes Bacterial agent 27 ≥90% ≥62 ≥40% 21 / 293 Yes Bacterial agent 28 ≥10% ≤8 ≤3% 0 No Bacterial agent 29 ≤50% ≥17 ≤5% 0 No Bacterial agent 30 ≥9% ≥49 ≤5% 0 No Bacterial agent 31 ≤50% ≥27 ≤3% 0 No Bacterial agent 32 ≥7% ≥37 ≤4% 0 No Bacterial agent 33 ≤50% ≥33 ≤4% 0No Bacterial agent 34 ≥10% ≥54 ≤9% 0 No Bacterial agent 35 ≤50% ≥23 ≤2% 0 No Bacterial agent 36 ≥9% 0 ≤3% 0 No Bacterial agent 37 ≤50% 0 ≤6% 0 No Bacterial agent 38 ≥10% ≥23 ≤4% 0 No Bacterial agent 39 ≤50% ≥19 ≤7% 0 No Bacterial agent 40 ≥5% ≥36 ≤3% 0 No Bacterial agent 41 ≥90% ≥62 ≥40% 18 / 193 Yes Bacterial agent 42 ≥90% ≥71 ≥40% 18 / 196 Yes Bacterial agent 43 ≥90% ≥68 ≥40% 19 / 252 Yes Bacterial agent 44 ≥90% ≥63 ≥40% 14 / 170 Yes

[0088] Example 6: Determination of the Effect of Microbial Agents on Toxic Control and Nitrogen Fixation in Other Leguminous Crops

[0089] The sampling method and steps similar to those in Examples 4 and 5 were used to determine the effects of the microbial agent on pea, broad bean, cowpea, alfalfa and other legume crops in terms of toxicity control and nitrogen fixation, and the results similar to those in Tables 4 and 5 were obtained.

[0090] Example 7: Selection of microbial agents for toxicity control and nitrogen fixation

[0091] According to the results in Tables 1-5 above, agents 1-27 and 41-44 simultaneously possess the following characteristics: 1) The agents contain at least four gene sequences from DNA sequences 1-12, and these genes may have no more than 10% base variation in different strains; 2) The agents have a significant inhibitory effect on the expression of the Aspergillus flavus PAB-01 protein reported in the literature, or have an inhibitory effect on one or more pathogenic factors / toxins of soil-borne plant pathogens such as Penicillium, Aspergillus, Fusarium, Sclerotinia, Pseudomonas solanacearum, Phytophthora, Sclerotinia, Pythium, and Rhizoctonia; 3) Although not rhizobia themselves, the agents can simultaneously regulate and increase the abundance of rhizobia in the rhizosphere of legume crops, increase the number of nodules in legume crops, and thus enhance the nitrogenase activity per plant; 4) The agents can promote early nodulation and prolong the duration of nodulation and nitrogen fixation in legume crops such as peanuts and soybeans.

[0092] The microbial agent of the present invention was tested by knocking out DNA sequences 1 to 12. The results showed that when at least four gene sequences remained after the knockout, the microbial agent still exhibited the coupled effects of controlling toxicity and nitrogen fixation, improving quality and increasing yield, and further promoting early nodulation and prolonged nodulation and nitrogen fixation in legumes. However, when three or fewer gene sequences were removed, the microbial agent no longer exhibited the coupled effects of controlling toxicity and nitrogen fixation, improving quality and increasing yield, and promoting early nodulation and prolonged nodulation and nitrogen fixation in legumes.

[0093] In summary, the above-mentioned bacterial agents 1-27 and 41-44 can be selected as toxicity-control and nitrogen-fixing coupled microbial agents.

[0094] The above-mentioned microbial agent combinations are merely examples to facilitate understanding of the technical solutions of the present invention and are not intended to limit the scope of protection of the present invention. Those skilled in the art will appreciate that other variations or modifications can be made based on the above description. It is not necessary and impossible to enumerate all embodiments here. Obvious variations or modifications derived therefrom remain within the scope of protection of the present invention.

[0095] Part II: Application of Microbial Agents for Toxic Control and Nitrogen Fixation

[0096] Example 8: Application of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Improving the Safety Level of Leguminous Crop Products

[0097] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as examples, the use of toxic control and nitrogen fixation coupled with microbial agents to improve the safety level of leguminous crop products is described.

[0098] 1. Application on peanuts: Mix the above-mentioned microbial agents 1, 9, and 43 with peanut seeding base fertilizer respectively and apply them to the field through a seed drill. The application rate of microbial agents is 80 billion to 100 billion viable bacteria per mu. At the same time, set up a plot without any of the above microbial agents as a control. The rest are managed in the conventional field. After the above-mentioned peanuts are harvested, peanut samples are collected using the national standard sampling method. (1) The abundance of aflatoxin-producing fungi in these samples is determined by the classic colony counting method. The reduction rate of the abundance of aflatoxin-producing fungi in peanut fruit is calculated, which is the control effect on the aflatoxin-producing fungi. (2) After the samples are placed under the same conditions for 6 months, the aflatoxin contamination level is determined by the national standard liquid chromatography-mass spectrometry method, and the control effect on aflatoxin is calculated. These test results show that the control effect of microbial agents 1, 9, and 26 on aflatoxin-producing fungi in field peanut fruit is more than 60%, and the control effect on aflatoxin in peanuts is more than 80%. The above results show that the application of toxic control and nitrogen fixation coupled with microbial agents significantly improved the safety level of field peanuts.

[0099] 2. Application on Soybeans: Before soybean sowing, agents 1, 9, and 26 were applied to the fields via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu. A control plot was established without any of the agents, while all other plots were managed using conventional field management. After the soybeans were harvested, soybean samples were collected using national standard sampling methods. The abundance of aflatoxin in soybeans was determined using the same method used for peanuts. The results showed that the abundance of aflatoxin in soybeans was reduced by over 65%, significantly reducing the risk of aflatoxin contamination in post-harvest soybeans and improving their quality and safety. These results demonstrate that the application of a combination of toxin control and nitrogen fixation coupled with microbial agents significantly improves the safety of soybeans in the field.

[0100] 3. The application on other leguminous crops has achieved similar results as the above-mentioned application on soybeans.

[0101] Example 9: Application of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Promoting Nodulation and Nitrogen Fixation in Leguminous Crops

[0102] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to promote nodulation and nitrogen fixation in leguminous crops is described.

[0103] 1. Application on Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative application rate of 80 to 100 billion viable bacteria per mu (approximately 1.5 to 2.5 acres) was achieved. A control plot was established without any of the agents, while all other plots were managed using conventional methods. After the peanuts emerged, continuous monitoring of root nodules was conducted. These surveys showed that during the flowering phase, the number of root nodules in the peanuts treated with agents 1, 9, and 26 increased by more than threefold, and the nitrogenase activity per plant increased by more than tenfold. These results demonstrate that the application of microbial agents, coupled with toxicity control and nitrogen fixation, significantly promoted nodulation and nitrogen fixation in peanuts in the field.

[0104] 2. Application on Soybeans: Before soybean sowing, microbial agents 1, 9, and 26 were applied to the fields via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million saplings). A control plot was established without any of the agents, while all other plots were managed using conventional methods. Root nodulation was continuously monitored after soybean seedlings emerged. These surveys showed that during the flowering stage, the number of root nodules in soybeans treated with agents 1, 9, and 26 increased by more than 2.7 times, and the nitrogenase activity per soybean plant increased by more than 10 times. These results demonstrate that the application of microbial agents, coupled with toxicity control and nitrogen fixation, significantly promoted soybean nodulation and nitrogen fixation in the field.

[0105] 3. When applied to other leguminous crops, it achieved similar effects in promoting nodulation and nitrogen fixation as in peanuts and soybeans.

[0106] Example 10: Application of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Improving the Yield of Leguminous Crops

[0107] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to improve the yield of leguminous crops is described.

[0108] 1. Application on Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative application rate of 80 to 100 billion viable bacteria per mu (approximately 1.5 to 2.5 acre) was used as a control. Conventional field management was used for all other plots. At harvest time, one mu (approximately 1 acre) of peanuts was harvested from both the treated and control plots. Calculations showed that the yield increase in the plots treated with agents 1, 9, and 26 was over 15%, achieving significant yield increases. These results demonstrate that the application of microbial agents coupled with toxicity control and nitrogen fixation significantly increased field peanut yields.

[0109] 2. Application on Soybeans: Before soybean sowing, microbial agents 1, 9, and 26 were evenly applied to the fields via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu. A control plot was established without any of the agents, while all other plots were managed conventionally. At harvest time, soybeans were harvested from one mu of land in each of the treated and control plots. Calculations showed that peanut yield increases in the plots treated with agents 1, 9, and 26 were all above 13%, achieving significant yield increases. These results demonstrate that the application of microbial agents coupled with toxicity control and nitrogen fixation significantly improved soybean yields per unit area in large fields.

[0110] 3. Application on Peas: Before sowing peas, agents 1, 9, and 26 were evenly applied to the field by hand broadcasting, with a cumulative application rate of 80 to 100 billion viable bacteria per mu. A control plot was established without any of the agents, while all other plots were managed using conventional methods. Yields were measured by weighing the pea seedlings and green pea pods at harvest. Treatment with agents 1, 9, and 26 resulted in yield increases exceeding 10%, demonstrating significant yield increases. These results demonstrate that the application of microbial agents coupled with toxicity control and nitrogen fixation significantly increased the yield per unit area of ​​pea seedlings and pods in the field.

[0111] 4. Application on other legume crops: Application of agents 1, 9, and 26 on legume crops such as broad beans yielded similar results to those on soybeans. Application of agents 1, 9, and 26 on kidney beans and cowpeas yielded similar results to those on peas. Application of agents 1, 9, and 26 on alfalfa yielded similar results to those on peas.

[0112] Example 11: Use of a microbial agent coupled with toxicity control and nitrogen fixation - recruiting indigenous rhizobia to increase the abundance of rhizobia in the rhizosphere soil of leguminous crops

[0113] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to improve the yield of leguminous crops is described.

[0114] 1. Application on Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative bacterial count of 80 to 100 billion per mu (approximately 100 to 100 billion) was achieved. A control plot was established without any of the agents, while all others were managed conventionally. Rhizosphere soil samples were collected during the fruiting phase and before harvest, and metagenomic sequencing and big data analysis were performed. These results showed that the abundance of rhizobia in the rhizosphere soil of peanuts treated with agents 1, 9, and 26 was significantly higher than in the control group, increasing by 10% to 20%. These results demonstrate that the application of microbial agents, coupled with toxicity control and nitrogen fixation, has a significant effect in recruiting indigenous rhizobia, thereby increasing the abundance of rhizobia in the rhizosphere soil of legume crops.

[0115] 2. When applied to other legume crops such as soybeans, peas, broad beans, cowpeas, and alfalfa, it achieved similar effects as the above-mentioned application on peanuts, which was to recruit indigenous rhizobia and thus increase the abundance of rhizobia in the rhizosphere soil of legume crops.

[0116] Example 12: Application of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Promoting Early Nodulation and Prolonging Nodulation and Nitrogen Fixation Time in Leguminous Crops

[0117] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the purpose of coupling toxicity control and nitrogen fixation with microbial agents to promote early nodulation of leguminous crops and prolong the nodulation and nitrogen fixation time is described.

[0118] 1. Application on Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative application rate of 80 to 100 billion viable bacteria per mu (approximately 100 to 100 billion) was achieved. A control plot was established without any of the agents, while all others were managed under conventional practices. Root nodulation was continuously monitored after the peanuts emerged. These results showed that root nodules formed approximately 7 days after emergence in peanuts treated with agents 1, 9, and 26, more than 20 days earlier than the conventional estimate of 30 days. New nodules were still observed during the full-fruiting stage, with nodules of older, middle-aged, and young plants coexisting in the field. Fresh, active nodules with nitrogenase activity were still observed at harvest, disproving the conventional wisdom that no new nodules form after the seed-filling stage and that all nodules are depleted by harvest. This significantly extends the duration of nodulation and nitrogen fixation. The above results show that the application of toxic control and nitrogen fixation coupled with microbial agents can achieve early nodulation of peanut legume crops and extend the nodulation and nitrogen fixation time.

[0119] 2. Application on Soybeans: Before soybean sowing, microbial agents 1, 9, and 26 were applied to the fields via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu. Plots not treated with any of the agents served as controls, while all other plots were managed under conventional practices. Root nodulation was continuously monitored after soybean seedlings emerged. These results showed that soybeans treated with microbial agents 1, 9, and 26 developed root nodules approximately 9 days after seedling emergence, approximately 20 days earlier than the conventional wisdom of around 30 days. New nodules were still observed during the grain-filling stage, with nodules from older, middle-aged, and young plants coexisting. Fresh, active nodules with nitrogenase activity were still observed at harvest, disproving the conventional wisdom that soybeans no longer develop new nodules after the grain-filling stage and that all nodules are depleted by harvest. This significantly extends the duration of nodulation and nitrogen fixation. The above results show that the application of toxic control and nitrogen fixation coupled with microbial agents can achieve early nodulation and prolong the nodulation and nitrogen fixation time.

[0120] 3. When applied to other leguminous crops, it achieved similar effects as the above-mentioned application on peanuts and soybeans, promoting early nodulation of leguminous crops and prolonging the time of nodulation and nitrogen fixation.

[0121] Example 13: Application of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Preventing Deficiency and Premature Aging of Leguminous Crops During Maturity

[0122] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to prevent premature aging of leguminous crops during maturity is described.

[0123] 1. Application on Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative application rate of 80 to 100 billion viable bacteria per mu (approximately 100 to 100 million acre) was achieved. A control plot was established without any of the agents, while all other plots were managed using conventional methods. Continuous monitoring of peanut leaf photosynthetic rates and senescence was conducted during the maturity phase. These results showed that the leaf photosynthetic rates of peanuts treated with agents 1, 9, and 26 increased by over 30% compared to the control, demonstrating a significant effect in preventing premature aging due to nutrient depletion. These results demonstrate that the application of microbial agents, coupled with toxicity control and nitrogen fixation, can effectively prevent premature aging of leguminous crops during the maturity phase.

[0124] 2. Application on Soybeans: Before soybean sowing, microbial agents 1, 9, and 26 were applied to the fields via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu. A control plot was established without any of the agents, while all other plots were managed conventionally. Root nodulation was continuously monitored after soybean seedlings emerged. These surveys revealed that soybeans treated with agents 1, 9, and 26 experienced a 20% increase in leaf photosynthetic rate, demonstrating a significant protective effect against premature aging due to nutrient depletion. These results demonstrate that the combination of toxicity control and nitrogen fixation with microbial agents resulted in earlier nodulation and a prolonged period of nodulation and nitrogen fixation.

[0125] 3. When applied to other leguminous crops, it has achieved similar effects as its application to peanuts and soybeans in preventing premature aging during maturity.

[0126] Example 14: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Increasing the Number of Leguminous Crops' Pods

[0127] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to increase the number of leguminous crop pods is described.

[0128] 1. Application on Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative application rate of 80 to 100 billion viable bacteria per mu (approximately 1.5 to 2.5 acres) was achieved. A control plot was established without any of the agents, while all other plots were managed using conventional methods. Peanut pod counts were surveyed at harvest time. These survey results showed that the number of viable pods in the peanuts treated with agents 1, 9, and 26 increased by over 15% compared to the control, significantly increasing pod production. These results demonstrate that the application of microbial agents combined with toxicity control and nitrogen fixation can significantly increase peanut pod production.

[0129] 2. Application on Soybeans: Before soybean sowing, microbial agents 1, 9, and 26 were applied to the fields via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million sq ft). A control plot was established without any of the agents, while all other plots were managed conventionally. Pod counts were surveyed at harvest time. These survey results showed that soybean pod counts increased by over 10% in the areas treated with agents 1, 9, and 26, significantly increasing pod number. These results demonstrate that the application of microbial agents for toxicity control and nitrogen fixation can significantly increase soybean pod number.

[0130] 3. When applied to other leguminous crops such as peas, broad beans, kidney beans, and cowpeas, it achieved a similar effect as the above-mentioned application on peanuts and soybeans, which significantly increased the number of pods.

[0131] Example 15: Use of a microbial agent coupled with toxicity control and nitrogen fixation to increase leguminous crop pod plumpness and reduce pod shrinkage rate

[0132] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to increase the plumpness of leguminous crops pods and reduce the pod shrinkage rate is described.

[0133] 1. Application on Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative application rate of 80 to 100 billion viable bacteria per mu (approximately 1.5 to 2.5 acres) was achieved. A control plot was established without any of the agents, while all other plots were managed using conventional methods. At harvest, the peanut pods were surveyed and the pod shriveling rate calculated. These survey results showed that the application of agents 1, 9, and 26 significantly improved pod plumpness and reduced pod shriveling rate by over 10%. These results demonstrate that the application of microbial agents combining toxicity control and nitrogen fixation significantly increases peanut pod plumpness and reduces pod shriveling rate.

[0134] 2. Application on Soybeans: Before soybean sowing, microbial agents 1, 9, and 26 were applied to the fields via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu. A control plot was established without any of the agents, while all other plots were managed conventionally. At harvest, soybean pods were surveyed and pod shriveling rates were calculated. These survey results showed that soybeans treated with agents 1, 9, and 26 showed significantly improved pod plumpness and a reduction in pod shriveling rates of over 10%. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly increases soybean pod plumpness and reduces pod shriveling rates.

[0135] 3. When applied to other leguminous crops such as peas, broad beans, kidney beans, and cowpeas, it achieved similar effects as the above-mentioned application on peanuts and soybeans, increasing the fullness of peanut pods and reducing the rate of shrunken pods.

[0136] Example 16: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Promoting Early Flowering and Pod Setting in Leguminous Crops

[0137] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to promote early flowering and pod setting in leguminous crops is described.

[0138] 1. Application on Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative application rate of 80 to 100 billion viable bacteria per mu (approximately 1.5 to 2.5 acres) was achieved. A control plot was established without any of the agents, while all other plots were managed conventionally. After the peanuts emerged, continuous surveys were conducted to analyze the flowering and pod setting times of the treated and control peanuts. These survey results showed that peanuts treated with agents 1, 9, and 26 flowered and podded at least three days earlier. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly promotes earlier flowering and pod setting in peanuts.

[0139] 2. Application on Soybeans: Before soybean sowing, microbial agents 1, 9, and 26 were applied to the fields via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu. A control plot was established without any of the agents, while all other plots were managed conventionally. After soybean seedlings emerged, continuous surveys were conducted to analyze the flowering and pod setting times of the treated and control soybeans. These surveys showed that soybeans treated with agents 1, 9, and 26 flowered and podded at least two days earlier. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly promotes earlier flowering and pod setting in soybeans.

[0140] 3. When applied to other leguminous crops such as peas, broad beans, kidney beans, and cowpeas, similar effects of early flowering and early pod setting as those mentioned above were achieved in peanuts and soybeans.

[0141] Example 17: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Alleviating the Occurrence of Peanut Fruit Rot

[0142] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the incidence of peanut fruit rot is described.

[0143] Microbial agents 1, 9, and 26 were mixed with peanut seeding base fertilizer and applied to the field via a seed drill at a cumulative rate of 80 to 100 billion viable bacteria per mu. A control plot was established without any of the agents, while all other plots were managed conventionally. At harvest, the incidence of peanut fruit rot was statistically analyzed compared to the control plots. These findings showed that the incidence of peanut fruit rot in the plots treated with agents 1, 9, and 26 was reduced by over 70% compared to the control group. These results demonstrate that the application of microbial agents, coupled with toxicity control and nitrogen fixation, significantly reduces peanut fruit rot.

[0144] Example 18: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Alleviating the Occurrence of Bacterial Wilt in Leguminous Crops

[0145] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the incidence of bacterial wilt in leguminous crops is described.

[0146] 1. Application to Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative application rate of 80 to 100 billion viable bacteria per mu (approximately 1.5 to 2.5 acres) was achieved. A control plot was established without any of the agents, while all other plots were managed using conventional methods. After the peanuts emerged, continuous surveys were conducted to statistically analyze the incidence of peanut bacterial wilt compared to the control plots. These survey results showed that the incidence of peanut bacterial wilt in the plots treated with agents 1, 9, and 26 was reduced by over 60% compared to the control plots. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly reduces the incidence of peanut bacterial wilt.

[0147] 2. Application in other leguminous crops: Application in other leguminous crops such as soybeans, peas, broad beans, kidney beans, and cowpeas has achieved similar effects as the above-mentioned application in peanuts in reducing the incidence of root rot in leguminous crops.

[0148] Example 19: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Reducing Powdery Mildew in Leguminous Crops

[0149] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the incidence of powdery mildew in leguminous crops is described.

[0150] 1. Application on Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative application rate of 80 to 100 billion viable bacteria per mu (approximately 1.5 to 2.5 acres) was achieved. A control plot was established without any of the agents, while all other plots were managed conventionally. After the peanuts emerged, continuous surveys were conducted to statistically analyze the incidence of peanut fruit rot compared to the control plots. These survey results showed that the incidence of powdery mildew in peanuts treated with agents 1, 9, and 26 was reduced by 73% to 81% compared to the control plots. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly reduces the incidence of powdery mildew in peanuts.

[0151] 2. Application in other leguminous crops: Application in other leguminous crops such as soybeans, peas, broad beans, kidney beans, and cowpeas has achieved similar effects as the above-mentioned application in peanuts in reducing the incidence of powdery mildew in leguminous crops.

[0152] Example 20: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Reducing the Occurrence of Leaf Spot Disease in Leguminous Crops

[0153] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the incidence of leaf spot disease in leguminous crops is described.

[0154] 1. Application to Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative application rate of 80 to 100 billion viable bacteria per mu (approximately 1.5 to 2.5 acres) was achieved. A control plot was established without any of the agents, while all other plots were managed using conventional methods. After the peanuts emerged, continuous surveys were conducted to determine the incidence of peanut leaf spot compared to the control plots. These survey results showed that the incidence of peanut leaf spot in the plots treated with agents 1, 9, and 26 was reduced by over 56% compared to the control plots. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly reduces the incidence of peanut leaf spot.

[0155] 2. Application in other leguminous crops: Application in other leguminous crops such as soybeans, peas, broad beans, kidney beans, and cowpeas has achieved similar effects as the above-mentioned application in peanuts in reducing the incidence of leaf spot disease in leguminous crops.

[0156] Example 21: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Reducing the Occurrence of Root Nematode Disease in Leguminous Crops

[0157] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the incidence of root nematode disease in leguminous crops is described.

[0158] 1. Application to Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative application rate of 80 to 100 billion viable bacteria per mu (approximately 1.5 to 2.5 acres) was achieved. A control plot was established without any of the agents, while all other plots were managed using conventional methods. After the peanuts emerged, continuous surveys were conducted to assess the incidence of root nematodes in the treated and control plots. These survey results showed that the incidence of root nematodes in peanuts treated with agents 1, 9, and 26 was reduced by over 63% compared to the control plot. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly reduces root nematode disease in peanuts.

[0159] 2. Application in other leguminous crops: It has been applied to other leguminous crops such as soybeans, peas, broad beans, kidney beans, and cowpeas, and has achieved similar effects as the above-mentioned application in peanuts in reducing the incidence of root nematode diseases in leguminous crops.

[0160] Example 22: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Alleviating the Occurrence of Root Rot in Leguminous Crops

[0161] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the incidence of root rot in leguminous crops is described.

[0162] 1. Application to Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative application rate of 80 to 100 billion viable bacteria per mu (approximately 1.5 to 2.5 acres) was achieved. A control plot was established without any of the agents, while all other plots were managed conventionally. After the peanuts emerged, continuous surveys were conducted to statistically analyze the incidence of peanut root rot compared to the control plots. These survey results showed that the incidence of peanut root rot in the plots treated with agents 1, 9, and 26 was reduced by over 60% compared to the control plots. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly reduces the incidence of peanut root rot.

[0163] 2. Application on Soybeans: Mix the aforementioned agents 1, 9, and 26 with soybean seeding base fertilizer and apply them to the field via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu. A control plot without any of the agents was established, while all other plots were managed conventionally. After soybean seedlings emerged, continuous surveys were conducted to statistically analyze the incidence of soybean root rot compared to the control plots. These survey results showed that peanut root rot incidence in the plots treated with agents 1, 9, and 26 was 60% to 70% lower than in the control group. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly reduces soybean root rot.

[0164] 3. Application in other leguminous crops: Application in other leguminous crops such as peas, broad beans, kidney beans, cowpeas, etc. has achieved similar effects in reducing root rot as the above-mentioned application in peanuts and soybeans.

[0165] Example 23: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Reducing the Occurrence of Phytophthora in Leguminous Crops

[0166] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the incidence of blight in leguminous crops is described.

[0167] 1. Application on Soybeans: Mix the aforementioned agents 1, 9, and 26 with soybean seeding base fertilizer and apply them to the field using a seed drill at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million sq m). A control plot without any of the agents should be established, while all other plots should be managed conventionally. After soybean seedlings emerge, conduct ongoing surveys to compare the incidence of soybean Phytophthora blight with that of the control plots. These survey results show that soybeans treated with agents 1, 9, and 26 showed a reduction of over 62% in Phytophthora blight compared to the control plots. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly reduces the incidence of soybean Phytophthora blight.

[0168] 2. Application in other leguminous crops: Application in other leguminous crops such as peanuts, peas, broad beans, kidney beans, and cowpeas has achieved similar effects as the above-mentioned application in soybeans in reducing the incidence of blight in leguminous crops.

[0169] Example 24: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Reducing the Occurrence of Sclerotinia Blight in Leguminous Crops

[0170] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the incidence of sclerotinia disease in leguminous crops is described.

[0171] 1. Application on Soybeans: Mix the aforementioned agents 1, 9, and 26 with soybean seeding base fertilizer and apply them to the field using a seed drill at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million sq m). A control plot without any of the agents should be established, while all other plots should be managed conventionally. After soybean seedlings emerge, conduct ongoing surveys to compare the incidence of sclerotinia rot in the control plots. These survey results show that the incidence of soybean sclerotinia rot in the soybean plots treated with agents 1, 9, and 26 was reduced by over 60% compared to the control plots. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly reduces the incidence of soybean sclerotinia rot.

[0172] 2. Application in other leguminous crops: Application in other leguminous crops such as peanuts, peas, broad beans, kidney beans, cowpeas, etc. has achieved similar effects as the above-mentioned application in soybeans in reducing the incidence of sclerotinia disease in leguminous crops.

[0173] Example 25: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Alleviating Downy Mildew in Leguminous Crops

[0174] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the incidence of downy mildew in leguminous crops is described.

[0175] 1. Application on Soybeans: Mix the aforementioned agents 1, 9, and 26 with soybean seeding base fertilizer and apply them to the field using a seed drill at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million sq ft). Set up plots without any of the agents as controls, while all other plots follow conventional field management. After soybean seedlings emerge, conduct ongoing surveys to compare the incidence of downy mildew with the control plots. These survey results show that soybeans treated with agents 1, 9, and 26 experienced a reduction of over 60% in downy mildew compared to the control plots. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly reduces soybean downy mildew.

[0176] 2. Application in other leguminous crops: Application in other leguminous crops such as peanuts, peas, broad beans, kidney beans, and cowpeas has achieved similar effects as the above-mentioned application in soybeans in reducing the incidence of downy mildew in leguminous crops.

[0177] Example 26: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Alleviating the Occurrence of Fusarium Wilt in Leguminous Crops

[0178] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the incidence of wilt disease in leguminous crops is described.

[0179] 1. Application on Soybeans: Mix the aforementioned agents 1, 9, and 26 with soybean seeding base fertilizer and apply them to the field using a seed drill at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million sq ft). A control plot without any of the agents should be established, while all other plots should be managed conventionally. After soybean seedlings emerge, conduct ongoing surveys to compare the incidence of soybean wilt with the control plots. These survey results show that soybean wilt incidence in the plots treated with agents 1, 9, and 26 was reduced by nearly 60% compared to the control plots. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly reduces soybean wilt.

[0180] 2. Application in other leguminous crops: Application in other leguminous crops such as peanuts, peas, broad beans, kidney beans, cowpeas, etc. has achieved similar effects as the above-mentioned application in soybeans in reducing the incidence of wilt disease in leguminous crops.

[0181] Example 27: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Reducing the Occurrence of Sclerotinia rot in Leguminous Crops

[0182] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the incidence of white rot in leguminous crops is described.

[0183] 1. Application on Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative application rate of 80 to 100 billion viable bacteria per mu (approximately 1.5 to 2.5 acres) was achieved. A control plot was established without any of the agents, while all other plots were managed conventionally. After the peanuts emerged, continuous surveys were conducted to determine the incidence of white rot in the peanuts compared to the control plots. These survey results showed that the incidence of white rot in the peanuts treated with agents 1, 9, and 26 was reduced by over 66% compared to the control plots. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly reduces the incidence of white rot in peanuts.

[0184] 2. Application in other leguminous crops: It has been applied to other leguminous crops such as soybeans, peas, broad beans, kidney beans, and cowpeas, and has achieved similar effects as its application in peanuts in reducing the incidence of white rot in leguminous crops.

[0185] Example 28: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Reducing the Incidence of Soybean Greening

[0186] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the severity of soybean greening is described.

[0187] Microbial agents 1, 9, and 26 were mixed with soybean seeding base fertilizer and applied to the field via seed drill at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 1.5 to 2.5 acres). A control plot was established without any of the agents, while all other plots were managed conventionally. After soybean seedlings emerged, continuous surveys were conducted to assess the incidence of greening in the treated and control soybeans. These survey results showed that the incidence of greening in soybeans treated with agents 1, 9, and 26 was reduced by over 35% compared to the control group. These results demonstrate that the application of microbial agents, coupled with toxicity control and nitrogen fixation, significantly reduces the incidence of greening in soybeans.

[0188] Example 29: Use of Toxin Control and Nitrogen Fixation Coupled Microbial Agents - Reducing the Incidence of Corn Ear Rot and Fusarium Toxins

[0189] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the incidence of corn ear rot and Fusarium toxins is described.

[0190] Microbial agents 1, 9, and 26 were mixed with corn seeding fertilizer and applied to the field via seed drill at a cumulative rate of 80 to 100 billion viable bacteria per mu. Alternatively, these agents were sprayed by drone during the early silking stage. Plots not treated with any of these agents served as controls, while all other plots were managed using conventional methods. Surveys were conducted at harvest time, and statistical analysis was performed to compare the incidence of ear rot and Fusarium toxins in the control plots. These survey results showed that the incidence of ear rot in the plots treated with agents 1, 9, and 26 was reduced by more than 50% compared to the control, and the incidence of Fusarium toxins in the plots treated with agents 1, 9, and 26 was reduced by more than 70% compared to the control. These results demonstrate that the application of microbial agents, coupled with toxicity control and nitrogen fixation, significantly reduces the incidence of ear rot and Fusarium toxins in corn.

[0191] Example 30: Use of Toxin Control and Nitrogen Fixation Coupled with Microbial Agents - Reducing the Abundance of Wheat Gibberellic Acid and Alleviating the Occurrence of Gibberellic Acid

[0192] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the abundance of wheat fusarium and the degree of fusarium toxin occurrence is described.

[0193] Microbial agents 1, 9, and 26 were mixed with wheat seeding base fertilizer and applied to the fields via seed drill at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million saplings). These agents were sprayed by drone during the wheat flowering period. Plots not treated with any of the agents served as controls, while all other plots were managed conventionally. Surveys were conducted at harvest time to compare the abundance of wheat fusarium head blight and the incidence of gibberellins. These survey results showed that the abundance of wheat fusarium head blight in the plots treated with agents 1, 9, and 26 was reduced by over 50% compared to the control, and the incidence of gibberellins in the plots treated with agents 1, 9, and 26 was reduced by over 60% compared to the control. These results demonstrate that the application of microbial agents, coupled with toxicity control and nitrogen fixation, significantly reduces the abundance of wheat fusarium head blight and mitigates the incidence of gibberellins.

[0194] Example 31: Application of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Promoting Carbon Emission Reduction in Leguminous Crops and Benefiting Soil Improvement

[0195] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to promote carbon emission reduction in leguminous crops is described.

[0196] 1. Application to Peanuts: Mix the aforementioned agents 1, 9, and 26 with peanut seeding base fertilizer, respectively—using a 20% and 30% reduction in nitrogen use compared to conventional controls. Apply the agents to the field using a seed drill, achieving a cumulative application rate of 80 to 100 billion viable bacteria per mu. A plot without any of the agents was established as a control, while all other plots were managed conventionally. After the peanuts emerged, continuous surveys were conducted to assess the carbon reduction levels of the treated and control peanuts. These survey results showed that peanuts treated with agents 1, 9, and 26 reduced carbon dioxide emissions by over 20% and significantly increased yields by approximately 10%. These results demonstrate that the application of microbial agents, coupled with toxicity control and nitrogen fixation, significantly reduces carbon emissions during peanut production.

[0197] 2. Application on Soybeans: Before soybean sowing, microbial agents 1, 9, and 26 were applied to the fields via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million sq m). Nitrogen fertilizer was reduced by 20% and 30% in the treatment groups compared to the control group's conventional fertilization level. Plots not treated with any of the agents served as controls, while all other plots were managed under conventional management. After soybean seedlings emerged, continuous surveys were conducted to assess carbon emission reductions in the treatment and control soybeans. These survey results showed that soybeans treated with agents 1, 9, and 26 reduced carbon dioxide emissions by over 20% and significantly increased soybean yields by over 10%. These results demonstrate that the application of microbial agents, coupled with toxicity control and nitrogen fixation, significantly reduces carbon emissions in soybean production.

[0198] 3. Its application on other leguminous crops such as peas, broad beans, kidney beans, and cowpeas has achieved similar effects in promoting carbon emission reduction as its application on peanuts and soybeans.

[0199] Example 32: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Promoting the Increase of Total Biomass of Leguminous Crops

[0200] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to promote the increase of the total biomass of leguminous crops is described.

[0201] 1. Application to Peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill. A cumulative application rate of 80 to 100 billion viable bacteria per mu (approximately 1.5 to 2.5 acres) was achieved. A control plot was established without any of the agents, while all other plots were managed conventionally. A survey was conducted at harvest time to determine the total biomass of the treated and control peanut plants. These survey results showed that the total biomass of the peanut plants treated with agents 1, 9, and 26 increased by more than 20%. These results demonstrate that the application of microbial agents combining toxicity control and nitrogen fixation significantly increases the total biomass of peanut plants.

[0202] 2. Application on Soybeans: Before soybean sowing, microbial agents 1, 9, and 26 were applied to the fields via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million sq ft). A control plot was established without any of the agents, while all other plots were managed conventionally. A survey was conducted at maturity, and the total biomass of the treated and control soybean plants was calculated. These results showed that the total biomass of soybeans treated with agents 1, 9, and 26 increased by over 15%. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly increases the total biomass of soybean plants.

[0203] 3. When applied to other leguminous crops such as peas, broad beans, kidney beans, and cowpeas, it achieved similar effects as the above-mentioned application on peanuts and soybeans in promoting the increase of the total biomass of leguminous crop plants.

[0204] Example 33: Application of Toxic Control and Nitrogen Fixation Coupled Microbial Agents - Reducing the abundance of harmful organisms such as Aspergillus terreus and Fusarium in the rhizosphere of leguminous crops, which is beneficial to improving the soil microbial population structure

[0205] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce the abundance of pests such as Aspergillus terreus and Fusarium in the rhizosphere of leguminous crops is described.

[0206] 1. Application on Peanuts: Inoculants 1, 9, and 26 were mixed with peanut seeding fertilizer and applied to the field via seed drill at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million acre). A control plot was established without any of the above agents, while all other plots were managed conventionally. At harvest, rhizosphere soil samples were collected. Metagenomic and big data analysis was used to analyze the abundance of pests such as Aspergillus and Fusarium in the rhizosphere soil compared with the control samples. These results showed that the abundance of Aspergillus and Fusarium in the rhizosphere of peanuts treated with agents 1, 9, and 26 decreased by 56% to 72%. These results demonstrate that the application of microbial agents for toxicity control and nitrogen fixation can significantly reduce the abundance of pests such as Aspergillus and Fusarium in the rhizosphere of leguminous crops.

[0207] 2. Application on Soybeans: Inoculants 1, 9, and 26 were mixed with soybean seeding fertilizer and applied to the field via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million acre). A control plot was established without any of the above agents, while all other plots were managed conventionally. Rhizosphere soil samples were collected at harvest time. Metagenomic and big data analysis was used to analyze the abundance of pests such as Aspergillus and Fusarium in the rhizosphere soil compared to the control plots. These results showed that the abundance of Aspergillus and Fusarium in the rhizosphere of soybeans treated with agents 1, 9, and 26 decreased by 56% to 72%. These results demonstrate that the application of microbial agents for toxicity control and nitrogen fixation can significantly reduce the abundance of pests such as Aspergillus and Fusarium in the rhizosphere of legume crops.

[0208] 3. When applied to other legume crops such as peas, broad beans, kidney beans, and cowpeas, it achieved similar effects as the above-mentioned application on peanuts and soybeans, significantly reducing the abundance of pests such as Aspergillus terreus and Fusarium in the rhizosphere of legume crops.

[0209] Example 34: Application of Toxic Control and Nitrogen Fixation Coupled Microbial Agents - Reducing Peanut Surface Spots and Improving Commercial Quality

[0210] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to reduce spots on the surface of peanuts is described.

[0211] Application on peanuts: Mix the aforementioned agents 1, 9, and 26 with the peanut seeding base fertilizer and apply them to the field using a seed drill, with a cumulative application rate of 80 to 100 billion viable bacteria per mu. A control plot without any of the agents was established, while all other plots were managed conventionally. At harvest, surveys were conducted to determine the presence of black spots on the shells of both treated and control peanuts. These survey results showed that the presence of black spots on the shells of peanuts treated with agents 1, 9, and 26 was reduced by over 80%. These results demonstrate that the application of microbial agents coupled with toxicity control and nitrogen fixation significantly reduces peanut surface spots, improving market value and competitiveness, and increasing profitability.

[0212] Example 35: Use of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Promoting Crop Resistance to Continuous Cropping

[0213] Continuous cropping of crops for many years often leads to the accumulation of crop pathogens, resulting in weak seedlings, diseased seedlings, and seedling shortages. Taking the above-mentioned microbial agents 1, 9, and 26 as examples, this article describes the use of toxicity control and nitrogen fixation coupled with microbial agents to promote crop resistance to continuous cropping.

[0214] 1. Application in Peanuts: A trial demonstrating continuous cropping resistance was conducted in fields of peanuts planted continuously for many years in Xingcheng, Liaoning Province. Agents 1, 9, and 26 were mixed with peanut seeding base fertilizer and applied via seed drill to the continuously cropped fields at a cumulative rate of 80 to 100 billion viable bacteria per mu. Plots not treated with any of the agents served as controls, while all other plots were managed using conventional methods. Regular surveys were conducted after peanut sowing. These surveys showed that peanut seedlings treated with agents 1, 9, and 26 exhibited stronger seedlings and a significantly lower number of diseased plants compared to the control group. The disease rate in the control group was over 10 times that of the treated groups. These results demonstrate that the application of microbial agents, coupled with toxicity control and nitrogen fixation, significantly enhances crop resistance to continuous cropping.

[0215] 2. Application in Soybeans: A trial demonstrating continuous cropping resistance was conducted in soybean fields planted for multiple years. Microbial agents 1, 9, and 26 were mixed with soybean seeding base fertilizer and applied via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu. A control plot was established without any of the agents, while all other plots were managed conventionally. Regular surveys were conducted after soybean sowing. These surveys showed that soybean seedlings treated with agents 1, 9, and 26 exhibited stronger seedlings and a significantly lower number of diseased plants compared to the control group. The disease rate in the control group was over six times that of the treated groups. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly enhances crop resistance to continuous cropping.

[0216] 3. Its application on other crops such as peas, broad beans, kidney beans, cowpeas, potatoes, etc. has achieved a similar significant effect in promoting the resistance of crops to repeated cropping as its application on peanuts and soybeans.

[0217] Example 36: Application of Toxic Control and Nitrogen Fixation Coupled with Microbial Agents - Promoting Soybean Yield Increase in Saline-Alkali Land

[0218] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of toxic control and nitrogen fixation coupled with microbial agents to promote soybean production in saline-alkali soil is described.

[0219] The pilot demonstration was conducted in saline-alkali soil with a pH range of 8.2 to 9.2. Before soybean planting, microbial agents 1, 9, and 26 were applied by drone to the fields at a cumulative rate of 80 to 100 billion viable bacteria per mu. A control plot was established without any of the agents, while all other plots were managed conventionally. Yield surveys were conducted after the soybean harvest. These surveys showed that soybean yields per unit area increased by over 15% in the areas treated with agents 1, 9, and 26. These results demonstrate that the application of microbial agents, combined with toxicity control and nitrogen fixation, significantly increases soybean yields in saline-alkali soil.

[0220] Example 26 7: Use of the Toxic Control and Nitrogen Fixation Coupled Microbial Agent - Preparation of Toxic Control and Nitrogen Fixation Coupled Microbial Compound Fertilizer

[0221] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of the toxic-control and nitrogen-fixing coupled microbial agents in the preparation of toxic-control and nitrogen-fixing coupled microbial compound fertilizers is described.

[0222] Based on the amount of compound fertilizer used per mu of land and a dosage of at least 100 billion viable bacteria of the toxic-control and nitrogen-fixing coupled microbial agent, compound fertilizer was mixed with toxic-control and nitrogen-fixing coupled microbial agents 1, 9, and 26, respectively, at a ratio of 10 g to 100 g of bacterial powder per kilogram of compound fertilizer. The toxic-control and nitrogen-fixing coupled microbial agents were then physically adsorbed and fixed onto the compound fertilizer to create three toxic-control and nitrogen-fixing coupled microbial compound fertilizers: 1, 9, and 26. These compound fertilizers are inorganic and can be purchased commercially or prepared using common ratios using inorganic fertilizers such as nitrogen, phosphate, and potassium fertilizers.

[0223] The three toxic-control and nitrogen-fixing coupled microbial compound fertilizers, 1, 9, and 26, were used as seed fertilizers in leguminous crops such as peanuts and soybeans at sowing. Fields treated with the same fertilizers served as controls, and ongoing surveys were conducted after sowing. These survey results showed that the three toxic-control and nitrogen-fixing coupled microbial compound fertilizers, 1, 9, and 26, significantly promoted nodulation and nitrogen fixation, increasing nodule number by 3.5 to 13 times, increasing nitrogenase activity per plant by more than 10 times, and increasing yield per unit area by more than 12%, achieving significant yield increases and demonstrating the key characteristics of the toxic-control and nitrogen-fixing coupled microbial agents described in the aforementioned examples.

[0224] The above results indicate that the toxic-control and nitrogen-fixing coupled microbial agent can be used to prepare the toxic-control and nitrogen-fixing coupled microbial compound fertilizer, and it has the effects of both compound fertilizer and toxic-control and nitrogen-fixing coupled microbial agent.

[0225] Example 38: Use of Toxic Control and Nitrogen Fixation Coupled Microbial Agents - Preparation of Toxic Control and Nitrogen Fixation Coupled Microbial-Organic Fertilizer

[0226] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of the toxic-control and nitrogen-fixing coupled microbial agents in the preparation of toxic-control and nitrogen-fixing coupled microbial-bioorganic fertilizer is described.

[0227] Based on the amount of bio-organic fertilizer used per mu of land and a dosage of at least 80 billion viable bacteria of the toxic-control and nitrogen-fixing coupled microbial agent, mix the bio-organic fertilizer with toxic-control and nitrogen-fixing coupled microbial agents 1, 9, and 26, respectively, at a ratio of 10 g to 100 g of bacterial powder per kilogram of bio-organic fertilizer. Then, physically adsorb and fix the toxic-control and nitrogen-fixing coupled microorganisms onto the bio-organic fertilizer to create three toxic-control and nitrogen-fixing coupled microbial fertilizers: 1, 9, and 26. These bio-organic fertilizers can be purchased commercially.

[0228] The three toxic-control and nitrogen-fixing coupled microbial-organic fertilizers (1, 9, and 26) were used as seed fertilizers in leguminous crops such as peanuts and soybeans at sowing. Fields treated with the same bio-organic fertilizers served as controls, and ongoing surveys were conducted after sowing. These survey results showed that the three toxic-control and nitrogen-fixing coupled microbial-organic fertilizers (1, 9, and 26) significantly promoted nodulation and nitrogen fixation, increasing nodule number by more than threefold, increasing nitrogenase activity per plant by more than tenfold, and increasing yield per unit area by 10% to 25%, achieving significant yield increases. These results demonstrate the key characteristics of the toxic-control and nitrogen-fixing coupled microbial agents described in the aforementioned examples.

[0229] The above results indicate that the toxic-control and nitrogen-fixing coupled microbial agent can be used to prepare the toxic-control and nitrogen-fixing coupled microbial-bioorganic fertilizer, and has the effects of both the bio-organic fertilizer and the toxic-control and nitrogen-fixing coupled microbial agent.

[0230] Example 39: Use of Toxic Control and Nitrogen Fixation Coupled Microbial Agents - Preparation of Toxic Control and Nitrogen Fixation Coupled Microbial-Trace Element Fertilizer

[0231] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of the toxic-control and nitrogen-fixing coupled microbial agents in the preparation of toxic-control and nitrogen-fixing coupled microbial-trace element fertilizers is described.

[0232] Based on the required trace element fertilizer (referred to as micro-fertilizer) per mu (approximately 1.5 acres) and a dosage of at least 80 billion viable bacteria of the toxic-control and nitrogen-fixing coupled microbial agent, mix the trace element fertilizer with 1, 9, and 26 toxic-control and nitrogen-fixing coupled microbial agents, respectively. Specifically, add at least 8g of powdered or liquid toxic-control and nitrogen-fixing coupled microbial agents (with a viable bacteria count of at least 80 billion) per mu (approximately 1.5 acres). Mix thoroughly using conventional physical methods to create the three toxic-control and nitrogen-fixing coupled microbial-trace element fertilizers 1, 9, and 26. These trace element fertilizers can be purchased commercially.

[0233] The three toxic-control and nitrogen-fixing coupled microbial-trace element fertilizers, 1, 9, and 26, were used as seed fertilizers in leguminous crops such as peanuts and soybeans at sowing. Fields treated with the same bio-organic fertilizers served as controls, and ongoing surveys were conducted after sowing. These surveys showed that the three toxic-control and nitrogen-fixing coupled microbial-trace element fertilizers, 1, 9, and 26, significantly promoted nodulation and nitrogen fixation, increasing nodule number by more than 2.8 times, increasing nitrogenase activity per plant by more than 10 times, and increasing yield per unit area by more than 9%, achieving significant yield increases. These results demonstrate the key characteristics of the toxic-control and nitrogen-fixing coupled microbial agents described in the aforementioned examples.

[0234] The above results indicate that the toxic-control and nitrogen-fixing coupled microbial agent can be used to prepare the toxic-control and nitrogen-fixing coupled microbial-trace element fertilizer, and it has the effects of both the trace element fertilizer and the toxic-control and nitrogen-fixing coupled microbial agent.

[0235] Example 40: Use of Toxic Control and Nitrogen Fixation Coupled Microbial Agents - Preparation of Toxic Control and Nitrogen Fixation Coupled Microbial Moisturizing Fertilizer

[0236] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of the toxic-control and nitrogen-fixing coupled microbial agents in the preparation of the toxic-control and nitrogen-fixing coupled microbial-water-retaining fertilizer is described.

[0237] Based on the water-retaining agent dosage per mu (approximately 1.5 acres) and the toxin-control and nitrogen-fixing coupled microbial agent with a count of at least 80 billion viable bacteria, mix the water-retaining agent with the toxin-control and nitrogen-fixing coupled microbial agents 1, 9, and 26, respectively. Specifically, add at least 8g of these powdered agents or liquid agents with a count of at least 80 billion viable bacteria per mu (approximately 1.5 acres). Mix the water-retaining agent using conventional physical methods or allow the toxin-control and nitrogen-fixing coupled microbial agents to be adsorbed and fixed onto the water-retaining agent to create the three toxin-control and nitrogen-fixing coupled microbial fertilizers 1, 9, and 26. These water-retaining agents can be purchased commercially.

[0238] The three toxic-control and nitrogen-fixing coupled microbial agents (1, 9, and 26) were used as seed fertilizers in leguminous crops such as peanuts and soybeans at sowing. Fields treated with the same water-retaining agent served as controls. Continued post-sowing surveys were conducted. These surveys showed that the three toxic-control and nitrogen-fixing coupled microbial agents (1, 9, and 26) significantly promoted nodulation and nitrogen fixation, increasing nodule number by more than threefold, increasing nitrogenase activity per plant by 10-fold, and increasing yield per unit area by more than 10%, achieving significant yield increases. These results demonstrate the key characteristics of the toxic-control and nitrogen-fixing coupled microbial agents described in the aforementioned examples.

[0239] The above results indicate that the toxic-control and nitrogen-fixing coupled microbial agent can be used to prepare the toxic-control and nitrogen-fixing coupled microbial-water-retaining fertilizer, and has the effects of both a water-retaining agent and a toxic-control and nitrogen-fixing coupled microbial agent.

[0240] Example 41: Use of Toxic Control and Nitrogen Fixation Coupled Microbial Agents - Preparation of Toxic Control and Nitrogen Fixation Coupled Microbial Inorganic and Organic Compound Fertilizer

[0241] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of the toxic-control and nitrogen-fixing coupled microbial agents in the preparation of the toxic-control and nitrogen-fixing coupled microbial inorganic-organic compound fertilizer is described.

[0242] Based on the dosage of inorganic-organic compound fertilizer and a toxic-control and nitrogen-fixing coupled microbial agent with a count of at least 80 billion viable bacteria per mu of land, the inorganic-organic compound fertilizer is mixed with toxic-control and nitrogen-fixing coupled microbial agents 1, 9, and 26, respectively. Specifically, at least 8g of powdered or liquid toxic-control and nitrogen-fixing coupled microbial agents with a count of at least 80 billion viable bacteria per mu of inorganic-organic compound fertilizer are added. The toxic-control and nitrogen-fixing coupled microorganisms are then adsorbed and fixed to the inorganic-organic compound fertilizer using conventional physical methods, resulting in the three toxic-control and nitrogen-fixing coupled microbial inorganic-organic compound fertilizers 1, 9, and 26. These inorganic-organic compound fertilizers can be purchased commercially or prepared by purchasing the inorganic compound fertilizer and bio-organic fertilizer separately and blending them according to conventional methods.

[0243] The three toxic-control and nitrogen-fixing coupled microbial inorganic-organic compound fertilizers, 1, 9, and 26, were used as seed fertilizers in leguminous crops such as peanuts and soybeans at sowing. Fields treated with the same inorganic-organic compound fertilizers served as controls, and ongoing surveys were conducted after sowing. These survey results showed that the three toxic-control and nitrogen-fixing coupled microbial inorganic-organic compound fertilizers, 1, 9, and 26, significantly promoted nodulation and nitrogen fixation, increasing nodule number by more than 3.3 times, increasing nitrogenase activity per plant by more than 9 times, and increasing yield per unit area by more than 10%, achieving significant yield increases. These results demonstrate the key characteristics of the toxic-control and nitrogen-fixing coupled microbial agents described in the aforementioned examples.

[0244] The above results indicate that the toxic-control and nitrogen-fixing coupled microbial agent can be used to prepare the toxic-control and nitrogen-fixing coupled microbial inorganic-organic compound fertilizer, and it has the effects of both the inorganic-organic compound fertilizer and the toxic-control and nitrogen-fixing coupled microbial agent.

[0245] Example 42: Use of Toxic Control and Nitrogen Fixation Coupled Microbial Agents - Preparation of Toxic Control and Nitrogen Fixation Coupled Microbial-Rhizobium Fertilizer

[0246] Taking the application of the above-mentioned microbial agents 1, 9, and 26 as an example, the use of the toxic-control and nitrogen-fixing coupled microbial agents in the preparation of toxic-control and nitrogen-fixing coupled microbial-rhizobium fertilizers is described.

[0247] Based on the dosage of rhizobium fertilizer and a dosage of a toxic-control and nitrogen-fixing coupled microbial agent with a count of no less than 80 billion viable bacteria per mu of land, mix the rhizobium fertilizer with the toxic-control and nitrogen-fixing coupled microbial agents 1, 9, and 26, respectively. Specifically, add no less than 8g of the powdered or liquid agents with a count of no less than 80 billion viable bacteria per mu of rhizobium fertilizer. Then, mix them by conventional physical mixing to create the three toxic-control and nitrogen-fixing coupled microbial agents - rhizobium fertilizers 1, 9, and 26. These rhizobium fertilizers, also known as rhizobium agents, can be purchased commercially or isolated from fresh leguminous nodules and then cultured.

[0248] The three toxic-control and nitrogen-fixing coupled microbial-rhizobium fertilizers, 1, 9, and 26, were used as seed fertilizers in the production of leguminous crops such as peanuts and soybeans at sowing. Fields treated with the same rhizobium fertilizers served as controls, and ongoing surveys were conducted after sowing. These survey results showed that the three toxic-control and nitrogen-fixing coupled microbial inorganic-organic compound fertilizers, 1, 9, and 26, significantly promoted nodulation and nitrogen fixation, increasing nodule number by more than 1.7 times, increasing nitrogenase activity per plant by more than 5 times, and increasing yield per unit area by more than 7%, achieving significant yield increases, demonstrating the key characteristics of the toxic-control and nitrogen-fixing coupled microbial agents described in the above examples.

[0249] The above results indicate that the toxic-control and nitrogen-fixing coupled microbial agent can be used to prepare the toxic-control and nitrogen-fixing coupled microbial-rhizobium fertilizer, and has the effects of both rhizobium fertilizer and toxic-control and nitrogen-fixing coupled microbial agent.

Claims

1. A microbial agent for controlling toxicity and fixing nitrogen, characterized by: It is a microbial composition with a coupled effect of toxicity control and nitrogen fixation, which has the function of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules in leguminous crops. It contains all the gene sequences in the nucleotide sequences shown in SEQ ID NOs. 1 to 12 or four or more of them.

2. The microbial agent for controlling toxicity and fixing nitrogen according to claim 1, characterized in that: The toxic control and nitrogen fixation coupled microbial agent is a composition of more than three kinds of microorganisms, containing 4, 5, 6, 7, 8 or 9 or more gene sequences of the nucleotide sequences shown in SEQ ID NO. 1 to 12.

3. The toxicity-controlling and nitrogen-fixing coupled microbial agent according to claim 1 or 2, characterized in that: The microbial sources include, but are not limited to, Bacillus amyloliquefaciens, Bacillus subtilis, Bacillus licheniformis, Brevibacillus laterosporus, Bacillus mucilaginosus, Bacillus velezensis, Bacillus siamensis, Paenibacillus polymyxa, Paenibacillus timonensis, Pseudomonas fluorescens, Pseudomonas mendocina, Enterobacter ludwigii, Microbacterium proteolyticum, Leclercia adcarboxglata, and Serratia marcescens. marcescens, Empedobacters sp., Priestia priestiamegaterium, Stenotrophomonas maltophilia bacteria.

4. The microbial agent for controlling toxicity and fixing nitrogen according to claim 1, characterized in that: The poison-controlling and nitrogen-fixing coupled microbial agent has an inhibitory effect on aflatoxin and / or toxins.

5. The microbial agent for controlling toxicity and fixing nitrogen according to claim 1, characterized in that: The toxic-control and nitrogen-fixing microbial agent has a significant inhibitory effect on the expression of Aspergillus flavus PAB-01 protein, with an inhibition rate of more than 90%. The amino acid sequence of the PAB-01 protein is shown in SEQ ID No.

13.

6. The microbial agent for controlling toxicity and fixing nitrogen according to claim 1, characterized in that: The toxic control and nitrogen fixation coupled microbial agent has an inhibitory effect on one or more pathogenic factors / toxins of soil-borne plant pathogens such as Penicillium, Aspergillus other than Aflatoxin, Fusarium, Sclerotium sclerotiorum, Pseudomonas solanacearum, Phytophthora, Sclerotinia sclerotiorum, Pythium, and Rhizoctonia solani.

7. The microbial agent for controlling toxicity and fixing nitrogen according to claim 1, characterized in that: The gene sequences shown in SEQ ID NOs. 1 to 12 may vary to a certain extent in different strains. When the degree of variation is small, not exceeding 10% base variation, and the corresponding biological activity function is present, the functional equivalents of the DNA sequences shown in SEQ ID NOs. 1 to 12 are constituted. The microbial composition contains all the gene sequences in the nucleotide sequences shown in SEQ ID NOs. 1 to 12, or 4 or more of the gene sequences, or the functional equivalents of these gene sequences, and has a coupled effect of controlling toxicity and fixing nitrogen, and regulates and increases the abundance of rhizobia in the rhizosphere of leguminous crops and increases the number of nodules in leguminous crops, and thus constitutes the coupled microbial agent for controlling toxicity and fixing nitrogen according to claim 1.

8. The microbial agent for controlling toxicity and fixing nitrogen according to claim 1, characterized in that: The toxic control and nitrogen fixation coupled microbial agent is a composition of three or more of the following microorganisms: Bacillus laterosporus with a deposit number of CCTCC NO: M 20231807, Bacillus timondii with a deposit number of CCTCC NO: M 20231809, Bacillus subtilis with a deposit number of CCTCC NO: M 20231811, Stenotrophomonas maltophilia with a deposit number of CCTCC NO: M 20231813, Bacillus amyloliquefaciens with a deposit number of CCTCC NO: M 20231816, Bacillus mucilaginosus with a deposit number of CCTCC NO: M 20231817, and Enterobacter ludwigii BG10-1 with a deposit number of CCTCC NO: M 2016014. Or it is a combination of one or more microorganisms selected from Bacillus laterosporus with a deposit number of CCTCC NO: M 20231807, Bacillus timondii with a deposit number of CCTCC NO: M 20231809, Bacillus subtilis with a deposit number of CCTCC NO: M 20231811, Stenotrophomonas maltophilia with a deposit number of CCTCC NO: M 20231813, Bacillus amyloliquefaciens with a deposit number of CCTCC NO: M 20231816, Bacillus mucilaginosus with a deposit number of CCTCC NO: M 20231817, and Enterobacter ludwigii BG10-1 with a deposit number of CCTCC NO: M 2016014, and other microorganisms, so that the combined microbial agent meets the requirements of the composition comprising the bacterial strain of SEQ ID NO: M 20231807, Bacillus timondii with a deposit number of CCTCC NO: M 20231809, Bacillus subtilis with a deposit number of CCTCC NO: M 20231811, Bacillus subtilis with a deposit number of CCTCC NO: M 20231813, Bacillus amyloliquefaciens with a deposit number of CCTCC NO: M 20231816, Bacillus mucilaginosus with a deposit number of CCTCC NO: M 20231817, and Enterobacter ludwigii BG10-1 with a deposit number of CCTCC NO: M 2016014. All gene sequences in the nucleotide sequences shown in NO.1~12 or 4 or more gene sequences therein or their functional equivalents have a coupled effect of toxic control and nitrogen fixation, have the effect of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules of leguminous crops, and constitute the toxic control and nitrogen fixation coupled microbial agent of claim 1.

9. The microbial agent for controlling toxicity and fixing nitrogen according to claim 1, characterized in that: The toxic control and nitrogen fixation coupled microbial agent is a combination of one or more of Bacillus laterosporus of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, Enterobacter Ludwigii BG10-1 of CCTCC NO: M 2016014, and the following four strains: Bacillus timundi of CCTCC NO: M 20231809, Bacillus subtilis of CCTCC NO: M 20231811, Stenotrophomonas maltophilia of CCTCC NO: M 20231813, and Bacillus mucilaginosus of CCTCC NO: M 20231817.

10. The microbial agent for controlling toxicity and fixing nitrogen according to claim 1, characterized in that: The toxic control and nitrogen fixation coupled microbial agent is a combination of Bacillus laterosporus of CCTCC NO: M 20231807, Bacillus amyloliquefaciens of CCTCC NO: M 20231816, Enterobacter Ludwigii BG10-1 of CCTCC NO: M 2016014, and other bacteria, so that the combined microbial agent meets the requirements of containing all gene sequences in the nucleotide sequences shown in SEQ ID NOs. 1 to 12 or 4 or more gene sequences therein or their functional equivalents, has a toxic control and nitrogen fixation coupled effect, has the effect of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules of leguminous crops, and constitutes the toxic control and nitrogen fixation coupled microbial agent of claim 1.

11. The microbial agent for controlling toxicity and fixing nitrogen according to claim 1, characterized in that: The proportion of live bacteria count of any strain in the toxic control and nitrogen fixation coupled microbial agent is greater than or equal to 1%.

12. The use of the toxic control and nitrogen fixation coupled microbial agent according to claim 1 in crop production is any of the following: for improving the quality and safety level of leguminous crop products; for promoting nodulation and nitrogen fixation of leguminous crops; for increasing the yield of leguminous crops; for recruiting indigenous rhizobia and increasing the abundance of rhizobia in the rhizosphere soil of leguminous crops; for promoting early nodulation of leguminous crops and prolonging the nodulation and nitrogen fixation time; for preventing leguminous crops from premature aging due to lack of fertilizer during maturity; for increasing the number of leguminous crop pods; for increasing the plumpness of leguminous crop pods and reducing the pod shrinkage rate; for promoting early nodulation of leguminous crops. Flowering and early pod formation; used to reduce the occurrence of peanut fruit rot; used to reduce the occurrence of bacterial wilt in legume crops; used to reduce the occurrence of powdery mildew in legume crops; used to reduce the occurrence of leaf spot in legume crops; used to reduce the occurrence of root nematode disease in legume crops; used to reduce the occurrence of root rot in legume crops; used to reduce the occurrence of root nematode disease in legume crops; used to reduce the occurrence of blight in legume crops; used to reduce the occurrence of sclerotinia rot in legume crops; used to reduce the occurrence of downy mildew in legume crops; used to reduce the occurrence of wilt in legume crops; used to reduce the occurrence of white rot in legume crops; used to reduce the degree of soybean greening; Used to reduce the incidence of corn ear rot and fusarium toxins; Used to reduce the abundance of wheat fusarium and alleviate the occurrence of fusarium toxins; used to promote carbon emission reduction of legume crops, which is beneficial to soil improvement; used to promote the increase of total biomass of legume crops; used to reduce the abundance of pests such as Aspergillus terreus and Fusarium in the rhizosphere of legume crops, which is beneficial to improve the soil microbial population structure; used to reduce spots on the surface of peanuts and increase marketability; used to promote soybean production in saline-alkali land.

13. The poison-controlling and nitrogen-fixing coupled microbial agent according to claim 1 is used to prepare a poison-controlling and nitrogen-fixing coupled microbial compound fertilizer, a poison-controlling and nitrogen-fixing coupled microbial organic fertilizer, a poison-controlling and nitrogen-fixing coupled microbial micro-fertilizer, a poison-controlling and nitrogen-fixing coupled microbial moisturizing fertilizer, a poison-controlling and nitrogen-fixing coupled microbial inorganic-organic compound fertilizer, a poison-controlling and nitrogen-fixing coupled microbial-rhizobium fertilizer, or a poison-controlling and nitrogen-fixing coupled microbial-seed dressing agent / seed dressing agent / seed soaking agent, and is used in production to simultaneously exert the effects of the poison-controlling and nitrogen-fixing coupled microbial agent.

14. A poison-controlling nitrogen-fixing coupled microbial compound fertilizer, a poison-controlling nitrogen-fixing coupled microbial organic fertilizer, a poison-controlling nitrogen-fixing coupled microbial micro-fertilizer, a poison-controlling nitrogen-fixing coupled microbial moisturizing fertilizer, a poison-controlling nitrogen-fixing coupled microbial inorganic-organic compound fertilizer, a poison-controlling nitrogen-fixing coupled microbial-rhizobium fertilizer, or a poison-controlling nitrogen-fixing coupled microbial-seed dressing agent / seed dressing agent / seed soaking agent obtained by using the poison-controlling nitrogen-fixing coupled microbial agent according to claim 1.

15. A method for preparing a poison-controlling nitrogen-fixing coupled microbial compound fertilizer, a poison-controlling nitrogen-fixing coupled microbial organic fertilizer, a poison-controlling nitrogen-fixing coupled microbial micro-fertilizer, a poison-controlling nitrogen-fixing coupled microbial moisturizing fertilizer, a poison-controlling nitrogen-fixing coupled microbial inorganic-organic compound fertilizer, a poison-controlling nitrogen-fixing coupled microbial-rhizobium fertilizer, or a poison-controlling nitrogen-fixing coupled microbial-seed coating agent / seed dressing agent / seed soaking agent using the poison-controlling nitrogen-fixing coupled microbial agent according to claim 1, characterized in that: Method for preparing a toxic-control and nitrogen-fixing coupled microbial compound fertilizer: according to the conventional dosage of compound fertilizer and the dosage of the toxic-control and nitrogen-fixing coupled microbial agent with a viable count of not less than 80 billion per mu of land, the compound fertilizer and the toxic-control and nitrogen-fixing coupled microbial agent are mixed in proportion, and then the toxic-control and nitrogen-fixing coupled microorganisms are adsorbed and fixed onto the compound fertilizer particles by physical methods to prepare the toxic-control and nitrogen-fixing coupled microbial compound fertilizer; Method for preparing toxic-control and nitrogen-fixing coupled microbial organic fertilizer: according to the conventional amount of organic fertilizer per mu of land and the amount of toxic-control and nitrogen-fixing coupled microbial agent with a viable count of not less than 80 billion, the organic fertilizer and the toxic-control and nitrogen-fixing coupled microbial agent are mixed in proportion, and then the toxic-control and nitrogen-fixing coupled microorganisms are adsorbed and fixed on the organic fertilizer particles by physical methods to prepare the toxic-control and nitrogen-fixing coupled microbial compound fertilizer; Method for preparing microbial fertilizers for toxin-control and nitrogen-fixing coupling: according to the conventional dosage of trace element fertilizers per mu of land and the dosage of microbial agent for toxin-control and nitrogen-fixing coupling with no less than 80 billion viable bacteria, the trace element fertilizers and the microbial agent for toxin-control and nitrogen-fixing coupling are proportioned, and then mixed by physical methods to adsorb and fix the microorganisms for toxin-control and nitrogen-fixing coupling onto the trace element fertilizer particles to prepare the microbial fertilizer for toxin-control and nitrogen-fixing coupling; Method for preparing a poison-controlling and nitrogen-fixing coupled microbial moisturizing fertilizer: according to the conventional dosage of water-retaining agent and the dosage of the poison-controlling and nitrogen-fixing coupled microbial agent with a viable count of not less than 80 billion per mu of land, the trace element fertilizer and the poison-controlling and nitrogen-fixing coupled microbial agent are mixed in proportion, and then the poison-controlling and nitrogen-fixing coupled microbial agent is mixed by conventional physical methods / or the poison-controlling and nitrogen-fixing coupled microbial agent is adsorbed and fixed on the water-retaining agent to prepare the poison-controlling and nitrogen-fixing coupled microbial moisturizing fertilizer; A method for preparing a toxic-control and nitrogen-fixing coupled microbial rhizobium fertilizer: according to the conventional amount of rhizobium fertilizer and the amount of the toxic-control and nitrogen-fixing coupled microbial agent with a viable count of not less than 80 billion per mu of land, the rhizobium fertilizer and the toxic-control and nitrogen-fixing coupled microbial agent are proportioned, and then mixed by conventional physical methods, or the two agents are mixed and then fixed on carrier particles to prepare the toxic-control and nitrogen-fixing coupled microbial rhizobium fertilizer; Method for preparing toxic-control and nitrogen-fixing coupled microbial organic-inorganic compound fertilizer: according to the conventional dosage of compound fertilizer per mu of land and the dosage of toxic-control and nitrogen-fixing coupled microbial agent with a viable count of not less than 80 billion, the organic-inorganic compound fertilizer and the toxic-control and nitrogen-fixing coupled microbial agent are mixed, and then the toxic-control and nitrogen-fixing coupled microorganisms are adsorbed and fixed on the organic and inorganic particles by physical methods to prepare the toxic-control and nitrogen-fixing coupled microbial organic-inorganic compound fertilizer; The method for preparing quality-improving and nitrogen-fixing coupled microorganisms-seed dressing agent / seed mixing agent / seed soaking agent: according to the amount of seed dressing agent or seed mixing agent or seed soaking agent per mu of land and the amount of quality-improving and nitrogen-fixing coupled microbial agent of not less than 80 billion viable bacteria, the seed dressing agent or seed mixing agent or seed soaking agent and the toxicity-controlling and nitrogen-fixing coupled microbial agent are proportioned, and then the toxicity-controlling and nitrogen-fixing coupled microbial agent and the seed dressing agent or seed mixing agent or seed soaking agent are evenly mixed through conventional physical mixing to prepare the toxicity-controlling and nitrogen-fixing coupled microorganisms-seed dressing agent / seed mixing agent / seed soaking agent.

16. The use of the toxic-control and nitrogen-fixing coupled microbial compound fertilizer or the toxic-control and nitrogen-fixing coupled microbial organic fertilizer or the toxic-control and nitrogen-fixing coupled microbial micro-fertilizer or the toxic-control and nitrogen-fixing coupled microbial moisturizing fertilizer or the toxic-control and nitrogen-fixing coupled microbial inorganic-organic compound fertilizer or the toxic-control and nitrogen-fixing coupled microbial-rhizobium fertilizer or the toxic-control and nitrogen-fixing coupled microbial-seed dressing agent / seed dressing agent / seed soaking agent configured according to claim 15 in crop production as described in any of the following: for improving the quality and safety level of leguminous crop products; for promoting nodulation and nitrogen fixation in leguminous crops; for increasing the yield per unit area of leguminous crops; for recruiting indigenous rhizobia to increase the abundance of rhizobia in the rhizosphere soil of leguminous crops; for promoting early nodulation of leguminous crops and prolonging the time of nodulation and nitrogen fixation; for preventing Premature aging of leguminous crops due to lack of fertilizer during maturity; used to increase the number of leguminous crop pods; used to increase the plumpness of leguminous crop pods and reduce the shrunken pod rate; used to promote early flowering and early pod setting of leguminous crops; used to reduce the occurrence of peanut fruit rot; used to reduce the occurrence of bacterial wilt of leguminous crops; used to reduce the occurrence of powdery mildew of leguminous crops; used to reduce the occurrence of leaf spot of leguminous crops; used to reduce the occurrence of root nematode disease of leguminous crops; used to reduce the occurrence of root rot of leguminous crops; used to reduce the occurrence of blight of leguminous crops; used to reduce the occurrence of sclerotinia disease of leguminous crops; used to reduce the occurrence of downy mildew of leguminous crops; used to reduce the occurrence of wilt of leguminous crops; used to reduce the occurrence of white rot of leguminous crops; used to reduce the degree of soybean green disease; Used to reduce the incidence of corn ear rot and fusarium toxins; Used to reduce the abundance of wheat fusarium and alleviate the occurrence of fusarium toxins; used to promote carbon emission reduction of legume crops, which is beneficial to soil improvement; used to promote the increase of total biomass of legume crops; used to reduce the abundance of pests such as Aspergillus terreus and Fusarium in the rhizosphere of legume crops, which is beneficial to improve the soil microbial population structure; used to reduce spots on the surface of peanuts and increase marketability; used to promote soybean production in saline-alkali land.

17. A method for crop production, characterized in that: A toxic-control and nitrogen-fixing coupled microbial agent is selected so that, upon analysis and determination, the microbial agent contains all or four or more gene sequences in the nucleotide sequences shown in SEQ ID NOs. 1 to 12, and has a toxic-control and nitrogen-fixing coupled effect, and has the effect of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules in leguminous crops; the toxic-control and nitrogen-fixing coupled microbial agent is applied to crops to control toxicity, fix nitrogen, improve quality and increase yield.

18. The method for crop production according to claim 17, characterized in that: The gene sequences shown in SEQ ID NO.1-12 may have no more than 10% base variation in different strains. When they have more than 90% identity with the corresponding DNA sequences 1-12 and have corresponding biological activity functions, they are equivalent to the corresponding gene sequences in the DNA sequences 1-12 shown in SEQ ID No.1-12, and are called functional equivalents of the DNA sequences shown in SEQ ID No.1-12. They contain all the gene sequences in the nucleotide sequences shown in SEQ ID NO.1-12 or 4 or more gene sequences therein or functional equivalents of these gene sequences, and have the coupled effect of controlling toxicity and fixing nitrogen, regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules in leguminous crops, and constitute the coupled microbial agent for controlling toxicity and fixing nitrogen according to claim 1, and the coupled microbial agent for controlling toxicity and fixing nitrogen is used for controlling toxicity and fixing nitrogen in crops, improving quality and increasing yield.