Arc microbial agent for quality improvement, nitrogen fixation and green yield increase, and use thereof

Abstract

The present invention belongs to the field of microorganisms. Specifically, disclosed are an ARC microbial agent for quality improvement, nitrogen fixation and green yield increase, and the use thereof. The ARC microbial agent for quality improvement, nitrogen fixation and green yield increase has the effects of improving quality, fixing nitrogen and increasing yield, thereby improving the quality of a leguminous crop product. The ARC microbial agent has the effects of regulating and improving the abundance of rhizobia in the rhizosphere of the leguminous crop and increasing the number of nodules in the leguminous crop; and contains gene sequences as shown in SEQ ID NOs: 1-4. The ARC microbial agent can be used in the production of leguminous crops such as soybeans, peanuts, peas, broad beans and cowpeas, thereby realizing quality improvement, nitrogen fixation and green yield increase. In addition, the ARC microbial agent is simple to use, has significant social and ecological benefits, and is easy to popularize and apply.

Description

ARC microbial agent for improving quality, fixing nitrogen and increasing green yield and its application Technical Field

[0001] The present invention belongs to the field of microorganisms, and in particular relates to a green ARC microbial agent for improving nitrogen fixation and increasing yield and its use. Background Art

[0002] Legume crops, including soybeans, peanuts, peas, broad beans, cowpeas, and alfalfa, are important sources of food and feed for humans. Global agriculture today faces numerous challenges in its pursuit of green and high-quality development. The development of the legume industry faces two common challenges: First, legumes like peanuts and soybeans are susceptible to contamination by highly toxic and carcinogenic aflatoxins, which not only reduce quality and yield, but also pose a serious threat to human health and safety. Aflatoxin B1, for example, is 10 times more toxic than potassium cyanide and is classified as a Class I carcinogen by the World Health Organization's International Agency for Research on Cancer. It contributes to 28.2% of liver cancer worldwide. Improving the quality, safety, and performance of legume products has long been a hot topic in the high-quality development of the legume agriculture sector.

[0003] Secondly, while legume crops like soybeans and peanuts form symbiotic nodules and fix nitrogen with soil rhizobia, these nodules are naturally few in number, their nitrogen fixation duration is short (typically, no nodules form and fix nitrogen in the first month of crop growth, and nodules begin to decay during the pod-filling or fruit-full stages), and their efficiency is low. 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 that defies the AON theory) is elusive. Improving the nodulation and nitrogen fixation efficiency of legumes like peanuts and soybeans remains a hotly debated and challenging issue internationally.

[0004] To address these challenges, the inventors' team, after over 20 years of continuous research, have successfully developed the ARC microbial agent, a green, yield-enhancing, nitrogen-fixing agent. This agent improves the quality of legume crops like soybeans and peanuts while simultaneously inducing efficient nodulation and nitrogen fixation, doubling their yields. The agent is simple to use, low-cost, and highly effective, boasting 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 demonstrates its enormous potential for application and has been demonstrated and validated in field trials across major soybean, peanut, and pea producing regions nationwide. This agent holds significant promise for boosting soybean oilseed production capacity and promoting green, low-carbon, and efficient production in my country. Summary of the Invention

[0005] In response to the shortcomings of the existing technology, the present invention provides an ARC microbial agent for improving quality, fixing nitrogen and increasing green yield. The agent is applied to the production of leguminous crops such as soybeans, peanuts, peas, broad beans, and cowpeas, achieving quality improvement, nitrogen fixation and green yield increase. It is simple to use, has significant social and ecological benefits, and is easy to promote and apply.

[0006] In order to solve the above technical problems, the technical solution adopted by the present invention is:

[0007] The ARC microbial agent for improving quality, fixing nitrogen, and increasing yield in a green manner has the effects of improving quality, fixing nitrogen, and increasing yield, thereby improving the quality of legume crop products. It has the functions of regulating and increasing the abundance of rhizobia in the rhizosphere of legume crops and increasing the number of nodules in legume crops. It contains all four gene sequences shown in SEQ ID No. 1-4.

[0008] According to the above scheme, the quality-enhancing, nitrogen-fixing, green, and yield-increasing ARC microbial agent can improve the quality of leguminous crop products by increasing the content of one or more nutritional functional components such as protein, glutamic acid, peanut resveratrol, and soy isoflavones.

[0009] According to the above scheme, the ARC microbial agent for improving quality, fixing nitrogen, and increasing green yield has a mitigating effect, that is, a control effect, on one or more soil-borne diseases such as peanut fruit rot, soybean greening, white rot of legumes, bacterial wilt, root rot, and sclerotinia. The agent has a harm reduction effect on one or more soil-borne diseases such as peanut fruit rot, soybean greening, white rot of legumes, bacterial wilt, root rot, and sclerotinia, with a reduction rate of at least 30%, and preferably over 80%.

[0010] According to the above scheme, the quality-improving, nitrogen-fixing, green, and yield-increasing ARC microbial agent can reduce the spots on the peanut fruit surface.

[0011] According to the above scheme, the green ARC microbial agent for improving nitrogen fixation and increasing yield significantly reduces the level of aflatoxin and improves the quality and safety level.

[0012] While the aforementioned ARC microbial agent for improving nitrogen and green yields is not itself a rhizobium, it can simultaneously regulate and increase the abundance of rhizobia in the rhizosphere of legume crops, increase the number of nodules formed in legume crops, and enhance nitrogenase activity per plant. This agent can promote early nodulation and prolong the duration of nodulation and nitrogen fixation in legume crops such as peanuts and soybeans. It can also increase the yield per unit area and the total biomass of legume crops.

[0013] According to the above scheme, the DNA sequence genes shown in SEQ ID No. 1-4 may have a certain degree of variation 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 greater than 90%, preferably greater than 95%, and more preferably greater than 99%, and when they have corresponding biological activities, these are called functional equivalents of the DNA sequences shown in SEQ ID No. 1-4, and containing these sequences is equivalent to containing the corresponding sequences of DNA sequences 1 to 4. When a microbial composition contains the DNA sequences shown in SEQ ID NO. 1 to 4 or their functional equivalents and has the effects of improving nitrogen fixation and increasing yield, and has the effects of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules in leguminous crops, it constitutes the ARC microbial agent for improving nitrogen fixation and increasing yield of the present invention.

[0014] The aforementioned DNA sequences 1-4 are specific sequences obtained after comparison with Genbank genome data. They are directly or indirectly related to the functions of the microbial agent of this patent. When containing all the gene sequences of the aforementioned DNA sequences 1-4, they have the effects of improving nitrogen fixation and increasing yield, regulating and increasing the abundance of rhizobia in the rhizosphere of legume crops and increasing the number of nodules in legume crops, constituting the ARC microbial agent of the present invention for improving nitrogen fixation and increasing yield. These genes may vary to a certain extent in different strains. If the degree of variation is small, such as no more than 10% base variation, and the corresponding biological activity is maintained, containing them is equivalent to containing the gene sequences shown in SEQ ID Nos. 1-4.

[0015] The ARC microbial agent for improving quality, fixing nitrogen and increasing yield in the present invention is a microbial composition. Through the synergistic effect of the various microorganisms in the microbial composition, the agent exerts the effect of improving quality and fixing nitrogen. When used in crop production, the agent has the effects of improving quality, fixing nitrogen and increasing yield. It has the effects of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules in leguminous crops. Although the agent itself is not a rhizobium, it can 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 individual plants; it can improve the yield level and total biomass of leguminous crop production; it can promote early nodulation of peanuts and soybeans, prolong the nodulation and nitrogen fixation time, and can also reduce spots on the surface of peanut fruits, significantly reduce the level of aflatoxin, and improve the quality and safety level.

[0016] The aforementioned ARC microbial agent for improving nitrogen fixation and increasing yield can be prepared by the following method: combining the microorganisms in the aforementioned microbial agent and fermenting the agent. The fermentation pathway can employ conventional fermentation pathways using bacteria disclosed in existing technologies, including those described in existing literature.

[0017] According to the above scheme, preferably, the ARC microbial agent for improving quality, fixing nitrogen and increasing green yield is a combination of three or more microorganisms.

[0018] According to the above scheme, the ARC microbial agent for improving nitrogen fixation and increasing green production of the present invention can be, but is not limited to, a composition of the following four microorganisms: Bacillus laterosporus with a deposit number of CCTCC NO: M 20231601, Bacillus amyloliquefaciens with a deposit number of CCTCC NO: M 20231598, Enterobacter Ludwigii with a deposit number of CCTCC NO: M 20231595, and Bacillus mucilaginosus with a deposit number of CCTCC NO: M 20231602.

[0019] Bacillus lateralis, deposit date is September 4, 2023, deposit number is CCTCC NO: M 20231601, classification name is: Brevibacterium lateralisAR1007, deposit unit name is China Center for Type Culture Collection, address is Wuhan University, Wuhan, China.

[0020] Bacillus amylolyticus, deposit date is September 4, 2023, deposit number is CCTCC NO: M 20231598, classification name is: Bacillus amylolyticusAR1004, deposit unit name is China Center for Type Culture Collection, address is Wuhan University, Wuhan, China.

[0021] Enterobacter ludwigii, deposited on September 4, 2023, with the deposit number CCTCC NO: M 20231595, the classification name: Enterobacter ludwigiiAR1001, the depository is China Center for Type Culture Collection, and the address is Wuhan University, Wuhan, China.

[0022] Bacillus mucilaginosus, deposit date is September 4, 2023, deposit number is CCTCC NO: M 20231602, classification name is: Bacillus mucilaginosusAR1008, deposit unit name is China Center for Type Culture Collection, address is Wuhan University, Wuhan, China.

[0023] According to the above scheme, the above-mentioned ARC microbial agent for improving nitrogen fixation and coupling green yield increase can be a combination of one or more microorganisms selected from Bacillus laterosporus with a deposit number of CCTCC NO: M 20231601, Bacillus amyloliquefaciens with a deposit number of CCTCC NO: M 20231598, Enterobacter Ludwigii with a deposit number of CCTCC NO: M 20231595, and Bacillus mucilaginosus with a deposit number of CCTCC NO: M 20231602, and other microorganisms, so that the combined microbial agent satisfies the requirement of containing all gene sequences in the nucleotide sequences shown in SEQ ID NOs. 1 to 4 or their functional equivalents, has the effect of coupling improving nitrogen fixation and quality improvement, has the effect of regulating and increasing the abundance of rhizobia in the rhizosphere of legume crops, and increasing the number of nodules of legume crops, has the effect of improving nitrogen fixation and increasing yield, has the effect of regulating and increasing the abundance of rhizobia in the rhizosphere of legume crops, and increasing the number of nodules of legume crops, and constitutes the ARC microbial agent for improving nitrogen fixation and green yield increase of the present invention.

[0024] According to the above scheme, in the above-mentioned ARC microbial agent for improving quality, fixing nitrogen and increasing green production, the proportion of the number of live bacteria of any one strain in the microbial agent, that is, the mixed microbial composition, is greater than or equal to 1%.

[0025] The above-mentioned ARC microbial agent for improving quality, fixing nitrogen and increasing green yield can be used in the production of leguminous crops as follows: to improve the quality and safety level of leguminous crop products; to promote nodulation and nitrogen fixation of leguminous crops; to increase the yield level of leguminous crops; to recruit indigenous rhizobia and increase the abundance of rhizobia in the rhizosphere soil of leguminous crops; to promote early nodulation of leguminous crops and prolong the nodulation and nitrogen fixation time; to prevent leguminous crops from premature aging due to lack of fertilizer during maturity; to increase the number of leguminous crop pods; to increase the fullness of leguminous crop pods and reduce the pod shrinkage rate; to promote early flowering and early pod setting of leguminous crops; to reduce the occurrence of peanut fruit rot; to reduce the It is used to reduce the occurrence of bacterial wilt in leguminous crops; it is used to reduce the occurrence of powdery mildew in leguminous crops; it is used to reduce the occurrence of leaf spot in leguminous crops; it is used to reduce the occurrence of root nematode disease in leguminous crops; it is used to reduce the occurrence of root rot in leguminous crops; it is used to reduce the occurrence of root nematode disease in leguminous crops; it is used to reduce the occurrence of sclerotinia disease in leguminous crops; it is used to reduce the occurrence of downy mildew in leguminous crops; it is used to reduce the occurrence of wilt in leguminous crops; it is used to reduce the occurrence of white rot in leguminous crops; it is used to reduce the incidence of soybean green disease; it is used to promote carbon emission reduction in leguminous crops, which is beneficial to soil improvement; it is used to promote the increase of total biomass of leguminous crops; it is used to reduce the surface spots of peanut fruits and increase marketability; it is used to promote the increase of soybean production in saline-alkali land.

[0026] The third aspect of the present invention provides the above-mentioned quality-improving and nitrogen-fixing green yield-increasing ARC microbial agent for preparing quality-improving and nitrogen-fixing coupled microbial compound fertilizer, which is used in production to simultaneously play the role of quality-improving and nitrogen-fixing green yield-increasing ARC microbial agent; or is used to prepare quality-improving and nitrogen-fixing coupled microbial organic fertilizer, which is used in production to simultaneously play the role of quality-improving and nitrogen-fixing green yield-increasing ARC microbial agent; or is used to prepare quality-improving and nitrogen-fixing coupled microbial fertilizer, which is used in production to simultaneously play the role of quality-improving and nitrogen-fixing green yield-increasing ARC microbial agent; or is used to prepare quality-improving and nitrogen-fixing coupled microbial moisturizing fertilizer, which is used in production to simultaneously play the role of quality-improving and nitrogen-fixing green yield-increasing ARC microbial agent; or is used to prepare quality-improving and nitrogen-fixing coupled microbial inorganic-organic compound fertilizer, which is used in production to simultaneously play the role of quality-improving and nitrogen-fixing green yield-increasing ARC microbial agent.

[0027] The present invention further provides a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial compound fertilizer, a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial organic fertilizer, a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial micro-fertilizer, a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial moisturizing fertilizer, a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial inorganic-organic compound fertilizer, or a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial-rhizobium fertilizer, which is obtained by configuring the above-mentioned quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial agent.

[0028] The fourth aspect of the present invention provides a method for preparing a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial compound fertilizer, a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial organic fertilizer, a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial micro-fertilizer, a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial moisturizing fertilizer, a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial inorganic-organic compound fertilizer, or a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial-rhizobium fertilizer using the above-mentioned quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial agent:

[0029] in:

[0030] Method for preparing the quality-enhancing, nitrogen-fixing, green and yield-increasing ARC microbial compound fertilizer: according to the conventional dosage of compound fertilizer and the dosage of the quality-enhancing, nitrogen-fixing, green and yield-increasing ARC microbial agent per mu of land with a viable count of not less than 80 billion, the compound fertilizer and the quality-enhancing, nitrogen-fixing, green and yield-increasing ARC microbial agent are mixed in proportion, and then the quality-enhancing, nitrogen-fixing, green and yield-increasing ARC microbial compound fertilizer is adsorbed and fixed to the compound fertilizer particles by physical methods to prepare the quality-enhancing, nitrogen-fixing, green and yield-increasing ARC microbial compound fertilizer;

[0031] Method for preparing the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial organic fertilizer: according to the conventional amount of organic fertilizer and the amount of the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial agent with a viable count of not less than 80 billion per mu of land, the organic fertilizer and the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial agent are mixed in proportion, and then the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial agent is adsorbed and fixed on the organic fertilizer particles by physical methods to prepare the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial compound fertilizer;

[0032] Method for preparing ARC microbial fertilizer for improving quality, fixing nitrogen and increasing green yield: according to the conventional dosage of trace element fertilizer and the dosage of ARC microbial agent for improving quality, fixing nitrogen and increasing green yield with a count of not less than 80 billion viable bacteria per mu of land, the trace element fertilizer and the microbial agent for improving quality, fixing nitrogen and increasing green yield are mixed, and then the ARC microorganisms for improving quality, fixing nitrogen and increasing green yield are adsorbed and fixed on the micro-fertilizer particles by physical methods to prepare the micro-fertilizer for improving quality, fixing nitrogen and increasing green yield;

[0033] Method for preparing the quality-improving nitrogen-fixing coupled microbial moisturizing fertilizer: according to the conventional dosage of water-retaining agent and the dosage of the quality-improving 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 quality-improving nitrogen-fixing green yield-increasing ARC microbial agent are mixed in proportion, and then the quality-improving nitrogen-fixing green yield-increasing ARC microbial moisturizing fertilizer is prepared by mixing by conventional physical methods / or adsorbing and fixing the quality-improving nitrogen-fixing green yield-increasing ARC microbial moisturizing fertilizer by the water-retaining agent;

[0034] Method for preparing the ARC microbial rhizobium fertilizer for improving quality, fixing nitrogen and increasing green yield: according to the conventional amount of rhizobium fertilizer and the amount of the ARC microbial agent for improving quality, fixing nitrogen and increasing green yield of not less than 80 billion viable bacteria per mu of land, the rhizobium fertilizer and the ARC microbial agent for improving quality, fixing nitrogen and increasing green yield are proportioned, and then mixed by conventional physical methods to prepare the ARC microbial rhizobium fertilizer for improving quality, fixing nitrogen and increasing green yield;

[0035] The method for preparing the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial organic-inorganic compound fertilizer is as follows: according to the conventional dosage of compound fertilizer per mu of land and the dosage of the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial agent of not less than 80 billion viable bacteria, the organic-inorganic compound fertilizer and the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial agent are mixed, and then the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial agent is adsorbed and fixed on the organic and inorganic particles through physical methods to prepare the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial organic-inorganic compound fertilizer.

[0036] The present invention also provides any of the following uses of the ARC microbial compound fertilizer for improving nitrogen and green yield increasing, or the ARC microbial organic fertilizer for improving nitrogen and green yield increasing, or the ARC microbial micro-fertilizer for improving nitrogen and green yield increasing, or the ARC microbial moisturizing fertilizer for improving nitrogen and green yield increasing, or the ARC microbial inorganic-organic compound fertilizer for improving nitrogen and green yield increasing, or the ARC microbial-rhizobium fertilizer for improving nitrogen and green yield increasing in crop production obtained by compounding the above-mentioned ARC microbial agent for improving nitrogen and green yield increasing, in crop production: 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 nodulation and nitrogen fixation time; for preventing premature aging of leguminous crops due to lack of fertilizer during maturity; for increasing the pod growth of leguminous crops It is used to increase the number of fruits; to increase the fullness of legume pods and reduce the pod shrinkage rate; to promote early flowering and early pod setting of legume crops; to reduce the occurrence of peanut fruit rot; to reduce the occurrence of bacterial wilt of legume crops; to reduce the occurrence of powdery mildew of legume crops; to reduce the occurrence of leaf spot of legume crops; to reduce the occurrence of root nematode disease of legume crops; to reduce the occurrence of root rot of legume crops; to reduce the occurrence of root nematode disease of legume crops; to reduce the occurrence of sclerotinia disease of legume crops; to reduce the occurrence of downy mildew of legume crops; to reduce the occurrence of wilt of legume crops; to reduce the occurrence of white rot of legume crops; to reduce the incidence of soybean green disease; to promote carbon emission reduction of legume crops, which is beneficial to soil improvement; to promote the increase of total biomass of legume crops; to reduce the surface spots of peanut fruits and increase marketability; to promote the increase of soybean yield in saline-alkali land.

[0037] A sixth aspect of the present invention provides a method for crop production, comprising selecting a microbial agent so that the microbial agent contains a gene sequence as shown in SEQ ID NOs. 1 to 4 after analysis and determination, and has the effect of improving the quality and fixing nitrogen, thereby improving the quality of legume crop products, and has the effect of regulating and increasing the abundance of rhizobia in the rhizosphere of legume crops and increasing the number of nodules in legume crops; applying the microbial agent to crops is used to improve the quality and fix nitrogen, thereby increasing green yields.

[0038] According to the above scheme, the DNA sequence genes shown in SEQ ID Nos. 1-4 may have a certain degree of variation 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, when the identity is greater than 90%, preferably greater than 95%, and more preferably greater than 99%, and when the corresponding biological activity is present, these are functional equivalents of the DNA sequences shown in SEQ ID Nos. 1-4, and containing these sequences is equivalent to containing the corresponding sequences of DNA sequences 1 to 4. When containing the gene sequences shown in SEQ ID Nos. 1 to 4 or their functional equivalents and having the quality-improving and nitrogen-fixing effects described in the ARC microbial agent for improving quality, fixing nitrogen, and increasing green yield, it can improve the quality of legume crop products, regulate and increase the abundance of rhizobia in the rhizosphere of legume crops, and increase the number of nodules in legume crops, it constitutes the ARC microbial agent for improving quality, fixing nitrogen, and increasing green yield of the present invention, and can be used for improving quality, fixing nitrogen, and increasing green yield of crops.

[0039] In the above scheme, the ARC microbial agent for improving quality, fixing nitrogen and increasing yield in a green way can improve the quality of leguminous crop products by increasing the content of one or more nutritional functional components such as protein, glutamic acid, peanut resveratrol, and soy isoflavones.

[0040] In the above solution, the ARC microbial agent for improving quality, fixing nitrogen, and increasing yield has a mitigating effect, that is, a control effect, on one or more soil-borne diseases such as peanut fruit rot, soybean greening, white rot of legumes, bacterial wilt, root rot, and sclerotinia. The agent has a harm reduction effect on one or more soil-borne diseases such as peanut fruit rot, soybean greening, white rot of legumes, bacterial wilt, root rot, and sclerotinia, with a reduction rate of at least 30%, and preferably over 80%.

[0041] In the above scheme, the quality-improving, nitrogen-fixing, green, yield-increasing ARC microbial agent can reduce the spots on the peanut fruit surface.

[0042] In the above scheme, the green ARC microbial agent for improving nitrogen fixation and increasing yield significantly reduces the level of aflatoxin and improves the quality and safety level.

[0043] In the above scheme, preferably, the ARC microbial agent for improving quality, fixing nitrogen and increasing green production is a combination of three or more microorganisms.

[0044] In the above scheme, the ARC microbial agent for improving nitrogen fixation and increasing green production can be, but is not limited to, a composition of the following four microorganisms: Bacillus laterosporus with a deposit number of CCTCC NO: M 20231601, Bacillus amyloliquefaciens with a deposit number of CCTCC NO: M 20231598, Enterobacter Ludwigii with a deposit number of CCTCC NO: M 20231595, and Bacillus mucilaginosus with a deposit number of CCTCC NO: M 20231602.

[0045] In the above scheme, the ARC microbial agent for improving nitrogen fixation and coupling green yield increase can be a combination of one or more microorganisms selected from Bacillus laterosporus with a deposit number of CCTCC NO: M 20231601, Bacillus amyloliquefaciens with a deposit number of CCTCC NO: M 20231598, Enterobacter Ludwigii with a deposit number of CCTCC NO: M 20231595, and Bacillus mucilaginosus with a deposit number of CCTCC NO: M 20231602, and other microorganisms, so that the combined microbial agent satisfies the requirement of containing all gene sequences in the nucleotide sequences shown in SEQ ID NOs. 1 to 4 or their functional equivalents, has the effect of coupling improving nitrogen fixation, regulating and increasing the abundance of rhizobia in the rhizosphere of legume crops, and increasing the number of nodules of legume crops. It has the effect of improving nitrogen fixation and increasing yield, regulating and increasing the abundance of rhizobia in the rhizosphere of legume crops, and increasing the number of nodules of legume crops, constituting the ARC microbial agent for improving nitrogen fixation and green yield increase of the present invention.

[0046] In the above scheme, in the ARC microbial agent for improving quality, fixing nitrogen and increasing green yield, the proportion of viable bacteria of any one strain in the microbial agent, i.e., the mixed microbial composition, is greater than or equal to 1%.

[0047] The beneficial effects of the present invention are:

[0048] 1. The ARC microbial inoculant, a green, yield-enhancing, nitrogen-fixing agent, can be used to increase the production of legumes such as peanuts and soybeans. This single technology improves the quality of legume products, further mitigates legume crop diseases, / or improves the quality and safety of crop products, and / or reduces peanut fruit spotting, while simultaneously inducing efficient nodulation and nitrogen fixation in soybeans and peanuts, significantly increasing yields. 2. It is simple to use, low-cost, and highly effective. 3. It is of great significance for promoting the expansion of soybean oilseed production capacity, high-quality development, and green, low-carbon, and efficient production in my country. Modes for Carrying Out the Invention

[0049] Example 1 Preparation of microorganisms and their compositions

[0050] Whole plants of peanuts, soybeans, peas, broad beans, cowpeas, and alfalfa, along with rhizosphere soil samples, were ground and mixed. Strains were isolated using conventional bacterial isolation methods and subsequently identified using conventional 16S rDNA analysis. Bacillus amyloliquefaciens, Brevibacillus laterosporus, Bacillus mucilaginosus, and Enterobacter ludwigii were obtained through these procedures. Details are shown in Table 1.

[0051] Table 1. Some of the strains isolated and identified from a mixture of major legume crops such as peanuts and soybeans are shown below:

[0052] Strain code or deposition number Strain name Strain code or deposition number Strain name CCTCC NO: M 20231601 Brevibacillus laterosporus strain 06 Brevibacillus laterosporus CCTCC NO: M 20231598 Bacillus amyloliquefaciens strain 07 Bacillus mucilaginosus CCTCC NO: M 20231602 Bacillus mucilaginosus strain 08 Enterobacter ludwigii CCTCC NO: M 20231595 Enterobacter ludwigii strain 09 Enterobacter ludwigii strain 05 Bacillus amyloliquefaciens strain 10 Bacillus mucilaginosus

[0053] The 10 microbial strains in Table 1 were amplified one by one by conventional bacterial culture medium amplification method to prepare batches of fermentation broth or bacterial powder of the 10 strains.

[0054] A bacterial agent obtained by mixing the fermentation broth or bacterial powder of the four strains of Bacillus laterosporus with a deposit number of CCTCC NO: M 20231601, Bacillus amyloliquefaciens with a deposit number of CCTCC NO: M 20231598, Enterobacter Ludwigii with a deposit number of CCTCC NO: M 20231595, and Bacillus mucilaginosus with a deposit number of CCTCC NO: M 20231602, and a bacterial agent obtained by mixing the fermentation broth or bacterial powder of the four strains of Bacillus laterosporus with a deposit number of CCTCC NO: M 20231601, Bacillus amyloliquefaciens with a deposit number of CCTCC NO: M 20231598, Enterobacter Ludwigii with a deposit number of CCTCC NO: M 20231595, Some strains of Bacillus subtilis 20231602 were mixed with other strains to form the ARC microbial composition for improving nitrogen fixation and green production, namely, ARC microbial agents 1 to 12 for improving nitrogen fixation and green production, and the combination information of the microbial agents formed by mixing some other strains is shown in Table 2, wherein the proportion of the viable count of any one strain in each microbial combination is greater than or equal to 1%.

[0055] Table 2. Microbial composition and bacterial agent information

[0056] Microbial agent number Microbial agent composition and proportion of viable bacteria count of its constituent strains (%) Microbial agent number Microbial agent composition and proportion of viable bacteria count of its constituent strains (%) Composition of agent 1: CCTCC NO: M 20231601 / CCTCC NO: M 20231598 / CCTCC NO: M 20231595 / CCTCC NO: M 20231602 Ratio: 33 / 33 / 1 / 33 Composition of agent 7: Strain 09 / CCTCC NO: M 20231598 / Strain 10 / CCTCC NO: M 20231601 Ratio: 33 / 1 / 33 / 33 Composition of agent 2: CCTCC NO: M 20231601 / CCTCC NO: M 20231598 / CCTCC NO: M 20231595 / CCTCC NO: M 20231602 Ratio: 1 / 33 / 33 / 33 Bacterial agent 8 composition: strain 09 / CCTCC NO: M 20231598 / strain 10 / CCTCC NO: M 20231601 Ratio: 1 / 33 / 33 / 33 Bacterial agent 3 composition: CCTCC NO: M 20231601 / CCTCC NO: M 20231598 / CCTCC NO: M 20231595 / CCTCC NO: M 20231602 Ratio: 33 / 33 / 33 / 1 Bacterial agent 9 composition: strain 05 / strain 06 / strain 07 / strain 08 Ratio: 33 / 33 / 33 / 1 Bacterial agent 4 composition: CCTCC NO: M 20231601 / CCTCC NO: M 20231598 / CCTCC NO: M 20231595 / CCTCC NO: M 20231602 Ratio: 33 / 1 / 33 / 33 Bacterial agent 10 Composition: Strain 05 / Strain 06 / Strain 07 / Strain 08 Ratio: 33 / 1 / 33 / 33 Bacterial agent 5 Composition: Strain 09 / CCTCC NO: M 20231598 / Strain 10 / CCTCC NO: M 20231601 Ratio: 33 / 33 / 33 / 1 Bacterial agent 11 Composition: Strain 05 / Strain 06 / Strain 07 / Strain 08 Ratio: 33 / 33 / 1 / 33 Bacterial agent 6 Composition: Strain 09 / CCTCC NO: M 20231598 / Strain 10 / CCTCC NO: M 20231601 Ratio: 33 / 33 / 1 / 33 Bacterial agent 12 Composition: Strain 05 / Strain 06 / Strain 07 / Strain 08 Ratio: 1 / 33 / 33 / 33

[0057] Example 2: Sequencing of microbial agents

[0058] A sufficient number of samples were taken from the microorganisms or microbial compositions, i.e., the bacterial agents, listed in Table 2 of Example 1. 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 document. The analysis results are shown in Table 1.

[0059] The results of the above determination and homology analysis showed that bacterial agents 1 to 4 in Table 2 contained all four gene sequences in DNA sequences 1 to 4 as shown in SEQ ID No. 1-4, with a homology of 100%; bacterial agents 5 to 12 contained all four genes in DNA sequences 1 to 4, but there were sequence variations, and the homology of their gene sequences with DNA sequences 1 to 4 was 90.2-100%, respectively.

[0060] Example 3: Determination of the effect of microbial inoculants on improving the quality and fixing nitrogen on peanuts

[0061] Taking peanuts as an example, the steps for determining the quality-enhancing and nitrogen-fixing effects of the above-mentioned microbial agents are as follows.

[0062] The microbial agents 1 to 12 in Table 2 of Example 1 above were applied to the field along with the peanut sowing base fertilizer, or during the peanut growing season, with an application rate of greater than or equal to 80 billion viable bacteria per mu. A plot not applied with any of the above microbial agents was set up as a control, and conventional field management was used for all other plots. Continuous investigations were conducted after emergence; during the first 30 days after emergence, an investigation was conducted on whether nodules had formed in advance; during the flowering and needle-setting period, an investigation was conducted on the number, weight, and nitrogenase activity of nodules per plant; and 1 to 15 days before harvest, an investigation was conducted on the nodulation of peanut roots during maturity, i.e., whether the nodulation time had been extended. The field disease reduction effect, i.e., disease prevention and control effect, was investigated from the seedling stage to the harvest stage; and after harvest, an investigation was conducted on the total plant biomass, peanut yield, peanut quality, including protein, glutamic acid, peanut resveratrol, and the abundance of aflatoxin, to determine whether the peanuts had improved quality and safety.

[0063] The above survey results are shown in Table 3.

[0064] According to the results in Table 3 above, agents 1-12 simultaneously possess the following characteristics: 1) they contain all four genes in DNA sequences 1-4, and these genes may have no more than 10% base variation in different strains; 2) the agents can improve the quality of legume crop products by increasing the content of one or more nutritional functional components such as protein, glutamic acid, and peanut resveratrol; 3) the agents have a mitigating effect on one or more soil-borne diseases such as peanut fruit rot, white rot, and bacterial wilt, or have a control effect, or reduce peanut fruit surface spots, or significantly reduce aflatoxin levels, thereby improving quality and safety; 4) although they are 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 increase the nitrogenase activity per plant; 5) the agents can promote early nodulation and prolong the time of nodulation and nitrogen fixation in legume crops such as peanuts; and 6) the agents can increase the yield and total biomass of legume crops.

[0065] Table 3. Results of the determination of the effect of microbial agents on improving the quality of peanuts and fixing nitrogen

[0066] Does the microbial composition, i.e., the agent number, improve quality? Does it have a preventive and control effect on peanut fruit rot, white rot, bacterial wilt, and other diseases? Or does it reduce peanut fruit surface spots? Or does it significantly reduce aflatoxin levels and improve quality and safety levels? Does it increase the abundance of rhizobia in the rhizosphere, double the promotion of nodulation and nitrogen fixation, advance nodulation, and prolong the time of nodulation and nitrogen fixation? Does it significantly increase yield and total biomass? Agent 1 Yes Yes Yes Yes Agent 2 Yes Yes Yes Yes Agent 3 Yes Yes Yes Yes Agent 4 Yes Yes Yes Yes Agent 5 Yes Yes Yes Yes Agent 6 Yes Yes Yes Yes Agent 7 Yes Yes Yes Yes Agent 8 Yes Yes Yes Yes Agent 9 Yes Yes Yes Yes Agent 10 Yes Yes Yes Yes Agent 11 Yes Yes Yes Agent 12 Yes Yes Yes Yes

[0067] The above-mentioned fungal agent can improve the quality of peanut products of leguminous crops - increase the content of one or more nutritional functional ingredients in peanut products such as protein, glutamic acid, peanut resveratrol, etc.: Peanut quality is improved: the protein content is increased by more than 0.2%, and preferably can reach more than 3%, the total content of essential amino acids for the human body such as leucine is increased by 0.15 mg / kg, and preferably, can reach more than 0.2 mg / kg, and the peanut resveratrol content is increased by more than 0.2 mg / kg, and more preferably, can reach more than 0.4 mg / kg.

[0068] Example 4: Determination of the Effect of Soybean Microbial Agent on Quality Improvement and Nitrogen Fixation

[0069] Taking soybean as an example, the steps for determining the quality improvement and nitrogen fixation effects of the above-mentioned microbial agents are described as follows.

[0070] Microbial agents 1-12 from Table 2 of Example 1 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 plot not treated with any of the microbial agents was set up as a control, while all other plots were managed using conventional field management. Continued surveys were conducted after seedling emergence. Early nodulation was investigated within the first 30 days after seedling emergence. Nodule number, nodule weight, and nitrogenase activity per plant were investigated during flowering and podding. Root nodulation during the mature soybean period was investigated 1-15 days before harvest to determine whether the nodulation time was prolonged. Field disease mitigation, i.e., disease control efficacy, was investigated from the seedling stage to the harvest stage. After harvest, total plant biomass, soybean yield, soybean quality, including protein, glutamic acid, soy isoflavones, and the abundance of aflatoxin were investigated to determine whether the soybeans improved quality and safety.

[0071] The above survey results are shown in Table 4. Combined with the sequencing results of Example 1, it is shown that microbial agents 1-12 simultaneously possess the following characteristics: 1) they contain all four genes in DNA sequences 1-4, and these gene sequences may have no more than 10% base variation among different strains; 2) the microbial agents can improve the quality of legume crop products by increasing the content of one or more nutritional functional components such as protein, glutamic acid, and soy isoflavones; 3) the microbial agents have a mitigating effect on one or more soil-borne diseases such as peanut fruit rot, soybean green disease, white rot of legumes, bacterial wilt, root rot, and sclerotinia, or have a control effect, or reduce peanut fruit surface spots, or significantly reduce aflatoxin levels, thereby improving quality and safety; 4) although not rhizobia themselves, the microbial 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 increase the nitrogenase activity per plant; 5) the microbial agents can promote early nodulation and prolong the time of nodulation and nitrogen fixation in legume crops such as peanuts and soybeans; and 6) the microbial agents can increase the yield and total biomass of legume crops.

[0072] Table 4. Results of microbial inoculants on soybean quality improvement and nitrogen fixation

[0073] Does the microbial composition, i.e., the agent number, improve the quality? Does it have a preventive and control effect on soil-borne diseases such as soybean greening, root rot, and sclerotinia, or significantly improve the quality and safety level? Does it double the promotion of nodulation and nitrogen fixation, early nodulation, and extended nodulation and nitrogen fixation time? Does it significantly increase yield and total biomass? Agent 1 Yes Yes Yes Yes Agent 2 Yes Yes Yes Yes Agent 3 Yes Yes Yes Yes Agent 4 Yes Yes Yes Yes Agent 5 Yes Yes Yes Yes Agent 6 Yes Yes Yes Yes Agent 7 Yes Yes Yes Yes Agent 8 Yes Yes Yes Yes Agent 9 Yes Yes Yes Yes Agent 10 Yes Yes Yes Yes Agent 11 Yes Yes Yes Yes Agent 12 Yes Yes Yes Yes

[0074] The above-mentioned microbial agent can improve the quality of soybean products of leguminous crops - increasing the content of one or more nutritional functional ingredients in soybean products such as protein, glutamic acid, and soy isoflavones: the protein content is increased by more than 0.3%, and preferably can reach more than 3%; the content of essential amino acids for the human body such as leucine is increased by more than 0.2 mg / kg in total, and preferably can reach more than 0.3 mg / kg; the soy isoflavone content is increased by more than 50 mg / kg, and preferably can reach more than 200 mg / kg.

[0075] Example 5: Determination of the Effect of Microbial Agents on Quality Improvement and Nitrogen Fixation of Other Leguminous Crops

[0076] The sampling method and steps similar to those in Examples 3 and 4 were used to determine the effect of microbial agents on the quality improvement and nitrogen fixation of other legume crops such as peas, broad beans, cowpeas, and alfalfa, and the results were similar to those of peanuts and soybeans mentioned above, and similar to the results in Tables 3 and 4.

[0077] Part 2: Application of ARC microbial agents for improving nitrogen fixation and increasing green production

[0078] Example 6: Application of ARC Microbial Agent for Green Yield Improvement and Nitrogen Fixation - Improving the Quality and Safety of Leguminous Crop Products

[0079] 1. Application on peanuts: Mix the above-mentioned microbial agents 1 to 8 with the peanut seeding base fertilizer respectively and apply them to the field through a seed drill. The application rate of the 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-mentioned microbial agents as a control. The rest are managed by conventional field management. After the soybeans 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 soybeans is calculated, which is the control effect on aflatoxin-producing fungi. (2) The content of isoflavones, protein, and glutamic acid in soybeans is determined by the standard method, and the quality improvement level is calculated. (3) 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 showed that agents 1-8 were all over 60% effective against aflatoxin-producing fungi in field peanuts and over 75% effective against aflatoxin in peanuts. They also significantly increased the resveratrol, protein, and essential amino acid content in peanuts: protein content increased by over 0.2%, the combined content of essential amino acids such as leucine increased by 0.15 mg / kg, and resveratrol content in peanuts increased by over 0.2 mg / kg. These results indicate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, has significantly improved the quality and safety of field peanuts.

[0080] 2. Application on Soybeans: Before soybean sowing, agents 1-8 were applied to the field using drones at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million saplings). Plots not treated with any of the agents served as controls, while all other plots were managed conventionally. After harvest, soybean samples were collected using national standard sampling methods and assayed for isoflavone, protein, and amino acid content using standard methods to calculate the improvement in soybean quality. These assays showed that the application of agents 1-8 significantly increased the isoflavone, protein, and essential amino acid content of soybeans grown in the field: protein content increased by over 0.3%, the total content of essential amino acids, including leucine, increased by over 0.2 mg / kg, and isoflavone content increased by over 50 mg / kg. These results demonstrate that the application of the ARC microbial agent, a quality-enhancing, nitrogen-fixing, and green yield-increasing agent, significantly improved soybean quality in the field.

[0081] 3. The application on other leguminous crops has achieved similar results as the above-mentioned application on soybeans.

[0082] Example 7: Use of ARC microbial agent for improving nitrogen fixation and increasing yield in green manner - promoting nodulation and nitrogen fixation in leguminous crops

[0083] 1. Application on Peanuts: Mix the aforementioned agents 1-8 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 investigations were conducted on the nodulation of peanut roots. These investigations revealed that during the flowering phase, the number of root nodules in peanuts treated with agents 1-8 increased by more than 2.6 times, and the nitrogenase activity per peanut plant increased by more than 8.5 times. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-improving, and yield-enhancing agent, significantly promoted the nodulation and nitrogen fixation of peanuts in the field.

[0084] 2. Application on Soybeans: Before soybean sowing, agents 1-8 were applied to the fields via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million hectares). 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 revealed that during the flowering phase, the number of root nodules in soybeans treated with agents 1-8 increased by over 2.5 times, and the nitrogenase activity per soybean plant increased by over 7.9 times. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-improving, and yield-enhancing agent, significantly promoted soybean nodulation and nitrogen fixation in the field.

[0085] 3. When applied to other leguminous crops, it achieved similar effects in promoting nodulation and nitrogen fixation as in peanuts and soybeans.

[0086] Example 8: Application of ARC microbial agent for improving quality, fixing nitrogen and increasing yield in green way - increasing the yield of leguminous crops

[0087] 1. Application on Peanuts: Mix the aforementioned agents 1-8 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 time, peanuts were harvested from one mu (approximately 1.5 acres) of fields treated with the agents and the control plots. Calculations showed that the yield increase in the fields treated with agents 1-8 was over 10% to 15%, achieving a significant yield increase. These results demonstrate that the application of the ARC microbial agent, a quality-enhancing, nitrogen-fixing, green yield-increasing agent, significantly increased the yield per unit area of ​​peanuts in the field.

[0088] 2. Application on Soybeans: Before soybean sowing, the aforementioned agents 1-8 were evenly applied to the field 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 treatment and control plots. Calculations showed that peanut yield increases in the fields treated with agents 1-8 ranged from 15% to 20%, achieving significant yield increases. These results demonstrate that the application of the ARC microbial agent, a quality-enhancing, nitrogen-fixing, green, and yield-increasing agent, significantly increased soybean yields per unit area in the field.

[0089] 3. Application on Peas: Before sowing peas, the aforementioned agents 1-8 were evenly applied to the field by hand broadcasting, with a cumulative application rate of 80 to 100 billion viable bacteria per mu. A plot without any of the agents was set up as a control, while all other plots were managed using conventional methods. Yields were measured by weighing the pea seedlings and green pea pods at harvest time. The yield increases for both pea seedlings and pea pods treated with agents 1-8 ranged from 7% to 15%, achieving significant yield increases. These results demonstrate that the application of the ARC microbial agent, a quality-enhancing, nitrogen-fixing, green yield-increasing agent, significantly increased the yield per unit area of ​​pea seedlings and pea pods in the field.

[0090] 4. Application on other legume crops: Application of agents 1-8 on legume crops such as broad beans yielded similar results to those on soybeans. Application of agents 1-8 on kidney beans and cowpeas yielded similar results to those on peas. Application of agents 1-8 on alfalfa also yielded similar results to those on peas.

[0091] Example 9: Use of ARC Microbial Agent for Green Yield Enhancement and Nitrogen Fixation - Recruiting Indigenous Rhizobia to Increase Rhizobium Abundance in the Rhizosphere Soil of Leguminous Crops

[0092] 1. Application on Peanuts: Mix the aforementioned agents 1-8 with the peanut seeding base fertilizer and apply them to the field using a seed drill. The application rate should be 80 to 100 billion viable bacteria per mu. A control plot should be established where none of the agents were applied. All other plots should be managed conventionally. Peanut rhizosphere soil samples were collected during the fruiting period and before harvest, and metagenomic sequencing and big data analysis were performed. These findings showed that the abundance of rhizobia in the rhizosphere soil of peanuts treated with agents 1-8 was significantly higher than that in the control group, increasing by 10% to 25%. These results indicate that the application of the ARC microbial agent, which improves nitrogen fixation and increases yield, has a significant effect in recruiting indigenous rhizobia, thereby increasing the abundance of rhizobia in the rhizosphere soil of leguminous crops.

[0093] 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.

[0094] Example 10: Use of ARC microbial agent for improving quality, fixing nitrogen and increasing yield: promoting early nodulation of leguminous crops and extending the time of nodulation and nitrogen fixation

[0095] 1. Application to Peanuts: Mix the aforementioned agents 1-8 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 others were managed under conventional practices. Root nodulation was continuously monitored after the peanuts emerged. These results showed that root nodules formed approximately 7 to 9 days after emergence in peanuts treated with agents 1-8, more than 20 days earlier than the conventional estimate of 30 days. New nodules were still observed during the full-fruiting stage, with the presence of old, middle-aged, and young nodules, as well as infant nodules, 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 ARC microbial agent for improving quality, fixing nitrogen and increasing green yield has achieved early nodulation of peanut legume crops and extended the nodulation and nitrogen fixation time.

[0096] 2. Application in Soybeans: Before soybean sowing, the aforementioned agents 1-8 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. Continuous investigations of soybean root nodulation were conducted after seedling emergence. These investigations revealed that soybeans treated with agents 1-8 developed root nodules approximately 6-8 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, viable 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 have decayed by harvest. These results demonstrate that the application of the ARC microbial agent, designed to improve quality, fix nitrogen, and increase yield, has achieved earlier nodulation and prolonged the duration of nodulation and nitrogen fixation.

[0097] 3. When applied to other leguminous crops, the above-mentioned microbial agents 1 to 8 achieved similar effects as those achieved in peanuts and soybeans, namely, promoting early nodulation and extending the nodulation and nitrogen fixation time of leguminous crops.

[0098] Example 11: Application of ARC microbial agent for improving quality, fixing nitrogen and increasing yield in green manner - preventing premature aging of leguminous crops during maturity

[0099] 1. Application on Peanuts: Mix the aforementioned agents 1-8 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 million acre) should be achieved. A control plot without any of the above agents should be established, while all other plots should be managed conventionally. Once the peanuts reach maturity, continuous monitoring of leaf photosynthetic rates and senescence should be conducted. These results show that leaf photosynthetic rates in peanuts treated with agents 1-8 increased by 20% to 45% compared to the control, demonstrating a significant effect in preventing premature aging due to nutrient deficiency. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, yield-enhancing agent, can effectively prevent premature aging due to nutrient deficiency in leguminous crops during maturity.

[0100] 2. Application on Soybeans: Before soybean sowing, agents 1-8 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 methods. After soybean seedlings emerged, continuous investigations were conducted on soybean root nodulation. These findings revealed that soybeans treated with agents 1-8 experienced increases in leaf photosynthetic rates of 25% to 50% or more, demonstrating a significant protective effect against premature aging caused by nutrient depletion. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, yield-enhancing agent, achieved earlier nodulation and prolonged the duration of nodulation and nitrogen fixation.

[0101] 3. Application in other leguminous crops: The application of fungal agents 1 to 8 in other leguminous crops such as peas, broad beans, and cowpeas has achieved similar effects as their application in peanuts and soybeans in preventing premature aging during maturity.

[0102] Example 12: Use of ARC microbial agent for improving quality, fixing nitrogen and increasing yield in green manner - increasing the number of leguminous crop pods

[0103] 1. Application on Peanuts: Mix the aforementioned agents 1-8 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 1-8 increased by 15% to 25% compared to the control, significantly increasing pod count. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, can significantly increase peanut pod count.

[0104] 2. Application on Soybeans: Before sowing, agents 1-8 were applied to the fields using drones at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 to 100 million active bacteria per acre). 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 10% to 20% or more in the soybeans treated with agents 1-8, significantly increasing pod count. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, can significantly increase soybean pod count.

[0105] 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.

[0106] Example 13: Use of the ARC microbial agent for improving quality, fixing nitrogen, and increasing yield – increasing leguminous crop pod plumpness and reducing pod shrinkage

[0107] 1. Application on Peanuts: Mix the aforementioned agents 1-8 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 1-8 significantly improved pod plumpness and reduced pod shriveling rate by over 7%. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly increased peanut pod plumpness and reduced pod shriveling rate.

[0108] 2. Application on Soybeans: Before soybean sowing, agents 1-8 were applied to the field using drones 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 calculated. These survey results showed that soybeans treated with agents 1-8 showed significantly improved pod plumpness and a reduction in pod shriveling rates of over 6%. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly increased soybean pod plumpness and reduced pod shriveling rates.

[0109] 3. The application of microbial agents 1 to 8 on other leguminous crops such as peas, broad beans, kidney beans, and cowpeas achieved similar effects as their application on peanuts and soybeans, increasing the plumpness of peanut pods and reducing the rate of shrunken pods.

[0110] Example 14: Use of the ARC microbial agent for improving quality, fixing nitrogen, and increasing yield in a green manner - promoting early flowering and pod formation in leguminous crops

[0111] 1. Application to Peanuts: Mix the aforementioned agents 1-8 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 1-8 flowered and pod set at least two days earlier. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly promotes earlier flowering and pod setting in peanuts.

[0112] 2. Application on Soybeans: Before soybean sowing, agents 1-8 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 survey results showed that soybeans treated with agents 1-8 flowered and pod set at least two days earlier. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly promotes earlier flowering and pod setting in soybeans.

[0113] 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.

[0114] Example 15: Use of ARC Microbial Agent for Green Yield Improvement and Nitrogen Fixation - Alleviating Peanut Fruit Rot

[0115] The aforementioned agents 1-8 were mixed with peanut seeding 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 others 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 1-8 was reduced by 50% to 70% compared to the control group. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly reduces peanut fruit rot.

[0116] Example 16: Use of ARC Microbial Agent for Green Yield Improvement and Nitrogen Fixation - Alleviating the Occurrence of Bacterial Wilt in Leguminous Crops

[0117] 1. Application to Peanuts: Mix the aforementioned agents 1-8 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, ongoing 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 peanuts treated with agents 1-8 was reduced by 55% to 73% compared to the control group. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly reduces the incidence of peanut bacterial wilt.

[0118] 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.

[0119] Example 17: Use of ARC Microbial Agent for Green Yield Improvement and Nitrogen Fixation - Reducing Powdery Mildew in Leguminous Crops

[0120] 1. Application to Peanuts: Mix the aforementioned agents 1-8 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 million acre) was achieved. A control plot was established without any of the agents, while all other plots were managed conventionally. After the peanuts emerged, ongoing 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 1-8 was reduced by over 50% compared to the control group. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly reduces the incidence of powdery mildew in peanuts.

[0121] 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.

[0122] Example 18: Use of ARC Microbial Agent for Green Yield Improvement and Nitrogen Fixation - Reducing the Occurrence of Leaf Spot Disease in Leguminous Crops

[0123] 1. Application to Peanuts: Mix the aforementioned agents 1-8 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, ongoing surveys were conducted to statistically analyze the incidence of peanut leaf spot compared to the control plots. These survey results showed that the incidence of peanut leaf spot in the peanuts treated with agents 1-8 was reduced by 55% to 75% compared to the control group. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly reduces the incidence of peanut leaf spot.

[0124] 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.

[0125] Example 19: Use of ARC Microbial Agent for Green Yield Improvement and Nitrogen Fixation - Reducing the Occurrence of Root Nematode Disease in Leguminous Crops

[0126] 1. Application to Peanuts: Mix the aforementioned agents 1-8 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, ongoing surveys were conducted to statistically analyze the incidence of root nematodes in the peanuts compared to the control plots. These survey results showed that the incidence of root nematodes in the peanuts treated with agents 1-8 was reduced by 50% to 65% compared to the control plots. These results demonstrate that the application of the ARC microbial agent, which improves quality, fixes nitrogen, and increases yield, significantly reduces root nematode disease in peanuts.

[0127] 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.

[0128] Example 20: Use of ARC Microbial Agent for Green Yield Improvement and Nitrogen Fixation - Alleviating the Occurrence of Root Rot in Leguminous Crops

[0129] 1. Application to Peanuts: Mix the aforementioned agents 1-8 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, ongoing surveys were conducted, and statistical analysis was performed to compare the incidence of peanut root rot with the control plots. These survey results showed that the incidence of peanut root rot in the peanuts treated with agents 1-8 was reduced by over 55% compared to the control plots. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly reduces peanut root rot.

[0130] 2. Application on Soybeans: Mix the aforementioned agents 1-8 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 should be established, while all other plots should be managed conventionally. After soybean seedlings emerge, conduct ongoing surveys to statistically analyze the incidence of soybean root rot compared to the control plots. These survey results show that peanut root rot incidence in the plots treated with agents 1-8 was reduced by over 55% compared to the control plots. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly reduces soybean root rot.

[0131] 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.

[0132] Example 21: Use of ARC Microbial Agent for Green Yield Improvement and Nitrogen Fixation - Reducing the Occurrence of Sclerotinia Blight in Leguminous Crops

[0133] 1. Application on Soybeans: Mix the aforementioned agents 1-8 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 hectares). Set up plots without any of the agents as controls, while all other plots receive conventional field management. After soybean seedlings emerge, conduct ongoing surveys to statistically analyze the incidence of sclerotinia rot in soybeans compared to the control. These survey results show that soybeans treated with agents 1-8 experienced a 50% to 65% reduction in phytophthora blight compared to the control group. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly reduces soybean sclerotinia rot.

[0134] 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.

[0135] Example 22: Use of ARC Microbial Agent for Green Yield Improvement and Nitrogen Fixation - Alleviating Downy Mildew in Leguminous Crops

[0136] 1. Application on Soybeans: Mix the aforementioned agents 1-8 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 hectares). Set up plots without any of the agents as controls, while all other plots receive conventional field management. After soybean seedlings emerge, conduct ongoing surveys to statistically analyze the incidence of downy mildew in soybeans compared to the control. These survey results show that soybeans treated with agents 1-8 experienced a reduction of over 60% in downy mildew compared to the control group. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly reduces soybean downy mildew.

[0137] 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.

[0138] Example 23: Use of ARC Microbial Agent for Green Yield Improvement and Nitrogen Fixation - Alleviating the Occurrence of Fusarium Wilt in Leguminous Crops

[0139] 1. Application on Soybeans: Mix the aforementioned agents 1-8 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 to 100 million acre). Set up plots without any of the agents as controls, while all other plots receive conventional field management. After soybean seedlings emerge, conduct ongoing surveys to statistically analyze the incidence of soybean wilt compared to the control. These survey results show that soybeans treated with agents 1-8 experienced a reduction of nearly 60% in soybean wilt compared to the control group. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly reduces soybean wilt.

[0140] 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.

[0141] Example 24: Use of ARC Microbial Agent for Improving Nitrogen Fixation and Green Yield Increase - Reducing the Occurrence of Sclerotium rot in Leguminous Crops

[0142] 1. Application to Peanuts: Mix the aforementioned agents 1-8 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 million acre) was achieved. A control plot was established without any of the agents, while all other plots were managed conventionally. After the peanuts emerged, ongoing surveys were conducted to statistically analyze 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 1-8 was reduced by over 65% compared to the control plots. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly reduces the incidence of white rot in peanuts.

[0143] 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.

[0144] Example 25: Use of ARC Microbial Agent for Quality Improvement, Nitrogen Fixation, and Green Yield Increase - Reducing the Incidence of Soybean Greening

[0145] The aforementioned microbial agents 1-8 were mixed with soybean 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 (approximately 1.5 to 2.5 acres). A control plot was established without any of the aforementioned agents, while all other plots were managed conventionally. After soybean seedlings emerged, continuous surveys were conducted to determine the incidence of soybean greening compared to the control plots. These survey results showed that soybean greening in the plots treated with agents 1-8 was reduced by 35% to 55% compared to the control group. These results demonstrate that the application of the ARC microbial agent, a green, yield-enhancing, and nitrogen-fixing agent, significantly reduces the incidence of soybean greening.

[0146] Reduce the incidence of corn ear rot and Fusarium toxins

[0147] Example 26: Application of ARC Microbial Agent for Green Yield Increase and Quality Improvement of Nitrogen Fixation - Promoting Carbon Emission Reduction in Leguminous Crops and Benefiting Soil Improvement

[0148] 1. Application to Peanuts: Mix the aforementioned agents 1-8 with a base fertilizer for peanut sowing—reducing the nitrogen content of the base fertilizer by 20% and 30% compared to the conventional control. Apply the agents to the field using a seed drill, at a cumulative rate of 80 to 100 billion viable bacteria per mu. A plot without any of the agents was set up as a control, while all other plots were managed conventionally. Continued surveys were conducted after the peanuts emerged, and the carbon reduction levels of the treated and control peanuts were statistically analyzed. These survey results showed that peanuts treated with agents 1-8 experienced a 20% to 30% reduction in carbon dioxide emissions, while also significantly increasing peanut yields by over 10%. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly reduces carbon emissions during peanut production.

[0149] 2. Application on Soybeans: Before soybean sowing, agents 1-8 were applied to the fields via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million saplings). Nitrogen fertilizer was reduced by 20% and 50% in the treatment groups compared to the control group's conventional fertilization level. Plots not receiving any of the agents served as controls, while all other plots were managed conventionally. After soybean seedlings emerged, continuous surveys were conducted to assess carbon emission reductions in the treated and control soybeans. These survey results showed that soybeans treated with agents 1-8 experienced a 20% to 50% reduction in carbon dioxide emissions, while also significantly increasing soybean yields by 7% to 22%. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly reduces carbon emissions during soybean production.

[0150] 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.

[0151] Example 27: Use of ARC microbial agent for improving quality, fixing nitrogen and increasing yield in a green way - promoting the increase of total biomass of leguminous crops

[0152] 1. Application on Peanuts: Mix the aforementioned agents 1-8 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. 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 1-8 increased by more than 16%. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly increases the total biomass of peanut plants.

[0153] 2. Application on Soybeans: Before soybean sowing, agents 1-8 were applied to the fields via drone at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 million acres). A control plot was established without any of the agents, while all other plots were managed conventionally. A survey was conducted at soybean maturity, and the total biomass of the treated and control soybean plants was calculated. These survey results showed that soybeans treated with agents 1-8 had an increase in total plant biomass of over 10%. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly increases soybean plant biomass.

[0154] 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.

[0155] Example 28: Use of ARC Microbial Agent for Green Yield Improvement and Nitrogen Fixation - Reducing the abundance of harmful organisms such as Aspergillus terreus and Fusarium in the rhizosphere of leguminous crops, which is beneficial for improving the soil microbial population structure

[0156] 1. Application on Peanuts: Mix the aforementioned agents 1-8 with the peanut seeding base fertilizer and apply them to the field via a seed drill at a cumulative rate of 80 to 100 billion viable bacteria per mu (approximately 100 to 100 million active bacteria per mu). A control plot without any of the above agents was established, 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 findings showed that the abundance of Aspergillus and Fusarium in the rhizosphere of peanuts treated with agents 1-8 decreased by between 47% and 67%. These results demonstrate that the application of the ARC microbial agent, which improves quality, fixes nitrogen, and increases yield, significantly reduces the abundance of pests such as Aspergillus and Fusarium in the rhizosphere of leguminous crops.

[0157] 2. Application on Soybeans: Agents 1-8 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 hectares). A control plot was established without any of the 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 samples. These results showed that the abundance of Aspergillus and Fusarium in the rhizosphere of soybeans treated with agents 1-8 decreased by between 45% and 69%. These results demonstrate that the application of the ARC microbial agent, which improves quality, fixes nitrogen, and increases yield, significantly reduces the abundance of pests such as Aspergillus and Fusarium in the rhizosphere of legume crops.

[0158] 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.

[0159] Example 29: Application of ARC microbial agent for improving quality, fixing nitrogen and increasing yield: reducing surface spots on peanuts and improving marketability

[0160] Application on peanuts: Mix the aforementioned agents 1-8 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 others were managed using conventional field management. At harvest, surveys were conducted to determine the presence of black spots on the peanut shells of the treated and control peanuts. These survey results showed that the presence of black spots on the peanut shells of the peanuts treated with agents 1-8 was reduced by over 66%. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly reduces surface spots on peanut shells, enhancing marketability and competitiveness, and increasing profitability.

[0161] Example 30: Use of ARC microbial agent for improving quality, fixing nitrogen and increasing yield in green manner - promoting crop resistance to repeated cropping

[0162] Continuous planting of crops for many years often leads to the accumulation of crop pathogens, resulting in weak seedlings, diseased seedlings, and missing seedlings. Taking the application of the above-mentioned microbial agents 1 to 8 as an example, the use of ARC microbial agents for improving quality, fixing nitrogen, and increasing green yield to promote crop resistance to continuous planting is described.

[0163] 1. Application in Peanuts: A trial demonstrating resistance to repeated cropping was conducted in fields in Xingcheng, Liaoning Province, where peanuts had been planted continuously for many years. The aforementioned microbial agents 1-8 were mixed with peanut sowing base fertilizer and applied to the fields using a seeder. The application rate was 80 to 100 billion viable bacteria per mu. A control plot was set up where none of the above agents were applied, while all other plots were managed using conventional methods. Regular and ongoing surveys were conducted after the peanuts were sown. These survey results showed that peanuts treated with agents 1-8 had stronger seedlings and a significantly lower number of diseased plants compared to the control group. The disease rate in the control group was over 4.5 times that of the treated group. These results indicate that the application of the ARC microbial agent, which improves quality, fixes nitrogen, and increases yield, significantly promotes resistance to repeated cropping.

[0164] 2. Application in Soybeans: Fields planted with soybeans for multiple years were selected for a trial demonstrating resistance to repeated cropping. Agents 1-8 were mixed with soybean seeding base fertilizer and applied via drone. The application rate reached 80 to 100 billion viable bacteria per mu (approximately 100 million sq ft). Plots not treated with any of the agents served as controls, while all other plots were managed using conventional methods. Regular and ongoing surveys were conducted after soybean sowing. These surveys showed that soybean seedlings treated with agents 1-8 were more robust and had 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 group. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and yield-enhancing agent, significantly enhances crop resistance to repeated cropping.

[0165] 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.

[0166] Example 31: Application of ARC Microbial Agent for Quality Improvement, Nitrogen Fixation, and Green Yield Increase - Promoting Soybean Yield Increase in Saline-Alkali Land

[0167] The pilot demonstration was conducted in saline-alkali soil with a pH range of 8.1 to 9.2. Before soybean sowing, agents 1 to 8 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 at harvest time. These surveys showed that soybean yields increased by 10% to 35% in the soybeans treated with agents 1 to 8. These results demonstrate that the application of the ARC microbial agent, a green, nitrogen-fixing, and quality-enhancing ARC microbial agent, significantly boosts soybean yields in saline-alkali soil.

[0168] Example 32: Use of ARC microbial agent for improving quality and fixing nitrogen and increasing yield in a green way - Preparation of microbial compound fertilizer for improving quality and fixing nitrogen

[0169] Based on the amount of compound fertilizer used per mu of land and a minimum of 100 billion viable bacteria count of the ARC Microbial Agent for Quality-Improving, Nitrogen-Fixing, and Green Yield-Increasing, the compound fertilizer was mixed with ARC Microbial Agents 1-8. The quality-improving, nitrogen-fixing, and green yield-increasing microorganisms were then adsorbed and fixed to the compound fertilizer using conventional physical methods to create eight types of quality-improving, nitrogen-fixing, and coupled microbial compound fertilizers 1-8. These compound fertilizers are inorganic and can be purchased commercially or prepared using inorganic fertilizers such as nitrogen, phosphorus, and potassium fertilizers according to common mixing ratios.

[0170] The eight quality-enhancing, nitrogen-fixing coupled microbial compound fertilizers 1-8 were used as seed fertilizers in the production of leguminous crops such as peanuts and soybeans at the time of sowing. Fields treated with the same compound fertilizers served as controls, and ongoing surveys were conducted after sowing. These survey results showed that the eight quality-enhancing, nitrogen-fixing coupled microbial compound fertilizers 1-8 significantly promoted nodulation and nitrogen fixation, increasing nodule number by 2.3 to 13 times, increasing nitrogenase activity per plant by more than 5.65 times, and increasing yield per unit area by more than 6.5% to 27%, achieving significant yield increases and demonstrating the key characteristics of the quality-enhancing, nitrogen-fixing, green, and yield-increasing ARC microbial inoculants described in the above examples.

[0171] The above results show that the ARC microbial agent for improving nitrogen fixation and increasing green yield can be used to prepare a microbial compound fertilizer for improving nitrogen fixation and coupling with quality improvement, and it has the effects of both compound fertilizer and ARC microbial agent for improving nitrogen fixation and increasing green yield.

[0172] Example 33: Use of ARC Microbial Agent for Quality Improvement and Nitrogen Fixation to Increase Green Yield - Configuration of Quality Improvement and Nitrogen Fixation Coupled Microorganism-Organic Fertilizer

[0173] Based on the amount of bio-organic fertilizer used per mu of land and the amount of ARC microbial agent for improving quality, nitrogen fixation, and green yield increase of at least 80 billion viable bacteria, the bio-organic fertilizer and ARC microbial agents 1 to 8 for improving quality, nitrogen fixation, and green yield increase are respectively mixed. The quality-improving, nitrogen-fixing coupled microorganisms are then adsorbed and fixed to the bio-organic fertilizer using conventional physical methods to create eight types of quality-improving, nitrogen-fixing coupled microorganisms-bio-organic fertilizers 1 to 8. The above bio-organic fertilizers can be purchased commercially.

[0174] The eight quality-enhancing, nitrogen-fixing coupled microorganisms and bio-organic fertilizers 1-8 were used as seed fertilizers in the production of leguminous crops such as peanuts and soybeans. Fields treated with the same bio-organic fertilizers served as controls, and ongoing surveys were conducted after sowing. These survey results showed that the eight quality-enhancing, nitrogen-fixing coupled microorganisms and bio-organic fertilizers 1-8 significantly promoted nodulation and nitrogen fixation, increasing nodule number by more than 2-fold, nitrogenase activity per plant by more than 5-fold, and yield per unit area by 6.5% to 37%, achieving significant yield increases. These results demonstrate the key characteristics of the quality-enhancing, nitrogen-fixing, and green yield-increasing ARC microbial inoculants described in the above examples.

[0175] The above results show that the ARC microbial agent for improving nitrogen fixation and increasing green yield can be used to prepare the coupled microbial-bioorganic fertilizer for improving nitrogen fixation and improving quality, and it has the effects of both bio-organic fertilizer and ARC microbial agent for improving nitrogen fixation and increasing green yield.

[0176] Example 34: Use of ARC Microbial Agent for Quality Improvement and Nitrogen Fixation to Increase Green Yield - Preparation of Quality Improvement and Nitrogen Fixation Coupled Microbial-Trace Element Fertilizer

[0177] Taking the application of the above-mentioned microbial agents 1 to 8 as an example, the use of the ARC microbial agent for improving nitrogen fixation and increasing yield in the preparation of quality-improving nitrogen fixation coupled microbial-trace element fertilizer is described.

[0178] Based on the micronutrient fertilizer dosage per mu (approximately 1 acre) and a minimum viable count of 80 billion ARC microbial agents for improving nitrogen fixation and increasing yield, mix the trace element fertilizers with ARC microbial agents 1 to 8 for improving nitrogen fixation and increasing yield, respectively. Mix the mixture using conventional physical methods to create eight types of coupled microbial and trace element fertilizers for improving nitrogen fixation and increasing yield. These trace element fertilizers can be purchased commercially.

[0179] The eight nitrogen-enhancing, coupled microbial-trace element fertilizers 1-8 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 eight nitrogen-enhancing, coupled microbial-trace element fertilizers 1-8 significantly promoted nodulation and nitrogen fixation, increasing nodule number by more than 2-fold, nitrogenase activity per plant by more than 5-fold, and yield per unit area by more than 7% to 20%, achieving significant yield increases. These results demonstrate the key characteristics of the ARC microbial inoculants described in the aforementioned examples for improving nitrogen, fixing, and increasing yields.

[0180] The above results show that the ARC microbial agent for improving nitrogen fixation and increasing green yield can be used to prepare a quality-improving nitrogen fixation coupled microbial-trace element fertilizer, and it has the effects of both trace element fertilizer and quality-improving nitrogen fixation and green yield-increasing ARC microbial agent.

[0181] Example 35: Use of ARC microbial agent for improving quality, fixing nitrogen and increasing yield in a green way - Preparation of microbial fertilizer for improving quality, fixing nitrogen and coupling it with moisturizing fertilizer

[0182] Based on the water-retaining agent dosage and the use of at least 80 billion viable bacteria of the ARC microbial agent for improving quality, nitrogen, and green yield, per mu of land, mix the water-retaining agent with ARC microbial agents 1-8 for improving quality, nitrogen, and green yield, respectively. Mix the water-retaining agent using conventional physical methods, or adsorb and immobilize the quality-improving, nitrogen-fixing coupled microorganisms onto the water-retaining agent to create eight types of quality-improving, nitrogen-fixing coupled microorganism-water-retaining fertilizers 1-8. The water-retaining agents are commercially available.

[0183] The eight nitrogen-enhancing microbial-water-retaining fertilizers (1-8) 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, and ongoing surveys were conducted after sowing. These survey results showed that the eight nitrogen-enhancing microbial-water-retaining fertilizers significantly promoted nodulation and nitrogen fixation, increasing nodule number by more than 2.6 times, increasing nitrogenase activity per plant by 5 times, and increasing yield per unit area by 7.5% to more than 25%, achieving significant yield increases. These results demonstrate the key characteristics of the ARC microbial agent for nitrogen-enhancing, green, and yield-increasing quality described in the examples above.

[0184] The above results show that the ARC microbial agent for improving nitrogen fixation and increasing green yield can be used to prepare the quality-improving nitrogen fixation coupled microbial-water-retaining fertilizer, and has the effects of both water-retaining agent and ARC microbial agent for improving nitrogen fixation and increasing green yield.

[0185] Example 36: Use of ARC microbial agent for improving nitrogen fixation and green yield increase - Preparation of microbial inorganic and organic compound fertilizer for improving nitrogen fixation

[0186] Based on the dosage of inorganic-organic compound fertilizer and a dosage of no less than 80 billion viable bacteria of the quality-enhancing, nitrogen-fixing, green and yield-increasing ARC microbial agent per mu of land, the inorganic-organic compound fertilizer and quality-enhancing, nitrogen-fixing, green and yield-increasing ARC microbial agent 1-8 are respectively mixed in proportion. The quality-enhancing, nitrogen-fixing coupled microorganisms are then adsorbed and fixed to the inorganic-organic compound fertilizer using conventional physical methods to prepare eight types of quality-enhancing, nitrogen-fixing coupled microbial inorganic-organic compound fertilizers 1-8. The above-mentioned inorganic-organic compound fertilizers can be purchased from the market, or the inorganic compound fertilizer and bio-organic fertilizer can be purchased separately and then blended according to conventional methods.

[0187] The eight quality-enhancing, nitrogen-fixing coupled microbial inorganic-organic compound fertilizers 1-8 were used as seed fertilizers in the production of leguminous crops such as peanuts and soybeans at the time of 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 eight quality-enhancing, nitrogen-fixing coupled microbial inorganic-organic compound fertilizers 1-8 significantly promoted nodulation and nitrogen fixation, increasing nodule number by more than 2.0 times, increasing nitrogenase activity per plant by more than 5 times, and increasing yield per unit area by more than 8% to 18%, achieving significant yield increases. These results demonstrate the key characteristics of the quality-enhancing, nitrogen-fixing, green, and yield-increasing ARC microbial inoculants described in the above examples.

[0188] The above results show that the ARC microbial agent for improving nitrogen fixation and increasing green yield can be used to prepare the inorganic-organic compound fertilizer coupled with improving nitrogen fixation, and it has the effects of both inorganic-organic compound fertilizer and ARC microbial agent for improving nitrogen fixation and increasing green yield.

[0189] Example 37: Use of ARC Microbial Agent for Quality Improvement, Nitrogen Fixation, and Green Yield Increase - Preparation of Quality Improvement, Nitrogen Fixation, and Coupled Microbial-Rhizobium Fertilizer

[0190] Based on the dosage of rhizobium fertilizer and a minimum of 80 billion viable bacteria per mu of land for the ARC microbial agent for improving nitrogen fixation and green yield, the rhizobium fertilizer was mixed with ARC microbial agents 1 to 8 for improving nitrogen fixation and green yield, and then physically mixed to create eight types of nitrogen fixation-enhancing coupled microbial-rhizobium fertilizers 1 to 8. These rhizobium fertilizers, also known as rhizobium agents, can be purchased commercially or isolated from fresh leguminous nodules and then cultured.

[0191] The eight nitrogen-enhancing, coupled microbial-rhizobium fertilizers 1-8 were used as seed fertilizers in 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 eight nitrogen-enhancing, coupled microbial inorganic-organic compound fertilizers 1-8 significantly promoted nodulation and nitrogen fixation, increasing nodule number by more than three times, nitrogenase activity per plant by more than three times, and yield per unit area by more than 6% to 26%, achieving significant yield increases. These findings demonstrate the key characteristics of the ARC microbial inoculants for nitrogen-enhancing, green, and yield-increasing properties described in the aforementioned examples.

[0192] The above results show that the ARC microbial agent for improving nitrogen fixation and increasing green yield can be used to prepare the coupled microbial-rhizobium fertilizer for improving nitrogen fixation and having the effects of both rhizobium fertilizer and ARC microbial agent for improving nitrogen fixation and increasing green yield.

Claims

1. ARC microbial agent for improving nitrogen fixation and increasing yield, characterized by: It has the effects of improving the quality of legume crops, fixing nitrogen and increasing yield, thereby improving the quality of legume products. It has the effects of regulating and increasing the abundance of rhizobia in the rhizosphere of legume crops and increasing the number of nodules in legume crops. It contains the gene sequences shown in SEQ ID No. 1-4.

2. The green ARC microbial agent for improving nitrogen fixation and increasing yield according to claim 1, characterized in that: The quality-enhancing, nitrogen-fixing, green, and yield-increasing ARC microbial agent can improve the quality of leguminous crop products by increasing the content of one or more nutritional functional components such as protein, glutamic acid, peanut resveratrol, and soy isoflavones.

3. The green ARC microbial agent for improving nitrogen fixation and increasing yield according to claim 1, characterized in that: The ARC microbial agent for improving quality, fixing nitrogen and increasing green yield has an alleviating effect on one or more soil-borne diseases such as peanut fruit rot, soybean green disease, white rot of leguminous crops, bacterial wilt, root rot, and sclerotinia rot; and / or reduce peanut fruit surface spots; and / or reduce aflatoxin levels and improve quality and safety levels.

4. The green ARC microbial agent for improving nitrogen fixation and increasing yield according to claim 1, characterized in that: The DNA sequence genes shown in SEQ ID No. 1-4 may have a certain degree of variation in different strains. When the degree of variation is small, not exceeding 10% of the base variation and having the corresponding biological activity function, it constitutes a functional equivalent of the DNA sequence shown in SEQ ID No. 1-4. The microbial composition contains the DNA sequence shown in SEQ ID NO. 1~4 or its functional equivalent, and has the effects of improving nitrogen quality and increasing yield, and has the effects of regulating and increasing the abundance of rhizobia in the rhizosphere of leguminous crops and increasing the number of nodules of leguminous crops, it constitutes the ARC microbial agent for improving nitrogen quality and increasing yield according to claim 1.

5. The green ARC microbial agent for improving nitrogen fixation and increasing yield according to claim 1, characterized in that: The ARC microbial agent for improving nitrogen fixation and increasing yield is a composition of the following four microorganisms: Bacillus laterosporus with a deposit number of CCTCC NO: M 20231601, Bacillus amyloliquefaciens with a deposit number of CCTCC NO: M 20231598, Enterobacter ludwigii with a deposit number of CCTCC NO: M 20231595, and Bacillus mucilaginosus with a deposit number of CCTCC NO: M 20231602.

6. The green ARC microbial agent for improving nitrogen fixation and increasing yield according to claim 1, characterized in that: The ARC microbial agent for improving quality, fixing nitrogen and increasing yield is a composition of the following four microorganisms: one or more strains of Bacillus laterosporus with a deposit number of CCTCC NO: M 20231601, Bacillus amyloliquefaciens with a deposit number of CCTCC NO: M 20231598, Enterobacter ludwigii with a deposit number of CCTCC NO: M 20231595, and Bacillus mucilaginosus with a deposit number of CCTCC NO: M 20231602, and a combination of other microorganisms, so that the combined microbial agent contains the DNA sequences shown in SEQ ID NOs. 1 to 4 or their functional equivalents, has the effects of improving quality, fixing nitrogen and increasing yield, thereby improving the quality of legume crop products, and has the effects of regulating and increasing the abundance of rhizobia in the rhizosphere of legume crops and increasing the number of nodules in legume crops. The ARC microbial agent for improving quality, fixing nitrogen and increasing yield according to claim 1 is thus constituted.

7. The green ARC microbial agent for improving nitrogen fixation and increasing yield according to claim 1, characterized in that: In the microbial agent, the proportion of viable bacteria of any one strain in the microbial agent is greater than or equal to 1%.

8. The green ARC microbial agent for improving nitrogen fixation and increasing yield according to claim 1, characterized in that: The ARC microbial agent for improving quality, fixing nitrogen and increasing green yield is a combination of three or more microorganisms.

9. the quality-improving and nitrogen-fixing green production-increasing ARC microbial agent according to any one of claims 1-8 is used for configuring the quality-improving and nitrogen-fixing coupled microbial compound fertilizer, is used in production to play the quality-improving and nitrogen-fixing green production-increasing ARC microbial agent effect simultaneously; or is used for configuring the quality-improving and nitrogen-fixing coupled microbial organic fertilizer, is used in production to play the quality-improving and nitrogen-fixing green production-increasing ARC microbial agent effect simultaneously; or is used for configuring the quality-improving and nitrogen-fixing coupled microbial fertilizer, is used in production to play the quality-improving and nitrogen-fixing green production-increasing ARC microbial agent effect simultaneously; or is used for configuring the quality-improving and nitrogen-fixing coupled microbial moisturizing fertilizer, is used in production to play the quality-improving and nitrogen-fixing green production-increasing ARC microbial agent effect simultaneously; or is used for configuring the quality-improving and nitrogen-fixing coupled microbial inorganic-organic compound fertilizer, is used in production to play the quality-improving and nitrogen-fixing green production-increasing ARC microbial agent effect simultaneously.

10. A quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial compound fertilizer, a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial organic fertilizer, a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial micro-fertilizer, a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial moisturizing fertilizer, a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial inorganic-organic compound fertilizer, or a quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial-rhizobium fertilizer, which is configured using the quality-improving, nitrogen-fixing, green-yield-increasing ARC microbial agent according to claim 1.

11. A method for preparing the quality-improving, nitrogen-fixing, green, and yield-increasing ARC microbial compound fertilizer, the quality-improving, nitrogen-fixing, green, and yield-increasing ARC microbial organic fertilizer, the quality-improving, nitrogen-fixing, green, and yield-increasing ARC microbial micronutrient fertilizer, the quality-improving, nitrogen-fixing, green, and yield-increasing ARC microbial moisturizing fertilizer, the quality-improving, nitrogen-fixing, green, and yield-increasing ARC microbial inorganic-organic compound fertilizer, or the quality-improving, nitrogen-fixing, green, and yield-increasing ARC microbial-rhizobium fertilizer according to claim 10: in: Method for preparing the quality-enhancing, nitrogen-fixing, green and yield-increasing ARC microbial compound fertilizer: according to the conventional dosage of compound fertilizer and the dosage of the quality-enhancing, nitrogen-fixing, green and yield-increasing ARC microbial agent per mu of land with a viable count of not less than 80 billion, the compound fertilizer and the quality-enhancing, nitrogen-fixing, green and yield-increasing ARC microbial agent are mixed in proportion, and then the quality-enhancing, nitrogen-fixing, green and yield-increasing ARC microbial compound fertilizer is adsorbed and fixed to the compound fertilizer particles by physical methods to prepare the quality-enhancing, nitrogen-fixing, green and yield-increasing ARC microbial compound fertilizer; Method for preparing the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial organic fertilizer: according to the conventional amount of organic fertilizer and the amount of the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial agent with a viable count of not less than 80 billion per mu of land, the organic fertilizer and the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial agent are mixed in proportion, and then the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial agent is adsorbed and fixed on the organic fertilizer particles by physical methods to prepare the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial compound fertilizer; Method for preparing ARC microbial fertilizer for improving quality, fixing nitrogen and increasing green yield: according to the conventional dosage of trace element fertilizer and the dosage of ARC microbial agent for improving quality, fixing nitrogen and increasing green yield with a count of not less than 80 billion viable bacteria per mu of land, the trace element fertilizer and the microbial agent for improving quality, fixing nitrogen and increasing green yield are mixed, and then the ARC microorganisms for improving quality, fixing nitrogen and increasing green yield are adsorbed and fixed on the micro-fertilizer particles by physical methods to prepare the micro-fertilizer for improving quality, fixing nitrogen and increasing green yield; Method for preparing the quality-improving nitrogen-fixing coupled microbial moisturizing fertilizer: according to the conventional dosage of water-retaining agent and the dosage of the quality-improving 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 quality-improving nitrogen-fixing green yield-increasing ARC microbial agent are mixed in proportion, and then the quality-improving nitrogen-fixing green yield-increasing ARC microbial moisturizing fertilizer is prepared by mixing by conventional physical methods / or adsorbing and fixing the quality-improving nitrogen-fixing green yield-increasing ARC microbial moisturizing fertilizer by the water-retaining agent; Method for preparing the ARC microbial rhizobium fertilizer for improving quality, fixing nitrogen and increasing green yield: according to the conventional amount of rhizobium fertilizer and the amount of the ARC microbial agent for improving quality, fixing nitrogen and increasing green yield of not less than 80 billion viable bacteria per mu of land, the rhizobium fertilizer and the ARC microbial agent for improving quality, fixing nitrogen and increasing green yield are proportioned, and then mixed by conventional physical methods to prepare the ARC microbial rhizobium fertilizer for improving quality, fixing nitrogen and increasing green yield; The method for preparing the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial organic-inorganic compound fertilizer is as follows: according to the conventional dosage of compound fertilizer per mu of land and the dosage of the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial agent of not less than 80 billion viable bacteria, the organic-inorganic compound fertilizer and the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial agent are mixed, and then the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial agent is adsorbed and fixed on the organic and inorganic particles through physical methods to prepare the quality-improving, nitrogen-fixing, green and yield-increasing ARC microbial organic-inorganic compound fertilizer.

12. Application of the green ARC microbial compound fertilizer for improving nitrogen fixation and increasing yield, the green ARC microbial organic fertilizer for improving nitrogen fixation and increasing yield, the green ARC microbial micro-fertilizer for improving nitrogen fixation and increasing yield, the green ARC microbial moisturizing fertilizer for improving nitrogen fixation and increasing yield, the green ARC microbial inorganic-organic compound fertilizer for improving nitrogen fixation and increasing yield, or the green ARC microbial-rhizobium fertilizer for improving nitrogen fixation and increasing yield obtained according to claim 11 in crop production: for improving the quality and safety level of leguminous crop products; for promoting nodulation and nitrogen fixation of leguminous crops; for increasing 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 premature aging of leguminous crops due to lack of fertilizer during maturity; for increasing the number of leguminous crop pods; for increasing leguminous crops. It can improve the fullness of crop pods and reduce the pod shrinkage rate; promote early flowering and pod setting of leguminous crops; reduce the occurrence of peanut fruit rot; reduce the occurrence of bacterial wilt of leguminous crops; reduce the occurrence of powdery mildew of leguminous crops; reduce the occurrence of leaf spot of leguminous crops; reduce the occurrence of root nematode disease of leguminous crops; reduce the occurrence of root rot of leguminous crops; reduce the occurrence of root nematode disease of leguminous crops; reduce the occurrence of sclerotinia disease of leguminous crops; reduce the occurrence of downy mildew of leguminous crops; reduce the occurrence of wilt of leguminous crops; reduce the occurrence of white rot of leguminous crops; reduce the occurrence of soybean green disease; promote carbon emission reduction of leguminous crops, which is beneficial to soil improvement; promote the increase of total biomass of leguminous crops; reduce the surface spots of peanut fruits and increase marketability; promote the increase of soybean yield in saline-alkali land.

13. A method for crop production, characterized in that: The microbial agent is selected so that it contains the gene sequence shown in SEQ ID NO. 1 to 4 after analysis and determination, and has the effect of improving the quality and fixing nitrogen, thereby improving the quality of legume crop products, and has the effect of regulating and increasing the abundance of rhizobia in the rhizosphere of legume crops and increasing the number of nodules in legume crops; the microbial agent is applied to crops to improve the quality and fix nitrogen, and increase yield in a green way.

14. The method according to claim 13, wherein: The DNA sequence genes shown in SEQ ID No. 1-4 may have a certain degree of variation in different strains. When the degree of variation is small, not exceeding 10% base variation, and has corresponding biological activity functions, the functional equivalents of the DNA sequences shown in SEQ ID No. 1-4 are formed, containing the gene sequences shown in SEQ ID NO. 1~4 or their functional equivalents, and have the effects of improving nitrogen quality, fixing nitrogen and increasing yield. They have the effects of 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 ARC microbial agent for improving nitrogen quality, fixing nitrogen and increasing yield in a green way according to claim 1, which can be used for improving nitrogen quality and increasing yield in a green way.