56 results about "Nitrogenase" patented technology

The invention discloses a pseudomonas for antimony detoxification and autotrophic nitrogen fixation and application of the pseudomonas. The strain is separated from the rhizosphere of a pioneer plant in antimony-polluted soil of a tin mine in cold water river city, Hunan province, and is identified to be capable of quickly oxidizing high-toxicity antimony (III) into low-toxicity antimony (V) through autotrophic metabolism in a high-antimony environment, so that antimony detoxification is realized, and the detoxification rate is 108.6 (mu mol / (L.day)). The strain has autotrophic nitrogen fixation activity and can achieve autotrophic growth under the oligotrophic condition of a mining area, and the nitrogenase activity is larger than or equal to 223.4 (nmol C2H2 / (g.h)). Particularly, the antimony detoxification and nitrogen fixation capabilities of the strain have a synergistic effect, so that the strain has unique advantages in the field of heavy metal antimony pollution remediation, and dual effects of antimony pollution treatment and soil improvement can be synchronously realized; the development and application prospects are good.

The invention discloses an autotrophic nitrogen fixation Klebsiella strain with antimony oxidation and growth promoting capabilities and application thereof. The strain is different from traditional heterotrophic nitrogen-fixing bacteria depending on an organic carbon source, nitrogen fixation can be carried out by taking carbon dioxide as a main carbon source under an oligotrophic condition, Sb (III) oxidation is taken as a main energy acquisition way, autotrophic metabolism is realized, and plant growth can also be promoted. Through identification, Sb (III) can be oxidized into Sb (V) in three days in a high-antimony environment, and the oxidation rate reaches 89.11 (mu mol / L / day); the activity of nitrogenase is greater than or equal to 167.4 (nmolC2H2 / (g.h)). The strain has unique advantages in the field of in-situ remediation of antimony-polluted tailings and other oligotrophic matrixes, and can synchronously realize multiple effects of antimony pollution treatment, soil nitrogen supply and quality improvement. The strain is safe to human and livestock, has no environmental pollution problem, is simple in culture condition and easy to preserve, and has good development and application prospects.

According to the high-temperature-resistant nitrogen-fixing bacteria microcapsule fertilizer and the preparation method thereof, aiming at the problems of thallus inactivation and nutrient loss in a high-temperature environment, a sodium alginate-chitosan double-layer embedding technology is combined with magnetic nanoparticle modification, and a stable double-layer structure is formed by optimizing the ratio of an embedding material, so that the high-temperature-resistant nitrogen-fixing bacteria microcapsule fertilizer is prepared. The survival rate and activity of the nitrogen-fixing bacteria at high temperature are remarkably improved, and meanwhile, the functions of intelligent nutrient release and magnetic recovery are achieved. The fertilizer is prepared from composite microcapsules, amino acid, humic acid and other raw materials according to a specific ratio through magnetic field orientation treatment, multi-stage mixing, atomization granulation and magnetic field screening processes, the particle size is controlled to be 0.5-3.35 mm, and the shelf life is prolonged to 12 months. The method has the technical effects that trehalose-glycerol / sorbitol synergistically forms a glassy protective film to maintain the stability of thalli, gamma-aminobutyric acid activates nitrogenase gene expression, magnetic nanoparticles construct a photosynthetic symbiotic system to optimize rhizosphere oxygen supply, and silanized FeO nanoparticles fix heavy metal ions through electrostatic adsorption, so that the heavy metal ions in the thalli are effectively adsorbed, and the heavy metal ions in the thalli are effectively adsorbed. And the humic acid buffers the pH fluctuation of the soil, so that the nitrogen fixation efficiency is finally improved by 30-50%, and the crop yield is increased by 15-25%.

The invention discloses bacillus safensis and application thereof to synergistically promoting soybean root nodule formation and yield increase with pseudomonas, the bacillus safensis is preserved in China General Microbiological Culture Collection Center (CGMCC), the preservation number is CGMCC No.34634, the preservation date is May 22, 2025, the preservation number is CGMCC No.34634, the preservation number is CGMCC No.34634, the preservation number is CGMCC No.34634, and the preservation number is CGMCC No.34634. The preservation address is Institute of Microbiology, Chinese Academy of Sciences, No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing. According to the method disclosed by the invention, the increase of nodules and the increase of nitrogenase activity can be promoted by co-inoculating the bacillus safensis and the pseudomonas, and meanwhile, the increase of the yield of a biological membrane is also promoted by co-inoculating.

Heterologous synthesis of a nitrogen-fixing system in a non-diazotrophic organism is a long-sought-after goal because of the significance of nitrogenase for areas related to agronomy, energy and environment. Two simplified nitrogenase analogs from Azotobacter vinelandii in Escherichia coli were successfully heterologously synthesized. One analog comprise the reductase component (NifH) and, in place of the catalytic component (NifDK), a cofactor maturase (NifEN); whereas the other analog comprised only NifEN. Metal, EPR and activity analyses verified the cluster composition and functional competence of the heterologously expressed NifH and NifEN. Moreover, NMR, nanoSIMS and growth experiments illustrated the abilities of both systems to mimic the nitrogenase in reducing N2 and incorporating the reduced N into the cellular mass. These results firmly established NifEN / NifH or NifEN as a simplified nitrogenase analog that could be engineered or expanded to facilitate future transgenic expression of nitrogenase and development of nitrogenase-based biotechnological applications.

The invention discloses a gene GmSCAMP3 for regulating and controlling the symbiotic nitrogen fixation capacity of soybean root nodules and application of the gene GmSCAMP3. Specifically, the invention provides a protein for regulating and controlling the symbiotic nitrogen fixation capacity of soybean root nodules, and the amino acid sequence of the protein is as shown in SEQ ID NO: 2. The invention also provides a method for improving soybean yield or changing soybean root nodule symbiotic nitrogen fixation capacity. The method comprises the step of introducing the polynucleotide encoding the protein into soybean cells or tissues to obtain transgenic soybeans. The invention finds that the nitrogenase activity of the Gmscamp3 mutant root nodule is only 65.79% of that of a wild type root nodule, which indicates that the GmSCAMP3 protein plays an important role in the process of regulating and controlling the symbiotic nitrogen fixation capacity of the soybean root nodule.

The invention relates to the technical field of biology, and discloses application of PHR2 in regulating nodulation and nitrogen fixation processes of leguminous plants under phosphorus starvation stress, and the application is any one of the following applications: P1, application in enhancing nitrogen fixation capability of the leguminous plants under the phosphorus starvation stress; p2, application in promoting formation of root nodules with nitrogen fixation capacity of leguminous plants under phosphorus starvation stress; p3, application of improving activity of nitrogenase in leguminous plant root nodules under phosphorus starvation stress; p4, application in leguminous plant breeding. The PHR2 protein and related biological materials thereof can be used for regulating and controlling the nitrogen fixation capacity of leguminous plants under phosphorus starvation stress.

The invention relates to an artificial efficient nitrogen fixation system and a preparation method and application thereof. The artificial efficient nitrogen fixation system contains recombinant engineering bacteria constructed by an artificial combination module and nitrogen fixation chassis microorganisms; wherein the artificial combination module sequentially comprises a constitutive promoter element Pc and a coding gene of methyltransferase selected from nitrogen-fixing pseudomonas stutzeri A1501 in the direction from upstream to downstream. According to the recombinant engineering bacteria constructed by introducing the artificial combination module into the nitrogen-fixing chassis microorganism, the nitrogenase activity of the nitrogen-fixing chassis microorganism can be remarkably improved, the nitrogen-fixing capacity of the nitrogen-fixing chassis microorganism is remarkably improved, and the recombinant engineering bacteria have a relatively good application prospect in the aspect of promoting plant growth.

The invention belongs to the technical field of soil improvement, and particularly relates to stenotrophomonas maltophilia C-GD and application thereof in nitrogen fixation. The nitrogenase activity of the stenotrophomonas maltophilia C-GD is 45.90 (ng / g / 24h), and the stenotrophomonas maltophilia C-GD shows relatively good nitrogen fixation activity and can be used for nitrogen fixation of soil. The invention provides a strain of Stenotrophomonas maltophilia C-GD with a nitrogen fixation function, and after the Stenotrophomonas maltophilia C-GD is prepared into a microbial inoculum to be applied, the key nutrient indexes of organic matters, total nitrogen, total phosphorus, available phosphorus, alkali-hydrolyzable nitrogen and the like in soil can be greatly improved.

The invention discloses application of a GmNSE1 gene in regulation and control of soybean root nodule nitrogen fixation efficiency. The application of the GmNSE1 gene or a biological material related to the GmNSE1 gene in at least one of the following aspects: A1) regulating and controlling the nitrogen fixation efficiency of soybean root nodules or preparing a product for regulating and controlling the nitrogen fixation efficiency of the soybean root nodules; a2) cultivating soybeans with improved nitrogen fixation efficiency or preparing products for improving the nitrogen fixation efficiency of the soybeans; a3) preparing transgenic soybeans; the nucleotide sequence of the GmNSE1 gene is as shown in SEQ ID NO. 1. The specificity of the GmNSE1 gene is expressed in a root nodule infection area, the GmNSE1 gene has E3 ubiquitin ligase enzyme activity in vitro, and compared with a control group, the nitrogenase activity of a soybean plant with the overexpressed GmNSE1 gene is remarkably improved, which shows that the GmNSE1 gene plays an important role in the process of regulating and controlling the symbiotic nitrogen fixation capacity of soybean root nodules, and the GmNSE1 gene has a good application prospect in the field of soybean root nodule symbiotic nitrogen fixation. The method has application prospects in research on nitrogen fixation mechanisms of leguminous crops and agricultural production.

The invention discloses application of a GmNSE1 gene in regulation and control of soybean root nodule nitrogen fixation efficiency. The application of the GmNSE1 gene or a biological material related to the GmNSE1 gene in at least one of the following aspects: A1) regulating and controlling the nitrogen fixation efficiency of soybean root nodules or preparing a product for regulating and controlling the nitrogen fixation efficiency of the soybean root nodules; a2) cultivating soybeans with improved nitrogen fixation efficiency or preparing products for improving the nitrogen fixation efficiency of the soybeans; a3) preparing transgenic soybeans; the nucleotide sequence of the GmNSE1 gene is as shown in SEQ ID NO. 1. The specificity of the GmNSE1 gene is expressed in a root nodule infection area, the GmNSE1 gene has E3 ubiquitin ligase enzyme activity in vitro, and the activity of a mutant with the GmNSE1 gene knocked out is remarkably reduced compared with that of wild nitrogenase, so that the GmNSE1 gene plays an important role in the process of regulating and controlling the symbiotic nitrogen fixation capacity of soybean root nodules; the method has application prospects in research on nitrogen fixation mechanisms of leguminous crops and agricultural production.

The invention discloses ammonium-tolerant nitrogen-fixing paenibacillus and application thereof, and belongs to the technical field of agriculture. The bacterial strain is preserved in Guangdong Microbial Culture Collection Center on September 19, 2025, the preservation number is GDMCC 66998, and the bacterial strain is named Paenibacillus sp. By taxonomy. The ammonium-tolerant nitrogen-fixing paenibacillus from the red soil rice field is obtained through screening, the strain still shows high nitrogenase activity under the condition of 80-100 mM NH4 < + >, and the limitation that traditional nitrogen-fixing bacteria are prone to being inhibited by ammonium is broken through. In addition, the strain can significantly improve the germination rate of rice seeds, promote the root length, plant height and biomass accumulation of rice seedlings, and show a good growth promotion effect. Due to the characteristic of forming spores, the strain can also be prepared into a microbial fertilizer preparation which is used for nitrogen fertilizer replacement and synergism in a rice field ecological system.

The invention discloses a microbial agent composition capable of promoting growth and yield increase of soybeans in saline soil and application of the microbial agent composition. A microbial agent is an enterobacter strain THD10 or a complex microbial agent formed by the enterobacter strain THD10. The microbial agent provided by the invention can significantly improve the plant height, fresh weight, dry weight, nodulation number and nitrogenase activity of soybeans. In saline soil, compared with treatment without inoculation of the microbial inoculum, inoculation treatment can significantly improve the soybean biomass, and the nitrogen fixation efficiency of a single soybean plant is improved; the SOD, CAT and POD contents of soybean leaves are increased, and the malondialdehyde content of the leaves is reduced; the method can improve the soybean yield and promote the improvement of soybean quality indexes, has very important significance for enlarging the soybean cultivation area and improving the per unit yield in China, and has important guidance and reference significance for cultivation of other crops in saline soil.

The invention relates to a method for promoting growth, nodulation and nitrogen fixation of indigofera pseudotinctoria by utilizing rhizobium D303 and a bacterial solution thereof. The biological preservation number of the rhizobium sp. D303 is CCTCC (China Center for Type Culture Collection) NO: M20252803. Pot culture tie-back tests prove that the indigofera pseudotinctoria seedlings inoculated with the rhizobium are obviously superior to a control group in the aspects of plant height, root length, overground dry weight, underground dry weight, root nodule number, root nodule weight, root nodule diameter, nitrogenase activity, antioxidant enzyme activity and the like. The rhizobium strain provided by the invention has efficient nodulation and nitrogen fixation capability and excellent growth promoting characteristic, obviously improves the growth speed, nodulation capability and nitrogen fixation efficiency of the indigofera pseudotinctoria, reduces the dependence on chemical nitrogen fertilizer, and has a broad application prospect in development of special biological bacterial fertilizer for indigofera pseudotinctoria. The method has important ecological and economic benefits for promoting the application of the indigofera pseudotinctoria in ecological restoration and feed production, and is suitable for slope greening, water and soil conservation and barren mountain ecological restoration projects.

The invention relates to novel paenibacillus with biomacromolecule degradation capacity and nitrogen fixation and phosphorus solubilization effects and application of the novel paenibacillus, the strain is identified as a novel strain of paenibacillus through multi-phase classification, and the novel paenibacillus has the capacity of degrading biomacromolecules such as starch, cellulose, casein, chitin and lecithin. The method can be applied to degradation of related carbohydrates, proteins and lipid substances and preparation of biological enzymes; the bacillus subtilis has the advantages that the bacillus subtilis can be used for preparing the bacillus subtilis, has the phosphorus dissolving capacity of dissolving organic phosphorus and inorganic phosphorus, has nitrogenase and beta-1, 3-glucanase and the activity of generating IAA (Indoleacetic Acid), can play a probiotic role on crops by fixing nitrogen, dissolving phosphorus and inhibiting pathogenic bacteria, can grow in an environment with the salinity not higher than 4.5%, and has certain salt-tolerant activity. Due to multiple functions, the strain can be used for preparing and applying biological nitrogen fertilizers, phosphate fertilizers and crop probiotics, so that the strain plays an important role in crop growth and green agricultural production.

The invention belongs to the technical field of soil nitrogen fixation and environmental protection, and relates to a method for enhancing soil nitrogen fixation by using pure iron powder. A method for enhancing soil nitrogen fixation by using pure iron powder comprises the following steps: mixing rice soil and wheat straw powder to obtain a soil sample; the method comprises the following steps: mixing a soil sample with pure iron powder under a waterflooding condition; pure iron powder is added to regulate and control microbial ecology of rice soil biological nitrogen fixation, the soil iron reduction process is enhanced, the abundance of iron reducing bacteria in soil is improved, the relative contribution of iron reducing bacteria nitrogen fixation is enhanced to a certain extent, the diversity of iron reducing bacteria communities is reduced, the ecological niches of dominant groups are enhanced, and the ecological niches of the dominant groups are increased. Finally, the nitrogenase activity of the soil is enhanced, and the nitrogen fixation capacity of the rice soil is improved.

Disclosed is a method of producing a non-naturally occurring peptide for reducing nitrogen, the method comprising: a) identifying a first group of peptides each having a binding pocket similar to a naturally occurring iron-sulfur cluster cofactor binding pocket of a naturally occurring nitrogenase, wherein each of the first group of peptides is not a nitrogenase or a subunit or domain of a nitrogenase; b) selecting, from the first group of peptides, a second group of peptides each having at least one target amino acid in the binding pocket; c) selecting, from the second group of peptides, a third group of peptides each having at least one target property; and d) modifying the sequence and / or structure of each of the third group of peptides to improve binding of the iron-sulfur cluster cofactor in the binding pocket. Also disclosed is a non-naturally occurring peptide for reducing nitrogen, comprising: at least one modification to a naturally occurring peptide; and a binding pocket for binding an iron-sulfur cluster cofactor; wherein the naturally occurring peptide is not a nitrogenase or a subunit or domain of a nitrogenase; and wherein the at least one modification improves binding of the iron-sulfur cluster cofactor in the binding pocket.

The invention discloses an activating agent capable of being used for improving the nitrogen fixation activity of farmland soil in situ and a preparation method of the activating agent, and relates to the technical field of agricultural chemistry. The activating agent is prepared from the biochar, the sodium molybdate, the goethite and the rutile as raw materials and is used for improving the nitrogen fixation activity of the farmland soil in situ. On the premise of not adding exogenous strains, by activating existing indigenous nitrogen-fixing microbial communities in farmland soil and stimulating high-efficiency expression of related nitrogen-fixing functional genes in a soil environment, in-situ activation of the nitrogen-fixing potential of the soil is achieved, and the nitrogen-fixing performance of the soil is improved. The activator for in-situ improvement of the nitrogen fixation activity of the farmland soil has a synergistic effect on improving the content of ammonium nitrogen in the soil and the activity of nitrogenase, has the outstanding advantages of high in-situ adaptability, precise action targeting, sustained and lasting nutrition release, environment friendliness, low cost and the like, can remarkably reduce the use of chemical fertilizers, and has a good application prospect. And sustainable development of green agriculture is realized.

The application discloses application of Paenibacillus polymyxa in improving yield of pyrroloquinoline quinone (PQQ), and application of a living silk microorganism in improving nitrogen fixation capacity of Paenibacillus polymyxa. The application also discloses a co-cultivation system with high PQQ yield. The application further discloses the living silk microorganism with a preservation number of CCTCC NO: M 20241604. The application further discloses Paenibacillus polymyxa with a preservation number of CCTCC NO: M 20241603. The application further discloses a fermentation liquor prepared by using the co-cultivation system. The application further discloses application of the co-cultivation system and the fermentation liquor. The application can promote yield of PQQ produced by the living silk microorganism and improve nitrogenase activity of Paenibacillus polymyxa simultaneously in a bidirectional way by co-cultivating and fermenting the living silk microorganism and Paenibacillus polymyxa, and the co-cultivation fermentation liquor can improve crop nitrogen fixation, stress resistance, growth and yield.

The present invention relates to methods and means for producing nitrogenase polypeptides in the mitochondria of plant cells.

The invention relates to an application method of a special microbial agent for improving the nitrogen fixation efficiency of soybean root nodules, and belongs to the field of agricultural microorganism application. A core strategy of staged time sequence application is adopted: firstly, before sowing or during sowing, a first complex microbial inoculant containing proline secreting bacteria and indigenous infectious microbe inhibiting bacteria is applied into soil, and a pre-culture period is performed for 5-15 days; during the period, proline is converted into indole-3-acetic acid (IAA) by a fourth microorganism which synchronously exists or is introduced, so that a growth-promoting microenvironment is established at the rhizosphere, and meanwhile, a third microorganism inhibits harmful infectious microbes; next, bradyrhizobium or sinorhizobium is applied to the seedlings or rhizosphere. According to the time sequence design, an optimized colonization ecological niche with low competition and high chemotactic signals is created for target rhizobium through a synergistic mechanism of firstly improving the environment and then introducing strains. According to the method, the infection success rate of rhizobium can be remarkably increased, the number of root nodules and nitrogenase activity are increased, soybean root development is synchronously promoted, and the soybean symbiotic nitrogen fixation efficiency and yield are improved.

This invention belongs to the field of agricultural technology, specifically relating to a method for recommending nitrogen fertilizer for peanuts that coordinates yield and biological nitrogen fixation. The method includes determining the agronomic critical nitrogen application rate based on peanut yield and a model showing the relationship between nitrogen application rate and peanut yield; determining the nitrogen inhibition threshold based on peanut root nodule weight, root nodule nitrogenase capacity, and a model showing the relationship between nitrogen application rate and root nodule weight and root nodule nitrogen fixation capacity; and determining the nitrogen fertilizer application rate that is conducive to yield formation and fully utilizing root nodule nitrogen fixation capacity based on the agronomic critical nitrogen application rate and the nitrogen inhibition threshold. This method coordinates yield and biological nitrogen fixation, minimizing problems caused by excessive nitrogen fertilizer application, such as inhibited root nodule nitrogen fixation, decreased nitrogen use efficiency, increased production costs, and environmental pollution.

The invention discloses a magnesium modified wheat straw biochar-based slow-release rhizobium fertilizer and a preparation method thereof. The preparation method comprises the following steps: constructing hierarchical porous biochar MgWBC with magnesium-based active sites by adopting magnesium acetate impregnation and high-temperature pyrolysis; granular urea is used as a core, chitosan is used for packaging to realize directional loading and surface phase inversion film sealing of an MgWBC layer and rhizobium, and a composite structure with pH response capability is constructed. According to the method disclosed by the invention, the stress resistance and the directional colonization efficiency of the rhizobium are remarkably improved by utilizing the characteristic that the nitrogenase is activated by magnesium and the refuge effect of pores of the biochar. In a soybean-corn intercropping system, soybean nodulation and nitrogen fixation are enhanced and corn nutrition is met while nutrient slow release is realized through a synergistic mechanism of promoting nitrogen by magnesium. Experiments show that the yield of an intercropping system and the total nitrogen and effective magnesium level of soil can be remarkably improved, and the stability of soil aggregates is improved. The method is of great significance in promoting intercropping weight loss and efficiency improvement and cultivated land conservation.

The invention discloses bacillus subtilis and application of the bacillus subtilis and pseudomonas to synergistically promote soybean root nodule formation and yield increase, the bacillus subtilis is preserved in China General Microbiological Culture Collection Center (CGMCC), the preservation number is CGMCC No.34635, the preservation date is May 22, 2025, the preservation number is CGMCC No.34635, and the preservation number is CGMCC No.34635. The preservation address is Institute of Microbiology, Chinese Academy of Sciences, No.3, Yard 1, Beichen West Road, Chaoyang District, Beijing. According to the method disclosed by the invention, the increase of nodules and the increase of nitrogenase activity can be promoted by co-inoculating the bacillus subtilis and the pseudomonas, and meanwhile, the increase of the yield of a biological membrane is also promoted by co-inoculating.

The invention belongs to the technical field of microbial agents, and particularly relates to a charcoal-based nitrogen-fixing bacterium agent as well as a preparation method and application thereof. According to the invention, a biochar-humic acid-sodium alginate embedding mode is adopted, so that a porous protection cabin can be provided to adsorb plant rhizosphere nutrients. Meanwhile, a low-oxygen microcell is created to protect nitrogenase, physical protection is provided for thalli, and external environmental stress is buffered. Wherein humic acid is used as a high-quality organic carbon source and is beneficial to stimulating thalli to rapidly proliferate in embedded particles and rhizosphere, and a large number of active cells can be obtained in a short time. The biochar-based nitrogen-fixing bacterium agent (namely biochar-humic acid-sodium alginate microspheres) prepared by the invention has the advantages of simple and convenient preparation method, stable storage performance and the like, and compared with other biological agents, the biomass of the dried microbial agent reaches 2.2 * 10 < 10 > CFU / g, so that the biochar-based nitrogen-fixing bacterium agent has a good application prospect.

This invention discloses an autotrophic nitrogen-fixing Klebsiella pneumoniae strain with antimony oxidation and growth-promoting capabilities and its applications. This strain differs from traditional heterotrophic nitrogen-fixing bacteria that rely on organic carbon sources. It can fix nitrogen under oligotrophic conditions using carbon dioxide as the primary carbon source and Sb(III) oxidation as the main energy acquisition pathway, achieving autotrophic metabolism and promoting plant growth. Identification showed that in a high-antimony environment, it can oxidize Sb(III) to Sb(V) within three days, with an oxidation rate of 89.11 (μmol / L / day); nitrogenase activity ≥167.4 (nmolC2H2 / (g·h)). This strain has unique advantages in the in-situ remediation of oligotrophic substrates such as antimony-contaminated tailings, simultaneously achieving multiple effects of antimony pollution control and soil nitrogen replenishment and quality improvement. The bacterium is safe for humans and animals, poses no environmental pollution problems, has simple cultivation conditions, is easy to preserve, and has good prospects for development and application.

The invention belongs to the technical field of application of microorganisms, and particularly relates to a diazo nutritional type acetobacter gluconicum mutant, a nitrogen-fixing microbial agent, a preparation method of the nitrogen-fixing microbial agent and application of the nitrogen-fixing microbial agent. The mutant strain with high nitrogenase activity and genetic stability is obtained through ARTP mutagenesis screening, the nitrogen fixation effect is remarkably improved, and the nitrogenase activity of the mutant strain on a nitrogen-free culture medium reaches 368.84 nmol.mg <-1 >. H <-1 >. The nitrogen-fixing bacterial strain disclosed by the invention can effectively improve index values such as plant height, root length and plant weight of rice, has a good growth-promoting effect on various crops, and is a broad-spectrum growth-promoting nitrogen-fixing bacterial strain. The nitrogen-fixing bacterial strain can be rapidly cultured in a short time, and is suitable for industrial production. The diazo nutritional glucose acetobacter mutant is a high-enzyme-activity mutant strain obtained through ARTP mutagenesis and acetylene reduction method screening and has good nitrogen fixation capacity, the nitrogenase activity of the diazo nutritional glucose acetobacter mutant is improved by 3.1 times, and the hereditary stability of the diazo nutritional glucose acetobacter mutant is good after multiple times of subculture.

The invention discloses a nitrogen limitation-based clostridium pasteurianum-rhodobacter sphaeroides coupling fermentation hydrogen production method and application thereof. The method comprises the following steps: performing dark fermentation on activated clostridium pasteurianum in a nitrogen-free culture medium in an argon environment to produce hydrogen; and adjusting the pH value of the obtained culture solution to 7.0-8.0, selectively sterilizing, inoculating activated rhodobacter sphaeroides, and carrying out light fermentation in an argon environment to produce hydrogen. The clostridium pasteurianum fermentation liquor can be directly subjected to rhodobacter sphaeroides photofermentation hydrogen production culture only by simply adjusting the pH value without operations such as centrifugal removal of thalli. According to the method, excessive proliferation of thalli is inhibited through nitrogen limitation, the flow direction of a carbon source is saved, high expression of nitrogenase is induced, and a second efficient hydrogen production way except hydrogenase is opened up, so that the hydrogen yield is further increased while the cost of a nitrogen source is reduced. The hydrogen production yield reaches 9.81 mol H2 / mol glucose, which is the highest level at present.

The application discloses a method for producing hydrogen through Clostridium pasteurianum-Rhodobacter sphaeroides coupling fermentation based on nitrogen limitation and application thereof. In the method, activated Clostridium pasteurianum is subjected to dark fermentation for producing hydrogen in a nitrogen-free culture medium and in an argon environment; then, the pH value of the obtained culture solution is adjusted to 7.0-8.0, the activated Rhodobacter sphaeroides is inoculated after selective sterilization, and the light fermentation for producing hydrogen is carried out in the argon environment. In the application, the fermentation solution of Clostridium pasteurianum does not need to be subjected to centrifugal removal of bacterial bodies and the like, and only needs to be simply adjusted in pH value, so that the light fermentation for producing hydrogen of Rhodobacter sphaeroides can be directly carried out. In the application, the nitrogen limitation is used to inhibit excessive proliferation of bacterial bodies, save carbon source flow, induce high expression of nitrogen fixation enzyme, and open a second high-efficiency hydrogen production path in addition to hydrogenase, so that the hydrogen production yield is further improved while the cost of nitrogen source is reduced. The hydrogen production yield of the application reaches 9.81 mol H2 / mol glucose, which is the highest level at present.