Alkali-lake Dietzia sp. GD-1 and application thereof in carbon sequestration and phosphorus solubilization

Abstract

The invention relates to the technical field of microbial carbon sequestration, in particular to a Dietzia alkali GD-1 strain and application thereof in carbon sequestration and phosphorus solubilization. The invention provides a strain of Dietzia natronanolimnaea GD-1, and the preservation number of the Dietzia natronanolimnaea GD-1 is CGMCC (China General Microbiological Culture Collection Center) No.31728. The invention further provides a preparation method of the Dietzia natronanolimnaea strain. The alkali lake Dietzschia sp. GD-1 provided by the invention is a carbon sequestration bacterium separated from fixed inter-plate soil in a salinization photovoltaic field area, has the capability of immobilizing carbon dioxide in the atmosphere, also has the capability of dissolving phosphorus, provides a strain resource for improving the carbon sequestration and emission reduction capability of the salinization soil, and has a wide application prospect. Technical support is provided for deeply knowing the microorganism regulation process of the saline-alkali soil nutrient substance occurrence form, a foundation is laid for providing a photovoltaic field soil microorganism improvement technology, and high-quality development of the saline-alkali fragile area photovoltaic industry is promoted.

Description

Technical Field

[0001] The invention relates to the technical field of microbial carbon fixation, in particular to a strain of Dietzia alkaline lake GD-1 and application thereof in carbon fixation and phosphorus dissolution. Background Art

[0002] The development of new energy has become the core way to deal with climate change. However, more than 80% of my country's new energy is distributed in fragile areas (such as saline-alkali land). It is urgent to explore new models of organic integration of energy, ecological restoration, and characteristic industrial development. Studies have shown that functional microorganisms such as nitrogen-fixing bacteria and cellulose-degrading bacteria play an important role in improving saline-alkali land, restoring soil physical and chemical properties, and ecological environment. Improving the potential of soil carbon sequestration and carbon sink is an urgent need to restore the ecological function of saline-alkali land and mitigate climate change. However, there is still a lack of research on the isolation and screening of soil carbon-fixing microorganisms and their carbon sequestration ability. There is even less research on the precise identification and screening of soil carbon-fixing microorganisms in photovoltaic fields in saline-alkali land. To this end, the separation and screening of efficient carbon-fixing indigenous functional microorganisms in photovoltaic fields in saline-alkali land has important potential application value for stimulating and enhancing the synergistic benefits of indigenous microorganisms in improving soil and fixing carbon and increasing carbon sinks. Summary of the invention

[0003] In order to solve the above problems, the present invention provides a strain of Dietzia alkaline lake GD-1 and its application in carbon fixation and phosphorus solubilization. The Dietzia alkaline lake GD-1 provided by the present invention has carbon fixation ability and phosphorus solubilization ability, which is of great significance for improving the soil carbon fixation and sink potential, promoting the high-quality development of the photovoltaic industry in salinization-vulnerable areas, and alleviating global warming.

[0004] In order to achieve the above object, the present invention provides the following technical solutions:

[0005] The invention provides a strain of Dietzia natronolimnaea GD-1, whose preservation number is CGMCC No.31728.

[0006] The present invention provides a carbon-fixing bacterial agent, comprising the Dietzia alkaline lake GD-1 described in the above technical solution.

[0007] Preferably, the OD of Dietzia alkaline lake GD-1 in the carbon-fixing bacteria agent is 600 Value ≥ 0.6.

[0008] The present invention provides a method for preparing the carbon-fixing bacteria agent described in the above technical solution, comprising the following steps:

[0009] The alkaline lake Dietzia GD-1 described in the above technical solution is inoculated into a culture medium for cultivation to obtain the carbon-fixing bacterial agent; the culture medium is a culture medium containing a carbon source or a culture medium without a carbon source.

[0010] Preferably, the culture medium further comprises an electron donor; the electron donor comprises sodium nitrite and / or sodium thiosulfate.

[0011] Preferably, the medium not containing a carbon source comprises the following components at the following concentrations: Na 2 HPO 4 0.5 g / L, KH 2 PO 4 0.5 g / L, MgSO 4 0.3 g / L, CaCl 2 0.2 g / L, NH 4 Cl 0.5g / L, NaNO 3 0.25 g / L, NaCl 0.4 g / L, trace element solution 10 mL / L; the trace element solution includes the following components: FeCl 2 0.3 g / L, FeSO 4 7H 2 O0.3g / L, MnSO 4 ·H 2 O 0.15g / L, ZnSO 4 7H 2 O 0.25g / L, CoCl 2 0.2 g / L; the pH value of the culture medium not containing a carbon source is 7.0-7.4.

[0012] Preferably, the culture medium containing a carbon source is prepared by adding a carbon source to the culture medium not containing a carbon source; the added carbon source includes: sodium carbonate and / or sodium bicarbonate.

[0013] The present invention provides the use of the alkaline lake Dietzia GD-1 described in the above technical scheme or the carbon-fixing bacteria agent described in the above technical scheme or the carbon-fixing bacteria agent prepared by the preparation method described in the above technical scheme in carbon fixation and / or dissolution of insoluble inorganic phosphorus, and the carbon source for carbon fixation includes an inorganic carbon source.

[0014] Preferably, the inorganic carbon source includes one or more of carbon dioxide, sodium bicarbonate and sodium carbonate.

[0015] Preferably, the poorly soluble inorganic phosphorus comprises calcium phosphate.

[0016] Beneficial effects:

[0017] The present invention provides a strain of Dietzia alkali lake GD-1 with a preservation number of CGMCC No. 31728. The Dietzia alkali lake GD-1 provided by the present invention is a carbon-fixing bacterium isolated from the fixed inter-plate soil in the salinized photovoltaic field, has the ability to fix carbon dioxide, and has the ability to solubilize phosphorus, provides bacterial resources for improving the carbon fixation and emission reduction capacity of salinized soil, provides technical support for in-depth understanding of the microbial regulation process of the storage form of nutrients in salinized soil, lays the foundation for proposing soil microbial improvement technology in photovoltaic fields, and promotes the high-quality development of the photovoltaic industry in salinized vulnerable areas. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required to be used in the embodiments are briefly introduced below.

[0019] Figure 1 This is the bacterial morphology of the carbon-fixing bacterium Dietzia alkali-lakea GD-1;

[0020] Figure 2 This is the colony characteristic diagram of the carbon-fixing bacterium Dietzia alkaline lake GD-1;

[0021] Figure 3 This is the phylogenetic tree of the 16S rRNA gene of the carbon-fixing bacterium Dietzia alkaline lake GD-1;

[0022] Figure 4 The results are for the determination of RubisCO enzyme activity of the carbon-fixing bacterium Dietzia alkaline lake GD-1;

[0023] Figure 5 The results of the determination of carbon fixation ability of the carbon-fixing bacterium Dietzia alkaline lake GD-1 under different conditions;

[0024] Figure 6 It is the carbon fixation pathway of the carbon-fixing bacterium Dietzia alkalinica GD-1;

[0025] Figure 7 These are the results of determination of the effective phosphorus content of the carbon-fixing bacterium Dietrichia alkaline lake GD-1 after culture in a liquid culture medium containing insoluble phosphate salts.

[0026] Biological Deposit Description

[0027] Dietzia natronolimnaea GD-1, classified and named Dietzia natronolimnaea, was deposited in the China General Microbiological Culture Collection (CGMCC) on August 23, 2024. The deposit address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Building 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC No. 31728. DETAILED DESCRIPTION

[0028] The present invention provides a strain of Dietzia alkaline lake GD-1, with a deposit number of CGMCC No. 31728. The Dietzia alkaline lake GD-1 provided by the present invention is a carbon-fixing bacterium isolated from the soil between fixed panels in a salinized photovoltaic field, and the bacterial body is short rod-shaped ( Figure 1 ), the colonies produced on solid culture medium are small, round, orange-red, opaque, smooth and sticky, with neat edges and low protrusions ( Figure 2 ), has the ability to fix carbon dioxide, and at the same time has the ability to solubilize phosphorus, and can maintain a good growth rate and carbon fixation ability in extremely low-nutrient salt culture media. The addition of inorganic carbon sources (sodium carbonate, sodium bicarbonate) and / or electron donors (sodium nitrite, sodium thiosulfate) can increase its growth rate, thereby improving its carbon fixation and phosphorus solubilization capabilities, which is of great significance for enhancing the soil carbon sequestration potential, promoting the high-quality development of the photovoltaic industry in salinization-vulnerable areas, and alleviating global climate warming.

[0029] Based on the above advantages, the present invention provides a carbon-fixing bacterial agent, including the alkaline lake Dietzia GD-1 described in the above technical solution.

[0030] As an embodiment, the OD of Dietzia alkaline lake GD-1 in the carbon-fixing bacteria agent is 600 Value ≥ 0.6.

[0031] The present invention provides a method for preparing the carbon-fixing bacteria agent described in the above technical solution, comprising the following steps:

[0032] The alkaline lake Dietzia GD-1 described in the above technical solution is inoculated into a culture medium for cultivation to obtain the carbon-fixing bacterial agent; the culture medium is a culture medium containing a carbon source or a culture medium without a carbon source.

[0033] As an embodiment, the culture medium further comprises an electron donor; the electron donor comprises sodium nitrite and / or sodium thiosulfate. The electron donor can increase the growth rate of Dietzia alkaline lake GD-1.

[0034] As an embodiment, the medium without a carbon source comprises the following components at the following concentrations: Na 2 HPO 4 0.5g / L, KH 2 PO 4 0.5 g / L, MgSO 4 0.3 g / L, CaCl 2 0.2 g / L, NH 4 Cl 0.5g / L, NaNO 3 0.25 g / L, NaCl 0.4 g / L, trace element solution 10 mL / L; the trace element solution includes the following components: FeCl 20.3 g / L, FeSO 4 7H 2 O 0.3g / L, MnSO 4 ·H 2 O 0.15g / L, ZnSO 4 7H 2 O 0.25g / L, CoCl 2 0.2 g / L. The alkaline lake Dietzia GD-1 of the present invention can utilize carbon dioxide in the atmosphere, so it can reproduce and fix carbon dioxide in the atmosphere in a culture medium without a carbon source.

[0035] As an embodiment, the culture medium containing a carbon source is prepared by adding a carbon source to the culture medium not containing a carbon source; the carbon source of the culture medium containing a carbon source includes sodium carbonate and / or sodium bicarbonate. Exogenous addition of an inorganic carbon source can increase the growth rate of Dietzia alkaline lake GD-1.

[0036] As an embodiment, the culture time is 5 to 7 days; the culture temperature is 28°C.

[0037] The present invention provides the use of the alkaline lake Dietzia GD-1 described in the above technical scheme or the carbon-fixing bacteria agent described in the above technical scheme or the carbon-fixing bacteria agent prepared by the preparation method described in the above technical scheme in carbon fixation and / or dissolution of insoluble inorganic phosphorus, and the carbon source for carbon fixation includes an inorganic carbon source.

[0038] As an embodiment, the inorganic carbon source includes one or more of carbon dioxide, sodium bicarbonate and sodium carbonate. The Dietzella alkali lake GD-1 provided by the present invention has a higher carbon fixation potential in a soil environment with a high carbonate content, which helps to develop the application of Dietzella alkali lake GD-1 in special environments (such as soda saline-alkali land).

[0039] As an embodiment, the insoluble inorganic phosphorus may be calcium phosphate. The Dietzia alkaline lake GD-1 provided by the present invention can dissolve the insoluble inorganic phosphorus into soluble phosphate.

[0040] In order to further illustrate the present invention, a strain of Dietzia alkaline lake GD-1 provided by the present invention and its application in carbon fixation and phosphorus dissolution are described in detail below in conjunction with the accompanying drawings and examples, but they should not be construed as limiting the scope of protection of the present invention.

[0041] Example 1 Isolation and Identification of Carbon-Fixing Bacteria Dietzia natronolimnaea GD-1

[0042] The target soil is the soil between fixed panels in the salinized photovoltaic field in Daqing. The specific screening scheme is as follows:

[0043] 1. Collect soil from a depth of 0 to 20 cm, take 10.0 g of the collected soil, add it to 100 mL of sterilized water, and place it on a shaker for shaking (180 rpm, shaking for 30 min) to make a soil suspension. Then dilute the soil suspension in a 10-fold gradient to 10 -4 times, take 0.2 mL of soil dilution, apply it to solid culture medium without organic carbon source, and culture at 28°C for 7 days. The formula of the solid culture medium is: Na 2 HPO 4 0.5 g / L, KH 2 PO 4 0.5 g / L, MgSO 4 0.3 g / L, CaCl 2 0.2 g / L, NaHCO 3 0.5 g / L, NH 4 Cl 0.5g / L, NaNO 3 0.25 g / L, NaCl 0.4 g / L, trace element solution 10 mL / L, agar 20 g / L; the pH value of the solid culture medium is 7.0. The formula of the trace element solution is: FeCl 2 0.3 g / L, FeSO 4 7H 2 O 0.3g / L, MnSO 4 ·H 2 O 0.15g / L, ZnSO 4 7H 2 O 0.25g / L, CoCl 2 0.2 g / L.

[0044] 2. Pick different colonies with better morphology, better growth potential and faster growth rate from the solid culture medium without organic carbon source, and purify them multiple times on a new solid culture medium without organic carbon source by inoculation loop plate streak method until pure culture. Preliminary separation and purification of 25 strains of carbon-fixing bacteria were carried out by dilution plate method, and the dominant bacteria were selected for carbon fixation function rescreening (refer to Example 2), and the one with the best effect was selected as the research object, recorded as GD-1. Scanning electron microscopy was used to determine that the bacterial morphology of the strain was short rod-shaped ( Figure 1 ), the colonies produced on solid culture medium are small, round, orange-red, opaque, smooth and sticky, with neat edges and low protrusions ( Figure 2 ).

[0045] 3. The strain was further subjected to 16S rRNA gene sequencing: the genomic DNA of the strain was amplified by PCR using universal primers 27F (SEQ ID NO.1) and 1492R (SEQ ID NO.2), and the amplified product was sequenced. The sequencing result is shown in SEQ ID NO.3, which is as follows:

[0046] SEQ ID NO.1: 5'-AGTTTGATCMTGGCTCAG-3';

[0047] SEQ ID NO.2: 5'-GGTTACCTTGTTACGACTT-3';

[0048]

[0049] Then, the NCBI database was used to perform BLAST analysis based on the strain 16S rRNA gene sequence, and a phylogenetic tree was constructed ( Figure 3 The results showed that the 16S rRNA gene sequence of the strain was homologous to Dietzia sp. (100% homology to Dietzia natronolimnaea strain DSM 44860 and Dietzia cercidiphyllistrain X0053). Figure 3 It can be seen that the isolated strain is closest to the genetic evolution of the two. Combined with its bacterial morphology and colony characteristics, the strain was identified as Dietzia sp. Further, the whole genome sequencing was performed on it: the extracted whole genome was sent to Shanghai Paisonno Biotechnology Co., Ltd. for second-generation sequencing, and the obtained sequence results were compared with the whole genome sequence of the reference strain Dietzia natronolimnaea S-XJ-1 in the NCBI database for average nucleotide consistency (ANI) analysis. When the ANI value is greater than 95%, it can be considered that the two are the same species. The ANI result showed 98.30%, so the strain was identified as Dietzia natronolimnaea. The strain GD-1 was deposited in the General Microbiology Center of the China Microbiological Culture Collection Administration, and the deposit number is CGMCC No. 31728.

[0050] Example 2 Carbon-fixing enzyme activity of carbon-fixing bacterium Dietzia alkaline lake GD-1

[0051] 1. Materials and methods

[0052] 1.1 Test strain: Dietzia alkaline lake GD-1, isolated and identified in Example 1.

[0053] 1.2 Detection of RubisCO carbon fixation enzyme activity of Dietzia alkaline lake GD-1

[0054] Pick the GD-1 colony purified in Example 1, inoculate it into 10 mL of liquid culture medium without organic carbon source (same as Example 1), and culture it at 28°C, 160r / min for 5 days. Take the bacterial solution at the end of the culture for RubisCO enzyme activity determination, and use the ribulose diphosphate carboxylase / oxygenase (RuBisCO) kit provided by Beijing Biosen Biotechnology Co., Ltd. for determination. At the same time, use the BCA protein concentration determination kit to measure the total protein content of the bacterial solution, and standardize the RuBisCO enzyme activity determination results. For specific measurement methods, refer to the kit instructions. The determination results are shown in Figure 4Among them, Dietzias p. is Dietzias alkaline lake GD-1, and 1 to 4 are the other 4 carbon-fixing bacteria isolated and purified in Example 1.

[0055] 1.3 Data processing

[0056] The raw data of this experiment were preliminarily sorted using Microsoft Excel Office 2016 software. The experimental results were plotted using GraphpadPism 9.5. Figure 4 , where different lowercase letters indicate significant differences among different groups.

[0057] 2. Results and Analysis

[0058] The results showed that the RuBisCO enzyme activity of Dietzia alkaline lake GD-1 was about 41.35 U / mg·prot, which was much greater than that of the other four carbon-fixing bacteria, indicating that it had a good carbon fixation ability.

[0059] Example 3 Study on the carbon fixation ability of Dietzia alkaline lake GD-1 under different conditions

[0060] 1. Materials and methods

[0061] 1.1 Test strain: Dietzia alkaline lake GD-1, isolated and identified in Example 1.

[0062] Carbon-free liquid medium: Na 2 HPO 4 0.5 g / L, KH 2 PO 4 0.5 g / L, MgSO 4 0.3 g / L, CaCl 2 0.2g / L, NH 4 Cl 0.5g / L, NaNO 3 0.25 g / L, NaCl 0.4 g / L, trace element solution 10 mL / L; pH value is 7.0. The formula of the trace element solution is: FeCl 2 0.3 g / L, FeSO 4 7H 2 O 0.3g / L, MnSO 4 ·H 2 O 0.15g / L, ZnSO 4 7H 2 O0.25g / L, CoCl 2 0.2 g / L.

[0063] Na 2 CO 3 Liquid medium: Add Na to the carbon-free liquid medium2 CO 3 0.63 g / L, pH value is 7.0.

[0064] NaHCO 3 Liquid medium: Add NaHCO to carbon-free liquid medium 3 0.5 g / L, pH value is 7.0.

[0065] NaNO 2 Liquid medium: NaNO 2 4.6 g / L, Na 2 HPO 4 0.5 g / L, KH 2 PO 4 0.5 g / L, MgSO 4 0.3 g / L, CaCl 2 0.2 g / L, NaHCO 3 0.5 g / L, NH 4 Cl 0.5g / L, NaNO 3 0.25 g / L, NaCl 0.4 g / L; pH value is 7.0.

[0066] Na 2 S 2 O 3 Liquid medium: NaNO 2 NaNO in liquid medium 2 4.6 g / L replaced by Na 2 S 2 O 3 5g / L, pH value is 7.0.

[0067] 1.2 Study on the carbon fixation ability of Dietzia alkaline lake GD-1 under different inorganic carbon sources and different electron donors

[0068] The GD-1 colony purified in Example 1 was picked and inoculated into 5 mL of carbon-free liquid medium, and cultured at 28°C, 160 r / min, for 5 days. 1 mL of the bacterial solution at the end of the culture was taken, and the abundance of the strain was determined by flow cytometry. 400 μL was then inoculated into 5 mL of carbon-free liquid medium (denoted as Carbon-free), Na 2 CO 3 Liquid culture medium, NaHCO 3 Liquid culture medium, NaNO 2 Liquid medium and Na 2 S 2 O 3The strain abundance in the liquid culture medium was tested again after culturing at 28°C for 5 days. The specific method for measuring microbial abundance was as follows: SYBR-Green (10000×, Solebol) was diluted 100 times and mixed with propidium iodide (PI, Solebol) at a ratio of 50:1 (the volume of SYBR-Green and PI was 50:1) to obtain the fluorescent dye. After 1 mL of the bacterial solution was passed through a 300-mesh nylon sieve to remove large particles, 10 μL of fluorescent dye was added to stain in the dark for 15 minutes, and the number of viable bacteria was determined after vortexing.

[0069] 1.3 Data processing

[0070] The experimental data were analyzed in CytExpert SRT software and then preliminarily sorted using Microsoft ExcelOffice 2016. The experimental results were plotted using GraphpadPism 9.5.

[0071] 2. Results and Analysis

[0072] Since the only carbon sources available to the strain during the culture process are atmospheric carbon dioxide and inorganic carbon sources in the culture medium, the increase in the strain biomass comes from carbon fixation. The experimental results of the carbon fixation ability of Dietzia alkaline lake GD-1 under different conditions are shown in Figure 5 The results showed that in the presence of atmospheric carbon dioxide alone, the strain concentration of Dietzella alkaline lake GD-1 could increase by 5406.06 Events / μL per day, indicating that it has a good ability to fix atmospheric carbon dioxide. When an inorganic carbon source was added, the average daily growth rate of the strain increased slightly, indicating that its carbon fixation ability was improved, which may be the result of the strain fixing carbon through other pathways. However, its utilization of the two different forms of inorganic carbon sources was not much different, and its average daily growth rate was higher in NaHCO 3 、Na 2 CO 3 When an electron donor was added, the carbon fixation capacity of the strain was further improved. 2 S 2 O 3 The average daily growth rate was 6085.67 Events / μL, and NaNO 2 When NaNO 2 The effect on improving the carbon fixation ability of the strain is better, which may be due to NaNO 2It can provide more energy for the strain to fix carbon. The results show that Dietzella alkaline lake GD-1 can maintain a good growth rate and carbon fixation ability in extremely low-nutrient salt culture media, and the related ability can be driven by electron donors. At the same time, it has a higher carbon fixation potential in soil environments with high carbonate content, which is helpful for the development of strains in special environments (such as karst soils).

[0073] Example 4 Analysis of the carbon fixation pathway of the carbon-fixing bacterium Dietzia alkaline lake GD-1

[0074] 1. Materials and methods

[0075] The alkaline lake Dietzella GD-1 isolated and identified in Example 1 was sent to Shanghai Paisono Biotechnology Co., Ltd., and the Illumina Novasek platform was used for second-generation sequencing, and the whole genome framework map was assembled and spliced. The bacteria were annotated and functionally identified using bioinformatics software and databases, and the carbon fixation pathway of strain GD-1 was inferred through the KEGG database to provide a basis for studying the carbon fixation ability of the strain. The results are shown in Figure 6 , where the letters represent the key enzymes in the carbon fixation pathway, pps—phosphoenolpyruvate carboxylase; ppc—phosphoenolpyruvate carboxylase; pycA—pyruvate carboxylase; maeB—malic enzyme; mdh—malate dehydrogenase; Fum—fumaryl esterase; sdh / frd—succinate dehydrogenase; sucD—succinate CoA synthetase; korA—2-oxoglutarate ferredoxin oxidoreductase; icd—isocitrate dehydrogenase; ACO—heptenoate hydratase; gltA—citrate synthase.

[0076] 2. Results and Analysis

[0077] Based on the KEGG database, the CO 2 Fixation pathway. First, pyruvate is converted into phosphoenolpyruvate by phosphoenolpyruvate synthase and then fixes CO with the participation of phosphoenolpyruvate carboxylase. 2 Oxaloacetate is generated, and oxaloacetate can generate malic acid under the catalysis of malic dehydrogenase. On the one hand, malic acid can be directly converted into pyruvic acid under the catalysis of malic enzyme to complete the carbon fixation cycle. On the other hand, it can form fumarate under the action of fumarate esterase, and then be converted into succinate by succinate dehydrogenase, and further form succinate coenzyme A, and can fix two molecules of CO again in the process of forming isocitrate. 2 , then citrate is generated by aconitic acid under the catalysis of heptenoate hydratase, and finally converted into oxaloacetate to form a carbon fixation cycle ( Figure 6 ). At the same time, the results of whole genome analysis also found that Dietzia alkaline lake GD-1 also has other key carbon fixation enzymes, such as ketoglutarate: ferredoxin reductase, which further enhances its carbon fixation ability.

[0078] Example 5 Study on the phosphate solubilization ability of carbon-fixing bacteria Dietzia alkaline lake GD-1

[0079] 1. Materials and methods

[0080] 1.1 Test strain: Dietzia alkaline lake GD-1, isolated and identified in Example 1.

[0081] Pikovskaya liquid medium: yeast extract 0.5 g / L, glucose 10 g / L, Ca 3 (PO 4 ) 2 5g / L, (NH 4 ) 2 SO 4 0.5 g / L, KCl 0.2 g / L, Mg 3 (PO 4 ) 2 0.1g / L, MgSO 4 0.0001 g / L, FeSO 4 0.0001g / L; pH value is 7.0.

[0082] 1.2 Detection of phosphate solubilization ability of carbon-fixing bacteria Dietrichia alkali-lake GD-1

[0083] 5g / L KH 2 PO 4 The standard solution was diluted to 0, 2, 2.5, 250, 500, and 1000 mg / L concentrations. The absorbance was determined by the molybdenum antimony colorimetric method. The standard curve equation was y = 0.0028x + 0.048, R 2 =0.9996.

[0084] The colonies purified from the solid medium without an organic carbon source in Example 1 were inoculated into 10 mL of Pikovskaya liquid medium and cultured at 28 ° C and 160 r / min for 5 days. A control group without inoculation of strains was set up. At the end of the culture, the bacterial solution was centrifuged, 1 mL of sterile water was added to the bacterial precipitate, and the mixture was ground with a tissue disruptor and mixed with the supernatant, and NaHCO was added. 3 After shaking and filtering, the organic phosphorus content was determined by the molybdenum antimony colorimetric method. Figure 7 Among them, Dietzias p. is Dietzias alkaline lake GD-1, and 1 to 4 are the other 4 carbon-fixing bacteria isolated and purified in Example 1.

[0085] 1.3 Data processing

[0086] The experimental data were preliminarily sorted using Microsoft Excel Office 2016. The experimental results were plotted using GraphpadPism 9.5.

[0087] 2. Results and Analysis

[0088] By using the ability to dissolve tricalcium phosphate as an indicator, it was determined that Dietzia alkaline lake GD-1 had the ability to dissolve phosphate and achieved the utilization of insoluble phosphates. Its phosphate dissolving capacity was 426.61 mg / L, while the phosphate dissolving capacity of the other four carbon-fixing bacteria was as low as 115.18 mg / L and as high as 301.25 mg / L, all lower than that of Dietzia alkaline lake GD-1.

[0089] In summary, the alkaline lake Dietzia GD-1 isolated by the present invention has the ability to fix atmospheric carbon dioxide and at the same time has the ability to solubilize phosphorus, which provides bacterial resources for improving the carbon fixation and emission reduction capacity of salinized soils, provides technical support for in-depth understanding of the microbial regulation process of the storage form of nutrients in salinized soils, lays the foundation for proposing soil microbial improvement technology in photovoltaic fields, and promotes the high-quality development of the photovoltaic industry in salinized vulnerable areas.

[0090] Although the above embodiment describes the present invention in detail, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the protection scope of the present invention.

Claims

1. A strain of Dietzia natronolimnaea GD-1, with a deposit number of CGMCC No.31728.

2. A carbon-fixing bacterial agent, characterized in that: It includes the Dietzia alkaline lake GD-1 described in claim 1.

3. The carbon-fixing bacterial agent according to claim 2, characterized in that The OD of Dietzia alkaline lake GD-1 in the carbon-fixing bacteria agent 600 Value ≥ 0.

6.

4. The method for preparing the carbon-fixing bacteria agent according to claim 2 or 3, characterized in that: The following steps are involved: The alkaline lake Dietzia GD-1 described in claim 1 is inoculated into a culture medium for cultivation to obtain the carbon-fixing bacterial agent; the culture medium is a culture medium containing a carbon source or a culture medium without a carbon source.

5. The preparation method according to claim 4, characterized in that: The culture medium further comprises an electron donor; the electron donor comprises sodium nitrite and / or sodium thiosulfate.

6. The preparation method according to claim 4, characterized in that: The culture medium not containing a carbon source comprises components with the following concentrations: Na2HPO4 0.5 g / L, KH2PO4 0.5 g / L, MgSO4 0.3 g / L, CaCl2 0.2 g / L, NH4Cl 0.5 g / L, NaNO3 0.25 g / L, NaCl 0.4 g / L, and a trace element solution of 10 mL / L; the trace element solution comprises components with the following concentrations: FeCl2 0.3 g / L, FeSO4·7H2O 0.3 g / L, MnSO4·H2O 0.15 g / L, ZnSO4·7H2O 0.25 g / L, and CoCl2 0.2 g / L; the pH value of the culture medium not containing a carbon source is 7.0-7.

4.

7. The preparation method according to claim 4, characterized in that: The culture medium containing the carbon source is prepared by adding the carbon source to the culture medium not containing the carbon source; the added carbon source includes: sodium carbonate and / or sodium bicarbonate.

8. Use of the alkaline lake Dietzia GD-1 described in claim 1 or the above carbon-fixing bacteria agent described in claim 2 or 3, or the carbon-fixing bacteria agent prepared by the preparation method described in any one of claims 4 to 7 in carbon fixation and / or dissolution of insoluble inorganic phosphorus, wherein the carbon source for carbon fixation includes an inorganic carbon source.

9. The use according to claim 8, characterized in that: The inorganic carbon source includes one or more of carbon dioxide, sodium bicarbonate and sodium carbonate.

10. The use according to claim 8, characterized in that: The poorly soluble inorganic phosphorus includes calcium phosphate.

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