Kocuria rootrophila and application thereof
By screening and identifying Rhizophila ZMC73, the problem of difficulty in effectively synthesizing IAA in saline-alkali land and tolerating saline-alkali stress in the prior art is solved, and the effect of promoting crop growth and improving yield in saline-alkali land is achieved.
Patent Information
- Application Number
- CN202510375780.6
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-27
- Publication Date
- 2025-07-29
AI Technical Summary
The prior art is difficult to screen out plant proliferation strains that can both synthesize IAA and tolerate saline and alkali stress, resulting in limited growth and reduced yields of saline and alkaline crops.
Rhizophila ZMC73 was screened and identified, which is able to grow in a saline-alkali environment and secrete IAA efficiently. It is used to prepare microbial agents and fertilizers to promote crop growth.
Rhizophila ZMC73 significantly promotes the growth of soybeans, corn and wheat in salt stress environments, improves the plant's ability to resist saline-alkali stress and diseases, and improves soil quality.
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Figure CN120384017A_ABST
Abstract
Description
Technical Field
[0001] The present invention belongs to the technical field of microbiology and relates to a Kocuria rhizophila and its application. Background Art
[0002] Plant growth-promoting rhizobacteria (PGPR), as microorganisms that can promote plant growth through mechanisms such as nitrogen fixation, phosphorus solubilization, and synthesis of plant hormones (such as IAA), have received extensive attention. Especially in adverse environments such as saline-alkali lands, crop growth is severely inhibited, and traditional agricultural measures are difficult to effectively improve soil conditions. Therefore, screening and developing highly efficient salt-tolerant plant growth-promoting bacteria, especially strains that can synthesize IAA (indole-3-acetic acid), are of great significance for increasing crop yields, improving soil quality, and promoting sustainable agriculture.
[0003] As a key plant auxin, IAA can promote root development, enhance nutrient absorption, and improve the tolerance of plants to adversity. However, in agricultural production practices, there are often various different plots, such as saline-alkali lands. Poor tolerance of crops to saline-alkali lands often leads to a decrease in crop growth rate, a reduction in yield, or poor resistance to pests and diseases. Therefore, strains that only have the ability to produce IAA are difficult to meet the actual needs of agricultural production. It is urgent to further develop strains that not only have the ability to produce IAA but also have the ability to tolerate saline-alkali stress. Summary of the Invention
[0004] In order to better meet the growth requirements of crops in different plots, especially saline-alkali lands, and improve the growth of crops in this type of plot and the soil microenvironment. The present invention has screened out a Kocuria rhizophila strain that produces IAA and tolerates saline-alkali environments, and its preservation information is as follows:
[0005] Species name: Kocuria rhizophila
[0006] Latin name: Kocuria rhizophila
[0007] Strain number: ZMC73
[0008] Preservation institution: General Microbiological Center of the China Committee for Culture Collection of Microorganisms
[0009] Abbreviation of the preservation institution: CGMCC
[0010] Address of the preservation institution: Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing
[0011] Preservation date: March 3, 2025
[0012] Deposit number: CGMCC NO.33694.
[0013] Specifically, Kocuria rhizophila ZMC73 was isolated from the roots of Oxytropis glabra in the saline-alkali beach of Alar City, Xinjiang Uygur Autonomous Region. The colony of Kocuria rhizophila ZMC73 is round, with neat edges, smooth surface, convex, yellow or orange-yellow, and opaque.
[0014] Furthermore, the 16S rDNA nucleotide sequence of Kocuria rhizophila ZMC73 is shown as SEQ ID NO.1.
[0015] Furthermore, Kocuria rhizophila ZMC73 has the ability to secrete IAA.
[0016] Furthermore, Kocuria rhizophila ZMC73 is tolerant to salt stress, and the salt includes NaCl. The present invention has confirmed that Kocuria rhizophila ZMC73 can grow in LB medium with NaCl concentration of 1-9% (w / v).
[0017] Furthermore, Kocuria rhizophila ZMC73 is tolerant to alkali stress, and the alkali includes NaOH. The present invention has confirmed that Kocuria rhizophila ZMC73 can grow in the range of pH value of 8.0-10.0.
[0018] In the second aspect, the present invention claims the application of Kocuria rhizophila ZMC73.
[0019] Furthermore, in the above application, Kocuria rhizophila ZMC73 is used to prepare a microbial inoculant, or Kocuria rhizophila ZMC73 is used to prepare a microbial fertilizer. The functions of the microbial inoculant and the microbial fertilizer include: promoting plant growth, enhancing the ability of plants to resist saline-alkali stress, enhancing the ability of plants to resist diseases, improving the physical and chemical properties of the soil, regulating the soil microbial flora, etc. The active ingredients of the microbial inoculant and the microbial fertilizer contain the cells of Kocuria rhizophila ZMC73 and / or its fermentation products.
[0020] In the third aspect, the present invention claims a microbial inoculant, which includes the cells of Kocuria rhizophila ZMC73 and / or its fermentation products.
[0021] In the fourth aspect, the present invention claims a microbial fertilizer, which includes the cells of Kocuria rhizophila ZMC73 and / or its fermentation products.
[0022] Compared with the prior art, the present invention "a Rhizobium radiobacter and its application" has the following beneficial effects: A Rhizobium radiobacter ZMC73 was isolated from the roots of Oxytropis glabra plants in a saline-alkali beach. This strain has the ability to grow in a saline-alkali environment. Specifically, Rhizobium radiobacter ZMC73 can grow in an environment with a NaCl concentration of 1-9% (w / v) or within a pH range of 8.0-10.0. In addition, Rhizobium radiobacter ZMC73 can efficiently synthesize IAA, and its ability to secrete IAA under L-tryptophan induction is 402.21 mg / L. It has been experimentally confirmed that Rhizobium radiobacter ZMC73 can significantly promote the growth of soybeans, corn, and wheat in a salt-stress environment, and significantly improve or enhance indicators such as plant height, root length, and fresh weight of stems and leaves.
[0023] Based on the characteristics of Rhizobium radiobacter ZMC73 producing IAA and tolerating saline-alkali, this strain provides a strain resource for the preparation of microbial fertilizers or inoculants, alleviating the environmental hazards caused by the excessive application of chemical fertilizers; on the other hand, it provides a safe and effective microbial resource for microbial fertilizers or inoculants suitable for saline-alkali areas. This strain has broad application prospects in the fields of saline-alkali land agriculture, ecological restoration, and adversity crop breeding, and is expected to become an important tool for increasing crop yields and improving soil quality. Description of the Drawings
[0024] Figure 1 shows the colony growth of Rhizobium radiobacter ZMC73 on LB medium.
[0025] Figure 2 is the phylogenetic tree of Rhizobium radiobacter ZMC73.
[0026] Figure 3 is the qualitative determination test of the IAA secretion ability of Rhizobium radiobacter ZMC73.
[0027] Figure 4 is the standard curve for IAA quantitative determination.
[0028] Figure 5 shows the growth of soybean seedlings in each group.
[0029] Figure 6 is the column chart of the plant height of soybean seedlings in each group.
[0030] Figure 7 is the column chart of the root length of soybean seedlings in each group.
[0031] Figure 8 is the column chart of the fresh weight of stems and leaves of soybean seedlings in each group.
[0032] Figure 9 is the column chart of the fresh weight of roots of soybean seedlings in each group.
[0033] Figure 10It is a bar graph of the leaf area of soybean seedlings in each group.
[0034] Figure 11 It is the growth situation of wheat seedlings in each group.
[0035] Figure 12 It is a bar graph of the plant height (A), root length (B), fresh weight of stems and leaves (C), and fresh weight of roots (D) of wheat seedlings in each group.
[0036] Figure 13 The growth situation of maize seedlings in each group.
[0037] Figure 14 It is a bar graph of the plant height (A), root length (B), fresh weight of stems and leaves (C), and fresh weight of roots (D) of maize seedlings in each group.
[0038] Explanation of reference numerals: ZMC73 represents the plants treated with the suspension of Klebsiella radicincitans (negative control); CK represents the plants treated with sterile water (blank control); NaCl + ZMC73 represents the plants treated with the suspension of Klebsiella radicincitans ZMC73 and NaCl solution (experimental group); NaCl + CK represents the plants treated with NaCl and sterile water (positive control). Detailed implementation mode
[0039] The technical solutions of the present invention will be clearly and completely described below in conjunction with the embodiments. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the scope of protection of the present invention.
[0040] Example 1
[0041] This example describes the isolation, purification and colony growth morphology of Klebsiella radicincitans ZMC73.
[0042] LB solid medium: 10 g of tryptone, 5 g of yeast extract powder, 10 g of NaCl, 15 g of agar, 1000 mL of H2O, pH = 7.0 - 7.2, sterilized at 121 °C for 30 min.
[0043] LB liquid medium: 10 g of tryptone, 5 g of yeast extract powder, 10 g of NaCl, 1000 mL of H2O, pH = 7.0 - 7.2, sterilized at 121 °C for 30 min.
[0044] IAA fermentation medium: 10 g of tryptone, 5 g of yeast extract powder, 10 g of NaCl, 1 g of L-tryptophan, 1000 mL of H2O, pH = 7.0 - 7.2, sterilized at 121 °C for 30 min.
[0045] Strain origin: The rhizophilic Coxsackie strain ZMC73 was isolated from the roots of Oxytropis parviflora in the saline-alkali beach of Aral City, Xinjiang Uygur Autonomous Region.
[0046] Separation and purification method: Take the collected Oxytropis microcarpa plants, rinse with tap water to remove the soil on the root surface of the plant, cut the roots into several sections, place them in 2% sodium hypochlorite and 70% ethanol for 7 minutes each, then rinse with sterile water 4 times, then grind the roots in a sterile mortar and pestle, and prepare a series of dilution concentrations of grinding solution with sterile water, and select a dilution concentration gradient of 10 -3 and 10 -4 The grinding liquid was coated on a plate, and then spread on a solid LB medium containing 3% NaCl, and cultured in a constant temperature incubator at 28°C; single colonies with different morphologies and colors were picked and streaked on the plate multiple times, and subcultured for 2 to 3 generations to obtain a purified Kockruppenfungia rhizophila ZMC73 strain with consistent growth morphology.
[0047] Example 2
[0048] This example describes the taxonomic identification of Coxiella rhizophila ZMC73.
[0049] 1. Biological identification
[0050] After culturing K. rhizogenes ZMC73 on LB solid medium plates at 30°C for 2 days, the colonies are round, with neat edges, smooth and raised surfaces, yellow or orange-yellow, and opaque. Figure 1 shown.
[0051] 2. Systematic classification and identification
[0052] Using the colony PCR identification method, pick Kocuria rhizophila ZMC73 on the LB solid medium plate with an inoculation loop. Dilute the bacterial mass with 15 μL of sterile water, centrifuge at 12,000 r / min for 5 min, discard the supernatant, resuspend with 15 μL of sterile water, repeat centrifugation once, incubate in a water bath at 95 °C for 5 min, and then place on ice for 15 - 20 min. Use the treated bacterial suspension as the PCR identification template, and amplify the target gene fragment with the universal bacterial primers 27F and 1492R (27F: 5'-AGAGTTTGATCCTGGCTCAG-3'; 1492R: 5'-GGCTACCTTGTTACGACTT-3'). The PCR reaction system is as follows: 1 μL each of 27F and 1492R primers (10 mmol / L), 3 μL of Kocuria rhizophila ZMC73 bacterial suspension, 12.5 μL of 2×PCR mix, and 7.5 μL of sterile water, for a total of 25 μL; use the system without adding the bacterial suspension as a control. The amplification program is: 95 °C for 3 min; 95 °C for 1 min, 56 °C for 1 min, 72 °C for 1 min, for 25 cycles; 72 °C for 10 min. Submit the PCR product for sequencing (Sangon Biotech (Xi'an) Co., Ltd.), and the 16S rDNA of Kocuria rhizophila ZMC73 is shown as SEQ ID NO.1.
[0053] Perform a homology comparison of the obtained SEQ ID NO.1 with the nucleic acid data in Ezbiocloud (https: / / www.ezbiocloud.net / identify). The results show that the homology between Kocuria rhizophila ZMC73 and Kocuria rhizophila strain TA68 is 99.86%. Use the software MOLECULAR EVOLUTIONARY GENETIC ANALYSIS software (MEGA 7.0) to construct a phylogenetic tree. As Figure 2 shown, the sequences of Kocuria rhizophila ZMC73 and Kocuria rhizophila TA68 can form a stable evolutionary branch, so it is named Kocuria rhizophila ZMC73.
[0054] Example 3
[0055] This example describes the evaluation of the salt (NaCl) and alkali (NaOH) tolerance capabilities of Kocuria rhizophila ZMC73.
[0056] 1. Evaluation of salt tolerance
[0057] Pick a single colony of *Kocuria rhizophila* ZMC73 and inoculate it into LB liquid medium. Incubate it on a shaker at 30 °C and 150 r / min for 12 h to obtain a seed culture solution. Inoculate the seed culture solution into LB liquid media containing 1%, 3%, 5%, 7%, 9%, and 11% (w / v) NaCl respectively, with an inoculation amount of 1%. Incubate it with shaking at 30 °C for 48 h, and measure the absorbance value (OD 600 ) at 600 nm; use LB liquid media with different NaCl concentrations without inoculating the seed culture solution as the control group. The results show that *Kocuria rhizophila* ZMC73 can grow in LB media with NaCl concentrations of 1-9% (w / v).
[0058] 2. Evaluation of alkali tolerance
[0059] Pick a single colony of *Kocuria rhizophila* ZMC73 and inoculate it into LB liquid medium. Incubate it on a shaker at 30 °C and 150 r / min for 12 h to obtain a seed culture solution. Use 1 M NaOH or 1 M HCl to adjust the pH value of the LB liquid medium, and set the following pH gradients: 7.0, 8.0, 9.0, 10.0, 11.0, 12.0. Inoculate the seed culture solution into LB liquid media with different pH values, with an inoculation amount of 1%. Incubate it with shaking at 30 °C for 48 h, and measure OD 600 , and use LB liquid media with different pH values without inoculating the seed culture solution as the control group. The results show that *Kocuria rhizophila* ZMC73 can grow in the range of pH 8.0-10.0.
[0060] Example 4
[0061] This example describes the qualitative and quantitative determination of the IAA secretion ability of *Kocuria rhizophila* ZMC73.
[0062] Salkowski colorimetric reagent: Dissolve 12 g of FeCl3 in 300 mL of deionized water, slowly add 429.7 mL of concentrated H2SO4 (concentration ≥ 98%), and make up the volume to 1 L after cooling.
[0063] Prepare an IAA standard curve: Prepare 50 mL of a standard IAA solution with a concentration of 100 μg / mL, and dilute it to 6.25, 12.5, 25, and 50 μg / mL respectively. Mix the IAA solution and the salksowski colorimetric solution in a volume ratio of 1:1, and react at room temperature in the dark for 30 min; use distilled water mixed with an equal volume of the salksowski colorimetric solution as the control. Then use ultraviolet spectrophotometry to measure the OD value (OD 530 ) at 530 nm for each concentration. Plot the IAA concentration on the x-axis and OD 530 on the y-axis to obtain the IAA standard curve.
[0064] Quantitative determination of the IAA concentration in the bacterial solution: Pick a single colony of *Kocuria rhizophila* ZMC73 and inoculate it into LB liquid medium. Incubate it on a shaker at 30 °C and 150 r / min for 12 h to obtain the seed culture solution. Take 100 μL of the seed culture solution and transfer it into the IAA fermentation medium. Incubate it on a shaker at 30 °C and 180 r / min for 3 d, then take it out, centrifuge at 12000 r / min for 2 min, take 500 μL of the supernatant, mix it evenly with an equal volume of Salkowski colorimetric solution, and place it in the dark at room temperature for 30 min to measure the OD 530 . Use the fermentation medium mixed with an equal volume of Salkowski colorimetric solution as the control group. According to the OD 530 and the standard curve, calculate the corresponding IAA content.
[0065] Figure 3 is the qualitative determination test for the ability of *Kocuria rhizophila* ZMC73 to secrete IAA; Figure 4 is the standard curve for IAA quantitative determination. It can be seen from Figure 3 that *Kocuria rhizophila* ZMC73 has the ability to produce IAA. Detect its OD 530 absorbance at this wavelength, and then according to the absorbance value and the Figure 4 standard curve, calculate that the ability of *Kocuria rhizophila* ZMC73 to secrete IAA under L-tryptophan induction is 402.21 mg / L.
[0066] Example 5
[0067] This example describes the promotion of soybean growth by *Kocuria rhizophila* ZMC73 under salt stress conditions.
[0068] Preparation of sterile soybean seedlings: Soak soybean seeds in 2% sodium hypochlorite for 3 min, wash them 3 times with sterile water, then soak them in 70% ethanol for 2 min, wash them 5 times with sterile water, germinate them in clear water at 25 °C for 3 d, and transfer them to flower pots containing nutrient soil:vermiculite with a volume ratio of 2:1 (total volume 180 cm 3 ) for germination.
[0069] Preparation of the bacterial suspension: Pick the cells of *Kocuria rhizophila* ZMC73 and inoculate them into LB liquid medium. Incubate them at 37 °C and 150 r / min. When the OD 600 value is greater than 1.5, centrifuge at 5000 rmp / min for 5 min, discard the supernatant, add 30 mL of sterile water to wash the cells twice to remove the medium components, and prepare a bacterial suspension of 1.5×10 8 CFU / mL.
[0070] Inoculation: The prepared bacterial suspension was inoculated onto the roots of soybean seedlings with fully opened dicotyledons cultured aseptically. 2 mL was inoculated for each plant, and they were allowed to grow at 25 °C for 30 days. During this period, NaCl solution was added to each pot until the NaCl concentration in the pot reached 0.3‰, with pH = 8.0 - 8.5. Observe the growth status of soybeans. When they grew for 30 days, measure the plant height, root length, fresh weight of stems and leaves, fresh weight of roots, and leaf area of soybeans. Using only inoculated bacterial suspension as the negative control (ZMC73); using only adding NaCl solution as the positive control (CK-NaCl); using sterile water as the blank control (CK).
[0071] Figure 5 are the growth conditions of soybean seedlings in each group. It can be seen that there are differences in the growth conditions of soybean plants between the experimental group and the control group. Figures 6 - 10 are respectively the bar charts of the plant height, root length, fresh weight of stems and leaves, fresh weight of roots, and leaf area of soybean seedlings in each group. From Figures 6 - 10 it can be known that through the analysis of the biomass of soybean plants, the plant height and root length of soybean plants in the experimental group are higher than those in the control group, and the fresh weight of stems and leaves, fresh weight of roots, and leaf area are all significantly higher than those in the control group (P ≤ 0.05).
[0072] Example 6
[0073] This example describes the promotion of wheat growth by Kocuria rhizophila ZMC73 under salt stress conditions.
[0074] Preparation of sterile wheat seedlings: Soak wheat seeds in 5% sodium hypochlorite for 3 min, rinse with sterile water 3 times, then soak in 70% ethanol for 5 min, rinse with sterile water 5 times, germinate in clear water at 25 °C for 1 day, and transfer to flower pots containing nutrient soil: vermiculite with a volume ratio of 2:1 (total volume 180 cm 3 ) for germination.
[0075] Preparation of bacterial suspension: The same as Example 5.
[0076] Inoculation: The prepared bacterial suspension was inoculated onto the roots of wheat with leaf length of 4 - 5 cm cultured aseptically. 5 mL was inoculated for each pot, and they were allowed to grow at 25 °C for 30 days. During this period, NaCl solution was added to each pot until the NaCl concentration in the pot reached 0.5‰, with pH = 8.0 - 8.5. Observe the growth status of wheat. When they grew for 30 days, measure the plant height, root length, fresh weight of stems and leaves, fresh weight of roots of wheat, and analyze whether Kocuria rhizophila ZMC73 has a promoting effect on wheat growth under NaCl stress. Using only inoculated bacterial suspension as the negative control (ZMC73); using only adding NaCl solution as the positive control (CK-NaCl); using sterile water as the blank control (CK).
[0077] Figure 11 are the growth conditions of wheat seedlings in each group. It can be seen that there are differences in the growth conditions of wheat plants between the experimental group and the control group.Figure 12 It is a bar graph of plant height (A), root length (B), fresh weight of stems and leaves (C), and fresh weight of roots (D) of wheat seedlings in each group. From Figure 12 it can be seen that through the analysis of the biomass of wheat plants, the plant height, root length, and fresh weight of stems and leaves of wheat plants in the experimental group were significantly higher than those in the control group (P≤0.05), and the fresh weight of roots was higher than that in the control group.
[0078] Example 7
[0079] This example describes the promotion of maize growth by *Kocuria rhizophila* ZMC73 under salt stress conditions.
[0080] Preparation of sterile maize seedlings: Soak maize seeds in 2% sodium hypochlorite for 3 min, wash them 3 times with sterile water, then soak them in 70% ethanol for 5 min, wash them 5 times with sterile water, germinate them in clear water at 25℃ for 3 d, and transfer them to flower pots containing nutrient soil: vermiculite at a ratio of 2:1 (total volume 180 cm 3 ).
[0081] Preparation of bacterial suspension: The same as in Example 5.
[0082] Inoculation: Inoculate the prepared bacterial suspension onto the roots of maize at the two-leaf and one-heart stage (plant height 4 - 5 cm) grown under sterile conditions, inoculate 3 mL per pot, continue to grow at 25℃ for 30 d, add saline water to each pot during this period until the NaCl concentration in the pot reaches 0.2‰, pH = 8.0 - 8.5, observe the growth status of maize, measure the plant height, root length, fresh weight of stems and leaves, and fresh weight of roots of maize after growing for 30 d, and analyze whether *Kocuria rhizophila* ZMC73 has a promoting effect on maize growth under NaCl stress. Use only inoculated bacterial suspension as the negative control (ZMC73); use only added NaCl solution as the positive control (CK-NaCl); use sterile water as the blank control (CK).
[0083] Figure 13 It shows the growth of maize seedlings in each group, Figure 14 It is a bar graph of plant height (A), root length (B), fresh weight of stems and leaves (C), and fresh weight of roots (D) of maize seedlings in each group. From Figures 13 - 14 it can be seen that there are differences in the growth of maize plants between the experimental group and the control group. Through the analysis of the biomass of maize plants, the plant height, root length, and fresh weight of stems and leaves of maize plants in the experimental group were significantly higher than those in the control group (P≤0.05), and the fresh weight of roots was higher than that in the control group.
[0084] In summary, a strain of Rhizobium radiobacter ZMC73 was isolated from the roots of Oxytropis glabra in saline-alkali beaches. This strain has the ability to grow in saline-alkali environments and can also secrete IAA efficiently. It has been verified that Rhizobium radiobacter ZMC73 can promote the growth of soybeans, corn, and wheat in salt-stressed environments and help plants resist salt-stressed environments. Rhizobium radiobacter ZMC73 has good application prospects in the preparation of microbial fertilizers and microbial inoculants suitable for saline-alkali areas.
[0085] The above-described embodiments are only a part of the embodiments of the present invention, rather than all embodiments. The detailed description of the embodiments of the present invention is not intended to limit the scope of the present invention claimed, but merely represents selected embodiments of the present invention. All other embodiments obtained by relevant deductions and substitutions made by those of ordinary skill in the art under the premise of not making creative efforts based on the concept of the present invention fall within the scope of protection of the present invention.
Claims
1. A Rhizobium radiobacter, characterized in that, The Rhizobium radiobacter is deposited in the China General Microbiological Culture Collection Center with the deposit number of CGMCC NO. 33694.
2. The Rhodococcus rhodochrous according to claim 1, characterized in that, The nucleotide sequence of the 16S rDNA of the Rhizobium radiobacter is as shown in SEQ ID NO.
1.
3. The Rhodococcus rhodochrous according to claim 1, characterized in that, The Rhizobium radiobacter secretes IAA.
4. The Rhodococcus rhodochrous according to claim 1, characterized in that, The Rhizobium radiobacter is tolerant to salt stress, and the salt includes NaCl.
5. The Rhodococcus rhodochrous according to claim 1, characterized in that, The Rhizobium radiobacter is tolerant to alkali stress, and the alkali includes NaOH.
6. Use of the Rhizobium radiobacter according to claim 1.
7. The application according to claim 6, characterized in that, The Rhizobium radiobacter is used for preparing a microbial inoculant.
8. The application according to claim 6, characterized in that, The Rhizobium radiobacter is used for preparing a microbial fertilizer.
9. A microbial inoculant, characterized in that, The microbial inoculant comprises the cells of the Rhizobium radiobacter according to any one of claims 1 to 5 and / or its fermentation products.
10. A microbial fertilizer, characterized in that, The microbial fertilizer comprises the cells of the Rhizobium radiobacter according to any one of claims 1 to 5 and / or its fermentation products.