CN109097305B - Rhizobium and application thereof - Google Patents
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Abstract
The invention discloses Rhizobium sp.WP 1-3 with the preservation number of CGMCC No. 14472. In addition, the invention also discloses application of the rhizobium in soil potassium dissolving and difficult soluble phosphorus decomposition in soil. The rhizobium of the invention is an advantageous strain screened by taking potassium feldspar as a potassium source, the strain can convert insoluble potassium elements in aluminosilicate in soil into soluble components in the growth and propagation processes, and meanwhile, the source of phosphorus elements in soil can be increased by promoting the decomposition of insoluble phosphorus. In addition, the strain also has the function of fixing nitrogen, and provides nitrogen needed by plant growth and metabolism.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to rhizobium and application thereof.
Background
Potassium and phosphorus are important nutrient elements in plants, are necessary for biological nitrogen fixation, and are key components for promoting plant growth and metabolism. Potassium and phosphorus play an important role in regulating cell penetration, transporting substances, promoting enzyme activation, promoting photosynthesis, improving the assimilation rate of carbon dioxide, promoting the synthesis and transportation of substances in crops, enhancing plant resistance and tolerance and the like. Most of potassium and phosphorus in soil exist in minerals such as aluminosilicate, apatite and the like, the potassium and phosphorus existing in the form of mineral potassium and mineral phosphorus cannot be directly absorbed by plants, and the mode of supplementing plants with chemical potassium and chemical phosphorus commonly adopted in agriculture easily damages soil structures to cause environmental pollution. With the increase of the environmental importance of people, the characteristics of the biological fertilizer in the aspects of improving the sources of nutrient elements in soil, increasing the diversity of soil and improving the quality of agricultural products attract extensive attention.
The phosphate-solubilizing potassium-solubilizing bacteria are strains which can effectively decompose potassium-containing mineral substances and phosphorus-containing mineral substances in soil, can convert insoluble potassium and phosphorus in the soil into soluble nutrients, and increase the sources of potassium elements and phosphorus elements in the soil. The development and utilization of the microbial fertilizer prepared by the high-efficiency phosphate-solubilizing and potassium-solubilizing bacteria have very important significance for the development of agricultural life and production and the environmental protection.
Disclosure of Invention
The technical problem to be solved by the invention is to provide rhizobium capable of being used for soil potassium dissolving and indissolvable phosphorus decomposition in soil aiming at the defects of the prior art. The strain is derived from soil and is ecologically safe, the strain is a dominant strain screened by taking potassium feldspar as a potassium source, insoluble potassium elements in aluminosilicate in the soil can be converted into soluble components in the growth and propagation processes, and meanwhile, the source of phosphorus elements in the soil can be increased by promoting the decomposition of insoluble phosphorus. In addition, the strain also has the function of fixing nitrogen, and provides nitrogen needed by plant growth and metabolism.
In order to solve the technical problems, the invention adopts the technical scheme that: a Rhizobium, which is characterized in that the Rhizobium is Rhizobium sp.WP 1-3, the preservation number is CGMCC No.14472, the preservation date is 7 months and 27 days in 2017, the preservation unit is China general microbiological culture Collection center, and the address is as follows: the institute of microbiology, national academy of sciences No.3, Xilu No.1, Beijing, Chaoyang, Beijing.
1. The strain has the following properties:
morphological characteristics: the bacterial colony of the WP1-3 strain is circular, and the bacilliform bacteria are colorless and transparent, have neat edges, are wet and sticky and have high elasticity.
Physiological and biochemical characteristics: a gram-negative bacterium.
2. The screening method of the strain comprises the following steps:
step one, enrichment: collecting soil samples about 20g from 10cm below the ground surface of garden, white sand road side, Bowen road square and Ganjai village in Yanta district of Xian city, drying the collected soil samples in an oven, sieving the dried soil with a 100-mesh sieve, accurately weighing 1g of the soil sieved with the 100-mesh sieve in 9mL of sterile water in an aseptic manner, standing for 30min, and oscillating for 30min in a shaking table at room temperature to obtain the dilution of 10-1The soil suspension of (a); sucking out 1mL of the above soil suspension, adding 9mL of sterile water, and mixing to obtain a dilution of 10-2The soil suspension of (a); from a dilution of 10-2Sucking 1mL of the soil suspension, adding 9mL of sterile water, and fully stirringMixing to obtain dilution of 10-3The soil suspension of (a);
step two, primary screening and purification: taking the dilution degree in the step one as 10-3Coating 0.1mL of soil suspension in a potassium bacteria culture medium by adopting a coating plate method, placing the potassium bacteria culture medium in a constant-temperature incubator at 30 ℃ for inverted culture for 5-7 days, regarding strains with transparent rings around colonies as strains with potassium-dissolving capacity, selecting the strains with the transparent rings around the colonies, carrying out multiple streaking separation and purification on the potassium bacteria culture medium to obtain purified strains (10 strains in total) with consistent forms through purification, transferring the purified strains to a nutrient agar culture medium, and storing the strains in a refrigerator at 4 ℃ for later use; and marking the preserved strains as WP1-1, WP1-2, WP1-3, WP2-1, WP2-2, WP2-3, WP3-2, WP3-3, WP4-1 and WP4-2 respectively; the potassium bacteria culture medium comprises the following components: 10g of glucose, 5g of calcium carbonate, 0.2g of disodium hydrogen phosphate (anhydrous), 0.2g of manganese sulfate heptahydrate, 0.2g of calcium sulfate dihydrate, 0.2g of sodium chloride, 18g of agar and 10g of potassium feldspar, wherein distilled water is added to the mixture to reach the volume of 1000mL, and the pH value of a culture medium is 7.2; the nutrient agar culture medium comprises the following components: 10g of peptone, 3g of beef extract, 5g of sodium chloride and 15-20 g of agar, adding distilled water to 1000mL, and controlling the pH value of a culture medium to be 7.2-7.4;
step three, re-screening: activating the strain stored in the step two to obtain a bacterial suspension, dripping the bacterial suspension into a potassium bacteria culture medium, culturing for 5 days at 30 ℃, measuring the diameter D of a potassium-dissolving ring and the diameter D of a bacterial colony generated by the strain by taking 1 mu L of sterile water as a control, and preliminarily determining the potassium-dissolving capacity of the strain according to the D/D value, wherein the results are shown in Table 1.
TABLE 1 determination of Potassium-solubilizing Capacity of 10 strains by the transparent circle method
| The diameter of the potassium dissolving ring is D/mm | Diameter d/mm of bacterial colony | Ratio D/D | |
| WP1-1 strain | 67 | 57 | 1.17 |
| WP1-2 Strain | 61 | 50 | 1.22 |
| WP1-3 strain | 69 | 52 | 1.33 |
| WP2-1 strain | 70 | 57 | 1.23 |
| WP2-2 Strain | 63 | 55 | 1.15 |
| WP2-3 strain | 68 | 57 | 1.19 |
| WP3-2 strain | 65 | 54 | 1.30 |
| WP3-3 strain | 63 | 51 | 1.24 |
| WP4-1 strain | 66 | 55 | 1.20 |
| WP4-2 strain | 65 | 53 | 1.21 |
| Sterile water | 0 | 0 | 0 |
The result shows that the potassium dissolving ring of the WP1-3 strain in the 10 strains with the potassium feldspar decomposing capability obtained by primary screening is the largest, and the D/D value reaches 1.33, so that the potassium decomposing capability of the WP1-3 strain is the strongest.
3. The identification method of the strain comprises the following steps:
(1) morphological identification: activating the screened WP1-3 strain on a potassium bacteria culture medium, and observing the growth characteristics of bacterial colonies; single colonies were picked with the inoculating loop for gram staining, microscopic examination, and results recorded.
As a result: the bacterial colony of the WP1-3 strain is circular, and the bacilliform bacteria are colorless and transparent, have neat edges, are wet and sticky and have high elasticity.
(2) Physiological and biochemical identification: gram-negative bacteria.
(3) Amplification and sequencing of 16 SrDNA:
the identification of the strain is completed by the company of Biotechnology engineering (Shanghai).
The identification process of the strain comprises the steps of extracting strain DNA, carrying out PCR amplification, carrying out gel electrophoresis, purifying and recovering, connecting, preparing competent cells, converting a connecting product, extracting and sequencing plasmids, and comparing a purified 16SrDNA sequence on a ribosome database; wherein the primer sequence of PCR amplification is as follows:
an upstream primer: 5 '-AGTTTGATCMTGGCTCAG-3'
A downstream primer: 5'-GGTTACCTTGTTACGACTT-3'
In the reaction system of PCR amplification, the addition amount of each substance is as follows: 0.5 microliter of template, wherein the concentration of the genome DNA is 20 ng/nL-50 ng/nL; 2.5 μ L of 10 × buffer, 1 μ L of dNTP with the concentration of 2.5mM, 0.2 μ L of enzyme, 1 μ L of each F with the concentration of 10 μ M, and 1 μ L of R with the concentration of 10 μ M, and adding double distilled water to make the volume to be 25 μ L; and (3) amplification procedure: pre-denaturation at 94 ℃ for 4min and denaturation at 94 ℃ for 45 s; annealing at 55 ℃ for 45 s; extending for 1min at 72 ℃; after 30 cycles, the extension was carried out at 72 ℃ for 10min and the reaction was stopped at 4 ℃.
WP1-3 strain 16SrDNA sequence analysis and alignment: the fragment size of PCR amplified product is 1351pb, the sequence of partial gene of 16SrDNA (SEQ ID NO.3) obtained by sequencing has 99% homology with the sequence of related fragment of rhizobium published in GenBank.
And (3) comprehensively analyzing and identifying the physiological and biochemical identification result and the 16SrDNA sequence to identify the bacterial strain WP1-3 of the invention as Rhizobium sp.
The invention also provides application of the rhizobium in soil potassium dissolving.
Furthermore, the invention also provides application of the rhizobium in decomposing the insoluble phosphorus in soil.
Compared with the prior art, the invention has the following advantages:
1. the rhizobium WP1-3 strain is originated from soil, and is ecologically safe.
2. The rhizobium WP1-3 strain can be used for soil potassium dissolving and insoluble phosphorus decomposition in soil. The strain is an advantageous strain screened by taking potassium feldspar as a potassium source, insoluble potassium elements in aluminosilicate in soil can be converted into soluble components in the growth and propagation processes of the strain, and the potassium-dissolving capacity of rhizobium WP1-3 can reach 22.591mg/L through determination. The rhizobium of the invention can also increase the source of phosphorus element in soil by promoting the decomposition of insoluble organic phosphorus and inorganic phosphorus, and the determination result shows that the rhizobium WP1-3 of the invention has the inorganic phosphorus decomposition capacity of 4.443mg/L and the organic phosphorus decomposition capacity of 22.408mg/L, meets the requirement of organism growth and metabolism process, thereby playing the roles of promoting plant growth and improving yield.
3. The rhizobium WP1-3 strain disclosed by the invention has the functions of dissolving potassium in soil and decomposing insoluble phosphorus in soil, and also has the function of fixing nitrogen. The nitrogen fixation capacity of rhizobium WP1-3 of the invention reaches 38.521mg/L after 7 days of culture, and rich nitrogen can be provided for plant growth.
4. The rhizobium WP1-3 strain can provide efficient and stable strains and basic data for subsequent development of biofertilizer, biopesticide and the like.
The technical solution of the present invention is further described in detail by the following examples.
Detailed Description
Screening of strains:
step one, enrichment: collecting soil samples about 20g from 10cm below the ground surface of garden, white sand road side, Bowen road square and Ganjai village in Yanta district of Xian city, drying the collected soil samples in an oven, sieving the dried soil with a 100-mesh sieve, accurately weighing 1g of the soil sieved with the 100-mesh sieve in 9mL of sterile water in an aseptic manner, standing for 30min, and oscillating for 30min in a shaking table at room temperature to obtain the dilution of 10-1The soil suspension of (a); sucking out 1mL of the above soil suspension, adding 9mL of sterile water, and mixing to obtain a dilution of 10-2The soil suspension of (a); from a dilution of 10-2Sucking out 1mL of the soil suspension, adding 9mL of sterile water, and mixing to obtain a dilution of 10-3The soil suspension of (a);
step two, primary screening and purification: taking the dilution degree in the step one as 10-30.1mL of the soil suspension is coated in a potassium bacteria culture medium by a coating plate method and placed in a constant temperature incubator at 30 DEG CThe culture medium is internally inverted for 5 to 7 days, strains with transparent circles around colonies are regarded as strains with potassium-dissolving capacity, the strains with the transparent circles around the colonies are selected to be subjected to multiple streaking separation and purification in a potassium bacteria culture medium to obtain purified strains (10 strains in total) with consistent shapes through purification, the purified strains are transferred to a nutrient agar culture medium, and the strains are stored in a refrigerator at 4 ℃ for standby; and marking the preserved strains as WP1-1, WP1-2, WP1-3, WP2-1, WP2-2, WP2-3, WP3-2, WP3-3, WP4-1 and WP4-2 respectively; the potassium bacteria culture medium comprises the following components: 10g of glucose, 5g of calcium carbonate, 0.2g of disodium hydrogen phosphate (anhydrous), 0.2g of manganese sulfate heptahydrate, 0.2g of calcium sulfate dihydrate, 0.2g of sodium chloride, 18g of agar and 10g of potassium feldspar, wherein distilled water is added to the mixture to reach the volume of 1000mL, and the pH value of a culture medium is 7.2; the nutrient agar culture medium comprises the following components: 10g of peptone, 3g of beef extract, 5g of sodium chloride and 15-20 g of agar, adding distilled water to 1000mL, and controlling the pH value of a culture medium to be 7.2-7.4;
step three, re-screening: activating the strain stored in the step two to obtain a bacterial suspension, dripping the bacterial suspension into a potassium bacteria culture medium, culturing for 5 days at 30 ℃, measuring the diameter D of a potassium-dissolving ring and the diameter D of a bacterial colony generated by the strain by taking 1 mu L of sterile water as a control, and preliminarily determining the potassium-dissolving capacity of the strain according to the D/D value, wherein the results are shown in Table 1.
The result shows that the potassium dissolving ring of the WP1-3 strain in the 10 strains with the potassium feldspar decomposing capability obtained by primary screening is the largest, and the D/D value reaches 1.33, so that the potassium decomposing capability of the WP1-3 strain is the strongest.
Identification of the strains:
(1) morphological identification: activating the screened WP1-3 strain on a potassium bacteria culture medium, and observing the growth characteristics of bacterial colonies; single colonies were picked with the inoculating loop for gram staining, microscopic examination, and results recorded.
As a result: the bacterial colony of the WP1-3 strain is circular, and the bacilliform bacteria are colorless and transparent, have neat edges, are wet and sticky and have high elasticity.
(2) Physiological and biochemical identification: gram-negative bacteria.
(3) Amplification and sequencing of 16 SrDNA:
the identification of the strain is completed by the company of Biotechnology engineering (Shanghai).
The identification process of the strain comprises the steps of extracting strain DNA, carrying out PCR amplification, carrying out gel electrophoresis, purifying and recovering, connecting, preparing competent cells, converting a connecting product, extracting and sequencing plasmids, and comparing a purified 16SrDNA sequence on a ribosome database; wherein the primer sequence of PCR amplification is as follows:
an upstream primer: 5 '-AGTTTGATCMTGGCTCAG-3'
A downstream primer: 5'-GGTTACCTTGTTACGACTT-3'
In the reaction system of PCR amplification, the addition amount of each substance is as follows: 0.5 microliter of template, wherein the concentration of the genome DNA is 20 ng/nL-50 ng/nL; 2.5 μ L of 10 × buffer, 1 μ L of dNTP with the concentration of 2.5mM, 0.2 μ L of enzyme, 1 μ L of each F with the concentration of 10 μ M, and 1 μ L of R with the concentration of 10 μ M, and adding double distilled water to make the volume to be 25 μ L; and (3) amplification procedure: pre-denaturation at 94 ℃ for 4min and denaturation at 94 ℃ for 45 s; annealing at 55 ℃ for 45 s; extending for 1min at 72 ℃; after 30 cycles, the extension was carried out at 72 ℃ for 10min and the reaction was stopped at 4 ℃.
WP1-3 strain 16SrDNA sequence analysis and alignment: the fragment size of PCR amplified product is 1351pb, the sequence of partial gene of 16SrDNA (SEQ ID NO.3) obtained by sequencing has 99% homology with the sequence of related fragment of rhizobium published in GenBank.
And (3) comprehensively analyzing and identifying the physiological and biochemical identification result and the 16SrDNA sequence to identify the bacterial strain WP1-3 of the invention as Rhizobium sp.
The application in soil potassium dissolving and insoluble phosphorus decomposing of soil is as follows:
(1) determination of the ability of the WP1-3 Strain to decompose Potassium-containing minerals
The total amount of potassium element is determined by sodium tetraphenylborate colorimetry in the experiment.
Adding 150mL of potassium bacteria liquid culture medium into a 250mL triangular flask, sterilizing at high temperature and high pressure, inoculating WP1-3 strain into the potassium bacteria liquid culture medium according to the proportion of 3% -6%, taking inoculated isometric sterile water as a control, repeating the test for each group of three strains, carrying out shake culture for 5-7 days under the conditions of 30 ℃ and 180r/min, then placing the mixture into a centrifugal machine for centrifugation, wherein the rotation speed of the centrifugal machine is 2000r/min, the centrifugation time is 20min, taking the centrifuged supernatant, adding 1mL of formaldehyde-EDTA masking agent, shaking up, adding 1mL of sodium tetraphenylborate solution into the shaken up solution, continuing shaking up, standing for 15min, shaking up again, then placing the solution into a 1mL cuvette, measuring absorbance at the wavelength of 420nm and determining the content of potassium element, taking a control group added with isometric sterile water as a zero setting group, and finding a result is shown in Table 2, and knowing that the strain of rhizobium WP1-3 can decompose mineral containing potassium, the potassium-resolving power of the WP1-3 strain was 22.591 mg/L.
The potassium bacteria liquid culture medium comprises the following components: 0.75g of cane sugar, 10g of potassium feldspar, 0.15g of ammonium sulfate, 0.3g of disodium hydrogen phosphate (anhydrous) and distilled water till the volume is 1000 mL.
TABLE 2 measurement results of decomposition of potassium-containing minerals by the WP1-3 strain
(2) Determination of inorganic phosphorus decomposing ability of WP1-3 Strain
The content of phosphorus element is measured by molybdenum-antimony colorimetric method in the experiment.
Adding 150mL of inorganic phosphorus culture medium into a 250mL triangular flask, sterilizing at high temperature and high pressure, inoculating WP1-3 strain into the inorganic phosphorus culture medium according to the proportion of 3-6%, taking inoculated isometric sterile water as a control, repeating the test for each group of three strains, culturing in a shaker for 5-7 days under the conditions of 30 ℃ and 180r/min, placing in a centrifuge for centrifugation, wherein the rotation speed of the centrifuge is 2000r/min, the centrifugation time is 20min, adding 2-3 drops of dinitrophenol indicator into the centrifuged supernatant, adjusting the supernatant to be just yellowish by using 100g/L of sodium hydroxide solution, accurately adding 5mL of molybdenum-antimony anti-chromogenic agent, shaking uniformly, adding water for constant volume, placing at room temperature of more than 15 ℃ for 30min, placing in a 1mL cuvette, measuring absorbance at the wavelength of 700nm and determining the content of phosphorus element, taking the control group added with isometric sterile water as a zeroing group, as a result, as shown in Table 3, it was found that the ability of the Rhizobium WP1-3 strain to decompose inorganic phosphorus was 4.443 mg/L.
The inorganic phosphorus culture medium comprises the following components: 10g of glucose, 20g of agar, 0.3g of sodium chloride, 0.3g of magnesium sulfate, 0.3g of potassium chloride, 0.03g of manganese sulfate tetrahydrate, 12g of tricalcium phosphate, 0.03g of ferrous sulfate heptahydrate, 0.5g of ammonium sulfate and distilled water are added to reach the volume of 1000mL, and the pH value of the inorganic phosphorus culture medium is 7.2.
TABLE 3 determination of inorganic phosphorus decomposition by WP1-3 Strain
(3) Determination of the ability of the WP1-3 Strain to decompose organic phosphorus
The content of phosphorus element is measured by molybdenum-antimony colorimetric method in the experiment.
Adding 150mL of an organophosphorus culture medium into a 250mL triangular flask, sterilizing at high temperature and high pressure, inoculating WP1-3 strain into the organophosphorus culture medium according to the proportion of 3% -6%, taking inoculated isometric sterile water as a control, performing repeated tests of each group of three strains, performing shake culture for 5-7 days at 30 ℃ and 180r/min, then placing the mixture into a centrifuge for centrifugation, wherein the rotation speed of the centrifuge is 2000r/min, the centrifugation time is 20min, adding 2-3 drops of dinitrophenol indicator into the centrifuged supernatant, adjusting the solution to be just yellowish by using 100g/L of sodium hydroxide solution, accurately adding 5mL of molybdenum-antimony anti-chromogenic agent, shaking uniformly, adding water to fix the volume, placing the mixture into a 1mL cuvette, measuring the absorbance at the wavelength of 700nm and determining the content of phosphorus, taking a control group added with isometric sterile water as a zero adjustment group, the rhizobium WP1-3 strain has organophosphorus decomposing capacity up to 22.408 mg/L.
The organophosphorus culture medium comprises the following components: 3.506g of glucose, 0.83g of peptone, 0.5g of yeast extract, 0.35g of monopotassium phosphate, 0.25g of calcium carbonate, 16-20 g of agar, and adding distilled water to reach the constant volume of 1000 mL.
(4) WP1-3 Strain Nitrogen fixation ability test
In the experiment, the nitrogen fixation capacity of the strain is measured by adopting a micro Kjeldahl nitrogen determination method.
Activating a WP1-3 strain, placing the activated strain into 50mL of nitrogen-free liquid culture medium for shake culture, simultaneously adding equivalent inactivated strain and equivalent sterile water into 50mL of nitrogen-free liquid culture medium respectively for shake culture as a control, wherein the shake culture conditions are 30 ℃ and 160r/min, and measuring the nitrogen content in the culture medium by using a micro Kjeldahl method at the end of 3 days and 7 days of culture respectively, and the result shows that the nitrogen fixing capacity of the WP1-3 strain reaches 10.825mg/L after 3 days of culture; after 7 days of culture, the nitrogen fixing capacity of the WP1-3 strain reaches 38.521 mg/L.
The nitrogen-free liquid culture medium comprises the following components: 50g of sucrose, 2g of monopotassium phosphate, 2g of dipotassium phosphate, 0.5g of magnesium sulfate heptahydrate, 0.005g of ferric trichloride, 0.1g of calcium carbonate, and the volume of deionized water is up to 1000mL, and the pH value of the culture medium is 7.0-7.5.
The above description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and all simple modifications, changes and equivalent structural changes made to the above embodiment according to the technical spirit of the present invention still fall within the protection scope of the technical solution of the present invention.
Sequence listing
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tccagccatg ccgcgtgagt gatgaaggcc ttagggttgt aaagctcttt caccggagaa 360
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agtatggaag aggtaagtgg aattccgagt gtagaggtga aattcgtaga tattcggagg 600
aacaccagtg gcgaaggcgg cttactggtc cattactgac gctgaggtgc gaaagcgtgg 660
ggagcaaaca ggattagata ccctggtagt ccacgccgta aacgatgaat gttagccgtc 720
gggcagtata ctgttcggtg gcgcagctaa cgcattaaac attccgcctg gggagtacgg 780
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gtgtcgtgag atgttgggtt aagtcccgca acgagcgcaa ccctcgccct tagttgccag 1020
catttagttg ggcactctaa ggggactgcc ggtgataagc cgagaggaag gtggggatga 1080
cgtcaagtcc tcatggccct tacgggctgg gctacacacg tgctacaatg gtggtgacag 1140
tgggcagcga gacagcgatg tcgagctaat ctccaaaagc catctcagtt cggattgcac 1200
tctgcaactc gagtgcatga agttggaatc gctagtaatc gcagatcagc atgctgcggt 1260
gaatacgttc ccgggccttg tacacaccgc ccgtcacacc atgggagttg gttttacccg 1320
aaggtagtgc gctaaccgca aggaggtcag a 1351
Claims (3)
1. A Rhizobium sp (Rhizobium sp.) is characterized in that the Rhizobium sp.WP 1-3 is deposited in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No. 14472.
2. Use of the rhizobia of claim 1 for soil potassium solubilization.
3. Use of the rhizobia of claim 1 to decompose poorly soluble phosphorus in soil.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201810984088.3A CN109097305B (en) | 2018-08-28 | 2018-08-28 | Rhizobium and application thereof |
Applications Claiming Priority (1)
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| CN104911127A (en) * | 2015-06-01 | 2015-09-16 | 云南省烟草农业科学研究院 | Rhizobium and microbial inoculum, preparation method and application thereof |
| CN105950520A (en) * | 2016-07-18 | 2016-09-21 | 武汉市农业科学技术研究院作物科学研究所 | Rhizobium capable of efficiently solubilizing phosphorus and application of rhizobium |
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