CN109097305A - A kind of rhizobium and its application - Google Patents
A kind of rhizobium and its application Download PDFInfo
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Abstract
The invention discloses a kind of rhizobium Rhizobium sp.WP 1-3, deposit number is CGMCC No.14472.In addition, the application the invention also discloses the rhizobium in soil potassium decomposing and decomposition soil in insoluble phosphorus.Rhizobium of the invention are the dominant strains screened using potassium feldspar as potassium resource, the bacterial strain can convert the insoluble potassium element in soil in alumino-silicate to during growth and breeding soluble component, while can increase the source of P elements in soil by promoting the decomposition of insoluble phosphorus.Furthermore this bacterial strain also has the function of fixed nitrogen, provides required nitrogen for plant growth metabolism.
Description
Technical field
The invention belongs to microorganisms technical fields, and in particular to a kind of rhizobium and its application.
Background technique
Potassium and phosphorus are nutrients important in plant, necessary for biological nitrogen fixation, and promote plant growth metabolism
Key component.Potassium and phosphorus are adjusting Premeabilisation of cells, matter transportation, promotion enzyme activation, promotion photosynthesis, raising carbon dioxide
Assimilability, promote crop substance in vivo synthesis and transhipment and enhancing plant resistance to environment stress and restrain oneself ability etc. play it is important
Facilitation.Potassium and phosphorus in soil are present in mostly in the mineral such as alumino-silicate and apatite, with mineral K and mineral phosphorus
Potassium and phosphorus existing for form cannot directly be absorbed by plant, agriculturally generally use in the form of chemical potassium and chemical phosphorus to plant
The mode that body is supplemented is easily destroyed soil texture to cause environmental pollution.With people's mentioning to the attention degree of environment
Height, bio-feritlizer is the characteristics of improving Nutrient Elements in Soil source, increasing soil geological unit and improve in terms of quality of agricultural product
Cause extensive concern.
Dissolving phosphor and dissolving potassium bacterium is a kind of strain that can effectively decompose potassium-bearing mineral matter and phosphorus ore containing matter in soil, can will be native
The potassium of slightly solubility and phosphorus are converted into soluble nutrient in earth, increase the source of potassium element and P elements in soil.Development and utilization are high
Microbial manure made by dissolving phosphor and dissolving potassium bacterium is imitated, is had for agricultural life production development and environmental protection highly important
Meaning.
Summary of the invention
Technical problem to be solved by the present invention lies in view of the above shortcomings of the prior art, provide one kind to can be used for soil
Potassium decomposing and the rhizobium for decomposing insoluble phosphorus in soil.The bacterial strain is derived from soil, and ecological safety, which is using potassium feldspar as potassium
The dominant strain that source screens can convert the insoluble potassium element in soil in alumino-silicate during growth and breeding
For soluble component, while the source of P elements in soil can be increased by promoting the decomposition of insoluble phosphorus.Furthermore this bacterial strain
Also have the function of fixed nitrogen, provides required nitrogen for plant growth metabolism.
In order to solve the above technical problems, the technical solution adopted by the present invention is that: a kind of rhizobium, which is characterized in that described
Rhizobium are Rhizobium sp.WP 1-3, and deposit number is CGMCC No.14472, and preservation day is on July 27th, 2017, are protected
Hiding unit is China Committee for Culture Collection of Microorganisms's common micro-organisms center, address: BeiChen West Road, Chaoyang District, BeiJing City 1
Number No. 3 Institute of Microorganism, Academia Sinica of institute.
1, the bacterial strain has the property that
Morphological feature: WP 1-3 bacterial strain bacterium colony is rounded, and rod bacterium is colorless and transparent, neat in edge, moisten it is sticky, rich in bullet
Property.
Physiological and biochemical property: gramnegative bacterium.
2, the screening technique of the bacterial strain includes:
Step 1: enrichment: the ground by Xi'an Yanta District crape myrtle garden, white sand road roadside, blog article road square and the stockaded village Gan Jia
Each 20g of pedotheque or so is acquired below table at 10cm, the pedotheque of acquisition is put into baking oven and is dried, soil mistake will be dried
100 meshes accurately weigh soil that 1g is sieved with 100 mesh sieve in 9mL sterile water with sterile manner, stand after 30min in room temperature item
Shaking table vibrates 30min under part, and obtaining dilution is 10-1Soil supension；Be sucked out 1mL from above-mentioned soil supension, be added 9mL without
It in bacterium water, mixes well, it is 10 that dilution, which is made,-2Soil supension；It is 10 from dilution-21mL is sucked out in soil supension, adds
Enter in 9mL sterile water, mix well, it is 10 that dilution, which is made,-3Soil supension；
Step 2: primary dcreening operation and purifying: taking dilution in step 1 is 10-3Soil supension 0.1mL use spread plate method
It is coated in B. mucilaginocus culture medium, is placed in 30 DEG C of constant incubators and is inverted culture 5 days~7 days, periphery of bacterial colonies is occurred transparent
The bacterial strain of circle is considered as the bacterial strain for having ability of dissolving potassium, has the bacterial strain of transparent circle repeatedly to be crossed around picking in B. mucilaginocus culture medium
It isolates and purifies, the consistent purifying bacterial strain (totally 10 plants) in the form of purifying obtains, purifying bacterial strain is transferred to nutrient agar
On, it is saved in 4 DEG C of refrigerators stand-by；And by the bacterial strain of preservation be respectively labeled as WP1-1, WP1-2, WP1-3, WP2-1, WP2-2,
WP2-3, WP3-2, WP3-3, WP4-1 and WP4-2；The composition of the B. mucilaginocus culture medium are as follows: glucose 10g, calcium carbonate 5g, phosphorus
Sour disodium hydrogen (anhydrous) 0.2g, seven water manganese sulfate 0.2g, calcium sulphate dihydrate 0.2g, sodium chloride 0.2g, agar 18g, potassium feldspar
Distilled water is added to 1000mL, medium pH 7.2 in 10g；The composition of the nutrient agar are as follows: peptone 10g, ox
Distilled water is added to 1000mL in meat extract 3g, sodium chloride 5g, agar 15g~20g, and medium pH is 7.2~7.4；
Step 3: secondary screening: activating the bacterial strain saved in step 2 to obtain bacteria suspension, bacteria suspension is added drop-wise to B. mucilaginocus training
It supports in base, it is control with 1 μ L sterile water that 30 DEG C, which are cultivated 5 days, molten potassium loop diameter D and colony diameter d that bacterial strain generates are measured, and
The ability of dissolving potassium that bacterial strain is primarily determined according to the size of D/d value, the results are shown in Table 1.
1 transparent circle method of table measures the ability of dissolving potassium of 10 plants of bacterial strains
| Molten potassium loop diameter D/mm | Colony diameter d/mm | Ratio D/d | |
| WP 1-1 bacterial strain | 67 | 57 | 1.17 |
| WP 1-2 bacterial strain | 61 | 50 | 1.22 |
| WP 1-3 bacterial strain | 69 | 52 | 1.33 |
| WP 2-1 bacterial strain | 70 | 57 | 1.23 |
| WP 2-2 bacterial strain | 63 | 55 | 1.15 |
| WP 2-3 bacterial strain | 68 | 57 | 1.19 |
| WP 3-2 bacterial strain | 65 | 54 | 1.30 |
| WP 3-3 bacterial strain | 63 | 51 | 1.24 |
| WP 4-1 bacterial strain | 66 | 55 | 1.20 |
| WP 4-2 bacterial strain | 65 | 53 | 1.21 |
| Sterile water | 0 | 0 | 0 |
The molten potassium circle of WP 1-3 bacterial strain is most in 10 plants of bacterial strains with decomposing of potassium feldspar ability that primary dcreening operation obtains as the result is shown
Greatly, D/d value reaches 1.33, therefore WP 1-3 bacterial strain ability of dissolving potassium is most strong.
3, the identification method of the bacterial strain are as follows:
(1) Morphological Identification: the WP 1-3 bacterial strain screened is activated on B. mucilaginocus culture medium, observes bacterial clump
Growth characteristics；Gram's staining is carried out with oese picking single colonie, microscopy records result.
As a result: WP 1-3 bacterial strain bacterium colony is rounded, and rod bacterium is colorless and transparent, neat in edge, moistens sticky, high resilience.
(2) Physiology and biochemistry is identified: Gram-negative bacteria.
(3) amplification and sequencing of 16SrDNA:
The identification of the strain is completed by Sangon Biotech (Shanghai) Co., Ltd..
The qualification process of strain includes the extraction of bacterial strain DNA, PCR amplification, gel electrophoresis, purification and recovery, connection, competence
The preparation of cell, connection product conversion, plasmid extracts and sequencing, and purified rear 16SrDNA sequence compares on ribosomes database
It is right；The wherein primer sequence of PCR amplification are as follows:
Upstream primer: 5 '-AGTTTGATCMTGGCTCAG-3 '
Downstream primer: 5 '-GGTTACCTTGTTACGACTT-3 '
In the reaction system of PCR amplification, the additional amount of each substance is as follows: 0.5 μ L of template, and wherein genomic DNA concentration is
20ng/nL~50ng/nL；2.5 μ L of 10x buffer, concentration are 1 μ L of dNTP, the 0.2 μ each 1 μ L of L, 10 μM of F of enzyme of 2.5mM,
10 μM of R1 μ L is added distilled water and is settled to 25 μ L；Amplification program: 94 DEG C of initial denaturations 4min, 94 DEG C of denaturation 45s；55 DEG C of annealing
45s；72 DEG C of extension 1min；In 72 DEG C of extension 10min after 30 circulations, reaction is terminated at 4 DEG C.
The analysis of WP1-3 bacterial strain 16SrDNA sequence and comparison: the clip size 1351pb of the product of PCR amplification, sequencing obtain
16SrDNA partial gene sequence (SEQ ID NO.3) and GenBank in the rhizobium associated clip sequence delivered have 99%
Homology.
Comprehensive Physiology and biochemistry qualification result and 16SrDNA sequence analyze and identify as a result, identifying bacterial strain WP 1-3 of the invention
For rhizobium (Rhizobium sp.).
The present invention also provides application of the above-mentioned rhizobium in soil potassium decomposing.
Further, the application the present invention also provides above-mentioned rhizobium in decomposing soil in insoluble phosphorus.
Compared with the prior art, the present invention has the following advantages:
1, rhizobium WP 1-3 bacterial strain of the invention is derived from soil, the bacterial strain ecological safety.
2, rhizobium WP 1-3 bacterial strain of the invention can be used for soil potassium decomposing and decompose insoluble phosphorus in soil.The present invention
Bacterial strain be the dominant strain screened using potassium feldspar as potassium resource, which can be by silicon in soil during growth and breeding
Insoluble potassium element in aluminate is converted into soluble component, passes through measurement, the ability of dissolving potassium of rhizobium WP1-3 of the invention
Up to 22.591mg/L.Rhizobium of the invention can also increase soil by promoting the organic phosphorus decomposition with Phos of slightly solubility
The source of middle P elements, it is 4.443mg/L that measurement result, which shows that rhizobium WP1-3 of the invention decomposes Phos ability, is decomposed
Organic phosphorus ability reaches 22.408mg/L, meets the needs of biology growing metabolic process, thus play promote plant growth and
Improve the effect of yield.
3, rhizobium WP 1-3 bacterial strain of the invention have the function of soil potassium decomposing and decompose soil in insoluble phosphorus with
Outside, also there is nitrogen fixation.The nitrogen fixing capacity of rhizobium WP1-3 of the invention after culture 7 days reaches 38.521mg/L, can be
Plant growth provides nitrogen abundant.
4, rhizobium WP1-3 bacterial strain of the invention can be provided for subsequent developments bio-feritlizer and biological pesticide etc. it is efficient,
Stable strain and basic data.
Below by embodiment, technical scheme of the present invention will be described in further detail.
Specific embodiment
The screening of bacterial strain:
Step 1: enrichment: the ground by Xi'an Yanta District crape myrtle garden, white sand road roadside, blog article road square and the stockaded village Gan Jia
Each 20g of pedotheque or so is acquired below table at 10cm, the pedotheque of acquisition is put into baking oven and is dried, soil mistake will be dried
100 meshes accurately weigh soil that 1g is sieved with 100 mesh sieve in 9mL sterile water with sterile manner, stand after 30min in room temperature item
Shaking table vibrates 30min under part, and obtaining dilution is 10-1Soil supension；Be sucked out 1mL from above-mentioned soil supension, be added 9mL without
It in bacterium water, mixes well, it is 10 that dilution, which is made,-2Soil supension；It is 10 from dilution-21mL is sucked out in soil supension, adds
Enter in 9mL sterile water, mix well, it is 10 that dilution, which is made,-3Soil supension；
Step 2: primary dcreening operation and purifying: taking dilution in step 1 is 10-3Soil supension 0.1mL use spread plate method
It is coated in B. mucilaginocus culture medium, is placed in 30 DEG C of constant incubators and is inverted culture 5 days~7 days, periphery of bacterial colonies is occurred transparent
The bacterial strain of circle is considered as the bacterial strain for having ability of dissolving potassium, has the bacterial strain of transparent circle repeatedly to be crossed around picking in B. mucilaginocus culture medium
It isolates and purifies, the consistent purifying bacterial strain (totally 10 plants) in the form of purifying obtains, purifying bacterial strain is transferred to nutrient agar
On, it is saved in 4 DEG C of refrigerators stand-by；And by the bacterial strain of preservation be respectively labeled as WP1-1, WP1-2, WP1-3, WP2-1, WP2-2,
WP2-3, WP3-2, WP3-3, WP4-1 and WP4-2；The composition of the B. mucilaginocus culture medium are as follows: glucose 10g, calcium carbonate 5g, phosphorus
Sour disodium hydrogen (anhydrous) 0.2g, seven water manganese sulfate 0.2g, calcium sulphate dihydrate 0.2g, sodium chloride 0.2g, agar 18g, potassium feldspar
Distilled water is added to 1000mL, medium pH 7.2 in 10g；The composition of the nutrient agar are as follows: peptone 10g, ox
Distilled water is added to 1000mL in meat extract 3g, sodium chloride 5g, agar 15g~20g, and medium pH is 7.2~7.4；
Step 3: secondary screening: activating the bacterial strain saved in step 2 to obtain bacteria suspension, bacteria suspension is added drop-wise to B. mucilaginocus training
It supports in base, it is control with 1 μ L sterile water that 30 DEG C, which are cultivated 5 days, molten potassium loop diameter D and colony diameter d that bacterial strain generates are measured, and
The ability of dissolving potassium that bacterial strain is primarily determined according to the size of D/d value, the results are shown in Table 1.
The molten potassium circle of WP 1-3 bacterial strain is most in 10 plants of bacterial strains with decomposing of potassium feldspar ability that primary dcreening operation obtains as the result is shown
Greatly, D/d value reaches 1.33, therefore WP 1-3 bacterial strain ability of dissolving potassium is most strong.
The identification of bacterial strain:
(1) Morphological Identification: the WP 1-3 bacterial strain screened is activated on B. mucilaginocus culture medium, observes bacterial clump
Growth characteristics；Gram's staining is carried out with oese picking single colonie, microscopy records result.
As a result: WP 1-3 bacterial strain bacterium colony is rounded, and rod bacterium is colorless and transparent, neat in edge, moistens sticky, high resilience.
(2) Physiology and biochemistry is identified: Gram-negative bacteria.
(3) amplification and sequencing of 16SrDNA:
The identification of the strain is completed by Sangon Biotech (Shanghai) Co., Ltd..
The qualification process of strain includes the extraction of bacterial strain DNA, PCR amplification, gel electrophoresis, purification and recovery, connection, competence
The preparation of cell, connection product conversion, plasmid extracts and sequencing, and purified rear 16SrDNA sequence compares on ribosomes database
It is right；The wherein primer sequence of PCR amplification are as follows:
Upstream primer: 5 '-AGTTTGATCMTGGCTCAG-3 '
Downstream primer: 5 '-GGTTACCTTGTTACGACTT-3 '
In the reaction system of PCR amplification, the additional amount of each substance is as follows: 0.5 μ L of template, and wherein genomic DNA concentration is
20ng/nL~50ng/nL；2.5 μ L of 10x buffer, concentration are 1 μ L of dNTP, the 0.2 μ each 1 μ L of L, 10 μM of F of enzyme of 2.5mM,
10 μM of R1 μ L is added distilled water and is settled to 25 μ L；Amplification program: 94 DEG C of initial denaturations 4min, 94 DEG C of denaturation 45s；55 DEG C of annealing
45s；72 DEG C of extension 1min；In 72 DEG C of extension 10min after 30 circulations, reaction is terminated at 4 DEG C.
The analysis of WP1-3 bacterial strain 16SrDNA sequence and comparison: the clip size 1351pb of the product of PCR amplification, sequencing obtain
16SrDNA partial gene sequence (SEQ ID NO.3) and GenBank in the rhizobium associated clip sequence delivered have 99%
Homology.
Comprehensive Physiology and biochemistry qualification result and 16SrDNA sequence analyze and identify as a result, identifying bacterial strain WP 1-3 of the invention
For rhizobium (Rhizobium sp.).
Application in soil potassium decomposing, decomposition soil in insoluble phosphorus:
(1) WP1-3 bacterial strain decomposes the ability measurement of potassium-bearing mineral
This experiment uses tetraphenylboron sodium colorimetric method for determining potassium element total amount.
150mL B. mucilaginocus fluid nutrient medium is added in 250mL triangular flask, through autoclave sterilization, then by WP 1-3
Bacterial strain is inoculated into B. mucilaginocus fluid nutrient medium according to 3%~6% ratio, to be inoculated with isometric sterile water for control, often
Three repetitions of group are tested, and are placed in centrifuge within shaking table culture 5 days~7 days under conditions of 30 DEG C, 180r/min and are centrifuged, wherein
Centrifuge speed is 2000r/min, centrifugation time 20min, and formaldehyde-EDTA screening agent 1mL is added in the supernatant after taking centrifugation,
It shakes up, the sodium tetraphenylborate solution that 1mL is added into the solution after shaking up continues to shake up, and shakes up after standing 15min, then sets again
In 1mL cuvette, absorbance is measured at 420nm wavelength and determines Determination of Potassium, pair of isometric sterile water is added
It is zeroing group according to group, the results are shown in Table 2, it is known that rhizobium WP1-3 bacterial strain can decompose potassium-bearing mineral, the potassium decomposing energy of WP1-3 bacterial strain
Power is 22.591mg/L.
The composition of the B. mucilaginocus fluid nutrient medium are as follows: sucrose 0.75g, potassium feldspar 10g, ammonium sulfate 0.15g, phosphoric acid hydrogen two
Sodium (anhydrous) 0.3g, adds distilled water to 1000mL.
The measurement result of table 2WP1-3 bacterial strain decomposition potassium-bearing mineral
(2) WP1-3 bacterial strain decomposes the ability measurement of Phos
This experiment measures phosphorus element content using molybdenum antimony resistance colorimetric method.
150mL Phos culture medium is added in 250mL triangular flask, through autoclave sterilization, then by WP 1-3 bacterial strain
It is inoculated into Phos culture medium according to 3%~6% ratio, to be inoculated with isometric sterile water as control, every group of three weights
Retrial is tested, and is placed in centrifuge within shaking table culture 5 days~7 days under conditions of 30 DEG C, 180r/min and is centrifuged, wherein centrifugal basket
Speed is 2000r/min, centrifugation time 20min, and~3 drop of the drop of dinitrophenol indicator 2 is added in the supernatant after taking centrifugation, and
Be adjusted to solution just with 100g/L sodium hydroxide solution and be in it is yellowish, it is accurate that the anti-color developing agent of 5mL molybdenum antimony is added, shake up, add water fixed
Appearance is placed on 15 DEG C of room temperature or more and places 30min, is subsequently placed in 1mL cuvette, and absorbance and true is measured at 700nm wavelength
Determine phosphorus element content, using the control group that isometric sterile water is added as zeroing group, the results are shown in Table 3, it is known that rhizobium WP1-3 bacterial strain
Decomposition Phos ability is 4.443mg/L.
The composition of the Phos culture medium are as follows: glucose 10g, agar 20g, sodium chloride 0.3g, magnesium sulfate 0.3g, chlorination
Potassium 0.3g, four water manganese sulfate 0.03g, tricalcium phosphate 12g, ferrous sulfate heptahydrate 0.03g, ammonium sulfate 0.5g add distilled water extremely
1000mL, the pH of the Phos culture medium are 7.2.
The measurement result of table 3WP1-3 bacterial strain decomposition Phos
(3) WP1-3 bacterial strain decomposes organic phosphorus ability measurement
This experiment measures phosphorus element content using molybdenum antimony resistance colorimetric method.
The organic phosphorus culture medium of 150mL is added in 250mL triangular flask, through autoclave sterilization, then by WP 1-3 bacterial strain
It is inoculated into organic phosphorus culture medium according to 3%~6% ratio, to be inoculated with isometric sterile water as control, every group of three weights
Retrial is tested, and is placed in centrifuge within shaking table culture 5 days~7 days under conditions of 30 DEG C, 180r/min and is centrifuged, wherein centrifugal basket
Speed is 2000r/min, centrifugation time 20min, and~3 drop of the drop of dinitrophenol indicator 2 is added in the supernatant after taking centrifugation, and
Be adjusted to solution just with 100g/L sodium hydroxide solution and be in it is yellowish, it is accurate that the anti-color developing agent of 5mL molybdenum antimony is added, shake up, add water fixed
Appearance is placed on 15 DEG C of room temperature or more and places 30min, is subsequently placed in 1mL cuvette, and absorbance and true is measured at 700nm wavelength
Determine phosphorus element content, the control group of isometric sterile water is added as zeroing group, which decomposes organic phosphorus energy
Power reaches 22.408mg/L.
The composition of the organic phosphorus culture medium are as follows: glucose 3.506g, peptone 0.83g, yeast extract 0.5g, biphosphate
Potassium 0.35g, calcium carbonate 0.25g, agar 16g~20g add distilled water to be settled to 1000mL.
(4) WP1-3 bacterial strain nitrogen fixing capacity is tested
This experiment measures bacterial strain nitrogen fixing capacity using micro-Kjeldahl.
It takes thallus to be placed in shaking table culture in 50mL nitrogen-free fluid nutrient medium after WP 1-3 actication of culture, while equivalent being taken to inactivate
Thallus and equivalent sterile water are separately added into shaking table culture in 50mL nitrogen-free fluid nutrient medium and are used as control, the item of the shaking table culture
Part is 30 DEG C, 160r/min, and containing in micro-Kjeldahl measurement culture medium is utilized at the end of culture in 3 days and 7 days
Nitrogen quantity, the results show that the nitrogen fixing capacity of WP 1-3 bacterial strain reaches 10.825mg/L after culture 3 days；After culture 7 days, WP 1-3 bacterium
The nitrogen fixing capacity of strain reaches 38.521mg/L.
The composition of the nitrogen-free fluid nutrient medium are as follows: sucrose 50g, potassium dihydrogen phosphate 2g, dipotassium hydrogen phosphate 2g, seven hydration sulphur
Sour magnesium 0.5g, ferric trichloride 0.005g, calcium carbonate 0.1g, deionized water are settled to 1000mL, and the pH of culture medium is 7.0~7.5.
The above is only presently preferred embodiments of the present invention, not does any restrictions to the present invention, all according to invention skill
Art any simple modification substantially to the above embodiments, change and equivalent structural changes, still fall within the technology of the present invention
In the protection scope of scheme.
Sequence table
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tccagccatg ccgcgtgagt gatgaaggcc ttagggttgt aaagctcttt caccggagaa 360
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gggcagtata ctgttcggtg gcgcagctaa cgcattaaac attccgcctg gggagtacgg 780
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aaggtagtgc gctaaccgca aggaggtcag a 1351
Claims (3)
1. a kind of rhizobium (Rhizobium sp.), which is characterized in that the rhizobium are Rhizobium sp.WP 1-3, are protected
It is hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.14472.
2. a kind of application of rhizobium as described in claim 1 in soil potassium decomposing.
3. a kind of application of rhizobium as described in claim 1 in decomposing soil in insoluble phosphorus.
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| CN110272848A (en) * | 2019-07-10 | 2019-09-24 | 西北农林科技大学 | One plant of rhizobium J16 and its application with potassium decomposing effect |
| CN110564637A (en) * | 2019-07-10 | 2019-12-13 | 西北农林科技大学 | Composite microbial inoculum for promoting wheat growth and application thereof |
| CN110713953A (en) * | 2019-11-15 | 2020-01-21 | 中国科学院南京土壤研究所 | Mesorhizobium strain with phosphate solubilizing property and application thereof |
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| CN110272848A (en) * | 2019-07-10 | 2019-09-24 | 西北农林科技大学 | One plant of rhizobium J16 and its application with potassium decomposing effect |
| CN110564637A (en) * | 2019-07-10 | 2019-12-13 | 西北农林科技大学 | Composite microbial inoculum for promoting wheat growth and application thereof |
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| CN109097305B (en) | 2021-06-04 |
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