CN116622551A - Efficient phosphorus-dissolving bacteria, and preparation method and application thereof - Google Patents
Efficient phosphorus-dissolving bacteria, and preparation method and application thereof Download PDFInfo
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- CN116622551A CN116622551A CN202310389007.6A CN202310389007A CN116622551A CN 116622551 A CN116622551 A CN 116622551A CN 202310389007 A CN202310389007 A CN 202310389007A CN 116622551 A CN116622551 A CN 116622551A
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- phosphorus
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- 238000002360 preparation method Methods 0.000 title abstract description 6
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 claims abstract description 56
- 229910052698 phosphorus Inorganic materials 0.000 claims abstract description 56
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- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 claims abstract description 24
- 241000520272 Pantoea Species 0.000 claims abstract description 18
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- 238000011081 inoculation Methods 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- RBTARNINKXHZNM-UHFFFAOYSA-K iron trichloride Chemical compound Cl[Fe](Cl)Cl RBTARNINKXHZNM-UHFFFAOYSA-K 0.000 description 1
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- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
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- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
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- 238000012360 testing method Methods 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
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- B09—DISPOSAL OF SOLID WASTE; RECLAMATION OF CONTAMINATED SOIL
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Abstract
The invention belongs to the technical field of microorganisms, and particularly relates to efficient phosphorus-dissolving bacteria, a preparation method and application thereof, wherein the phosphorus-dissolving bacteria are named as follows: pantoea cypripediP11, biological deposit number: cctccc NO: m2023159, the high-efficiency phosphorus-dissolving bacteria Pantoea cypripediP11 provided by the invention can convert insoluble phosphorus into soluble phosphorus, the content of the insoluble phosphorus can reach 663.59mg/L, the content of effective phosphorus in soil can be increased by 1.46 times, and compared with the phosphorus-dissolving bacteria Burkholderia ubonensis of the prior invention, the high-efficiency phosphorus-dissolving bacteria can produce indoleacetic acid and secrete siderophores, and promote plant root growth.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to efficient phosphorus dissolving bacteria and a preparation method and application thereof.
Background
Phosphorus is one of the essential elements for plant growth and development, and most of phosphorus in soil exists in the form of insoluble phosphorus salt and is difficult to be absorbed and utilized by plants. In order to ensure the phosphorus element required for plant growth, only phosphorus fertilizer can be frequently applied, however, heavy metal ions such as Ca in soil 2+ 、Al 3+ And Fe (Fe) 3+ The plasma can convert soluble phosphorus in the phosphate fertilizer into insoluble phosphorus, resulting in low phosphate fertilizer utilization. In addition, frequent application of phosphate fertilizer causes a series of soil problems such as soil hardening, acidification, unbalance of nutrient elements and flora, and the like, thereby deteriorating the ecological environment of the soil.
In order to solve the problem, the efficient phosphorus-dissolving bacteria are screened to dissolve indissolvable phosphorus in soil, so that the method is an important way for reducing the use amount of phosphate fertilizer and developing green agriculture. The method has the advantages that the indissolvable phosphorus is converted into the soluble phosphorus which can be absorbed and utilized by plants by utilizing the phosphorus-dissolving bacteria, so that the content of the effective phosphorus in the soil is increased, and the utilization efficiency of the phosphate fertilizer is improved, thereby reducing the accumulation of indissolvable phosphorus in the soil, relieving the problems of soil hardening and the like, simultaneously reducing the pollution of chemical fertilizer to the environment, and having important significance for sustainable development of agriculture.
Disclosure of Invention
In view of the above problems, in one aspect, the present invention provides a high-efficiency phosphorus-dissolving bacteria, where the phosphorus-dissolving bacteria is named: pantoea cypripedip11, biological deposit No.: cctccc No. M2023159.
In a second aspect, the invention also provides a fermentation product obtained by fermenting the phosphorus-dissolving bacteria Pantoea cypripedidi P11.
In a third aspect, the invention provides a microbial inoculum comprising the phosphorus-dissolving bacteria Pantoea cypripedii P11 or the fermentation product.
In a fourth aspect, the invention provides a phosphate solubilizing bacterial fertilizer, which contains the phosphate solubilizing bacteria Pantoea cypripedii P11 or the fermentation product.
In a fifth aspect, the invention provides a soil ecological environment regulator, which contains the phosphorus-dissolving bacteria Pantoea cypripedii P11 or the fermentation product.
In a sixth aspect the invention provides the use of a composition comprising said lysophosphate Pantoea cypripedii P11 or said fermentation product or said microbial inoculum.
Further, the method is applied to the production of indoleacetic acid and/or siderophores.
Further, including use in promoting plant growth and increasing plant yield; further, the concentration of the bacterial suspension in the phosphate solubilizing bacteria Pantoea cypripedii P11, the fermentation product and/or the bacterial agent is 10 8 cfu/mL。
Further, the method also comprises the application in dissolving phosphorus, improving the utilization rate of phosphate fertilizer and/or increasing the content of soluble phosphorus in soil.
Further, the use of the microbial inoculum in preventing and/or ameliorating soil problems caused by the application of phosphate fertilizer; the soil problems include soil hardening, acidification, nutrient element and flora imbalance.
In a seventh aspect, the invention provides a preparation method of the efficient phosphorus-dissolving bacteria Pantoea cypripedi P11, which comprises the following steps:
collecting rhizosphere soil samples from 5-10cm below the Zhong Mingzhen Jin Fengcun surface layer of the copper tomb city;
weighing 10g of soil sample into a 250mL triangular flask, adding 90mL of sterile water, and 180 r.min -1 Shaking for 30min to prepare soil suspension, and sequentially preparing 10 by 10-fold dilution method -2 、10 -3 And 10 -4 Is a soil dilution of (2);
respectively sucking 0.1mL of soil dilution liquid, coating the soil dilution liquid on an inorganic phosphorus solid culture medium, repeating each gradient three times, and culturing in an inverted manner in a 28 ℃ incubator;
after 3-5d, picking out a colony which has obvious phosphate-dissolving ring and has good growth vigor, transferring the colony to an inorganic phosphorus solid culture medium again by a streaking method, and carrying out subculture for 3 times and purifying again to obtain the efficient phosphate-dissolving bacteria Pantoea cyprip edii P.
Further, the formula of the inorganic phosphorus solid culture medium is as follows: agar 18.0g, glucose 10.0g,NaCl0.3g,KCl0.3g, (NH) 4 ) 2 SO 4 0.5g，MgSO 4 ·7H 2 O 0.3g，FeSO 4 ·4H 2 O 0.036g，MnSO 4 ·4H 2 O 0.03g，Ca 3 (PO 4 ) 2 5.0g, distilled water 1L and pH value 7.0-7.2.
The invention has the beneficial effects that:
the phosphorus-dissolving bacteria Pantoea cypripedi P11 provided by the invention can convert insoluble phosphorus into the highest content of the soluble phosphorus up to 663.59mg/L, can improve the effective phosphorus content of soil by 1.46 times, has stronger phosphorus-dissolving capability than the prior invention phosphorus-dissolving bacteria Burkholderia ubonensis (with the publication number of CN 111662846B), can produce indoleacetic acid and secrete siderophores, and can promote plant root growth.
Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The objectives and other advantages of the invention may be realized and attained by the structure particularly pointed out in the written description and claims hereof as well as the appended drawings.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions of the prior art, the following description will briefly explain the drawings used in the embodiments or the description of the prior art, and it is obvious that the drawings in the following description are some embodiments of the present invention, and other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 shows a morphology of P.lysogen Pantoea cypripedii P11 on LB medium in an example of the invention;
FIG. 2 shows a phylogenetic tree based on the l6S rDNA gene sequence of P.lysogen Pantoea cypripedii P in an embodiment of the invention;
FIG. 3 shows a morphology of phosphorus-dissolving bacteria Pantoea cypripedii P11 on inorganic phosphorus solid medium in an embodiment of the invention;
FIG. 4 shows a morphology of P.lysogen Pantoea cypripedii P spot-plated onto CAS medium in an embodiment of the invention.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The following describes the process and effect of obtaining the efficient phosphorus-dissolving bacteria Pantoea cypripedii P11 according to the present invention in detail with reference to specific examples.
Example 1 screening of Phosphomorph Pantoea cypripedii P11
1.1 soil sample collection: soil samples are collected under the surface layer of Zhong Mingzhen Jin Fengcun in the copper tomb city by 5-10cm;
1.2 preparation of culture medium:
preparing inorganic phosphorus liquid culture medium: glucose 10.0g,NaCl0.3g,KCl0.3g, (NH) 4 ) 2 SO 4 0.5g，MgSO 4 ·7H 2 O 0.3g，FeSO 4 ·4H 2 O 0.036g，MnSO 4 ·4H 2 O 0.03g，Ca 3 (PO 4 ) 2 5.0g, distilled water 1L and pH value 7.0-7.2. The inorganic phosphorus solid culture medium is added with 18.0g of agar.
Preparing an LB liquid culture medium: 10.0g of tryptone, 5.0g of yeast extract powder, 10.0g of NaCl, 1L of distilled water and pH value of 7.0-7.2. Agar 18.0g was added to the LB solid medium.
1.3 separation and purification of the phosphorus-dissolving bacteria: 10g of rhizosphere soil sample is weighed into a 250mL triangular flask, 90mL of sterile water is added, and 180 r.min -1 Shaking for 30min to prepare soil suspension, and sequentially preparing 10 by 10-fold dilution method -2 、10 -3 And 10 -4 Respectively sucking 0.1mL of soil diluent, coating the soil diluent on an inorganic phosphorus solid culture medium, repeating each gradient for three times, culturing in an inverted manner in a 28 ℃ incubator for 3-5 days, picking out bacterial colonies which have obvious phosphorus dissolving rings and good growth vigor, transferring the bacterial colonies to the inorganic phosphorus solid culture medium again by a streaking method, subculturing for 3 times, and finally purifying to obtain the phosphorus dissolving bacteria Pantoea cypripedii P with strong phosphorus dissolving capacity. The glycerol is used for seed preservation at-20 ℃ and-80 ℃ and preserved in China center for type culture Collection (eight-path No. 299 university of Wuhan in Wuchang district of Wuhan, hubei province, post code 430072) on 2 months and 20 days in 2023, wherein the preservation name is Pantoea cypripedii P, and the preservation number is CCTCC NO: M2023159.
Example 2 Pantoea cypripedii P11 identification
2.1 morphological characterization: gram-negative bacteria, which were milky yellow and opaque colonies on LB liquid medium, had smooth surfaces as shown in FIG. 1.
2.2 16S rDNA identification: PCR amplification was performed using bacterial universal primers 27F (5'-AGAGTTTGATCCTGGCTCAG-3') and 1492R (5'-TACGGCTACCTTGTTACGACTT-3'), and the resulting 16S rDNA sequence was uploaded to Genbank for sequence alignment, identified as strain Pantoea cypripedii, phylogenetic tree as in FIG. 2.
16S rDNA sequence > OP810910.1 Pantoea cypripedii P11 and 16S ribosomal RNA gene, and partial sequence is as follows:
TACCTGCAGTCGACGGTAGCACAGAGGAGCTTGCTCCTTGGGTGACGAGTGGCGGACGGGTGAGTAATGTCTGGGGATCTGCCCGATGGAGGGGGATAACTACTGGAAACGGTAGCTAATACCGCATAACGTCGCAAGACCAAAGTGGGGGACCTTCGGGCCTCACACCATCGGATGAACCCAGATGGGATTAGCTAGTAGGTGGGGTAACGGCTCACCTAGGCGACGATCCCTAGCTGGTCTGAGAGGATGACCAGCCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCCATGCCGCGTGTATGAAGAAGGCCTTCGGGTTGTAAAGTACTTTCAGCGGGGAGGAAGGGTGTGAAGTGAATAGCTTCATGCATTGACGTTACCCGCAGAAGAAGCACCGGCTAACTCCGTGCCAGCAGCCGCGGTAATACGGAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCACGCAGGCGGTCTGTCAAGTCGGATGTGAAATCCCCGGGCTTAACCTGGGAACTGCATTCGAAACTGGCAGGCTGGAGTCTCGTAGAGGGGGGTAGAATTCCAGGTGTAGCGGTGAAATGCGTAGAGATCTGGAGGAATACCGGTGGCGAAGGCGGCCCCCTGGACGAAGACTGACGCTCAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCGACTTGGAGGTTGTGCCCTTGAGGCGTGGCTTCCGGAGCTAACGCGTTAAGTCGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGATGCAACGCGAAGAACCTTACCTGGCCTTGACATCCACGGAATTNGGCAGAGATGCCTTAGTGCCTTCGGGAACCGTGAGACAGGTGCTGCATGGCTGTCGTCAGCTCGTGTTGTGAAATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTATCCTTTGTTGCCAGCGATTCGGTCGGGAACTCAAAGGAGACTGCCGGTGATAAACCGGAGGAAGGTGGGGATGACGTCAAGTCATCATGGCCCTTACGGCCAGGGCTACACACGTGCTACAATGGCGCATACAAAGAGAAGCGACCTCGCGAGAGCAAGCGGACCTCATAAAGTGCGTCGTAGTCCGGATTGGAGTCTGCAACTCGACTCCATGAAGTCGGAATCGCTAGTAATCGTAGATCAGAATGCTACGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCATGGGAGTGGGTTGCAAAAGAAGTAGGTAGCTTAACCTTCGGGAGGGCGCTACCAC
2.3 physiological Biochemical experiment identification
Table 1 Pantoea cypripedii P11 physiological and biochemical characterization (+Positive; -negative;)
| Physiological and biochemical test | Results | Physiological and biochemical test | Results |
| Lysine | - | Urease enzyme | - |
| Ornithine | - | Gas production | + |
| Xylose | + | MR | + |
| Raffinose | + | VP | - |
| Sorbitol | - | Indigo substrate | + |
| Adonis amurensis L | - | Dynamic force | + |
| Phenylalanine (Phe) | - | Hydrogen sulfide | - |
The results indicate that the phosphate solubilizing bacteria Pantoea cypripedii P can utilize xylose and raffinose, can not utilize lysine, ornithine, sorbitol, adonitol and phenylalanine, has no urease activity, can produce organic acid, can produce gas, can produce tryptophan enzyme to decompose tryptophan to produce indole, has motility and can not decompose sulfur-containing amino acid.
Example 3 determination of phosphorus-dissolving Capacity
3.1 Qualitative determination of Pantoea cypripedii P11 phosphorus-dissolving Capacity
The purified phosphate solubilizing bacteria Pantoea cypripedii P are smeared at the center of an inorganic phosphorus solid culture medium, and are cultured for 7 days at the temperature of 28 ℃, the growth of the phosphate solubilizing bacteria on the inorganic phosphorus solid culture medium is shown as a figure 3, the diameter (D) of a phosphate solubilizing ring of a bacterial colony and the diameter (D) of the bacterial colony are measured, the phosphate solubilizing ring and the diameter (D) of the bacterial colony are repeated for three times, D/D is an index for representing the relative phosphate solubilizing capacity of the phosphate solubilizing bacteria, and the size of the phosphate solubilizing capacity of the bacterial strain is primarily judged according to the size of the D/D value, wherein the larger the D/D value is, the stronger the phosphate solubilizing capacity is. The D/D value of the phosphorus-dissolving bacteria Pantoea cypripedii P is larger than 3, and the phosphorus-dissolving capacity is proved to be strong, and the specific results are shown in Table 2.
TABLE 2 phosphorus dissolving Capacity of phosphorus dissolving bacteria Pantoea cypripedii P11
| Strain | Diameter D (mm) of phosphorus dissolving ring | Colony diameter d (mm) | D/d |
| 1 | 25 | 6.9 | 3.623 |
| 2 | 27 | 8.0 | 3.375 |
| 3 | 23 | 6.1 | 3.770 |
| Average value of | 25 | 7.0 | 3.589 |
3.2 Quantitative determination of Pantoea cypripedii P11 phosphorus-dissolving Capacity
The phosphorus-dissolving bacteria Pantoea cypripedii P11 are inoculated into an inorganic phosphorus liquid culture medium, the inorganic phosphorus liquid culture medium without bacteria is used as a control group, shaking culture is carried out for 7d at 28 ℃, and the phosphorus-dissolving amount in the supernatant is detected every 24h by a molybdenum-antimony colorimetric method, so that the results are shown in Table 3.
TABLE 3 amount of dissolved phosphate group of Pantoea cypripedii P11 to tricalcium phosphate
| Days (days) | 1 | 2 | 3 | 4 | 5 | 6 | 7 |
| Soluble phosphorus content (mg/L) | 269.96 | 470.88 | 616.25 | 630.35 | 644.95 | 663.59 | 650.99 |
It can be seen that after the phosphorus-dissolving bacteria Pantoea cypripedii P11 is inoculated into the inorganic phosphorus liquid culture medium, the content of soluble phosphorus in the solution gradually rises, the solution is cultured for 4 days, the content of the soluble phosphorus reaches 630.35mg/L, the solution is stable, the content of the soluble phosphorus reaches the highest 663.59mg/L on the 6 th day, and the soluble phosphorus content reaches 650.99mg/L on the 7 th day.
Example 4 Pantoea cypripedii P11 determination of indoleacetic acid (IAA) production Capacity
Salkowski color solution: 15mL of 0.5mo1/L FeC1 3 300mL of concentrated sulfuric acid (specific gravity 1.84), 500mL of distilled water, and the mixture is stored in a dark place. To 1mL of the sample was added 4mL of the colorimetric solution at the time of use. The separated and purified lysophosphate Pantoea cypripedii P11 is inoculated into LB liquid medium containing L-tryptophan and cultured for 6d at 30 ℃. Centrifuging the bacterial suspension at 5000r/min for 10min, mixing 1ml supernatant with 4ml Salkowski colorimetric solution, standing in dark for 30min, and measuring at 530nmIs a solid phase, and is a liquid phase. 3 replicates were set and measured once for 24 hours. And calculating the IAA content in the phosphate solubilizing bacteria fermentation liquor according to a standard curve to obtain the IAA yield capability of the phosphate solubilizing bacteria, wherein the result is shown in Table 4.
TABLE 4 IAA production by P-lytic bacteria Pantoea cypripediiP11
| Days (days) | 1 | 2 | 3 | 4 | 5 | 6 | 7 |
| IAA(μg/ml) | 22.89 | 33.60 | 40.55 | 41.56 | 40.80 | 39.82 | 37.44 |
As can be seen, after the phosphorus-dissolving bacteria Pantoea cypripedii P11 is inoculated into LB liquid medium, the IAA content in the solution gradually rises, and the IAA content reaches 41.56 mug/ml at the highest after 4d cultivation.
Example 5 Pantoea cypripedii P11 siderophore production assay
Lysophosphoric Pantoea cypripedii P11 was inoculated onto CAS agar plates, producing yellow halos around the strain. CAS plate medium: solution a:60.5mg CAS,50mL distilled water, 10mL ferric chloride solution (containing 1mM FeCl) 3 ·6H 2 O,10mM HCl); solution B:72.9mg of HDTMA (cetyltrimethylammonium bromide), 40mL of distilled water; solution C: adding the solution A into the solution B, uniformly mixing, and sterilizing at 121 ℃ for 15min;2mL of 1mM calcium chloride solution, 2mL of 1mM magnesium sulfate solution, and pH value of the solution is adjusted to 6.8-7.0. Distilled water is added to the volume of 1000mL, 18g of agar is added, sterilization is carried out for 15min at 121 ℃,50mL of solution C is added when the temperature is reduced to below 60 ℃, and the flat plate is manufactured after uniform mixing. The determination method of the siderophore comprises the steps of applying the phosphate-dissolving bacteria Pantoea cypripedii P to the center of a CAS plate culture medium, placing the CAS plate culture medium in a 28 ℃ incubator for 48 hours, and observing and recording the color change around a colony. The ratio (D/D) of the diameter of the siderophore ring to the diameter of the colonies of the lysophosphoric bacteria was 3.5, as shown in FIG. 4.
Example 6 Pantoea cypripedii P11 evaluation of phosphorus solubilizing ability in soil
5.0g of soil is taken, 25mL of sterile water is added, sterilization is carried out for 30min at 121 ℃, after cooling, 5mL of phosphorus-dissolving bacteria Pantoea cypripedii P are inoculated, and simultaneously, 5mL of LB liquid medium is inoculated as a parallel control group, and three groups are parallel. Standing at 25deg.C, collecting supernatant after 7d, and determining soluble effective phosphorus content by Mo-Sb colorimetric method.
The content of soluble effective phosphorus in the non-inoculation control group is 48.01mg/kg, and the content of soluble effective phosphorus in the experimental group is 118.34mg/kg. Proved by the test, the phosphorus-dissolving bacteria Pantoea cypripedii P11 can effectively improve the content of soluble phosphorus in soil, and the increment reaches 70.33mg/kg, so that the soluble phosphorus in soil is improved by 1.46 times.
Example 7 Pantoea cypripedii P11 growth-promoting experiment
Taking full barley seeds, washing with sterile water for 5 times, respectively taking 50 seeds, placing in sterile water (control group) and 10 8 Soaking cfu/mL Pantoea cypripedii P11 bacterial suspension (experimental group) for 1min, and planting seeds in flowerpot filled with sterilized soilThe culture was performed at 28℃for 10 days, and the results are shown in Table 5.
TABLE 5 growth promoting effect of lysophosphate Pantoea cypripedii P11
| Control group | Experimental group | |
| Survival rate (%) | 48.0 | 72.0 |
| Bud length mean (cm) | 16.5 | 17.2 |
| Fresh weight average value (mg) | 229.62 | 311.74 |
| Dry weight average (mg) | 21.88 | 24.84 |
As can be seen from the data in table 5, lysophosphate Pantoea cypripedii P11 has a good pro-active effect on barley seeds.
Although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims (12)
1. The efficient phosphorus-dissolving bacteria is characterized by being named as: pantoeacypripedi P11, biological deposit number: CCTCCNO: m2023159.
2. A fermentation product obtained by fermenting the phosphorus-dissolving bacterium Pantoea cypripediP11 according to claim 1.
3. A microbial agent comprising the lysophosphoric Pantoea cypripediP according to claim 1 or comprising the fermentation product according to claim 2.
4. A phosphate solubilizing bacterial fertilizer comprising phosphate solubilizing bacteria Pantoea cypripediP11 according to claim 1 or comprising the fermentation product according to claim 2.
5. A soil ecological environment regulator comprising the phosphorus-solubilizing bacterium pantoea cypripedip11 according to claim 1 or the fermentation product according to claim 2.
6. Use of the phosphorus-solubilizing bacteria pantoea cypripedip11 of claim 1 or the fermentation product of claim 2 or the microbial inoculum of claim 3 in the production of indoleacetic acid and/or siderophores.
7. Use of the phosphorus-solubilizing bacteria pantoea pripedip11 of claim 1 or the fermentation product of claim 2 or the microbial inoculum of claim 3 for promoting plant growth and increasing plant yield.
8. The use according to claim 7, wherein the phosphorus-solubilizing bacteria Pantoea cypripediP, fermentation products and/or bacteria in the bacterial agentSuspension concentration of 10 8 cfu/ml。
9. Use of the phosphorus-solubilizing bacteria pantoea cypripedip11 of claim 1 or the fermentation product of claim 2 or the microbial inoculum of claim 3 for solubilizing phosphorus, increasing the utilization rate of phosphate fertilizer and/or increasing the content of soluble phosphorus in soil.
10. Use of the kresoxim-methyl of pantoea cypripedip11 of claim 1 or the fermentation product of claim 2 or the microbial inoculum of claim 3 for preventing and/or ameliorating soil problems caused by the application of phosphate fertilizer;
the soil problems include soil hardening, acidification, nutrient element and flora imbalance.
11. A method for preparing the efficient phosphorus-dissolving bacteria pantoea cypripedip11 as claimed in claim 1, which is characterized by comprising the following steps:
collecting rhizosphere soil samples from 5-10cm below the Zhong Mingzhen Jin Fengcun surface layer of the copper tomb city;
weighing 10g of soil sample into a 250mL triangular flask, adding 90mL of sterile water, and 180 r.min -1 Shaking for 30min to prepare soil suspension, and sequentially preparing 10 by 10-fold dilution method -2 、10 -3 And 10 -4 Is a soil dilution of (2);
respectively sucking 0.1mL of soil dilution liquid, coating the soil dilution liquid on an inorganic phosphorus solid culture medium, repeating each gradient three times, and culturing in an inverted manner in a 28 ℃ incubator;
after 3-5d, picking out a colony which has obvious phosphate-dissolving ring and has good growth vigor, transferring the colony to an inorganic phosphorus solid culture medium again by a streaking method, and carrying out subculture for 3 times and purifying again to obtain the efficient phosphate-dissolving bacteria Pantoeacyprip ediP.
12. The method for preparing the efficient phosphorus-dissolving bacteria Pantoea cypripedip11, which is characterized in that,
the formula of the inorganic phosphorus solid culture medium is as follows: 18.0g of agar, 10.0g of glucose, 0.3g of NaCl, 0.3g of KCl, (NH) 4 ) 2 SO 4 0.5g，MgSO 4 ·7H 2 O0.3g，FeSO 4 ·4H 2 O0.036g，MnSO 4 ·4H 2 O0.03g，Ca 3 (PO 4 ) 2 5.0g, distilled water 1L and pH value 7.0-7.2.
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| CN114164140B (en) * | 2021-10-28 | 2023-12-12 | 中国林业科学研究院华北林业实验中心 | Efficient phosphorus-dissolving bacteria MQR6 and fermentation product and application thereof |
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