CN108034609B - Paenibacillus strain T1-1 with phosphate solubilizing effect and application thereof - Google Patents
Paenibacillus strain T1-1 with phosphate solubilizing effect and application thereof Download PDFInfo
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- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
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- C02—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F—TREATMENT OF WATER, WASTE WATER, SEWAGE, OR SLUDGE
- C02F3/00—Biological treatment of water, waste water, or sewage
- C02F3/34—Biological treatment of water, waste water, or sewage characterised by the microorganisms used
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- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
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- C02F2101/105—Phosphorus compounds
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Abstract
The invention belongs to the technical field of microorganisms, and particularly discloses a paenibacillus strain T1-1 with a phosphate solubilizing effect and application thereof. The preservation number of the paenibacillus strain T1-1 is CGMCC No. 14927. The microbial agent or microbial fertilizer prepared from the bacillus-like strain can obviously promote the growth of vegetables, effectively reduce the usage amount of chemical phosphate fertilizer, and can be popularized and used in agricultural production.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to a paenibacillus strain with a phosphate solubilizing effect and application thereof.
Background
The phosphate-dissolving bacteria can secrete organic acid and phosphatase, the organic acid can convert mineral phosphorus into soluble dihydrogen phosphate and hydrogen phosphate ions, and the process is called degradation of the mineral phosphorus; the phosphatase promotes mineralization (hydrolysis) of phosphorus for effective utilization by plants. Therefore, the lysobacteria play an important role in promoting the absorption of the plant phosphorus. The microbial method is adopted to improve the effectiveness of phosphorus in soil, increase the nutrient level of soluble phosphorus in soil, promote the effective utilization of phosphorus by plants and improve the microbial environment of soil.
The Paenibacillus has the characteristics of long survival period, strong stress resistance and the like, is widely distributed in different soils and plant rhizomes, and can invade the interior of plant tissues. Many strains of this genus have received considerable attention from researchers due to their nitrogen fixation, phosphorus solubilizing ability, resistance to pathogenic bacteria, and growth promoting effects on plants.
The microbial fertilizer refers to a specific product which contains living microorganisms and provides nutrition to plants and regulates plant growth through specific actions of the microorganisms. The microbial fertilizer is one of the important directions of sustainable development of agriculture and forestry in the future. But the outstanding problems of single strain, single function, short shelf life and the like still need to be solved.
Disclosure of Invention
In order to solve the problems in the prior art, the invention aims to provide a paenibacillus strain T1-1 with a phosphate solubilizing effect and application of a microbial agent or bacterial fertilizer prepared by the same in planting of vegetables and the like.
In order to realize the purpose of the invention, the technical scheme of the invention is as follows:
in a first aspect, the invention provides a Paenibacillus strain T1-1 with a phosphate-solubilizing effect, which is identified as Paenibacillus sp, is preserved in the China general microbiological culture Collection center (CGMCC for short, with the address of No. 3 Homew No.1, the institute of microbiology, China academy of sciences, postal code: 100101), has a preservation date of 2017, 11 months and 17 days, and has a preservation number of CGMCC No. 14927.
The paenibacillus strain T1-1 provided by the invention grows well on LD solid culture medium, the colony is beige, the edge is smooth and has no wrinkles, the diameter is 1mm-1.5mm, the strain is gram-positive bacteria, the growth temperature is 25-55 ℃, the optimal growth temperature is 37 ℃, and the optimal pH value is 6.8-7.0. Under adverse conditions, the thalli are transformed to generate spores which have strong adverse environment stress resistance and can tolerate the high temperature of 100 ℃ in a short time.
To facilitate the analysis of the phosphate solubilizing ability of the strains, a laboratory phosphate solubilizing test was designed to study the solubilization of inorganic and organic phosphorus by the strains, as detailed in examples 2-3.
In a second aspect, the present invention provides a microbial inoculant or microbial fertilizer comprising paenibacillus strain T1-1.
Further, the microbial agent can be a fermentation liquid or a culture liquid of the paenibacillus strain T1-1, and the specific preparation method can be as follows:
inoculating Paenibacillus strain T1-1 into liquid culture medium, culturing at 30 deg.C in incubator, stopping culturing after 85% of thallus produces spores, and obtaining fermentation liquid or culture solution.
Wherein the formula of the liquid culture medium is as follows: 10g/L glucose, 5.0g/L Ca3PO4)2、5.0g/L MgCl2、0.25g/L MgSO4·7H2O、0.2g/L KCl、1g/L(NH4)2SO4pH 7.0. Sterilizing at 121 deg.C for 25 min.
When the Paenibacillus strain T1-1 is to be subjected to solid culture, the solid culture medium used may be one in which 14g of agar per liter of the liquid culture medium described above is added.
Further, the microbial fertilizer contains the microbial agent and an adsorption carrier.
The adsorption carrier consists of turfy soil, potassium humate, vermiculite and ground phosphate rock; preferably, the mass ratio of the components is 3-5: 0.5-1.5: 1-2: 0.1-0.5, more preferably in a mass ratio of 3.5:1:1.5: 0.3. When the adsorption carrier is prepared, the raw materials are mixed and crushed, and then are sieved by a 100-mesh sieve, sterilized and cooled for later use.
Preferably, the microbial fertilizer is prepared by mixing a microbial agent and an adsorption carrier according to the mass ratio of 1:1 and then drying.
Wherein the microbial agent is adjusted to have a colony concentration of not less than 10 before being mixed with the adsorption carrier8CFU/mL. In one embodiment of the invention, the microbial agent used in the preparation of the microbial fertilizer is 5 × 108CFU/mL。
The microbial fertilizer can be applied to planting gramineous plants such as wheat, corn, rice, cucurbitaceous cucumbers and the like.
In a third aspect, the invention provides an application of the paenibacillus strain T1-1 in preparing a phosphate solubilizing preparation.
And the application of the paenibacillus strain T1-1 as an active ingredient preparation in agricultural production.
And the application of the microbial agent or the microbial fertilizer in vegetable planting. The microbial fertilizer has a remarkable promoting effect on the growth of vegetables, can effectively reduce the usage amount of chemical phosphate fertilizer, and can be popularized and used in agricultural production.
Through detection, the paenibacillus strain T1-1 and the microbial agent or the microbial fertilizer especially prepared by the paenibacillus strain T1-1 are nontoxic to human beings and livestock and have no pathogenicity.
The raw materials or reagents involved in the invention are all common commercial products, and the operations involved are all routine operations in the field unless otherwise specified.
The invention has the beneficial effects that:
the paenibacillus strain T1-1 provided by the invention is separated from the rhizosphere soil of the gramineous plant corn, has strong environment adaptability and can generate spores in uncomfortable environment. Meanwhile, the results of laboratory phosphate solubilizing tests and plant tests show that the strain has dissolving capacity for various inorganic or organic phosphorus sources, so that the strain can be used for dissolving the phosphorus source in soil. On the other hand, according to a laboratory phosphorus dissolution test, the paenibacillus strain T1-1 has a good phosphorus source dissolving effect in an NBRIP liquid culture medium, and the fact shows that the paenibacillus strain T1-1 can be used for dissolving phosphorus elements in a liquid environment, such as dissolving a phosphorus source in an eutrophic water body.
The paenibacillus strain T1-1 separated from the rhizosphere soil of the gramineous plant corn is used for dissolving inorganic and organic phosphorus sources in the soil, has the characteristics of stable phosphate dissolving, high efficiency and broad spectrum, is favorable for plants to absorb phosphorus elements necessary for growth, and promotes the growth vigor of the plants; the microbial fertilizer prepared by mixing the strain fermentation liquor and a fertilizer matrix (an adsorption carrier) can quickly form a dominant flora after being applied to soil, has good fertilizer effect, can obviously improve the content of available phosphorus in the soil, and promotes the healthy growth of plants. The strain is used for dissolving inorganic and organic phosphorus sources in eutrophic water, and can promote aquatic plants to absorb redundant phosphorus elements in the water, thereby preventing and treating water eutrophication.
Detailed Description
The present invention is further illustrated by the following examples. It is to be understood that the following examples are given for illustrative purposes only and are not intended to limit the scope of the present invention. Various modifications and alterations of this invention will become apparent to those skilled in the art without departing from the spirit and scope of this invention.
The experimental procedures used in the following examples are all conventional procedures unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Example 1 preparation of microbial Agents
Inoculating Paenibacillus strain T1-1 into liquid culture medium, culturing at 30 deg.C in incubator, stopping culturing after 85% of thallus produces spores, and obtaining fermentation liquid or culture solution.
Wherein the formula of the liquid culture medium is as follows: 10g/L glucose, 5.0g/L Ca3(PO4)2、5.0g/L MgCl2、0.25g/L MgSO4·7H2O、0.2g/L KCl、1g/L(NH4)2SO4pH 7.0. Sterilizing at 121 deg.C for 25 min.
The concentration of the bacterial colony in the obtained fermentation liquid was 5X 108CFU/mL。
EXAMPLE 2 determination of phosphorus solubilizing ability in liquid Medium
100mL of NBRIP liquid medium (10g/L glucose, 5.0g/L MgCl) was poured into a 250mL Erlenmeyer flask2、0.25g/L MgSO4·7H2O、0.2g/L KCl、1g/L(NH4)2SO4pH 7.0), 5.0g/L of inorganic Ca was added thereto, respectively3(PO4)2、FePO4、AlPO4And 2.0mL/L of organic phosphorus (lecithin) as a phosphorus source.
The fermentation broth obtained in example 1 was inoculated, and shake-cultured (28 ℃ C., 160r/min) for 8d, in comparison with Escherichia coli JM 109. Centrifuging at 4 deg.C (10000 r/min) for 15min, collecting appropriate amount of supernatant, and measuring phosphorus content (determined by Mo-Sb colorimetry), and pH value.
The results show that the Paenibacillus strain T1-1 can dissolve different inorganic phosphorus and organic phosphorus and is Ca-tolerant3(PO4)2The phosphorus dissolving capacity of the fertilizer is very strong and reaches 378 mg/L. Meanwhile, the composite material also has a certain phosphorus dissolving capacity on organophosphorus lecithin.
EXAMPLE 3 determination of phosphorus solubilizing ability in solid Medium
The phosphorus solubilizing ability of Paenibacillus phosphate solubilizing strain T1-1 was determined using NBRIP solid medium.Activating the strain of Paenibacillus phosphate solubilizing T1-1 in LD solid culture medium, and culturing at 30 ℃ overnight; the activated single colony is treated with sterile toothpick containing Ca3(PO4)2The center of NBRIP solid medium was spotted and incubated at 30 ℃ for 7 days in triplicate. The colony diameter (D) and the diameter (D) of the phosphorus-solubilizing circle around the colony were measured.
Phosphorus Solubilization Index (PSI): PSI is D/D.
The results showed that Paenibacillus strain T1-1 contained Ca3(PO4)2The PSI of the solid NBRIP medium of (1) was 2.46.
EXAMPLE 4 preparation of microbial Fertilizer
1. Mixing turfy soil, potassium humate, vermiculite and ground phosphate rock according to a mass ratio of 3.5:1:1.5:0.3, crushing, sieving by a 100-mesh sieve, sterilizing and cooling to obtain a microbial fertilizer adsorption carrier (matrix) for later use;
2. and (3) mixing the fermentation liquor obtained in example 1 and the adsorption carrier obtained in the step 1 in a mass ratio of 1:1, mixing and drying to obtain the microbial fertilizer.
EXAMPLE 5 Effect of application of microbial Fertilizer
The wheat seeds are sterilized and then placed in a sterile plate paved with wet filter paper, and the sterilized wheat seeds are placed in a constant temperature box at 25 ℃ for germination acceleration. After germination, seedlings with consistent development are picked and transplanted into plastic pots, and three seedlings are planted in each pot. The microbial fertilizer was applied as a base fertilizer (0.4 g/pot) and the adsorption vehicle was applied as a blank control. The experiment was set up in triplicate. After the second week of transplantation, the roots and stems and leaves of the plants were collected by destructive sampling and the root length and stem length were recorded separately.
The results show that the plant height and fresh weight are respectively increased by 35% and 54% by using the microbial fertilizer compared with the blank control (Table 1), and the microbial fertilizer has a better plant growth promoting effect and can effectively reduce the usage amount of the phosphate fertilizer. Therefore, the strain has potential popularization market in the application of plants such as vegetables and the like, and is expected to obtain good economic benefit.
| Treatment of | Plant height (cm) | Fresh weight (g) |
| Control | 20.43b | 0.28b |
| Microbial fertilizer | 27.59a | 0.43a |
Data in the table were analyzed by statistical software SPSS, and different letters in the same column indicated significant differences at the p-0.05 level.
It should be understood that the technical solutions of the above embodiments, in which the amounts of reagents or raw materials used are proportionally increased or decreased, are substantially the same as those of the above embodiments.
Although the invention has been described in detail hereinabove with respect to a general description and specific embodiments thereof, it will be apparent to those skilled in the art that modifications or improvements may be made thereto based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.
Claims (10)
1. The Paenibacillus strain T1-1 with the phosphate solubilizing effect is characterized in that the classification name of the Paenibacillus strain T1-1 is Paenibacillus (Paenibacillus sp.), and the preservation number is CGMCC No. 14927.
2. A microbial agent comprising the strain of claim 1.
3. The microbial agent according to claim 2, wherein the preparation method comprises: inoculating paenibacillus strain T1-1 into a liquid culture medium, culturing in an incubator at 30 ℃, stopping culturing after 85% of thalli generate spores, and obtaining a fermentation liquid or a culture solution;
wherein the formula of the liquid culture medium is as follows: 10g/L glucose, 5.0g/L Ca3(PO4)2、5.0g/L MgCl2、0.25g/L MgSO4·7H2O、0.2g/L KCl、1g/L(NH4)2SO4Sterilizing at 121 deg.C for 25min and pH 7.0.
4. A microbial fertilizer comprising the strain of claim 1.
5. The microbial fertilizer according to claim 4, which is prepared by the following method: the number of colonies of the strain is not less than 108Mixing the CFU/mL fermentation liquor with an adsorption carrier in a mass ratio of 1-3: 1-3, and drying to obtain the CFU/mL fermentation liquid.
6. The microbial fertilizer according to claim 5, wherein the microbial agent is mixed with the adsorption carrier in a mass ratio of 1:1.
7. A microbial fertilizer according to claim 5 or 6, wherein the adsorption carrier consists of turfy soil, potassium humate, vermiculite and ground phosphate rock in a mass ratio of 3-5: 0.5-1.5: 1-2: 0.1-0.5.
8. A microbial fertilizer according to claim 7, wherein the adsorption carrier consists of turfy soil, potassium humate, vermiculite and ground phosphate rock in a mass ratio of 3.5:1:1.5: 0.3.
9. Use of the paenibacillus strain T1-1 of claim 1, or the microbial agent of claim 2 or 3, or the microbial fertilizer of any one of claims 4-8 for improving the yield and quality of crops.
10. Use of the paenibacillus strain T1-1 of claim 1, or the microbial agent of claim 2 or 3, for the preparation of a microbial fertilizer or a bio-organic fertilizer.
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| CN110684695B (en) * | 2019-11-11 | 2022-10-04 | 安顺学院 | Paenibacillus polymyxa QZY-1 and application thereof |
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