CN113812298A - By using phosphate solubilizing bacteria and vitamin B12Method for realizing rapid growth of poplar - Google Patents

By using phosphate solubilizing bacteria and vitamin B12Method for realizing rapid growth of poplar Download PDF

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Publication number
CN113812298A
CN113812298A CN202111091371.1A CN202111091371A CN113812298A CN 113812298 A CN113812298 A CN 113812298A CN 202111091371 A CN202111091371 A CN 202111091371A CN 113812298 A CN113812298 A CN 113812298A
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solubilizing bacteria
phosphate solubilizing
vitamin
phosphorus
soil
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CN202111091371.1A
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Chinese (zh)
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董磊
闫中园
王丽贤
单德江
翟冰
杨雷
吕浩
周劲男
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Zhongbang Landscape Planning And Design Co ltd
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Zhongbang Landscape Planning And Design Co ltd
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G17/00—Cultivation of hops, vines, fruit trees, or like trees
    • A01G17/005—Cultivation methods
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01C—PLANTING; SOWING; FERTILISING
    • A01C21/00—Methods of fertilising, sowing or planting

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  • Life Sciences & Earth Sciences (AREA)
  • Soil Sciences (AREA)
  • Environmental Sciences (AREA)
  • Botany (AREA)
  • Fertilizers (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The invention relates to the utilization of phosphate solubilizing bacteria and vitamin B12Method for realizing rapid growth of poplar by using phosphate solubilizing bacteria and vitamin B12Mixing and applying to soil for planting plants, wherein the phosphate solubilizing bacteria is Citrobacter freundii and can be used for adding vitamin B12Adding into liquid containing phosphate solubilizing bacteria, and mixing to obtain product containing phosphate solubilizing bacteria and vitamin B12The growth promoter according to (1) can be applied to soil in which plants are planted, phosphate solubilizing bacteria can be inoculated into a culture medium for culture, the liquid containing the phosphate solubilizing bacteria is prepared, and the growth promoter can be applied alone or together with a phosphate fertilizer. The invention can promote the absorption of plants such as poplar and the like to phosphorus elements in soil, thereby improving the survival ability of poplar.

Description

By using phosphate solubilizing bacteria and vitamin B12Method for realizing rapid growth of poplar
Technical Field
The invention relates to the utilization of phosphate solubilizing bacteria and vitamin B12A method for realizing the rapid growth of poplar belongs to the technical field of agricultural microorganisms.
Background
The poplar is one of three fast growing tree species in the world, and is an important tree species of protection forests, urban landscaping and industrial timberlands in China due to strong adaptability and wide application range. More nutrients are consumed when the poplar grows, but in recent years, nutrient elements in soil are insufficient due to the fact that the rotation cutting period of the poplar is shortened.
Phosphorus is a component of important compounds such as nucleotide, phospholipid, DNA, RNA, ATP and the like in plants, and participates in physiological and biochemical processes such as energy metabolism, carbohydrate metabolism, substance transformation, operation and the like in plants. The phosphorus in the soil usually exists in the form of an inorganic phosphorus compound and an organic phosphorus compound, and the two forms of phosphorus mostly exist in the form of insoluble or slightly soluble phosphorus, cannot be directly absorbed by plants, and is soluble phosphorus which is easily absorbed by the plants. Most of inorganic phosphorus in phosphate fertilizer applied to soil is easy to react with Ca2+、Mg2+、Mn2+、Fe3+The chelate is generated by complexing of metal ions and is difficult to be utilized by plants, and organic phosphorus is converted into available inorganic phosphorus through the action of microorganisms and can be absorbed and utilized by plants. Only 20% of soil phosphorus can be absorbed and utilized by plants, and due to low effectiveness and utilization rate, phosphorus becomes one of key elements for limiting plant growth.
A plurality of microorganisms for promoting the utilization of rhizosphere nutrients by plants are distributed in the soil, and the microorganisms comprise phosphate-solubilizing microorganisms, and the phosphate-solubilizing microorganisms can convert insoluble, slow-acting and mineral phosphorus compounds in the soil into simple inorganic phosphide which is easily absorbed and utilized by plants, so that the content of soluble phosphorus in the soil is increased for the plants to absorb and utilize.
On the other hand, vitamins are essential micronutrients from many photosynthetic plants and bacteria. Because vitamins are essential cofactors in various metabolic pathways and act as antioxidants, they need to be provided to organisms that cannot produce them.
In plant-microorganism interactions, vitamins can affect the proliferation of plant root growth-promoting bacteria in and around the root system. Vitamins come from root secretions or are produced by rhizobacteria and fungi, wherein some of the vitamins secreted by the plant roots are not completely synthesized by the plant, but are produced by microorganisms, which are absorbed and secreted by the plant roots.
Disclosure of Invention
The invention aims to utilize Citrobacter freundii (B.freundii)Citrobacter freundii) And trace amount of vitamin B12Promote the absorption of plants such as poplar to phosphorus element in soil, and further improve the survival ability of poplar.
The technical scheme of the invention is as follows: by using phosphate solubilizing bacteria and vitamin B12The method for realizing the rapid growth of poplar comprises the steps of mixing phosphate solubilizing bacteria and vitamin B12Mixed and applied to soil for planting plants, wherein the phosphate solubilizing bacteria are Citrobacter freundii (B)Citrobacter freundii）。
The plant may be any plant, in particular a tree, requiring phosphorus.
The plant can be a fast growing tree species such as poplar.
The soil may be soil lacking phosphorus capable of being assimilated by plants and containing phosphorus that is poorly assimilated by plants.
The phosphorus that is difficult to be absorbed by plants may be organic phosphorus that is difficult to be absorbed by plants and/or inorganic phosphorus that is difficult to be absorbed by plants.
The inorganic phosphorus hardly absorbed by plants includes Ca3(PO4)2、AlPO4And FePO4And the like.
The phosphorus that is difficult to be absorbed by the plant may be phosphorus in a phosphate fertilizer applied to the soil and/or phosphorus originally in the soil.
Preferably, vitamin B is added12Adding into liquid containing phosphate solubilizing bacteria, and mixing to obtain product containing phosphate solubilizing bacteria and vitamin B12The growth promoter (mixed liquid) of (1), the growth promoter being applied to the soil in which the plant is planted.
Preferably, the phosphate solubilizing bacteria are inoculated into a culture medium for culture, and the liquid containing the phosphate solubilizing bacteria is prepared from a culture solution.
Preferably, the medium is LB medium.
Preferably, the concentration of phosphate solubilizing bacteria in the growth promoter is 2 x 1010 CFU/ml (expressed as colony forming unit concentration).
Preferably, vitamin B in the growth promoter12The concentration was 0.05 g/l.
Preferably, inorganic phosphorus is further added to the growth promoter.
Preferably, the inorganic phosphorus (added to the growth promoter) is Ca3(PO4)2。
Preferably, the inorganic phosphorus (added to the growth promoter) is AlPO4。
Preferably, the inorganic phosphorus (added to the growth promoter) is FePO4。
The invention has the beneficial effects that: selecting Citrobacter freundii suitable for soil environment as phosphate solubilizing bacteria, and mixing with vitamin B12The citric acid fimbriatum is applied to soil for planting plants in a mixed mode, the citric acid fimbriatum is suitable for the soil environment, particularly can be used for the soil environment for planting plants, organic phosphorus and inorganic phosphorus which are difficult to be absorbed by the plants can be converted into forms which are easy to be absorbed by the plants in the soil environment, a required absorbable phosphorus source is provided for the growth of the plants, the obstruction caused by the fact that the phosphorus in the soil is difficult to be absorbed by the plants to the growth of the plants is overcome, the growth of the plants is facilitated, particularly for fast-growing trees such as poplar trees and the like, the demand for the phosphorus is relatively large, the phosphorus which is difficult to be absorbed by the soil is converted into the absorbable phosphorus, short plates which are relatively insufficient in the soil in a plurality of existing fields are effectively overcome, and the growth of the plants is promoted and guaranteed; using phosphorus and vitamin B12Synergistic effect on the reproduction and metabolism of phosphate solubilizing bacteria, vitamin B is added12，Provides favorable conditions for the propagation and metabolism of the phosphate solubilizing bacteria, and is favorable for ensuring the concentration and the activity of the phosphate solubilizing bacteria.
The invention is not only suitable for poplar, but also suitable for other plants needing phosphorus, and is mainly used for occasions with relative insufficient phosphorus absorbable in soil. The phosphate fertilizer, especially the phosphate fertilizer which is difficult to be directly absorbed by plants, can be mixed with the growth promoter or applied simultaneously, and the phosphate solubilizing bacteria provided by the invention can convert the phosphorus which is difficult to be absorbed in the phosphate fertilizer into a form which is easy to be absorbed, so that the utilization rate of phosphate in the phosphate fertilizer is improved, and a sufficient phosphorus source is provided for the propagation and metabolism of the phosphate solubilizing bacteria.
Drawings
FIG. 1 is the experimental result of the dissolving capacity of the phosphate solubilizing bacteria of the present invention for different inorganic insoluble phosphorus sources;
FIG. 2 shows the pair of Simultaneous phosphate solubilizing bacteria Ca according to the present invention3(PO4)2Experimental results of the dissolving ability of (a);
FIG. 3 shows the experimental results of the seedling height for the growth of poplar according to the present invention;
FIG. 4 shows the experimental results of the leaf width of the poplar growth according to the present invention;
FIG. 5 shows the results of the leaf number experiment of poplar growth according to the present invention.
Detailed Description
Referring to FIGS. 1-5, the phosphate solubilizing ability of phosphate solubilizing bacteria and the growth of poplar were tested.
One) experiment of dissolving ability of phosphate solubilizing bacteria to different inorganic insoluble phosphorus sources
Experimental articles:
LB culture medium: peptone 10g/l, yeast powder 5g/l, sodium chloride 5g/l, the pH of the medium was adjusted with NaOH to 7.4.
Phosphate solubilizing medium: 10g/l of glucose; (NH)4)2SO4 0.5 g/l；MgSO4·7H2O 0.3 g/l；MnSO4·4H2O 0.03 g/l；KCl 0.3 g/l；FeSO4·7H2O 0.03 g/l；Ca3(PO4)210g/l, vitamin B120.05g/l, the pH of the medium was adjusted to 7.4 with NaOH. The other inorganic insoluble phosphorus source is AlPO4And FePO4Replacement of Ca3(PO4)2。
The experimental steps are as follows:
phosphate solubilizing bacteriaCitrobacter freundiiInoculating into LB liquid culture medium, culturing at 30 deg.C and 150rpm for 24 hr, centrifuging 20ml of bacteria solution in 50ml sterile centrifuge tube at 8000rpm for 2min, discarding supernatant, and washing with 20ml sterile ultrapure water for 2 times to obtain seed solution.
Inoculating the seed solution into 100ml phosphate dissolving culture medium (3 different inorganic slightly soluble phosphorus sources) at a ratio of 2%, treating each slightly soluble phosphorus source with no seed solution as control, culturing at 30 deg.C and 150rpm for 2 days, centrifuging culture solution 8000rpm for 10min, collecting supernatant, filtering with 0.22 μm water phase filter membrane, and detecting the concentration of soluble phosphorus in the culture solution by spectrophotometry.
The experimental results are as follows: as shown in FIG. 1, phosphate solubilizing bacteriaCitrobacter freundiiHas dissolving capability to three different inorganic insoluble phosphorus sources, wherein Ca is contained3(PO4)2Has the strongest dissolving capacity.
II) phosphate solubilizing bacteria pair Ca at different times3(PO4)2Dissolving capacity of
Experimental articles:
LB culture medium: peptone 10g/l, yeast powder 5g/l, sodium chloride 5g/l, the pH of the medium was adjusted with NaOH to 7.4.
Phosphate solubilizing medium: 10g/l of glucose; (NH)4)2SO4 0.5 g/l；MgSO4·7H2O 0.3 g/l；MnSO4·4H2O 0.03 g/l；KCl 0.3 g/l；FeSO4·7H2O 0.03 g/l；Ca3(PO4)210g/l, vitamin B120.05g/l, the pH of the medium was adjusted to 7.4 with NaOH.
Phosphate solubilizing bacteriaCitrobacter freundiiInoculating into LB liquid culture medium, culturing at 30 deg.C and 150rpm for 24 hr, centrifuging 20ml of bacteria solution in 50ml sterile centrifuge tube at 8000rpm for 2min, discarding supernatant, and washing with 20ml sterile ultrapure water for 2 times to obtain seed solution.
The experimental steps are as follows:
inoculating the seed solution into 100ml of phosphate solubilizing culture medium at a ratio of 2%, performing control treatment with no seed solution, culturing at 30 deg.C and 150rpm for 3 parallel treatments, respectively sampling at 12h, 24h, 36h, 48h and 60h, centrifuging the culture solution at 8000rpm for 10min, collecting supernatant, filtering with 0.22 μm water phase filter membrane, and detecting the concentration of soluble phosphorus in the culture solution by spectrophotometry.
The experimental results are as follows: as shown in FIG. 2, phosphate solubilizing bacteriaCitrobacter freundiiCa can be dissolved from 12h3(PO4)2And reaches a maximum at 48h and then begins to fall.
Third) growth promotion of poplar seedlings
And (3) sowing the poplar seeds into seedling pots, compacting soil in 5 seeds per pot, watering, placing in a plant growth chamber for normal-temperature culture, and uniformly managing. Wherein, each pot is filled with 3kg of air-dried soil, and after the poplar seedlings come out of the soil, 1 healthy seedling is reserved in each pot for applying bacteria. Set 4 treatments: blank control (CK, no microbial inoculum), phosphate solubilizing bacteria (B) ((B))Citrobacter freundii) 10mL (concentration 108 cfu. mL)-1) Vitamin B1210mL (concentration 108 cfu. mL)-1) Phosphate solubilizing bacteria combined with vitamin B1210mL (concentration 108 cfu. mL)-1) Each treatment was repeated 6 times. After 6 months of culture, leaf width measurements were made on the height of the poplar seedling, the number of leaves and the 3 rd and 4 th leaves below the apical bud.
The results (average values) are shown in FIGS. 3, 4 and 5, and the P-solubilizing bacteria can be combined with vitamin B no matter the seedling height, leaf number or leaf width of poplar seedling12The growth promotion effect on poplar seedlings is the best, and then the phosphate solubilizing bacteria treatment and vitamin B treatment are carried out12It also has a small growth promoting effect on poplar seedlings.
For industrial implementation, the culture of the phosphate solubilizing bacteria can be based onCitrobacter freundiiThe characteristic of (2) is any schematic industrial culture method. The vitamin B12Any suitable form such as powder or solution may be used.
Considering that phosphate fertilizers now generally contain a higher proportion of phosphorus that is poorly absorbed by plants, the growth promoters of the present invention can be used with phosphate fertilizers, either mixed together prior to application or applied simultaneously to the soil in the same or adjacent areas depending on the application type adapted to each. For example, growth promoters and phosphate fertilizers are mixed into the planting soil of saplings.
The timing of the application of the growth promoters of the present invention depends on the actual plant demand for phosphorus. For example, planting soil mixed with a growth promoter may be filled into the tree hole at the time of planting. The planting soil can be mixed with a phosphate fertilizer, the mixing amount of the phosphate fertilizer can be reduced by about 50 percent compared with the conventional amount, and the phosphorus in the phosphate fertilizer is fully utilized by utilizing the action of phosphate-solubilizing bacteria. The amount of growth promoter to be used may be determined on the basis of the amount of soil to be planted or the amount of phosphate fertilizer to be mixed into the soil, for example, the growth promoter (2 x 10 in terms of concentration of phosphate solubilizing bacteria) is applied per kg of soil to be planted10 CFU/ml, the same applies hereinafter) may be 10-100 mg, or 200-2000 mg growth promoter per kg phosphate fertilizer may be applied. The application mode is that the fertilizer is embedded into the tree pit together with the phosphate fertilizer. The application amount, application timing and application mode of the growth promoting agent can be determined according to specific situations (for example, sampling analysis of the conditions of total phosphorus, soluble phosphorus and the like in soil) and growth requirements of plants, and the change rule of phosphorus form in soil after the growth promoting agent is applied under specific situations, the relation between the change rule and the application amount of the growth promoting agent and the like can also be obtained according to experiments so as to better grasp the use amount, the application timing and the like under the specific situations.
According toCitrobacter freundiiCharacteristics and general laws of microbial metabolism, it is expected that the phosphate solubilizing bacteria can perform the same or similar phosphate solubilizing action for most of the organic and inorganic phosphorus in other forms, particularly phosphorus used as a phosphate fertilizer.
For convenience, the present specification will refer to Citrobacter freundii: (Citrobacter freundii) Referred to as phosphate solubilizing bacteria.
The technical means disclosed by the invention can be combined arbitrarily to form a plurality of different technical schemes except for special description and the further limitation that one technical means is another technical means.

Claims (10)

1. By using phosphate solubilizing bacteria and vitamin B12The method for realizing the rapid growth of poplar is characterized in that phosphate solubilizing bacteria and vitamin B12Mixed and applied to soil for planting plants, and the phosphate solubilizing bacteria are Citrobacter freundii.
2. The method of claim 1, wherein vitamin B is administered12Adding into liquid containing phosphate solubilizing bacteria, and mixing to obtain product containing phosphate solubilizing bacteria and vitamin B12The growth promoter according to (1), which is applied to soil in which plants are planted.
3. The method according to claim 2, wherein the liquid containing the phosphate solubilizing bacteria is prepared by inoculating the phosphate solubilizing bacteria into a culture medium and culturing.
4. The method according to claim 3, wherein the medium is LB medium.
5. The method according to any of claims 2 to 4, characterized in that the concentration of phosphate solubilizing bacteria in the growth promoter is 2 x 1010 CFU/ml。
6. The method according to any of claims 2 to 4, characterized in that vitamin B in the growth promoter12The concentration was 0.05 g/l.
7. The method according to any of claims 2 to 4, wherein an inorganic phosphorus is added to the growth promoter.
8. The method of claim 7, wherein the inorganic material is selected from the group consisting ofPhosphorus is Ca3(PO4)2。
9. The method of claim 7, wherein the inorganic phosphorus is AlPO4。
10. The method of claim 7, wherein the inorganic phosphorus is FePO4。
CN202111091371.1A 2021-09-17 2021-09-17 By using phosphate solubilizing bacteria and vitamin B12Method for realizing rapid growth of poplar Pending CN113812298A (en)

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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1316513A (en) * 2000-04-03 2001-10-10 江农 Deeply fermented liquid culture medium for dephosphorizing bacteria
CN1331061A (en) * 2000-06-30 2002-01-16 李良发 Microbe fertilizer with nitrogen-fixating and P and K decomposing functions
JP2005110526A (en) * 2003-10-03 2005-04-28 Japan Science & Technology Agency Microorganism utilizing poorly soluble phytate in soil, phytase produced by the same and utilization thereof
CN101629147A (en) * 2009-06-30 2010-01-20 南京林业大学 Bacillus cereus and application thereof in promoting growth of poplars
CN101948361A (en) * 2010-09-29 2011-01-19 安茂生物科技有限公司 Special organic solid fertilizer for poplar, producing method and using method thereof
CN104371945A (en) * 2014-05-30 2015-02-25 西北大学 Citrobacter freundii with phosphorus-dissolving potassium-dissolving capability and application thereof
CN105062910A (en) * 2015-07-16 2015-11-18 南京林业大学 Poplar rhizosphere phosphate-solubilizing bacteria Pseudomonas frederiksbergensis JW-SD2 and application thereof

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1316513A (en) * 2000-04-03 2001-10-10 江农 Deeply fermented liquid culture medium for dephosphorizing bacteria
CN1331061A (en) * 2000-06-30 2002-01-16 李良发 Microbe fertilizer with nitrogen-fixating and P and K decomposing functions
JP2005110526A (en) * 2003-10-03 2005-04-28 Japan Science & Technology Agency Microorganism utilizing poorly soluble phytate in soil, phytase produced by the same and utilization thereof
CN101629147A (en) * 2009-06-30 2010-01-20 南京林业大学 Bacillus cereus and application thereof in promoting growth of poplars
CN101948361A (en) * 2010-09-29 2011-01-19 安茂生物科技有限公司 Special organic solid fertilizer for poplar, producing method and using method thereof
CN104371945A (en) * 2014-05-30 2015-02-25 西北大学 Citrobacter freundii with phosphorus-dissolving potassium-dissolving capability and application thereof
CN105062910A (en) * 2015-07-16 2015-11-18 南京林业大学 Poplar rhizosphere phosphate-solubilizing bacteria Pseudomonas frederiksbergensis JW-SD2 and application thereof

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Application publication date: 20211221