CN103361277A - Galactomyces geotrichum P14, application method thereof, and microbial inoculum prepared from same - Google Patents
Galactomyces geotrichum P14, application method thereof, and microbial inoculum prepared from same Download PDFInfo
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Abstract
The invention provides galactomyces geotrichum P14, an application method thereof, and a microbial inoculum prepared from the same, which belong to the technical field of micro-biofertilizer, and aim to realize high-efficiency phosphorus solubilization from the biological aspect by virtue of screened phosphorus solubilizing fungus. According to the microbial inoculum prepared from the galactomyces geotrichum P14, various insoluble phosphates in soil are converted to be in a water-soluble state with high efficiency, and then can be absorbed and utilized by plants. The microbial inoculum is suitable for reddened soil, moisture soil, brown soil and other types of soil, and has the effects of releasing insoluble phosphorus in the soil, preventing soil chemical fixation of phosphorus, improving the utilization efficiency of phosphorus fertilizer, improving the agronomic characters of crops, and increasing yield.
Description
Technical field
The invention belongs to soil microorganisms microbial inoculum field, relate to specifically the molten phosphorus geotrichum candidum of strain P14(Galactomyces geotrichum P14), contain preparation and the application of the phosphorus-solubilizing bacteria agent of this bacterium.
Background technology
At present, agriculture production increases day by day to the demand of phosphate fertilizer, and the area of the scarce phosphorus of China's soil accounts for 2/3 of total area under cultivation according to statistics.Main method is to rely on to use chemical phosphatic ferfilizer, but has caused the soil soil property to worsen, and fertility descends, and produces soil compaction, causes soil acidification, and objectionable impurities is to phenomenons such as soil pollutions.Especially " for the first time national Pollutant source investigation communique " result shows that agricultural has reached the historical high point to the destructiveness of physical environment, is mainly chemical fertilizer and agricultural chemicals etc. and spreads unchecked use.Want to solve the problem of agricultural pollution, improving energetically on the production efficiency basis of agricultural, also want simultaneously the consumption of large degree ground restriction chemical fertilizer and agricultural chemicals, these depend on the microbial bacteria fertilizer technology.
China's phosphate fertilizer resource relatively lacks, and the mode that satisfies agriculture production by a large amount of imports has improved production cost.Soil itself contains abundant phosphoric, but be that mineral phosphorus form exists more than 90%, it can not allow crop directly utilize, and use molten (solution) phosphobacterin material that can decompose indissoluble attitude phosphorus in the soil, making it form a phosphorus element supply area at crop rhizosphere, improve the supply of crop available phosphorus, is an important channel that effectively Soil Phosphorus is utilized, significant to our the present and following grain security, application prospect is very considerable simultaneously.
At first Staltrom and Sachett find soil molten (solution) phosphorus microorganism, and nineteen fifty Menkina isolates the bacillus megaterium of degrading organic phosphor and dissolving phosphoric acid DFP from soil.Jonson has studied the molten phosphorus effect of fungi Aspergillus niger the earliest.Simultaneously, scientist has carried out unremitting research to the ability of molten (solution) phosphorus activation of microorganism soil indissoluble phosphorus decomposing.Under the field condition, utilize
32The research of P isotopic dilution method finds that the phosphoric that absorbs with the wheat of visiing Lay Penicillium notatum (Penicillium bilaii) inoculation has 18% to come from the insoluble phosphorus ore, for the soil that not have to inoculate then plant can't utilize this phosphorus source.Its molten phosphorus characteristic is not subjected to the inhibition of soil available phosphorus simultaneously.Simultaneously, molten (solution) phosphorus microorganism has effect (Freitas etc., 1997 that promote plant growth and improve the crop economical character; Katiyar etc., 2003), promote in addition crop to inhale phosphorus, improve as output effect (Chabot, 1996; Peix etc., 2001), can improve output 15～18%(Wakelin etc., 2007 such as molten phosphorus Penicillium notatum inoculation wheat).The above results shows molten (solution), and the phosphorus bacterium has the huge applications potentiality for agricultural.
At first, USSR (Union of Soviet Socialist Republics) has been carried out the large-area applications (Cooper, 1959) of phosphorus decomposing bacillus megaterium, simultaneously other country also introduce for the production of.India has produced and has used the phosphorus decomposing microbial fertilizer since 20 century 70s, product has Phosphobacterin(Sharma and Singh, 1971), Microphos(Tomar, 1994), Phosphotika(Ahlawat and Rai, 1997) and Azophos(premalatha etc., 2004).Canadian Philom Bios company uses molten phosphorus Penicillium notatum to produce molten phosphorus microbial-bacterial fertilizer (trade(brand)name Jumpstart) on average to increase production 6～9%(Gleddie etc., 1991).At present application of China molten (solution) phosphorus bacterial strain maximum or eighties of last century screening 50～sixties (Ge Cheng, Wu Wei 1995; Jiang Ruibo, 2005).Commercially produce and fairly large use although molten (solution) phosphorus microbial fertilizer enters, really be used for the bacterial strain of agriculture production seldom.
According to incompletely statistics, the whole world filters out altogether 36 and belongs to, 92 kinds of molten (solution) phosphorus microorganisms, 60 kinds of bacterium wherein, 4 kinds of actinomycetes, 28 kinds of fungi.Molten (solution) phosphorus microbial resources abundant in the environment has formed contrast with at present less for the production of use.Therefore screening efficiently molten (solutions) phosphorus, strain excellent that adaptive capacity to environment is strong are still core, especially can significantly dissolve soil indissoluble phosphorus, improve efficiently molten (solution) phosphorus bacterial strain of the soil type of phosphate fertilizer utilising efficiency, adaptation different geographical.The dissolved phosphorus ability of the molten phosphorus geotrichum candidum P14 that the present invention relates to is significantly higher than visits Lay Penicillium notatum and phosphorus dissolvable penicillium oxalicum P 8 (Penicillium oxalicum P8).Molten phosphorus geotrichum candidum P14 bacterial strain can dissolve the multiple indissoluble phosphorus of soil such as tricalcium phosphate, octocalcium phosphate and phosphoric acid ten calcium, and it promotes crop to inhale the effect of phosphorus and growth.Stable under edaphic condition, also be not subjected to the impact of phosphate fertilizer.Simultaneously, it can strengthen the bioavailability of external source indissoluble phosphorus, reduces soil the chemistry of phosphorus is fixed.In the processes such as dissolving phosphoric acid DFP, octocalcium phosphate, it secretes organic acid, polypeptide and acid phosphatase, and the secretory volume height is high negative correlation with water-soluble phosphorus concentration.Molten phosphorus geotrichum candidum P14 dissolving effect of phosphorous is the common result who participates in of organic acid, polypeptide and acid phosphatase.Its many feature has superiority.
Summary of the invention
The purpose of this invention is to provide the molten phosphorus geotrichum candidum of strain P14(Galactomyces geotrichum P14) and use.
The purpose of this invention is to provide a kind of phosphorus-solubilizing bacteria agent that contains molten phosphorus geotrichum candidum P14.
The purpose of this invention is to provide the application of a kind of molten phosphorus geotrichum candidum P14.
The molten phosphorus geotrichum candidum of one strain P14(Galactomyces geotrichum P14), preserving number is: CGMCC No.7680.
Described molten phosphorus geotrichum candidum P14(Galactomyces geotrichum P14) application method is used for indissoluble attitude phosphorus is converted into soluble state phosphorus, perhaps for the preparation of the microbial inoculum that indissoluble attitude phosphorus is converted into soluble state phosphorus.
Described indissoluble attitude phosphorus comprises: Ca
3(PO
4)
2, Ca
8H
2(PO
4)
67H
2O, CaHPO
4, FePO
4, in bone meal and the Morocco's ground phosphate rock one or more.
A kind of phosphorus-solubilizing bacteria agent, described phosphorus-solubilizing bacteria agent comprises the component of following weight part:
1 part of molten phosphorus geotrichum candidum P14CGMCC No.7680 fermenting culture
Additive 8-16 part
Wherein, described additive is the peat composed of rotten mosses or diatomite, adsorb molten phosphorus geotrichum candidum P14 fermenting culture after, thalline content is not less than 10
7Cfu/g.
The preparation method of described phosphorus-solubilizing bacteria agent may further comprise the steps:
Molten phosphorus geotrichum candidum P14 inoculation on potato synthetic medium PDA, is cultivated 24h～28h for 26 ℃～29 ℃, then access the 500mL triangular flask, 26 ℃～29 ℃, liquid culture 36h～40h under 180rpm～200rpm; Then by volume 1%～2% inoculum size to be linked into liquid amount be in 60% the 50L seeding tank, at 180rpm～200rpm, pH7.0～pH7.4, under air flow 0.3vvm～0.5vvm, cultivate 24h～28h after, 5%～10% inoculum size is encased in the fermentor tank that liquid amount is 65% 1000L by volume again, at 180rpm～200rpm, pH7.0～pH7.4, under air flow 0.3vvm～0.5vvm, bacteria containing amount reaches 10 behind the cultivation 3d
9CFU/mL; After fermentation is finished, add the peat composed of rotten mosses or the diatomite of the good bacterium of death of monks or nuns according to the mass ratio of 1 ︰ 8～16, mixing is dried to moisture content 25～30%, and fineness is 60 orders～80 orders.
The composition of substratum in the fermentor tank:
Zulkovsky starch 10g/L; Semen Maydis powder 5g/L; Yeast powder 5g/L; Sucrose 10g/L; (NH
4)
2SO
40.5g/L; KCl0.3g/L; NaCl0.3g/L; KH
2PO
40.1g/L; MgSO
47H
2O0.3g/L; FeSO
40.03g/L; MnSO
44H
2O0.03g/L; Add water and be settled to 1L; PH7.0～7.4.
The triangular flask liquid nutrient medium forms: peeled potatoes 150g/L, Semen Maydis powder 10g/L, glucose 20g/L, KH
2PO
40.5g/L, MgSO
47H
2O0.5g/L; Add water and be settled to 1L; The pH nature.
Substratum forms in the seeding tank: yeast powder 0.5g/L; Glucose 10.0g/L; KH
2PO
41.0g/L; (NH
4)
2SO
40.5g/L; NaCl0.2g/L; MgSO
47H
2O0.03g/L; MnSO
40.03g/L; FeSO
40.003g/L; Add water and be settled to 1L; PH7.0～7.4.
The molten phosphorus geotrichum candidum of a strain of the present invention P14 is to separate from the red soil phosphorus ore soil property of Changsha, Hunan Liuyang Phosphate Mine periphery in 2010.
Molten phosphorus geotrichum candidum P14 provided by the present invention, it has following Microbiological Characteristics:
1, molten phosphorus geotrichum candidum P14 bacterium colony on PDA 28 ℃ the growth 3 days, diameter 20-30mm, thread, produce a large amount of conidium structures, the edge is bordering on white, middle little yellow, without transudate, reverse side is faint yellow.
2, conidium is cylindrical, common 3.5～5.4 μ m * 2.2～3.0 μ m.
3, the physio-biochemical characteristics of molten phosphorus geotrichum candidum P14 are as follows:
The physiology and morphology biochemical characteristic of table 1 P14 bacterial strain
4, the homology feature of molten phosphorus geotrichum candidum P14 is seen Fig. 1.
Molten phosphorus geotrichum candidum P14 of the present invention has the effect that improves soil dissolved phosphorus content, especially has multiple indissoluble effect of phosphorous in the dissolving soil, can prevent that the soil chemistry of phosphorus from fixing, and improves the function of phosphate fertilizer utilization efficiency.Be fit to the multiple soil types such as red soil, cinnamon soil, moisture soil.
Molten phosphorus geotrichum candidum P14 of the present invention is under the liquid culture condition, its molten (solution) phosphorus mechanism research is drawn: the content of its secretion organic acid, polypeptide, acid phosphatase is subjected to the remarkably influenced of water-soluble phosphorus concentration in the substratum, and the secretory volume height is high negative correlation with water-soluble available phosphate concentration.Therefore these secretory product and molten phosphorus are closely related.
Molten phosphorus geotrichum candidum P14 of the present invention and molten (solution) phosphobacterin thereof have following effect:
1., molten phosphorus geotrichum candidum P14 of the present invention, can dissolve soil indissoluble phosphorus.The stronger dissolving Ca of molten phosphorus geotrichum candidum P14 performance on solid medium
3(PO
4)
2, Ca
8H
2(PO
4)
67H
2O, CaHPO
4, FePO
4, bone meal and Morocco's ground phosphate rock (MPR) ability.
2., inoculate the available phosphorus content that molten phosphorus geotrichum candidum P14 can significantly increase soil, minimizing Phosphorus Fertilizer Rates, raising phosphate fertilizer utilization efficiency.
3., the molten phosphorus geotrichum candidum of soil inoculation P14 can significantly promote crop root to grow, improve the biomass of crop.
The molten phosphorus geotrichum candidum of the present invention P14(Galactomyces geotrichum P14) deposit number is CGMCC No.7680; June 4 2013 preservation time, address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, China Committee for Culture Collection of Microorganisms common micro-organisms center.
Description of drawings
Fig. 1 is molten phosphorus geotrichum candidum P14 homology feature;
Fig. 2 is molten phosphorus geotrichum candidum P14 strain morphology.
Embodiment
Be intended to further specify the present invention below in conjunction with embodiment, and unrestricted the present invention.
The preparation of embodiment 1 molten phosphorus geotrichum candidum bacterial strain P14 and phosphorus-solubilizing bacteria agent thereof
1, the composition of fermentation tank culture medium (g/L)
Zulkovsky starch 10g/L; Semen Maydis powder 5g/L; Yeast powder 5g/L; Sucrose 10g/L; (NH
4)
2SO
40.5g/L; KCl0.3g/L; NaCl0.3g/L; KH
2PO
40.1g/L; MgSO
47H
2O0.3g/L; FeSO
40.03g/L; MnSO
44H
2O0.03g/L; Add water and be settled to 1L; PH7.0～7.4.
The triangular flask liquid nutrient medium forms: peeled potatoes 150g/L, Semen Maydis powder 10g/L, glucose 20g/L, KH
2PO
40.5g/L, MgSO
47H
2O0.5g/L; Add water and be settled to 1L; The pH nature.
Substratum forms in the seeding tank: yeast powder 0.5g/L; Glucose 10.0g/L; KH
2PO
41.0g/L; (NH
4)
2SO
40.5g/L; NaCl0.2g/L; MgSO
47H
2O0.03g/L; MnSO
40.03g/L; FeSO
40.003g/L; Add water and be settled to 1L; PH7.0～7.4.
2, culturing process
Molten phosphorus geotrichum candidum P14 inoculation on potato synthetic medium (PDA), is cultivated 24h～28h for 26 ℃～29 ℃, then access the 500mL triangular flask, 26 ℃～29 ℃, liquid culture 36h～40h under 180rpm～200rpm.Then by volume 1%～2% inoculum size to be linked into liquid amount be in 60% the 50L seeding tank, at 180rpm～200rpm, pH7.0～pH7.4, under air flow 0.3vvm～0.5vvm, cultivate 24h～28h after, 5%～10% inoculum size is encased in the fermentor tank that liquid amount is 65% 1000L by volume again, at 180rpm～200rpm, pH7.0～pH7.4, under air flow 0.3vvm～0.5vvm, bacteria containing amount reaches 10 behind the cultivation 3d
9CFU/mL.After fermentation is finished, add the peat composed of rotten mosses or the diatomite of the good bacterium of death of monks or nuns according to the ratio of 1 ︰ 8～16, mixing is dried to moisture content 25～30%, and fineness is 60 orders～80 orders.Detect packing.
The molten phosphorus ability of embodiment 2 molten phosphorus geotrichum candidum P14
1, the effect of solubilizing phosphate under the solid culture condition
At selective medium---(substratum forms (g/L) to improvement Pikovskaya substratum: (NH
4)
2SO
40.5 NaCl 0.2, MgSO
47H
2O 0.03, and KCl 0.2, MnSO
40.03, FeSO
40.003, yeast powder 0.5, glucose 10.0, phosphorus source 5.0)---on, the pedotheque that will gather in the red soil phosphorus ore soil property of Changsha Liuyang Phosphate Mine periphery carries out stepwise dilution, with 10 of the soil solution
-4Spread plate has carried out the separation of molten (solution) phosphorus microorganism.Big or small initial gross separation according to molten phosphorus circle goes out phosphate-solubilizing bacteria and fungi 30 strains.Then, take tricalcium phosphate on the selective medium in unique phosphorus source, further compared the molten phosphorus ability of these molten (solution) phosphorus bacterial strains, the molten phosphorus ability of fungi obtains the stronger fungi P14 of the molten phosphorus ability of a strain at last greater than bacterium.In conjunction with bacterial strain P14 form, physiological and biochemical index and 18S rDNA sequential analysis, identify that it belongs to geotrichum candidum and belongs to (Galactomyces geotrichum), the molten phosphorus geotrichum candidum of called after P14.Itself and phosphorus dissolvable penicillium oxalicum P 8, visit Lay Penicillium notatum ATCC20851 and bacillus megaterium ATCC14581 molten phosphorus circle and colony growth diameter on the different phosphate sources nutrient agar and compare (table 2).The result shows that molten phosphorus geotrichum candidum P14 and the molten phosphorus characteristic of molten phosphorus penicillium oxalicum are better than visiing Lay Penicillium notatum and bacillus megaterium.
The molten phosphorus geotrichum candidum of table 2 is colony growth and molten phosphorus loop diameter (mm) on the different phosphate sources nutrient agar
Annotate: "-" represents without molten phosphorus circle
On the Pikovskaya improvement solid selective medium take difficultly-soluble phosphates as unique phosphorus source, cultivate after 3 days for 28 ℃, bacterial strain P14 and control strain phosphorus dissolvable penicillium oxalicum P 8, visit Lay Penicillium notatum ATCC20851 and bacillus megaterium ATCC14581, to bone meal, FePO
4, CaHPO
4, Ca
3(PO
4)
2, octocalcium phosphate, Morocco's ground phosphate rock (MPR) molten phosphorus ability compare, P14 shows stronger dissolving bone meal, FePO
4, CaHPO
4, Ca
3(PO
4)
2, octocalcium phosphate, Morocco's ground phosphate rock ability (table 3).Molten phosphorus ability is significantly higher than the control strain phosphorus dissolvable penicillium oxalicum P 8, visits Lay Penicillium notatum and bacillus megaterium.The molten phosphorus kind of bacterial strain P14 of the present invention is many, efficient is high, adaptable, have application potential.
The amount of phosphorus dissolved (P:mg/20mL) of the molten phosphorus geotrichum candidum plate culture medium 3d of table 3 inoculation
Annotate: "-" expression does not detect available phosphorus
2, the effect of solubilizing phosphate under the liquid culture condition
(substratum forms (g/L): (NH at the liquid nutrient medium take difficultly-soluble phosphates as unique phosphorus source
4)
2SO
40.5 NaCl 0.2, MgSO
47H
2O 0.03, and KCl 0.2, MnSO
40.03, FeSO
40.003, yeast powder 0.5, glucose 10.0, Morocco's ground phosphate rock (≤80 order) 5.0) in, 28 ℃ of contrasts of cultivating molten phosphorus geotrichum candidum P14, phosphorus dissolvable penicillium oxalicum P 8 and Bai Lai Penicillium notatum and not connecing bacterium, 180rpm, molten phosphorus ability measurement result is as shown in table 4.
Different phosphorus-solubilizing bacteria 3d conversion MPR are the quantity of water-soluble phosphorus under the table 4 liquid culture condition
Molten phosphorus geotrichum candidum P14, molten phosphorus penicillium oxalicum and Bai Lai Penicillium notatum have obvious effect of solubilizing phosphate, significant difference between bacterial strain (P＜0.05), ammonium nitrogen is for seasonable, molten phosphorus geotrichum candidum P14 is through the cultivation of 3d, water-soluble phosphorus in the 50mL substratum is 10.23mg, more molten phosphorus penicillium oxalicum and Bai Lai Penicillium notatum improve respectively 0.99mg and 5.09mg, and gap is remarkable.Nitric nitrogen is for seasonable, and the amount of phosphorus dissolved of molten phosphorus geotrichum candidum P14 is higher than ammonium nitrogen.Water-soluble phosphorus in the 50mL substratum is 12.11mg, accounts for 48.1% of total phosphorus in the ground phosphate rock, and more molten phosphorus penicillium oxalicum and Bai Lai Penicillium notatum improve respectively 2.94mg and 4.27mg.Under nitric nitrogen and ammonium nitrogen condition, the amount of phosphorus dissolved of molten phosphorus geotrichum candidum P14 all is significantly higher than other group.
Embodiment 3 molten phosphorus geotrichum candidum P14 are to the utilization of external source indissoluble phosphorus
Under condition of pot, adopt light loamy soil, corn is for supplying to study thing, studied molten phosphorus geotrichum candidum P14 to the sex effect of biologically effective of external source indissoluble phosphorus, test comprises tricalcium phosphate, octocalcium phosphate, tertiary iron phosphate, aluminum phosphate, bone meal, fluorapatite, Morocco's Rock Phosphate (72Min BPL) as supplying examination phosphorus source with indissoluble phosphorus, and effect of solubilizing phosphate is as shown in table 5.
Table 5 different phosphate sources is processed soil available phosphorus over time
Under this test conditions, the result shows that molten phosphorus geotrichum candidum P14 is to Ca
3(PO
4)
2, AlPO
4, MPR activation capacity stronger, to FePO
4Activation a little less than, having showed molten phosphorus geotrichum candidum P14 has certain selectivity to indissoluble phosphorus kind.Molten phosphorus geotrichum candidum P14 effect is lower, and it is the fastest that tricalcium phosphate discharges phosphoric, and bone meal, aluminum phosphate take second place.Almost do not show the effect that increases biological yield when not inoculating molten phosphorus geotrichum candidum, therefore molten phosphorus geotrichum candidum P14 has significance to the biological effectiveness that strengthens indissoluble phosphorus.
The effect of solubilizing phosphate of molten phosphorus geotrichum candidum P14 is mainly manifested between 10～60d, surpasses the 60d effect of solubilizing phosphate and descends.
Embodiment 4 molten phosphorus geotrichum candidum P14 are to the effect of the molten phosphorus of different soils
(No. 1 soil is light loamy soil, available phosphorus 13.5 μ g/g at three kinds of different soils environment; No. 2 soil are middle loamy texture moisture soil, available phosphorus 4.7 μ g/g; No. 3 soil are that middle loamy texture takes off moisture soil, available phosphorus 5.2 μ g/g) in, behind the molten phosphorus geotrichum candidum P14 microbial inoculum 180d of inoculation the present invention preparation, its available phosphorus significantly improves, and has the efficient phosphorus-dissolution effect.The result is as shown in table 6.
The different soils available phosphorus of the molten phosphorus geotrichum candidum P14 microbial inoculum of table 6 inoculation is (μ g/g) over time
The components such as the octocalcium phosphate of molten phosphorus geotrichum candidum P14 microbial inoculum in can activating soil, phosphoric acid ten calcium, aluminum phosphate.Take No. 1 soil as example, the front 30d of the soil available phosphorus content of interpolation microbial inoculum advances the speed the fastest, rises to 65.3 μ g/g from 13.5 μ g/g, and then speed slows down, and reaches steady state behind 60d.This moment, the available phosphorus content of soil was saturated, and plant constantly absorbs and can promote this bacterial strain to continue degraded indissoluble phosphorus, and lasting phosphorus source is provided.And its available phosphorus content of soil that does not add microbial inoculum does not have change basically.No. 2 and No. 3 soil also are analogues.Under different soil, the available phosphorus content that adds the microbial inoculum group all is significantly higher than un-added.Therefore this microbial inoculum is fit to polytype soil.
Embodiment 5 molten phosphorus geotrichum candidum P14 are to the crop yield effect
By the culturing process of embodiment 1, make microbial inoculum, under condition of pot, maize planting, peanut, (amount of application is 7.5kg/ mu, with P with microbial inoculum (amount of application 1g/kg soil) and inorganic phosphorus fertilizer
2O
5Meter) process, the result is as shown in table 7.The result shows that inorganic phosphorus fertilizer, inorganic phosphorus fertilizer+deactivation microbial inoculum are lower to increasing the biomass effect, and inorganic phosphorus fertilizer+microbial inoculum treatment group is better than above-mentioned two groups.This explanation live bacteria agent P14 competence exertion production-increasing function.But Different Crop is slightly variant, microbial inoculum on corn effect greater than peanut.
The molten phosphorus geotrichum candidum of table 7 P14 is on the impact of corn, peanut biomass (mg/ basin)
| Crop | Process | Available phosphorus (μ g/g) | Biomass (kg/ basin) |
| ? | P 2O 5 | 2.33 | 1.98 |
| Corn | P 2O5+ deactivation P14 | 2.45 | 2.04 |
| ? | P 2O 5+P14 | 3.92 | 2.36 |
| ? | P 2O 5 | 2.42 | 1.84 |
| Peanut | P 2O 5+ deactivation P14 | 2.37 | 2.01 |
| ? | P 2O 5+P14 | 3.84 | 2.24 |
Claims (9)
1. the molten phosphorus geotrichum candidum of strain P14(Galactomyces geotrichum P14), it is characterized in that preserving number is: CGMCC No.7680.
2. application method molten phosphorus geotrichum candidum P14(Galactomyces geotrichum P14 claimed in claim 1) is characterized in that, is used for indissoluble attitude phosphorus is converted into soluble state phosphorus.
3. application method molten phosphorus geotrichum candidum P14(Galactomyces geotrichum P14 according to claim 2) is characterized in that, for the preparation of the microbial inoculum that indissoluble attitude phosphorus is converted into soluble state phosphorus.
4. according to claim 2 or 3 described application methodes, it is characterized in that described indissoluble attitude phosphorus comprises: Ca
3(PO
4)
2, Ca
8H
2(PO
4)
67H
2O, CaHPO
4, FePO
4, in bone meal and the Morocco's ground phosphate rock one or more.
5. a phosphorus-solubilizing bacteria agent is characterized in that, described phosphorus-solubilizing bacteria agent comprises the component of following weight part:
1 part of molten phosphorus geotrichum candidum P14CGMCC No.7680 fermenting culture
Additive 8-16 part
Wherein, described additive is the peat composed of rotten mosses or diatomite, adsorb molten phosphorus geotrichum candidum P14 fermenting culture after, thalline content is not less than 10
7Cfu/g.
6. the preparation method of phosphorus-solubilizing bacteria agent claimed in claim 5 is characterized in that, may further comprise the steps:
Molten phosphorus geotrichum candidum P14 inoculation on potato synthetic medium PDA, is cultivated 24h～28h for 26 ℃～29 ℃, then access the 500mL triangular flask, 26 ℃～29 ℃, liquid culture 36h～40h under 180rpm～200rpm; Then by volume 1%～2% inoculum size to be linked into liquid amount be in 60% the 50L seeding tank, at 180rpm～200rpm, pH7.0～pH7.4, under air flow 0.3vvm～0.5vvm, cultivate 24h～28h after, 5%～10% inoculum size is encased in the fermentor tank that liquid amount is 65% 1000L by volume again, at 180rpm～200rpm, pH7.0～pH7.4, under air flow 0.3vvm～0.5vvm, bacteria containing amount reaches 10 behind the cultivation 3d
9CFU/mL; After fermentation is finished, add the peat composed of rotten mosses or the diatomite of the good bacterium of death of monks or nuns according to the mass ratio of 1 ︰ 8～16, mixing is dried to moisture content 25～30%, and fineness is 60 orders～80 orders.
7. preparation method according to claim 6 is characterized in that, the composition of substratum in the fermentor tank: Zulkovsky starch 10g/L; Semen Maydis powder 5g/L; Yeast powder 5g/L; Sucrose 10g/L; (NH
4)
2SO
40.5g/L; KCl0.3g/L; NaCl0.3g/L; KH
2PO
40.1g/L; MgSO
47H
2O0.3g/L; FeSO
40.03g/L; MnSO
44H
2O0.03g/L; Add water and be settled to 1L; PH7.0～7.4.
8. preparation method according to claim 6 is characterized in that, the triangular flask liquid nutrient medium forms: peeled potatoes 150g/L, Semen Maydis powder 10g/L, glucose 20g/L, KH
2PO
40.5g/L, MgSO
47H
2O0.5g/L; Add water and be settled to 1L; The pH nature.
9. preparation method according to claim 6 is characterized in that, substratum forms in the seeding tank: yeast powder 0.5g/L; Glucose 10.0g/L; KH
2PO
41.0g/L; (NH
4)
2SO
40.5g/L; NaCl0.2g/L; MgSO
47H
2O0.03g/L; MnSO
40.03g/L; FeSO
40.003g/L; Add water and be settled to 1L; PH7.0～7.4.
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