CN101195543B - Phytase microbial inoculum fertilizer, producing method and application thereof - Google Patents
Phytase microbial inoculum fertilizer, producing method and application thereof Download PDFInfo
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Abstract
The invention relates to a phytase microbial inoculum fertilizer and method for preparation and application, which comprises phytase strain and fermentation culture medium, wherein viable bacterial quantity in the microbial inoculum is not less than 0.1 billion /ml. The phytase microbial inoculum of the invention transfers phosphor phytate which is not adsorbed by plant in the soil or domestic fowl fecal into inorganic phosphorous nourishment which can be adsorbed by plant. The phytase microbial inoculum fertilizer can accelerate adsorption of N, P and K and growth and development of plant. The soil structure can be modified and the soil activity is adjusted through metabolic movement of microbe, further the soil property is improved and the soil fertility is increased. The microbial inoculum of the invention is high in active bacterial number, long in retention period, simple in production technique, easy to realize automatic production and the like, which is especially suitable for fertilization of economic plants such as vegetable, fruit tree, wheat, corn and the like.
Description
Technical field:
The invention belongs to ecological agriculture technical field, relate to microbial preparation, particularly a kind of phytase microbial inoculum fertilizer and production method and employing phytase microbial inoculum are used as bio-feritlizer.
Background technology:
China's agriculture production is extensive use of chemical fertilizer.China is the country that uses chemical fertilizer, agricultural chemicals quantity maximum in the world.Extensively not only caused soil pollution to fertilize, also increased the weight of water body organic contamination and eutrophication pollution, even had influence on underground water and air by agricultural run-off.The superelevation amount is used the Soil structure that chemical fertilizer also destroys the arable land.Reduce the quality of agricultural-food, the disease and pest incidence improves.Simultaneously, the exhaustion of mineral resource has been quickened in the extraordinary development of chemical fertilizers production.Day by day become a big problem that threatens sustainable development of agricultural production.
Correct guiding peasant rationally adjusts Fertilization Structure, the obvious free of contamination new-type fertilizer of development and popularization fertilizer efficiency, to realize the agriculture production increasing both production and income, to improve quality of agricultural product, recovery and raising soil Sustainable Production ability, is current agriculture production developing direction.
Phytase (myo-Inositol-hexaphosphate-phosphohyolrolase) can make phytic acid be degraded into inositol and phosphoric acid.Liu Lilis etc. have been reported screening study { Tianjin Normal University's journal, 2006,26 (2) 19-22} of phytase secretion bacterial strain.Chinese patent CN1552872A discloses the clone and the expression of a kind of heat-stable phytase and gene thereof.Liu Lili, Yang Wenbo etc. have reported that the phytase gene engineering bacterial strain can be made into microbiobacterial agent.{ clone and the expression of bacterium heat-stable phytase gene, microbiology circular, 2004,36 (4): 71-76}.But about phytase microbial inoculum fertilizer and production method and adopt phytase microbial inoculum to increase soil fertility, promote the application of plant-growth, do not see bibliographical information as yet.
Summary of the invention:
An object of the present invention is to provide a kind of phytase microbial inoculum fertilizer.
Another object of the present invention provides a kind of production method of phytase microbial inoculum fertilizer.
A further object of the present invention provides phytase microbial inoculum and is applied to increase soil fertility, promotes plant-growth and improves the method for crop yield.
Technology contents of the present invention is as follows:
A kind of phytase microbial inoculum fertilizer is characterized in that it is made up of phytase bacterial strain and fermention medium, viable count in the microbial inoculum 〉=100,000,000/mL.
Phytase microbial inoculum fertilizer of the present invention, phytase bacterial strain wherein (bacillus sp.) is that (phytase gene engineering strain SDLTP02 derives from the Ph D dissertation database to the phytase gene engineering strain: Liu Lili, 2000 grades, the microbiology specialty, resource bacterium and engineering research direction, thesis topic: the structure of dynamic (dynamical) research of the molten phosphorus of phosphorus bacteria and enzymolysis phytic acid genetic engineering bacterium, in April, 2004).
Each composition proportion of the fermention medium of phytase microbial inoculum fertilizer of the present invention is: bran wheat bran 20-40%, glucose 5-20%, (NH
4)
2HPO
45-20%, K
2HPO
40.1-1%, MgSO
47H
2O 0.1-1%, NaCl 2-10%, all the other are water.
Each composition proportion of preferred fermention medium is: wheat bran 35g, glucose 2g, (NH
4)
2HPO
410g, K
2HPO
40.2g, MgSO
47H
2O 0.1g, NaCl 2g, water 100mL.
The production method of phytase microbial inoculum fertilizer of the present invention is characterized in that the step that comprises:
(1) get phytase gene engineering strain SDLTP02 slant strains 2-4 articulating and go in the meat soup substratum, at 35-45 ℃, 24-36h is cultivated in concussion, obtains seed liquor;
(2) transfer in fermention medium with the inoculum size of 5-20% seed liquor, 35-45 ℃, cultivate 24-48h, make microbial inoculum, cryopreservation, standby.
Being prepared as of broth culture of the present invention:
Take by weighing peptone 5g, extractum carnis 3g, NaCl 5g and MnSO
4H
2O 5mg uses dissolved in distilled water, is settled to 1L, pH7.0-7.2.
The preparation method of fermention medium of the present invention is: take by weighing wheat bran 20-40%, glucose 5-20%, (NH
4)
2HPO
45-20%, K
2HPO
40.1-1%, MgSO
47H
2O 0.1-1%, NaCl 2-10%, all the other are water, pH7.0-7.2.
Preferred manufacturing procedure is: take by weighing wheat bran 35g, glucose 2g, (NH
4)
2HPO
410g, K
2HPO
40.2g, MgSO
47H
2O 0.1g, NaCl 2g, water 100mL.pH7.0-7.2。
The present invention further provides the application of phytase microbial inoculum fertilizer aspect the raising plant photosynthetic rate.
The present invention further provides the application of phytase microbial inoculum fertilizer aspect the promotion plant-growth.
The present invention also provides simultaneously phytase microbial inoculum fertilizer increasing plant to the application aspect nitrogen, phosphorus, the potassium absorption.
The present invention also describes the application of phytase microbial inoculum fertilizer aspect the raising plant biomass simultaneously in detail.
Plant of the present invention is: vegetables, fruit tree, wheat or corn.
Application method of the present invention is: the dry blending soil of packing in container and sieving, the sowing plant strain, regularly quantitatively water, then phytase microbial inoculum is imposed on 2～3cm place under the seed, the amount of application of phytase liquid bacterial agent: 50～200mL/kg, results after 3 months, determination test result respectively.
Experimental result confirms: the phytase microbial inoculum of using doses can not be by the phytate phosphorus of plant absorbing with the major part in the soil, and being converted into can be by the inorganic phosphorus of plant absorbing.Particularly phytase microbial inoculum can be applicable to economic plants such as wheat, corn, clover.After using microbial inoculum, the photosynthesis rate of stem and leaf of Wheat and corn is obviously strengthened; Improved the dry weight of stem and leaf of Wheat and maize seedling; Promoted the root activity of corn; Promoted the absorption of plant to nitrogen, phosphorus, potassium, the nitrogen of stem and leaf of Wheat and maize seedling, phosphorus, potassium content obviously increase; Improved the output of clover.
The present invention adopts the phytase gene engineering bacterial strain SDLTPO2 of Microbiological Lab of Tianjin Normal University preservation, and (phytase gene engineering strain SDLTP02 derives from the Ph D dissertation database: Liu Lili, 2000 grades, the microbiology specialty, resource bacterium and engineering research direction, thesis topic: the structure of dynamic (dynamical) research of the molten phosphorus of phosphorus bacteria and enzymolysis phytic acid genetic engineering bacterium, in April, 2004), relevant patent of invention (clone of heat-stable phytase gene and expression, the patent No.: CN1552872A) disclose the clone and the expression of a kind of heat-stable phytase and gene thereof.
Phytase microbial inoculum of the present invention is compared the positively effect that is had with the disclosed various microbial inoculums of prior art and is:
Phytase microbial inoculum of the present invention can not be by the phytate phosphorus of plant absorbing with the major part in the soil, and being converted into can be by the inorganic phosphorus of plant absorbing.Simultaneously can also promote the absorption of plant to nitrogen, phosphorus, potassium.The microbial growth Metabolic activity can improve the activity of soil.By the potential of hydrogen of fine setting soil, further improved the performance of soil, prolonged fertilizer efficiency.Particularly phytase microbial inoculum can be applicable to economic plants such as wheat, corn.
Phytase base microbial inoculum itself of the present invention does not contain environmentally harmful material, and cost of manufacture is low, is that a kind of to have fertilizer efficiency good, can improve the microbial fertilizer of soil activation, can create very large economic benefit.
Description of drawings
Fig. 1 phytase microbial inoculum is to the influence of stem and leaf of Wheat photosynthetic rate.
Fig. 2 phytase microbial inoculum is to the influence of stem and leaf of Wheat over-ground part dry weight.
Fig. 3 phytase microbial inoculum is to the influence of the full nitrogen of stem and leaf of Wheat over-ground part, phosphorus, potassium content.
Fig. 4 phytase microbial inoculum is to the influence of maize seedling photosynthetic rate.
Fig. 5 phytase microbial inoculum is to the influence of maize seedling over-ground part dry weight.
Fig. 6 phytase microbial inoculum is to the influence of the full nitrogen of maize seedling over-ground part, phosphorus, potassium content.
Embodiment:
The present invention will be further described below in conjunction with embodiment.These embodiment describe typical case of the present invention, but the present invention is not limited to this.What need be illustrated is:
(phytase gene engineering strain SDLTP02 derives from the Ph D dissertation database to phytase gene engineering strain SDLTP02 of the present invention: Liu Lili, 2000 grades, the microbiology specialty, resource bacterium and engineering research direction, thesis topic: the structure of dynamic (dynamical) research of the molten phosphorus of phosphorus bacteria and enzymolysis phytic acid genetic engineering bacterium, in April, 2004) by the preservation of Microbiological Lab of Tianjin Normal University.The public can make by oneself with reference to above-mentioned document.
Embodiment 1:
Get slant strains 3 articulatings and go in seed (meat soup) substratum of 100ml, 37 ℃, 24h is cultivated in concussion.The inoculum size of the seed liquor (20%, volume) of 20ml is transferred in the fermention medium of 100ml, 37 ℃, cultivate 48h, make microbial inoculum.Viable count in the microbial inoculum 〉=100,000,000/mL are preserved standby down for 4 ℃.
The wherein preparation of seed (meat soup) substratum: take by weighing peptone 5g, extractum carnis 3g, NaCl 5g and MnSO
4H
2O 5mg uses dissolved in distilled water, is settled to 1L, pH7.0-7.2.
The preparation of fermention medium: take by weighing wheat bran 35g, glucose 2g, (NH
4)
2HPO
410g, K
2HPO
40.2g, MgSO
47H
2O 0.1g, NaCl 2g, water 100mL, pH7.0-7.2.
The method that the viable count measuring method employing Shen Ping of microbial inoculum etc. provides (Shen Ping, Fan Xiurong, Li Guangwu. microbiology experiment (third edition). Beijing: Higher Education Publishing House, 2003).Measurement result: 1.5 hundred million/mL of viable count in the microbial inoculum.
Embodiment 2:
Get slant strains 4 articulatings and go in seed (meat soup) substratum of 100ml, 40 ℃, 36h is cultivated in concussion.The inoculum size of the seed liquor (15%, volume) of 15ml is transferred in the fermention medium of 100ml, 40 ℃, cultivate 24h, make microbial inoculum.Viable count in the microbial inoculum 〉=100,000,000/mL are preserved standby down for 4 ℃.
The wherein preparation of seed (meat soup) substratum: take by weighing peptone 5g, extractum carnis 3g, NaCl 5g and MgSO
47H
2O 5mg uses dissolved in distilled water, is settled to 1L, pH7.0-7.2.
The preparation of fermention medium: take by weighing wheat bran 40g, glucose 5g, (NH
4)
2HPO
415g, K
2HPO
40.4g, MgS047H20 0.3g and NaCl 8g, water 100mL, pH7.0-7.2.
The method that the viable count measuring method employing Shen Ping of microbial inoculum etc. provides (Shen Ping, Fan Xiurong, Li Guangwu. microbiology experiment (third edition). Beijing: Higher Education Publishing House, 2003).1.5 hundred million/mL of viable count in the microbial inoculum.
Embodiment 3
The phytase microbial inoculum that obtains with embodiment 1 carries out basin and carries the wheat test, carries out in Tianjin Normal University 1 day～June 1 March in 2006.Test in sowing on March 1, June 1 gathered in the crops.Test is 28.2 * 30cm with the basin specification.Every basin was adorned 3.0mm and was sieved dry blending soil 12.5kg, sowed 20, field planting 15 strains.Regularly quantitatively water each experimental group carrying out horizontal strict conformance.Phytase microbial inoculum imposes on 2～3cm place under the seed.
5 experimental group are established in test, and each experimental group is provided with 3 repetitions.Each experimental group is respectively:
Blank group (CK)
Phytase microbial inoculum (liquid) amount of application: 50mL, 100mL, 150mL, 200mL
(1) phytase microbial inoculum is to the influence of wheat photosynthetic rate
Respectively on April 1st, 2006 10:00a.m. and June 1 10:00a.m. measure the stem and leaf of Wheat photosynthetic rate.Condition determination is light intensity 700 μ molm
-2S
-1, 25 ℃ of room temperatures.Measurement result (measuring method is with reference to the photosynthetic instrument instrument of Li-6400 working instructions) is seen shown in Figure 1.Fig. 1 phytase microbial inoculum is to the influence of stem and leaf of Wheat photosynthetic rate.
As can be seen from Figure 1, use microbial inoculum after, the stem and leaf of Wheat photosynthesis rate was obviously strengthened in 2 months.And with the increase of microbial inoculum amount of application, the corresponding raising of stem and leaf of Wheat photosynthetic rate, and be significantly higher than control group.
(2) phytase microbial inoculum is to the influence of stem and leaf of Wheat over-ground part dry weight
The stem and leaf of Wheat dry weight on June 1st, 2006 measure (measuring method is seen: Zhang Zhiliang, Zhai Weijing. plant physiology experiment instructs (the 3rd edition), Higher Education Publishing House, the 3rd printing of Beijing: 2004-3 .).Phytase microbial inoculum is seen Fig. 2 and table 1 to the influence of stem and leaf of Wheat over-ground part dry weight, and Fig. 2 phytase microbial inoculum is to the influence of stem and leaf of Wheat over-ground part dry weight.
Table 1 phytase microbial inoculum is to the influence of stem and leaf of Wheat over-ground part dry weight
| Treatment capacity (mL) | 0-(CK) | 50 | 100 | 150 | 200 |
| Dried wheat seedling is resurveyed and is decided result (g) | 0.31 cC | 0.44 bAB | 0.50 bB | 0.53 aA | 0.37 cC |
Analyze from Fig. 2 and table 1, when using microbial inoculum 150mL, stem and leaf of Wheat over-ground part dry weight has extremely significantly (0.01 level) difference in control group.
(3) phytase microbial inoculum to the influence of the full nitrogen of stem and leaf of Wheat, full phosphorus, full potassium content (measuring method is seen: Peng Jianwei, Song Haixing. plant science experimental center, Plant Nutrition experimental technique: the Plant Nutrition experiment instruction).
Table 2 phytase microbial inoculum is to the influence of stem and leaf of Wheat over-ground part total nitrogen content
| Microbial inoculum amount of application (mL) | 0(CK) | 50 | 100 | 150 | 200 |
| The full nitrogen determination result of wheat seeding (% dry weight) | 2.12 bB | 1.60 cC | 2.00 bB | 1.70 cC | 2.96 aA |
Table 3 phytase microbial inoculum is to the influence of stem and leaf of Wheat over-ground part content of tatal phosphorus
| Microbial inoculum amount of application (mL) | 0(CK) | 50 | 100 | 150 | 200 |
| The full phosphorus measurement result of wheat seeding (% dry weight) | 0.51 cC | 0.54 bcBC | 0.57 bAB | 0.62 aA | 0.55 bBC |
Table 4 phytase microbial inoculum is to the influence of the full potassium content of stem and leaf of Wheat over-ground part
| Microbial inoculum amount of application (mL) | 0(CK) | 50 | 100 | 150 | 200 |
| The full potassium measurement result of wheat seeding (% dry weight) | 5.11 bB | 4.79 bcBC | 5.00 bB | 4.48 cC | 5.88 aA |
Fig. 3, table 2 shows that using microbial inoculum has improved the stem and leaf of Wheat total nitrogen content, and with the increase of handling level, total nitrogen content increases.Analyze according to Fig. 3 table 3, use microbial inoculum 100mL, 150mL respectively, the stem and leaf of Wheat content of tatal phosphorus is higher than contrast, and utmost point significant difference is arranged.Above-mentioned stem and leaf of Wheat phosphorus cellulose content increases, and the increase that this and stem and leaf of Wheat absorb the plain amount of phosphorus has certain relation.By Fig. 3 table 4 as can be seen, use microbial inoculum 200mL, the full potassium content of stem and leaf of Wheat is higher than contrast, and utmost point significant difference is arranged.
Embodiment 4
The phytase microbial inoculum that obtains with embodiment 1 carries out basin and carries the corn test, carries out in Tianjin Normal University 1 day～August 1 May in 2006.Test in sowing on May 1, August 1 gathered in the crops.Test is 28.2 * 30cm with the basin specification.Every basin was adorned 3.0mm sieve dry soil 12.5kg, sowed 10, field planting 5 strains.Regularly quantitatively water each experimental group carrying out horizontal strict conformance.Microbial inoculum imposes on 2～3cm place under the seed.
5 experimental group are established in test, and each experimental group is provided with 3 repetitions.Each experimental group is respectively:
Blank (CK)
Phytase microbial inoculum (liquid) amount of application (mL): 50,100,150,200
(1) phytase microbial inoculum is to the influence of maize seedling photosynthetic rate
Respectively on June 2nd, 2006 10:00a.m. and August 1 10:00a.m. measure the maize seedling photosynthetic rate.Condition determination is light intensity 700 μ molm
-2S
-1, 26 ℃ of room temperatures.Measurement result (measuring method is with reference to the photosynthetic instrument instrument of Li-6400 working instructions) is seen shown in Figure 4.
By Fig. 4 analysis, all variant in each experimental group.Wherein in the measured value on August 1, each level of microbial inoculum, photosynthesis rate is significantly higher than contrast.But photosynthetic rate on average shows and is more or less the same between each experimental group.
(2) phytase microbial inoculum is to the influence of maize seedling over-ground part dry weight
Table 5 phytase microbial inoculum is to the influence of maize seedling over-ground part dry weight
| Microbial inoculum amount of application (mL) | 0(CK) | 50 | 100 | 150 | 200 |
| Maize seedling dry weight measurement result (g) | 3.65 bB | 5.5946 aAB | 6.3528 aA | 5.4918 aAB | 7.1374 aA |
The maize seedling dry weight is measured on August 1st, 2006 and is carried out.By Fig. 5, table 5 is analyzed, and the microbial inoculum different levels is handled experiment, and maize seedling over-ground part dry weight all is significantly higher than control group.
(3) phytase microbial inoculum is to the influence of maize seedling root activity
The root system vigor is meant root system and respiratory intensity, receptivity, the relevant ability index of catalytic capability, and it can reflect the vigorous degree of growing of root system well.The measuring method employing TTC reduction method of root activity (chief editor of Laiyang Agricultural College plant physiology teaching and research room. plant physiology experiment instructs, 2005. Zhang Zhiliangs, the Qu Wei mountain valley with clumps of trees and bamboo. and plant physiology experiment instructs (third edition). Beijing: Higher Education Publishing House, 2003.).Under this test conditions, each experimental group maize seedling root system vitality test the results are shown in Table 6.Each experimental group is compared with control group, and utmost point significant difference is arranged.
Table 6 phytase microbial inoculum is to the influence of maize seedling root activity (tetrazole reduction intensity)
| Microbial inoculum amount of application (mL) | 0(CK) | 50 | 100 | 150 | 200 |
| Root activity test determination result (μ g/gh) | 21.54 bB | 34.21 abAB | 46.01 aA | 25.62 bB | 34.03 abAB |
(4) phytase microbial inoculum to the influence of the full nitrogen of maize seedling, full phosphorus, full potassium (measuring method is seen: Peng Jianwei, Song Haixing. plant science experimental center, Plant Nutrition experimental technique: Plant Nutrition experiment instruction.
Table 7 phytase microbial inoculum is to the influence of maize seedling over-ground part total nitrogen content
| The full nitrogen of maize seedling | 1.18 | 1.02 | 1.20 | 0.91 | 0.90 |
| Measurement result (% dry weight) | bAB | bcB | aA | cB | cB |
Table 8 phytase microbial inoculum is to the influence of maize seedling over-ground part content of tatal phosphorus
| Microbial inoculum amount of application (mL) | 0(CK) | 50 | 100 | 150 | 200 |
| The full phosphorus measurement result of maize seedling (% dry weight) | 2.28 cB | 2.79 aA | 2.52 bA | 2.80 aA | 2.74 abA |
Table 9 phytase microbial inoculum is to the influence of the full potassium content of maize seedling over-ground part
| Microbial inoculum amount of application (mL) | 0(CK) | 50 | 100 | 150 | 200 |
| The full potassium measurement result of maize seedling (% dry weight) | 0.29 aA | 0.35 aA | 0.32 aA | 0.32 aA | 0.35 aA |
By Fig. 6, table 7,8,9 as can be seen, when using microbial inoculum 100mL, the maize seedling total nitrogen content is compared with control group, there were significant differences, significant difference (0.05 level).Use that the maize seedling content of tatal phosphorus of each experimental group all is significantly higher than control group after the microbial inoculum.And difference is (0.01 level) extremely significantly.Each is handled to use microbial inoculum, complete comparing with control group, the maize seedling potassium content is improved, and the full potassium content of each treatment group, with the treatment capacity difference, maize seedling absorbs potassium element to be increased.
Embodiment 5
The phytase microbial inoculum that obtains with embodiment 1 carries out basin and carries the test of A Ergangjin alfalfa, carries out in Tianjin Normal University 1 day～July 1 May in 2006.Test in sowing on May 1, July 1 gathered in the crops.Test is 28.2 * 30cm with the basin specification.Every basin was adorned 3.0mm sieve dry soil 12.5kg.Sowing.Regularly quantitatively water each experimental group carrying out horizontal strict conformance.Microbial inoculum imposes on 2～3cm place under the seed.
5 experimental group are established in test, and each experimental group is provided with 3 repetitions.Each experimental group is respectively:
Blank (CK)
Phytase microbial inoculum (liquid) amount of application (mL): 50,100,150,200
Table 10 phytase microbial inoculum is to the influence of alfalfa ground top fresh weight
| Microbial inoculum amount of application (mL) | 0(CK) | 50 | 100 | 150 | 200 |
| Clover dry weight measurement result (g) | 46.4dC | 49.0dC | 60.9aA | 53.6cB | 58.0bA |
Table 11 phytase microbial inoculum is to the influence of alfalfa ground top dry weight
| Microbial inoculum amount of application (mL) | 0(CK) | 50 | 100 | 150 | 200 |
| Clover dry weight measurement result (g) | 8.0cB | 9.2bB | 11.7aA | 8.8bcB | 9.3bB |
Experimental result shows, carries out basin at phytase microbial inoculum and carries that phytase microbial inoculum can improve fresh weight and the dry weight of clover in the test of A Ergangjin alfalfa, promptly can improve the output of clover.
After the preferred embodiment that describes in detail, being familiar with this technology personage can be well understood to, can carry out various variations and modification not breaking away under above-mentioned claim and the spirit, all foundations technical spirit of the present invention all belongs to the scope of technical solution of the present invention to any simple modification, equivalent variations and modification that above embodiment did.And the embodiment that the present invention also is not subject in the specification sheets to be given an actual example.
Claims (9)
1. phytase microbial inoculum fertilizer is characterized in that it is made up of phytase bacterial strain and fermention medium, and wherein each composition proportion of fermention medium is: wheat bran 20-40%, glucose 5-20%, (NH
4)
2HPO
45-20%, K
2HPO
40.1-1%, MgSO
47H
2O 0.1-1%, NaCl 2-10%, all the other are water; Viable count in the microbial inoculum 〉=100,000,000/mL; Phytase bacterial strain wherein is phytase gene engineering strain SDLTP02.
2. phytase microbial inoculum fertilizer as claimed in claim 1, wherein each composition proportion of fermention medium is: wheat bran 35g, glucose 2g, (NH
4)
2HPO
410g, K
2HPO
40.2g, MgSO
47H
2O 0.1g, NaCl 2g, water 100mL.
3. the production method of a phytase microbial inoculum fertilizer is characterized in that the step that comprises:
(1) get phytase bacterial strain slant strains 2-4 articulating and go in the meat soup substratum, at 35-45 ℃, 24-36h is cultivated in concussion, obtains seed liquor;
(2) transfer in fermention medium with the inoculum size of 5-20% seed liquor, 35-45 ℃, cultivate 24-48h, make microbial inoculum, cryopreservation, standby; Phytase bacterial strain wherein is phytase gene engineering strain SDLTP02.
4. the production method of phytase microbial inoculum fertilizer as claimed in claim 3, being prepared as of fermention medium wherein: wheat bran 20-40%, glucose 5-20%, (NH
4)
2HPO
45-20%, K
2HPO
40.1-1%, MgSO
47H
2O 0.1-1% and NaCl 2-10%, all the other are water, pH 7.0-7.2.
5. the application of the described phytase microbial inoculum fertilizer of claim 1 aspect the raising plant photosynthetic rate.
6. the application of the described phytase microbial inoculum fertilizer of claim 1 aspect the promotion plant-growth.
7. the described phytase microbial inoculum fertilizer of claim 1 is increasing plant to the application aspect nitrogen, phosphorus, the potassium absorption.
8. the application of the described phytase microbial inoculum fertilizer of claim 1 aspect the raising plant biomass.
9. as each described application of claim 5-8, plant wherein is vegetables, fruit tree, wheat or corn.
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| WO2015119681A1 (en) * | 2014-02-04 | 2015-08-13 | University Of Florida Research Foundation, Inc. | Pteris vittata phytase nucleotide and amino acid sequences and methods of use |
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| CN102382782A (en) * | 2011-10-17 | 2012-03-21 | 天津师范大学 | Phytase microbial inoculum and preparation method thereof |
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| CN1284489A (en) * | 1999-08-16 | 2001-02-21 | 季中忱 | Fermenting agent for fast enzymolyizing and converting straw fertilizer and its preparation |
| CN1733902A (en) * | 2005-07-01 | 2006-02-15 | 南开大学 | Transgenic bacillus strain secreting phytase and its uses |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015119681A1 (en) * | 2014-02-04 | 2015-08-13 | University Of Florida Research Foundation, Inc. | Pteris vittata phytase nucleotide and amino acid sequences and methods of use |
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