CN107164288A - A kind of compound Soluble phosphorus microbial inoculum and its preparation method and application - Google Patents
A kind of compound Soluble phosphorus microbial inoculum and its preparation method and application Download PDFInfo
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- CN107164288A CN107164288A CN201710574017.1A CN201710574017A CN107164288A CN 107164288 A CN107164288 A CN 107164288A CN 201710574017 A CN201710574017 A CN 201710574017A CN 107164288 A CN107164288 A CN 107164288A
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Abstract
The invention belongs to microbial technology field, specifically related to a kind of compound Soluble phosphorus microbial inoculum, it is made up of phosphorus-solubilizing bacteria WS32 with phosphorus-solubilizing bacteria P1616 mixed fermentations, the Classification And Nomenclature of the phosphorus-solubilizing bacteria WS32 is pseudomonad (Pseudomonas sp.), it is deposited in China typical culture collection center on June 9th, 2017, deposit number is CCTCC NO:M 2017319, the phosphorus-solubilizing bacteria P1616 Classification And Nomenclature are bacillus (Bacillus sp.), are deposited in China typical culture collection center on June 2nd, 2017, and deposit number is CCTCC NO:M 2017298.The beneficial effects of the invention are as follows：1st, WS32 of the invention and P1616 and wheat have compatibility, can occur agglutinating reaction with wheat agglutinin, can settle down survival for a long time in wheat root, play the effect of wheat growth-promoting.2nd, composite bacteria agent of the invention has strong production organic acid and phosphatase ability, it is ensured that efficient phosphorus-dissolution, phosphorus decomposing effect.
Description
Technical field
The invention belongs to microbial technology field, and in particular to a kind of compound Soluble phosphorus microbial inoculum and its preparation method and application.
Background technology
One of nutriment, most important for plant growth necessary to phosphorus is plant growth, but the big portion in soil
Organophosphor and Phos is divided but can not to be directly absorbed and used by plants.The arable soil of China 74% lacks phosphorus, and 95% in soil
Phosphorus above is invalid phosphorus, and causing effective phosphor resource, there is a serious shortage in the supply.Phosphorus nutrients, but plant can be provided to plant using phosphate fertilizer
To this season utilization rate typically only 5%~25% of phosphate fertilizer, due to arable land long-term application of P fertilizer, most of phosphate fertilizer is as invalid
State is accumulated in arable soil, and ecological environment is impacted.
Soil Soluble phosphorus microorganism is not only able to the absorption for promoting plant to various nutrients, promotes the life of plant root
It is long, utilization rate of the plant to phosphorus is improved, improves plant nutrient condition, crop yield is improved, increases plant resistance against diseases, can also change
Kind soil texture, improves the content of organic matter, improves salt-soda soil, to cultivating and giving full play to soil ecology fertility, keep agricultural raw
State environment plays an important roll.Therefore, the research of soil Soluble phosphorus microorganism has good development prospect and application value.
The content of the invention
, can be efficiently by soil it is an object of the invention to provide a kind of compound Soluble phosphorus microbial inoculum in order to solve the above problems
In invalid phosphorus be converted to the high-quality phosphorus for being available for wheat directly to absorb.
The invention provides following technical scheme：
The compound Soluble phosphorus microbial inoculum of the present invention, is made up, the phosphorus-solubilizing bacteria of phosphorus-solubilizing bacteria WS32 with phosphorus-solubilizing bacteria P1616 mixed fermentations
WS32 Classification And Nomenclature is pseudomonad (Pseudomonas sp.), is deposited in Chinese Typical Representative culture guarantor on June 9th, 2017
Tibetan center, deposit number is CCTCC NO:M 2017319, depositary institution address is Wuhan University, the phosphorus-solubilizing bacteria P1616's
Classification And Nomenclature is bacillus (Bacillus sp.), is deposited in China typical culture collection center on June 2nd, 2017, is protected
It is CCTCC NO to hide numbering:M 2017298, depositary institution address is Wuhan University.
Strain morphology feature of the present invention：Phosphorus-solubilizing bacteria WS32 bacterial strains are Gram-negative bacteria, and bacterium colony is neat, in faint yellow, connects
Catalase test display is negative, and amylase test display is positive；Phosphorus-solubilizing bacteria P1616 is gram-positive bacteria, and bacterium colony is irregular, in white
Color, there is gemma, and catalase test display is positive, and amylase test display is positive.
The compound Soluble phosphorus microbial inoculum of the present invention is that liquid is combined Soluble phosphorus microbial inoculum or solid union Soluble phosphorus microbial inoculum.
The preparation method that liquid of the present invention is combined Soluble phosphorus microbial inoculum is as follows,
Phosphorus-solubilizing bacteria WS32 and phosphorus-solubilizing bacteria P1616 bacterium solution are pressed 1：In 0.5~2 volume ratio mixing, two of which bacterium solution
Bacterium number 1 × 108~9 × 108In the range of CFU/mL, then by the bacterium solution of mixing with 2~5% inoculum concentration access 80~
In 120mL beef extract-peptone fluid nutrient mediums, under conditions of 25~30 DEG C, 120~180r/min, concussion and cultivate 18~
48h, that is, be made liquid and be combined Soluble phosphorus microbial inoculum.
The preparation method of solid union Soluble phosphorus microbial inoculum of the present invention is as follows,
It is combined in above-mentioned liquid and dry 60~180g of peat mixings is added in Soluble phosphorus microbial inoculum, that is, solid union Soluble phosphorus is made
Microbial inoculum.
The total bacteria count that the liquid of the present invention is combined phosphorus-solubilizing bacteria WS32 and phosphorus-solubilizing bacteria P1616 in Soluble phosphorus microbial inoculum is 1 × 108~9
×108CFU/mL。
The compound Soluble phosphorus microbial inoculum of the present invention is promoting the growth of wheat, is promoting wheat to P acquisition and reduction fertilizer application
The application of amount.
The seed dressing of the invention that Soluble phosphorus microbial inoculum is combined for wheat or coating pour root or root dipping.
It is 1~4L that liquid of the present invention, which is combined every mu of wheat paddock amount of application of Soluble phosphorus microbial inoculum, and solid union Soluble phosphorus microbial inoculum is every mu small
Wheatland amount of application is 1~4kg.
WS32 16SrRNA is as shown in SEQ ID NO.1, and P1616 16SrRNA is as shown in SEQ ID NO.2.
The beneficial effects of the invention are as follows：
1st, WS32 of the invention and P1616 has compatibility with wheat, can be with wheat agglutinin generation agglutinating reaction, can be
Wheat root settles down survival for a long time, plays the effect of wheat growth-promoting.
2nd, composite bacteria agent of the invention has strong production organic acid and phosphatase ability, it is ensured that efficient phosphorus-dissolution, phosphorus decomposing effect.
3rd, being combined Soluble phosphorus microbial inoculum or solid union Soluble phosphorus microbial inoculum using the WS32 and P1616 of the present invention liquid prepared has
Significant effect of solubilizing phosphate, can dissolve indissoluble state Phos, can degrade indissoluble state organophosphor again, utilize WS32 and P1616 itself
Metabolic capability the inorganic phosphorus element and organic phosphorus element of not solvent are discharged, increase substantially in bacterium solution or quick-acting in soil
The content of phosphorus.
4th, compound Soluble phosphorus microbial inoculum application method of the invention is simple, can be using dressing seed, be coated, pouring the methods such as root, root dipping
Row inoculation.
5th, liquid of the invention is combined Soluble phosphorus microbial inoculum or the application dosage of solid union Soluble phosphorus microbial inoculum is low, and liquid is combined Soluble phosphorus
Every mu of wheat paddock amount of application of microbial inoculum is only 1~4L, and every mu of wheat paddock amount of application of solid union Soluble phosphorus microbial inoculum is only 1~4kg, with it
Its every mu of wheat paddock amount of application compares for 30~50kg heavy dose of biological fertilizer products, and life can be greatly lowered in the present invention
Produce cost, energy-conserving and environment-protective.
Brief description of the drawings
Fig. 1 is titanium pigment concentration in slightly solubility Phos tricalcium phosphate culture medium zymotic fluid；
Fig. 2 is phosphorus-solubilizing bacteria production activity of acid phosphatase in slightly solubility organophosphor Phytate Ca medium zymotic fluid
Fig. 3 is WS32 bacterium colony figure；
Fig. 4 is P1616 bacterium colony figure；
Fig. 5 is WS32 and P1616 bacterium solution volumes ratio is 1:1 bacterium colony figure；
Fig. 6 is WS32 and P1616 bacterium solution volumes ratio is 1:2 bacterium colony figure；
Fig. 7 is WS32 and P1616 bacterium solution volumes ratio is 2:1 bacterium colony figure.
Embodiment
The present invention is illustrated with reference to specific embodiment.
The screening of embodiment 1WS32 bacterial strains and P1616 bacterial strains
Filtered out using Selective agar medium binding lectin screening method from wheat rhizosphere has special affinity interaction with wheat
29 plants of wheat rhizosphere phosphorus-solubilizing bacteria；From different Soluble phosphorus culture mediums, Ca is added respectively3(PO4)2、FePO4、AlPO4、
CaHPO4·2H2The slightly solubility Phos such as O, and the slightly solubility organophosphor such as lecithin, phytic acid calcium；Surveyed using molybdenum antimony resistance colorimetric method
The concentration of titanium pigment in fixed each bacterial strain fermentation liquor, determines 29 plants of bacterial strains to the solvability of different shape indissoluble phosphorus, determines it
In 5 plants of bacterium effect of solubilizing phosphate it is preferable.This 5 plants of wheat efficient phosphorus-dissolution bacterium are mixed two-by-two in varing proportions respectively, its are determined molten
Phosphorus ability, it is found that wherein WS32 bacterial strains are optimal with P1616 bacterial strains effect of solubilizing phosphate, and two plants of bacterium are sequenced through 16SrRNA molecules and determined
WS32 is pseudomonas, and P1616 is bacillus.
Embodiment 2 is combined the phosphate solubilization of Soluble phosphorus microbial inoculum
By WS32 and P1616 bacterium solutions respectively by volume 1：0、0：1、2：1、1：1、1：In 2 mixing, two of which bacterium solution
Bacterium number 1 × 108~9 × 108In the range of CFU/mL, tricalcium phosphate culture medium, triangular flask training are accessed with 3% inoculum concentration
Support, condition of culture be 160r/min, 28 DEG C, culture 9 days；Determine solvable in zymotic fluid using molybdenum antimony resistance colorimetric method timing daily
The concentration of property phosphorus, as a result as shown in figure 1, in Fig. 1, W:P represents WS32：P1616, is combined effect of solubilizing phosphate of the phosphorus-solubilizing bacteria compared with single strain
Greatly improve.
Embodiment 3 is combined Soluble phosphorus microbial inoculum secretion organic acid and phosphatase ability
(1) Soluble phosphorus fungi degradation Phos principle is secretion organic acid, and it is soluble state phosphorus to dissolve indissoluble state Phos in soil, is supplied
Plant absorption is utilized.
Using tricalcium phosphate culture medium, high performance liquid chromatography surveys kinds of organic acids and concentration.Liquid submerged culture 7d, takes
Bacterium solution 1mL, centrifugation, then the upper liquid chromatogram after 0.22 μm of filter membrane vacuum filtration.Separation condition：Chromatographic column is Hypersil C18
(250nm × 4.6mm i d, 5 μm), mobile phase is 0.02 mol/L KH2PO4Cushioning liquid-methanol solution (99%:1%),
With 1mol/L phosphorus acid for adjusting pH to 2.60, ultraviolet detection wavelength 210nm, sample size 10 μ L, flow velocity 0.5mL/min, column temperature 30
DEG C, obtain table 1.
From table 1, the kinds of organic acids that compound phosphorus-solubilizing bacteria is produced in tricalcium phosphate culture medium is more, and concentration is bigger,
Be conducive to the dissolving to indissoluble state Phos.
The high performance liquid chromatography of table 1 is to strain secretes kinds of organic acids and the measure of concentration
(2) Soluble phosphorus fungi degradation organophosphor principle is produces phosphatase, by the enzyme degradation of phosphatase, by the difficulty in soil
Dissolubility organic phosphorus degrading is available phosphorus.
Measure to phosphatase activity in bacterium solution uses Phytate Ca medium, and disodium phenyl phosphate method is determined.As a result such as Fig. 2 institutes
Show.In Fig. 2, W:P represents WS32：P1616, is combined phosphorus-solubilizing bacteria activity of acid phosphatase highest, this accelerates the decomposition of phytic acid calcium,
Promote phosphorus element release, so as to improve wheat phosphorus nourishing.
The preparation of the liquid Soluble phosphorus microbial inoculum of embodiment 4 and the potted plant experiment for wheat
Liquid Soluble phosphorus microbial inoculum includes liquid and is combined Soluble phosphorus microbial inoculum, liquid WS32 Soluble phosphorus microbial inoculum and liquid P1616 phosphorus-solubilizing bacterias
Agent.
Liquid is combined the preparation of Soluble phosphorus microbial inoculum
By WS32 and P1616 bacterium solutions respectively by volume 2：1、1：1、1：Bacterium number in 2 mixing, two of which bacterium solution exists
1×108~9 × 108In the range of CFU/mL, and accessed with 3% inoculum concentration in 100mL beef extract-peptone fluid nutrient mediums,
Under the conditions of 28 DEG C, 160r/min, concussion and cultivate 30h is made liquid and is combined Soluble phosphorus microbial inoculum.
The preparation of liquid WS32 Soluble phosphorus microbial inoculums
By WS32, (bacterium number in bacterium solution is 1 × 108~9 × 108In the range of CFU/mL) 100mL is accessed with 3% inoculum concentration
In beef extract-peptone fluid nutrient medium, under the conditions of 28 DEG C, 160r/min, concussion and cultivate 30h is made liquid WS32 Soluble phosphorus
Microbial inoculum.
The preparation of liquid P1616 Soluble phosphorus microbial inoculums
By P1616, (bacterium number in bacterium solution is 1 × 108~9 × 108In the range of CFU/mL) accessed with 3% inoculum concentration
In 100mL beef extract-peptone fluid nutrient mediums, under the conditions of 28 DEG C, 160r/min, concussion and cultivate 30h is made liquid
P1616 Soluble phosphorus microbial inoculums.
Liquid Soluble phosphorus microbial inoculum is used for the potted plant experiment of wheat
Wheat seed vernalization, takes appropriate wheat seed water to rinse once, then with 70% alcohol-pickled 3min sterilizations simultaneously
Remove shrivelled seed, then with aseptic water washing 5 times, finally soaked with appropriate water, be put in vernalization in 25 DEG C of incubators, treat seed
Showing money or valuables one carries unintentionally to sow.Wheat is potted plant to set 3 treatment groups and 1 control group：Apply liquid and be combined Soluble phosphorus microbial inoculum treatment group (WS32
It is respectively 2 with P1616 bacterium solution volumes ratio:1,1:1,1:2), apply liquid WS32 Soluble phosphorus microbial inoculums treatment group, apply liquid P1616 Soluble phosphorus
Microbial inoculum treatment group and apply blank cultures control group.Three repeating samples of each treatment group.Treatment group is inoculated with liquid phosphorus-solubilizing bacteria per basin
Agent 5mL, control group only taps into 5mL blank cultures, and per 5 plants of basin final singling after seedling all sprouts, daily management liquid manure is adopted in 125d
Receive, determine the phosphorus content of Plant Height in Wheat, fresh weight, dry weight, aerial part blade phosphorus content and potted plant rear soil.
Result of the test is as shown in table 2：
Liquid is combined Plant Height in Wheat, fresh weight, dry weight, content of soil available phosphor, the blade content of tatal phosphorus of Soluble phosphorus microbial inoculum inoculation
The control increase by 6.1%~25.6%, 6.9%~8.9%, 15.1%~19.3%, 8.3% of liquid microbial inoculum is not connect respectively
~14.8%, 37.8%~49.7%, each parameter also has a more substantial increase compared with the wheat that single bacterium agent is inoculated with.Demonstrate and apply liquid
State, which is combined Soluble phosphorus microbial inoculum, can improve absorption of the wheat to phosphorus, wheat growth-promoting.
The liquid Soluble phosphorus microbial inoculum of table 2 is used for the potted plant experiment of wheat
The preparation of the solid-state Soluble phosphorus microbial inoculum of embodiment 5 and the potted plant experiment for wheat
Solid-state Soluble phosphorus microbial inoculum includes solid union Soluble phosphorus microbial inoculum, solid-state WS32 Soluble phosphorus microbial inoculum and solid-state P1616 phosphorus-solubilizing bacterias
Agent.
The preparation of solid union Soluble phosphorus microbial inoculum
By WS32 and P1616 bacterium solutions respectively by volume 2：1、1：1、1：Bacterium number in 2 mixing, two of which bacterium solution exists
1×108~9 × 108In the range of CFU/mL, and accessed with 3% inoculum concentration in 100mL beef extract-peptone fluid nutrient mediums,
Under the conditions of 28 DEG C, 160r/min, concussion and cultivate 30h adds dry peat 150g and mixed, that is, solid union Soluble phosphorus is made
Microbial inoculum.
The preparation of solid-state WS32 Soluble phosphorus microbial inoculums
By WS32, (bacterium number in bacterium solution is 1 × 108~9 × 108In the range of CFU/mL) 100mL is accessed with 3% inoculum concentration
In beef extract-peptone fluid nutrient medium, under the conditions of 28 DEG C, 160r/min, concussion and cultivate 30h adds dry peat 150g
Mix, that is, solid-state WS32 Soluble phosphorus microbial inoculums are made.
The preparation of solid-state P1616 Soluble phosphorus microbial inoculums
By P1616, (bacterium number in bacterium solution is 1 × 108~9 × 108In the range of CFU/mL) accessed with 3% inoculum concentration
In 100mL beef extract-peptone fluid nutrient mediums, under the conditions of 28 DEG C, 160r/min, concussion and cultivate 30h adds dry mud
Charcoal 150g is mixed, that is, solid-state P1616 Soluble phosphorus microbial inoculums are made.
Solid-state Soluble phosphorus microbial inoculum is used for the potted plant experiment of wheat
The potted plant experiment difference of the potted plant experiment of the wheat of the present embodiment and the wheat of embodiment 4 is, embodiment 4
Wheat potted plant experiment " treatment group is inoculated with liquid phosphorus-solubilizing bacteria agent 5mL per basin, and control group only taps into 5mL blank cultures ", this
The potted plant experiment of the wheat of embodiment " is inoculated with solid-state Soluble phosphorus microbial inoculum 5g per basin, control group is accessed through 121 DEG C, 20min inactivation treatments
Solid-state Soluble phosphorus microbial inoculum 5g ".
Result of the test is as shown in table 3
The Plant Height in Wheat of solid union microbial inoculum inoculation, fresh weight, dry weight, the control of blade content of tatal phosphorus more non-inoculum respectively
Increase by 6.3%~12.7%, 7.6%~9.3%, 13.1%~16.7%, 42.5%~52.9%, each parameter connects compared with single bacterium agent
The wheat planted also has a more substantial increase.Absorption of the wheat to phosphorus can be improved using solid union Soluble phosphorus microbial inoculum by demonstrating, and be promoted
Enter wheat growth.
The solid-state Soluble phosphorus microbial inoculum of table 3 is used for the potted plant experiment of wheat
The preferred embodiments of the present invention are the foregoing is only, are not intended to limit the invention, although with reference to foregoing reality
Apply example the present invention is described in detail, for those skilled in the art, it still can be to foregoing each implementation
Technical scheme described in example is modified, or carries out equivalent substitution to which part technical characteristic.All essences in the present invention
God is with principle, and any modification, equivalent substitution and improvements made etc. should be included in the scope of the protection.
SEQUENCE LISTING
<110>Agricultural University Of Anhui
<120>A kind of compound Soluble phosphorus microbial inoculum and its preparation method and application
<130> 2
<160> 2
<170> PatentIn version 3.3
<210> 1
<211> 1406
<212> DNA
<213>Pseudomonad（Pseudomonas sp.）
<400> 1
cacatgcaag tcgagcggta gagagaagct tgcttctctt gagagcggcg gacgggtgag 60
taatgcctag gaatctgcct ggtagtgggg gataacgttc ggaaacggac gctaataccg 120
catacgtcct acgggagaaa gcaggggacc ttcgggcctt gcgctatcag atgagcctag 180
gtcggattag ctagttggtg aggtaatggc tcaccaaggc gacgatccgt aactggtctg 240
agaggatgat cagtcacact ggaactgaga cacggtccag actcctacgg gaggcagcag 300
tggggaatat tggacaatgg gcgaaagcct gatccagcca tgccgcgtgt gtgaagaagg 360
tcttcggatt gtaaagcact ttaagttggg aggaagggtt gtagattaat actctgcaat 420
tttgacgtta ccgacagaat aagcaccggc taactctgtg ccagcagccg cggtaataca 480
gagggtgcaa gcgttaatcg gaattactgg gcgtaaagcg cgcgtaggtg gtttgttaag 540
ttggatgtga aatccccggg ctcaacctgg gaactgcatt caaaactgac tgactagagt 600
atggtagagg gtggtggaat ttcctgtgta gcggtgaaat gcgtagatat aggaaggaac 660
accagtggcg aaggcgacca cctggactaa tactgacact gaggtgcgaa agcgtgggga 720
gcaaacagga ttagataccc tggtagtcca cgccgtaaac gatgtcaact agccgttgga 780
agccttgagc ttttagtggc gcagctaacg cattaagttg accgcctggg gagtacggcc 840
gcaaggttaa aactcaaatg aattgacggg ggcccgcaca agcggtggag catgtggttt 900
aattcgaagc aacgcgaaga accttaccag gccttgacat ccaatgaact ttctagagat 960
agattggtgc cttcgggaac attgagacag gtgctgcatg gctgtcgtca gctcgtgtcg 1020
tgagatgttg ggttaagtcc cgtaacgagc gcaacccttg tccttagtta ccagcacgtt 1080
atggtgggca ctctaaggag actgccggtg acaaaccgga ggaaggtggg gatgacgtca 1140
agtcatcatg gcccttacgg cctgggctac acacgtgcta caatggtcgg tacagagggt 1200
tgccaagccg cgaggtggag ctaatcccat aaaaccgatc gtagtccgga tcgcagtctg 1260
caactcgact gcgtgaagtc ggaatcgcta gtaatcgcga atcagaatgt cgcggtgaat 1320
acgttcccgg gccttgtaca caccgcccgt cacaccatgg gagtgggttg caccagaagt 1380
agctagtcta accttcggga ggacgg 1406
<210> 2
<211> 1453
<212> DNA
<213>Bacillus（Bacillus sp.）
<400> 2
gccgtgcggg gtgctataat gcagtcgagc gaactgatta gaagcttgct tctatgacgt 60
tagcggcgga cgggtgagta acacgtgggc aacctgcctg taagactggg ataacttcgg 120
gaaaccgaag ctaataccgg ataggatctt ctccttcatg ggagatgatt gaaagatggt 180
ttcggctatc acttacagat gggcccgcgg tgcattagct agttggtgag gtaacggctc 240
accaaggcaa cgatgcatag ccgacctgag agggtgatcg gccacactgg gactgagaca 300
cggcccagac tcctacggga ggcagcagta gggaatcttc cgcaatggac gaaagtctga 360
cggagcaacg ccgcgtgagt gatgaaggct ttcgggtcgt aaaactctgt tgttagggaa 420
gaacaagtac gagagtaact gctcgtacct tgacggtacc taaccagaaa gccacggcta 480
actacgtgcc agcagccgcg gtaatacgta ggtggcaagc gttatccgga attattgggc 540
gtaaagcgcg cgcaggcggt ttcttaagtc tgatgtgaaa gcccacggct caaccgtgga 600
gggtcattgg aaactgggga acttgagtgc agaagagaaa agcggaattc cacgtgtagc 660
ggtgaaatgc gtagagatgt ggaggaacac cagtggcgaa ggcggctttt tggtctgtaa 720
ctgacgctga ggcgcgaaag cgtggggagc aaacaggatt agataccctg gtagtccacg 780
ccgtaaacga tgagtgctaa gtgttagagg gtttccgccc tttagtgctg cagctaacgc 840
attaagcact ccgcctgggg agtacggtcg caagactgaa actcaaagga attgacgggg 900
gcccgcacaa gcggtggagc atgtggttta attcgaagca acgcgaagaa ccttaccagg 960
tcttgacatc ctctgacaac tctagagata gagcgttccc cttcggggga cagagtgaca 1020
ggtggtgcat ggttgtcgtc agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag 1080
cgcaaccctt gatcttagtt gccagcattt agttgggcac tctaaggtga ctgccggtga 1140
caaaccggag gaaggtgggg atgacgtcaa atcatcatgc cccttatgac ctgggctaca 1200
cacgtgctac aatggatggt acaaagggct gcaagaccgc gaggtcaagc caatcccata 1260
aaaccattct cagttcggat tgtaggctgc aactcgccta catgaagctg gaatcgctag 1320
taatcgcgga tcagcatgcc gcggtgaata cgttcccggg ccttgtacac accgcccgtc 1380
acaccacgag agtttgtaac acccgaagtc ggtggagtaa ccgtaaggag ctagccgcct 1440
aagggacaga gtg 1453
Claims (9)
1. a kind of compound Soluble phosphorus microbial inoculum, it is characterised in that be made up of phosphorus-solubilizing bacteria WS32 with phosphorus-solubilizing bacteria P1616 mixed fermentations, described
Phosphorus-solubilizing bacteria WS32 Classification And Nomenclature is pseudomonad (Pseudomonas sp.), is deposited in Chinese Typical Representative training on June 9th, 2017
Thing collection is supported, deposit number is CCTCC NO:M2017319, the phosphorus-solubilizing bacteria P1616 Classification And Nomenclature are bacillus
(Bacillus sp.), is deposited in China typical culture collection center on June 2nd, 2017, and deposit number is CCTCC NO:
M2017298。
2. a kind of compound Soluble phosphorus microbial inoculum according to claim 1, it is characterised in that the compound Soluble phosphorus microbial inoculum is multiple for liquid
Close Soluble phosphorus microbial inoculum or solid union Soluble phosphorus microbial inoculum.
3. a kind of preparation method of compound Soluble phosphorus microbial inoculum as claimed in claim 2, it is characterised in that the liquid is combined Soluble phosphorus
The preparation method of microbial inoculum is as follows,
Phosphorus-solubilizing bacteria WS32 and phosphorus-solubilizing bacteria P1616 bacterium solution are pressed 1：Bacterium in 0.5~2 volume ratio mixing, two of which bacterium solution
Number is 1 × 108~9 × 108In the range of CFU/mL, the bacterium solution of mixing is then accessed into 80~120mL with 2~5% inoculum concentration
In beef extract-peptone fluid nutrient medium, under conditions of 25~30 DEG C, 120~180r/min, 18~48h of concussion and cultivate, i.e.,
The liquid is made and is combined Soluble phosphorus microbial inoculum.
4. the preparation method of a kind of compound Soluble phosphorus microbial inoculum according to claim 3, it is characterised in that the solid union is molten
The preparation method of phosphobacterin is as follows,
It is combined in the liquid and dry 60~180g of peat mixings is added in Soluble phosphorus microbial inoculum, that is, the solid union Soluble phosphorus is made
Microbial inoculum.
5. a kind of preparation method of compound Soluble phosphorus microbial inoculum according to claim 3, it is characterised in that the obtained liquid
Phosphorus-solubilizing bacteria WS32 and phosphorus-solubilizing bacteria P1616 total bacteria count are 1 × 10 in compound Soluble phosphorus microbial inoculum8~9 × 108CFU/mL。
6. a kind of compound Soluble phosphorus microbial inoculum described in claim 1, it is characterised in that the compound Soluble phosphorus microbial inoculum is promoting wheat
The application of growth, promotion wheat to P acquisition and reduction fertilizer application amount.
7. a kind of compound Soluble phosphorus microbial inoculum described in claim 1, it is characterised in that the compound Soluble phosphorus microbial inoculum is used for mixing for wheat
Plant or be coated or pour root or root dipping.
8. the preparation method of a kind of compound Soluble phosphorus microbial inoculum according to claim 3, it is characterised in that the liquid is combined molten
Every mu of wheat paddock amount of application of phosphobacterin is 1~4L.
9. the preparation method of a kind of compound Soluble phosphorus microbial inoculum according to claim 4, it is characterised in that the solid union is molten
Every mu of wheat paddock amount of application of phosphobacterin is 1~4kg.
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