CN104371946A - Pseudomonas vranovensis with phosphorus solubilizing capability and application thereof - Google Patents

Pseudomonas vranovensis with phosphorus solubilizing capability and application thereof Download PDF

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CN104371946A
CN104371946A CN201410237364.1A CN201410237364A CN104371946A CN 104371946 A CN104371946 A CN 104371946A CN 201410237364 A CN201410237364 A CN 201410237364A CN 104371946 A CN104371946 A CN 104371946A
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zsw
phosphorus
liquid
soil
pseudomonas
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朱晓丽
宋进喜
马俊杰
李贺
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Northwest University
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/38—Pseudomonas
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01C—PLANTING; SOWING; FERTILISING
    • A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01C—PLANTING; SOWING; FERTILISING
    • A01C21/00—Methods of fertilising, sowing or planting
    • 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
    • A01G7/00—Botany in general
    • A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/10—Animals; Substances produced thereby or obtained therefrom
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor

Abstract

The invention discloses pseudomonas vranovensis with phosphorus solubilizing capability, a preparation method and application thereof. Through an ultraviolet-ray plasma composite mutation technology for multiple composite mutation, a bacterial strain ZSW-3-5-1a with high solubilizing efficiency on phosphorus and good stability is selected and bred, has the classification name of pseudomonas vranovensis, is preserved in China General Microbiological Culture Collection Center, and the preservation number is CGMCC No. 8750. The bacterial strain has efficient solubilizing capability on calcium phosphate and calcium phytate, and also is capable of generating a certain amount of indoleacetic acid. By using a liquid containing the bacterium or a solid inocula to inoculate corn, soybean, mustard and other crops, utilization rate on phosphorus in soil is improved and crop output is improved.

Description

A kind of not village pseudomonas and application thereof with dissolving P capacity
Technical field
The present invention relates to not village pseudomonas and application thereof that a strain has dissolving P capacity, this bacterial strain ultraviolet-plasma body complex mutation obtains, and can improve the utilization ratio of soil phosphorus, improves crop yield, belongs to field of agricultural microbial technology.
Background technology
Phosphorus is one of large key element of plant nutrition three, and phosphorus is under-supply becomes one of most important factor of restriction crop yield and quality gradually.In soil, total phosphorous is sufficient, total phosphorus containing 0.02-0.5%, but there is (mainly calcium phosphate, aluminum phosphate, tertiary iron phosphate in the form greatly mainly with insoluble inorganic phosphate, in alkaline soils based on calcium phosphate), only have 1% to be that the titanium pigment salt form that can directly absorb with plant exists.For meeting plant growth needs, the whole world every year will to the nearly 3,000 ten thousand tons of phosphate fertilizer of soil application, but wherein 80% become the invalid phosphorus that plant is difficult to absorb due to absorption, deposition or fixed action, become the main body of soil phosphorus.According to statistics, China has the arable soil of 74 % to lack phosphorus, but a lot of Soil total nitrogen is but very high.Zhang Fusuo etc. are to the investigation of provinces and regions, China loess plateau 7, and soil available phosphorus content average out to 6mgP/Kg, full phosphorus amount then reaches 1230mgP/Kg, is 205 times of available phosphorus.According to statistics, China has 7880.9 × 10 from the phosphate fertilizer applying farmland accumulative 1949 to 1992 4t (P 2o 5), wherein nearly 6000 × 10 4t (P 2o 5) be accumulated in soil and be not utilized, its this season utilization ratio is only 5% ~ 25%.And the phosphorus element resting on soil easy with rainwater or irrigate loss enter water body, cause body eutrophication to pollute.Therefore, how to excavate the phosphorus base resource that soil is potential, improve the utilization ratio of phosphorus, be one of hot subject of studying of scientific worker always.
Soil phosphorus is divided into Inorganic phosphorus and the large class of organic phosphorus two.Mineral soil is based on inorganic phosphorus, and organophosphorus accounts for 20% ~ 50% of full phosphorus.Plant mainly utilizes the Inorganic phosphorus in soil, and it absorbs the chemical form mainly H of phosphorus element 2pO 4 -and HPO 4 2-, the phosphorus in soil is main to be existed with non-solvent.The factor affecting Soil Phosphorus utilization ratio is a lot, and wherein the utilization ratio of microorganism to Soil Phosphorus has the greatest impact.Research proves: there is a large amount of microorganisms in soil, certain micro-organisms can activate insoluble phosphorus by metabolite, increase available phosphorus content in soil, promote plant growth, the microorganism with this ability is called phosphate solubilizing bacteria or phosphorus-solubilizing bacteria (phosphate solubilizing microorganisms, PSM).Phosphate solubilizing bacteria is except insoluble in dissolving soil or insoluble phosphorus element, and part phosphate solubilizing bacteria also has potassium decomposing, produces the growth that indolylacetic acid, siderophore, synthesis acc deaminase etc. promote plants.The microbe species with dissolving P capacity is a lot, the phosphate solubilizing bacteria of current report has mainly contained bacillus, erwinia, Rhodopseudomonas, Agrobacterium, serratia, Penicillium and Flavobacterium etc., but very few to the report of the not village pseudomonas with dissolving P capacity.Chen Shouwen etc. [patent No.: CN101851596 B] report a strain and have the clostridium butylicum separating inorganic phosphorus ability, and it separates inorganic phosphorus ability is 204.70 ± 25.78mg/L, and separating phytic acid ca ability is 52.31 ± 12.60mg/L.The reports such as Lin Qimei: the bacterium phosphate solubilization that dissolved phosphorus breeze ability is stronger is 26.92-43.34mg/L, most of fungi is then 59.64-145.36 mg/L.As can be seen here, current China for the production of phosphorus decomposing bacterial strain, its dissolving P capacity is lower, and effect is unstable, in addition, the major part be separated from nature wild all there is poor growth in phosphate solubilizing bacteria, viability is weak, validity period is short, phosphorus decomposing and growth promotion ability is lower and require the shortcomings such as harsh to edatope, the needs as growth-promoting microbial fertilizer can not be met.For this reason, carry out the modification of wild-type strain, break the eubolism of wild-type strain, making it the target metabolic product (as improved phosphorus decomposing, ability of dissolving potassium etc.) required for producing, improving its resistance capacity etc., reach this object, major measure is exactly the seed selection needing to carry out bacterial classification, as carried out physics and chemistry mutagenesis etc.
At present, the modification of microorganism mainly contains mutagenesis and engineered method, and microbial fertilizer to be directly manured into soil by the thalline of living and in environment, the therefore application of clear and definite restriction gene engineering bacteria in the bacterial manure standard of China.Ray, nuclear radiation and chemomorphosis are the selection by mutation means generally used at present, but a kind of mutagenic compound of life-time service often make bacterial strain produce resistance, want to obtain larger variation and just must adopt new mutation source.Plasma radiation is direct sedimentary energy in biomolecules, causes transgenation, thus reaches the object of selection by mutation.The method mutagenesis is adopted to have the advantages such as mutation frequency is high, mutation spectrum is wide, physiological damage is little.In addition, using plasma and ultraviolet two kinds of mutagenic compound alternately and compound use, effectively can avoid the mutagenesis saturation effect using single mutagenic compound to produce.
Summary of the invention
An object of the present invention be for prior art exist deficiency, by ultraviolet-plasma body complex mutation obtain a strain have higher dissolving P capacity not village pseudomonas ( pseudomonas vranovensis), this bacterial strain can also produce a certain amount of indolylacetic acid;
Two of object of the present invention is to provide microbiobacterial agent containing above-mentioned not village pseudomonas and preparation method thereof;
Three of object of the present invention is to provide the application of above-mentioned not village pseudomonas in agriculture production.
Implementation procedure of the present invention is as follows:
Not village pseudomonas ( pseudomonas vranovensis) ZSW-3-5-1a, be deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica) on January 20th, 2014, CGMCC No. 8750.
The present invention, when specifically using, is be made into microbiobacterial agent.
Containing not village pseudomonas ( pseudomonas vranovensis) preparation method of ZSW-3-5-1a microbial inoculum, comprise the following steps:
1) strain fermentation
1. by frozen not village pseudomonas ZSW-3-5-1a quick-thawing under 37 DEG C of conditions, be equipped with in the test tube of 10-15ml liquid nutrient medium A according to the inoculum size access of 0.5-1%, 28-32 DEG C, quiescent culture 8-12 hour, obtain the primary seed solution of ZSW-3-5-1a;
Liquid nutrient medium A consists of: Tryptones 5 g, yeast powder 5 g, NaCl 10g, distilled water 1000ml, pH value 7.2;
2. secondary triangular flask liquid culture
Be equipped with in the triangular flask of 50-100ml liquid nutrient medium A by primary seed solution according to the inoculum size access of 3-5%, 28-32 DEG C, 150-200rpm cultivate 8-12 hour;
3. ferment tank
The one-level nutrient solution of ZSW-3-5-1a is accessed in the fermentor tank that fermention medium E is housed according to the inoculum size of 5-10% respectively, carry out fermentation culture, tank temperature 28-32 DEG C, cultivate pH6.8-7.5, within aerated culture 48-72 hour, obtain bacteria suspension, be liquid phosphorus decomposing microbial inoculum, living bacteria count can reach 2.1-5 × 10 9cFU/ml;
Fermention medium E consists of: glucose 10-15g, wheat bran 5-10g, dregs of beans 30-50g, Semen Maydis powder 3-5g, (NH 4) SO 40.2-0.4g, KH 2pO 45-10g, K 2hPO 40.1-0.2g, KCl 0.2-0.5 g, Ca 2(PO 4) 310-13 g, MgSO 47H 2o 0.25-0.5g, MgCl 26H 2o 5-10g, water 1000ml, pH 6.8 ~ 7.5;
2) straw powder, turfy soil, wheat bran and dregs of beans were pulverized 60 mesh sieves, the ratio uniform mixing being 20-30:20-30:5-10:30-50 according to mass ratio adds the fermention medium E of 30-50% again, after mixing, with cloth bag or high-temperature resistance plastice packed, 1-1.5Kg/ bag, 0.1MPa sterilizing 2-3 hour, takes out cooling and is fermentation solid state substrate;
3) by the bacteria suspension obtained in step 1) according to the dosage of 100-200ml/kg and step 2) in the fermentation solid state substrate of gained mix, the 28-32 DEG C of 5-7 days that ferments, namely obtain solid fungicide, cell density reaches 1-2 × 10 9cFU/g.
Present invention also offers the using method containing ZSW-3-5-1a microbial inoculum, concrete using method is:
Liquid bacterial agent soaks seed or plant root 2-5 hour when plant seed soaking or leaching root with 100-500 liquid bacterial agent diluent doubly, fill with root in nursery stage or vegetative period and adopt 1000-5000 liquid bacterial agent diluent 10-40ml/Kg soil dosage pouring diluent doubly, whole vegetative period fills with root 1-3 time; Solid-state microbial inoculum is as base manure and use of topdressing, and using dosage is 5-15g/Kg soil.
Described liquid bacterial agent diluent is that the liquid bacterial agent of step 1) gained and fermention medium C are mixed with gained according to the volume ratio of 1:50-100 or 1:1000-5000, and solid phosphorus decomposing microbial inoculum is the phosphorus decomposing solid fungicide of step 3) gained.
Compared with prior art, the present invention has following beneficial effect:
1) method of using plasma mutagenesis and ultraviolet mutagenesis repeatedly circular treatment, go down to posterity 15 stabilization characteristics of genetics, ensure that the stability of mutant strain;
2) phosphate solubilizing bacteria provided, compared with existing phosphate solubilizing bacteria, had both had higher solution inorganic phosphorus ability, and had had again certain solution organophosphorus and ability of dissolving potassium, can secrete indolylacetic acid, had ACC desaminase activity and waited the growth promoting plant, improve the output of farm crop;
3) the large production that microbial inoculum nutritional requirement of the present invention simply, is easily cultivated, growth cycle is short, can carry out mass-producing.
Accompanying drawing explanation
Fig. 1: 1A is the colonial morphology of ZSW-3-5-1a A on substratum; 1B is the colonial morphology of ZSW-3-5-1a on substratum B; 1C is the gramstaining photo of ZSW-3-5-1a;
Fig. 2 is the Stability Determination separating inorganic phosphorus ability in ZSW-3-5-1a succeeding generations;
Fig. 3 is ZSW-3-5-1a phylogeny figure;
Fig. 4 is ZSW-3-5-1a on the impact of soil quick-effective phosphor and quick-acting potassium content: wherein Fig. 4 A applies liquid or solid ZSW-3-5-1a microbial inoculum to the impact of soil effective K; Wherein Fig. 4 B applies liquid or solid ZSW-3-5-1a microbial inoculum to the impact of rapid available phosphorus in soil.
Embodiment
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.
The mutagenesis screening of embodiment 1 ZSW-3-5-1a
According to the bacterial strain that the screening method seed selection dissolving P capacity of phosphorus decomposing bacterial strain provided by the invention is strong, the step of preparation is as follows:
1) ZSW-3 with higher dissolving P capacity screened using laboratory from plant rhizosphere is as starting strain;
2) mutagenic and breeding
(1) single cell suspension of starting strain ZSW-3 is prepared
Be inoculated in by starting strain ZSW-3 in liquid nutrient medium A, 28-32 DEG C, 150-180rpm cultivate 8-12 hour, centrifugal, and with stroke-physiological saline solution washing, be placed in and be equipped with in the triangular flask of granulated glass sphere, vibration, makes it be dispersed into single celled bacteria suspension;
Described liquid nutrient medium A consists of: Tryptones 5-10 g, yeast powder 3-5 g, NaCl 5-10 g, distilled water 1000ml, pH value 6.8-7.5.
(2) ultraviolet mutagenesis
The bacteria suspension of step (1) gained is regulated concentration to 10 respectively 7cFU/ml, gets 0.1ml and coats on solid medium B, carry out ultraviolet mutagenesis, and the frequency of ultraviolet mutagenesis is 15W, and irradiation distance is 30cm, irradiation time 8min; 28 DEG C of quiescent culture 7 days, select single bacterium colony that 28 transparent circles are larger, carry out shaking flask to sieve again, the dissolving P capacity of each bacterial strain is measured with molybdenum blue colorimetric method, select the bacterial strain that 5 strain Decomposing phosphate activities are the highest, make shaking flask more respectively to sieve again, select the high and bacterial strain ZSW-3-5 of good stability of a strain Decomposing phosphate activity, make the mutagenesis of bacteria suspension for next step plasma body;
The step that described shake flask fermentation sieves again is: 28 the transparent circle diameter D first obtained above-mentioned separation and the higher inoculation of colony diameter d ratio, in the above-mentioned liquid nutrient medium A of 100ml, are cultivated 12 hours.Getting 5ml bacterium liquid is inoculated in the Erlenmeyer flask of the 250ml that 100ml liquid nutrient medium B is housed, 28 DEG C, 150rpm shaking table shaking culture 7 days;
Described liquid nutrient medium B consists of: glucose 10 g, Ca 3(PO 4) 25g, MgCl 26H 2o 5-8g, MgSO 47H 2o 0.25g, KCl 0.2 g, (NH 4) 2sO 40.1g, distilled water 1000mL, pH 7.0.
(3) plasma body mutagenesis
By the ZSW-3-5 bacterial strain of step (2) gained, make 10 7the bacteria suspension of CFU/ml, getting 0.1ml is spread evenly across in sterile petri dish, culture dish is put on the electrode below plasma, regulate the position of top electrode, make distance controlling between upper/lower electrode at about 3mm, regulating voltage is 3V, electric current is 0.5A, make air or argon gas discharging, obtain uniform air or argon medium barrier discharge plasma, discharge time is 3min.Immediately with stroke-physiological saline solution or phosphoric acid salt wash-out after mutagenesis, coat on solid medium B, then carry out shaking flask and sieve again, choose the strain bacterium ZSW-3-5-1 that a strain Decomposing phosphate activity is the highest, make the mutagenesis of bacteria suspension for next step; The same step of method (2) that described shaking flask is sieved again;
(4) the bacteria suspension viable count of step (3) gained is adjusted to 10 7cFU/ml, is cycled to repeat ultraviolet mutagenesis → plasma body mutagenesis 1-2 time, finally obtains the bacterial strain ZSW-3-5-1a that a strain Decomposing phosphate activity is the highest.It is separated calcium phosphate, Yelkin TTS and its corresponding original strain of phytic acid ca energy force rate and improves 129.1%, 11.01% and 79.01% respectively.ZSW-3-5-1a compares in table 1 with wild phosphorus decomposing bacterial strain ZSW-3 dissolving P capacity.
This bacterial strain has following characteristic: on solid medium A, colony colour is creamy white, circular protrusions, neat in edge, and the colonial morphology that ZSW-3-5-1a grows on solid medium A as shown in figure 1; On solid medium B, bacterium colony is less, circular, bacterium colony central flat shape, and faint yellow, have transparent circle, the colonial morphology that ZSW-3-5-1a grows on solid medium B as shown in fig. 1b; G -, tubbiness is shaft-like, aerobic or amphimicrobian, and its gramstaining photo is as shown in accompanying drawing 1C; Its suitableeest growth temperature 28 ~ 32 DEG C, the suitableeest pH value is 6.8 ~ 7.5; Separating inorganic phosphorus ability in liquid medium within B is 1030 ± 16.2 mg/l, can produce oxalic acid, lactic acid, citric acid and succsinic acid, the oxalic acid content in its fermented liquid is 32.14mg/l, and tartaric acid content is 218.23mg/L, malic acid content is 557.6mg/L, and lactic acid content is 5.57mg/L.Separating phytic acid ca ability in this bacterial strain liquid medium within C is 580 ± 10.12 mg/l, separates Yelkin TTS ability and reach 135 ± 5.42mg/l in liquid medium within D, and the ability that this bacterial strain produces indolylacetic acid is 44.6 ± 3.2mg/l, active without acc deaminase; This strain stability is good, and 9 its solution inorganic phosphorus abilities that go down to posterity are still comparatively stable, and it the results are shown in Figure 2.
Described solid medium A consists of: Tryptones 5-10 g, yeast powder 3-5 g, NaCl 5-10 g, distilled water 1000 mL, agar powder 15-20g, distilled water 1000ml, pH value 6.8-7.5.
Described solid medium B consists of: glucose 10-15 g, Ca 3(PO 4) 25-13 g, MgCl 26H 2o 5-8g, MgSO 47H 2o 0.25-0.5 g, KCl 0.2-0.5 g, (NH 4) 2sO 40.1-0.2g, agar powder 12-15 g, distilled water 1000mL, pH 6.8-7.5.
Described liquid nutrient medium C consists of: peptone 5-10g, extractum carnis 3-5g, NaCl5-10g, soybean lecithin 2-3g, distilled water 1000 ml, pH 6.8-7.0.
Described liquid nutrient medium D consists of: glucose: 10-15g, (NH 4) 2sO 40.5g, NaCl 0.3g, KCl 0.3g, MgSO 47H 2o 0.3g, FeSO 47H 2o 0.3g, MnSO 4h 2o 0.03g, phytic acid ca 5g, distilled water 1000 ml, pH 7.0-7.5.
The not village pseudomonas with dissolving P capacity provided by the invention is obtained by mutagenic breeding method, in order to identify the Phylogenetic of ZSW-3-5-1a, carried out part 16Sr RNA sequencing to it.
Concrete grammar is: adopt universal primer 27FP1(5 '-AGAGTTTGATCCTGGCTCAG-3) and 1429R(5 '-GGTTACCTTGTTACGACT T-3 ').PCR reaction system is: the dNTP 0.5 μ l of 10mMol, template DNA 1 μ l, 10 × PCR buffer 5 μ l, 25mMol MgCl 23 μ l, each 1 μ l of primer, TaqDNA polysaccharase 0.25 μ l, ddH 2o 37.5 μ l; Denaturation: 95 DEG C 3 minutes, circulation primary; Sex change: 95 DEG C 1 minute, anneal 55 DEG C 1 minute, extend 72 DEG C 2 minutes, circulate 35 times; Extend eventually: 72 DEG C 5 minutes.The agarose gel electrophoresis of 1.0% is separated, extracts target stripe, send Beijing three to win the order-checking of polygala root company.Sequencing result as shown in SEQ ID No:1,
CTGCAGTCGAGCGGATGAGAAGAGCTTGCTCTTCGATTCAGCGGCGGACGGGTGAGTAATACCTAGGAATCTGCCTGGTAGTGGGGGACAACGTTTCGAAAGGAACGCTAATACCGCATACGCCCTACGGGGGAAAGCAGGGGACCTTCGGGCCTTGCGCTATCAGATGAGCCTAGGTCGGATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGACGATCCGTAACTGGTCTGAGAGGATGATCAGTCACACTGGAACTGAGACACGGTCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGGACAATGGGCGAAAGCCTGATCCAGCCATGCCGCGTGTGTGAAGAAGGTCTTCGGATTGTAAAGCACTTTAAGTTGGGAGGAAGGGTACTTACCTAATACGTGAGTATTTTGACGTTACCGACAGAATAAGCACCGGCTAACTCTGTGCCAGCAGCCGCGGTAATACAGAGGGTGCAAGCGTTAATCGGAATTACTGGGCGTAAAGCGCGCGTAGGTGGTTCGTTAAGTTGGATGTGAAATCCCCGGGCTCAACCTGGGAACTGCATCCAAAACTGGCGAGCTAGAGTAGGGCAGAGGGTGGTGGAATTTCCTGTGTAGCGGTGAAATGCGTAGATATAGGAAGGAACACCAGTGGCGAAGGCGACCACCTGGGCTCATACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGTCAACTAGCCGTTGGAATCCTTGAGATTTTAGTGGCGCAGCTAACGCATTAAGTTGACCGCCTGGGGAGTACGGCCGCAAGGTTAAAACTCAAATGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAAACCTTACCAGGCCTTGACATGCAGAGAACTTTCCAGAGATGGATTGGTGCCTTCGGGAACTC
Choose above partial sequence input ncbi database and carry out sequence alignment, utilize MEGA 4.1 to process comparison result and set up phylogenetic tree (Fig. 3).This bacterial strain and not village pseudomonas have the homology of 99%, therefore identify ZSW-3-5-1a belong to not village pseudomonas ( pseudomonas vranovensis).
Embodiment 2 ZSW-3-5-1a separates calcium phosphate, Yelkin TTS and phytic acid ca ability and measures
Be dispensed in the triangular flask of 250ml by liquid nutrient medium B, C or D by every bottle of 100ml, sterilizing is for subsequent use.ZSW-3-5-1a is inoculated in 50ml liquid nutrient medium A, 28 DEG C, quiescent culture, after 36 hours, gets 100 μ l(liquid nutrient medium dilution A bacterium liquid, its OD600 is made to be 0.7) bacterium liquid is inoculated in above-mentioned liquid nutrient medium B, C or D, often group three is parallel, and 28 DEG C, 150rpm cultivates 7 days, get the bacterium liquid that above-mentioned shaking table is cultivated, centrifugal 10 minutes of 10000rpm, gets supernatant liquor, measures phosphorus content in supernatant liquor by the anti-method of molybdenum antimony.Experimental result shows: it is 1030 ± 16.2 mg/l that ZSW-3-5-1a separates calcium phosphate ability, and separating Yelkin TTS ability is 135 ± 5.42mg/l, and separating phytic acid ca can up to 580 ± 10.12 mg/l.
Described liquid nutrient medium A consists of: Tryptones 5 g, yeast powder 5 g, NaCl 10 g, distilled water 1000 ml, pH value 6.8-7.5.
Described liquid nutrient medium B consists of: glucose 10g, Ca 3(PO 4) 25 g, MgCl 26H 2o 5 g, MgSO 47H 2o 0.25 g, KCl 0.2 g, (NH 4) 2sO 40.1 g, distilled water 1000 ml, pH 7.0.
Described liquid nutrient medium C consists of: peptone: 10g, extractum carnis: 3g, NaCl:5g, soybean lecithin 2g, distilled water 1000 ml, pH 7.0.
Described liquid nutrient medium D consists of: glucose 10g, (NH 4) 2sO 40.5g, NaCl 0.3g, KCl 0.3g, MgSO 47H 2o 0.3g, FeSO 47H 2o 0.3g, MnSO4H 2o 0.03g, phytic acid ca 5g, distilled water 1000 ml, pH 7.0.
Embodiment 3 ZSW-3-5-1a produces the mensuration of indolylacetic acid (IAA) ability
The aseptic L-Trp that concentration is 200 μ gml-1 is added in the 250ml triangular flask that 100ml liquid nutrient medium A is housed, ZSW-3-5-1a is inoculated in 50ml liquid nutrient medium A, 28 DEG C, after 150rpm cultivates 20 hours, get 100 μ l(culture medium A dilution bacterium liquid, its OD600 is made to be 0.7) bacterium liquid is inoculated in the above-mentioned liquid nutrient medium A containing L-Trp, if three parallel, 28 DEG C, 150 rpm cultivate 72 hours, 4000 g, centrifugal 10 min, adopt the concentration of IAA in spectrophotometry supernatant liquor.Experimental result shows: it is 44.6 ± 3.2mg/l that ZSW-3-5-1a produces IAA concentration.
Embodiment 4 is containing the preparation of ZSW-3-5-1a microbial inoculum
Comprise the following steps:
1) strain fermentation
1. by frozen ZSW-3-5-1a quick-thawing under 37 DEG C of conditions, be equipped with in the test tube of 10-15ml liquid nutrient medium A according to the inoculum size access of 0.5%, 28 DEG C, quiescent culture 12 hours, obtains the primary seed solution of ZSW-3-5-1a;
2. secondary triangular flask liquid culture
By primary seed solution according to 5% inoculum size access be equipped with in the triangular flask of 100ml liquid nutrient medium A, 28 DEG C, 150rpm cultivates 12 hours;
3. ferment tank
By the one-level nutrient solution of ZSW-3-5-1a according to 5% inoculum size access in the fermentor tank that fermention medium E is housed respectively, carry out fermentation culture, tank temperature 28 DEG C, cultivate pH7.0, aerated culture 48 hours, obtain bacteria suspension, be liquid phosphorus decomposing microbial inoculum, living bacteria count can reach 5 × 10 9cFU/ml;
Described fermention medium E consists of: glucose 10g, wheat bran 5g, dregs of beans 30g, Semen Maydis powder 5g, (NH 4) SO 40.2g, KH 2pO 45g, K 2hPO 40.1g, KCl 0.2 g, Ca 2(PO 4) 310g, MgSO 47H 2o 0.25g, MgCl 26H 2o 5g, water 1000ml, p H 7.0;
2) straw powder, turfy soil, wheat bran and dregs of beans high speed disintegrator being pulverized, cross 60 mesh sieves, is the ratio uniform mixing of 30:30:10:30 according to mass percent; , then add fermention medium C according to the ratio that mass ratio is 30-50%, after mixing, with cloth bag or high-temperature resistance plastice packed, 1-1.5Kg/ bag, 0.1MPa sterilizing 2-3 hour, takes out cooling and is fermentation solid state substrate;
3) by the bacteria suspension obtained in step 1) according to the dosage of 100-200ml/kg and step 2) in the fermentation solid state substrate of gained mix, 28-32 DEG C ferments 5 days, and namely obtain solid fungicide, cell density reaches 1-2 × 10 9cFU/g.
Embodiment 5
Experiment uses the liquid bacterial agent of embodiment 4, and soil picks up from farmland, Chang'an county, Shaanxi Province comparatively deep soil, is 8.2 for examination soil pH, organic content is 10.81g/kg, total nitrogen content is 0.982g/kg, and available phosphorus contents is 17.42mg/kg, and quick-acting potassium content is 5.36mg/kg.Experiment 3 process are set altogether, each process do respectively 3 parallel.20g soil (above-mentioned soil natural air-dry after, grinding) is added respectively in each sterilized petri dishes.Process 1(CK1) be blank group, add 10ml fermention medium C in each plate respectively; Process 2(solid fungicide group) for adding ZSW-3-5-1a solid thalline group, specific practice is: add 10ml sterile distilled water in plate, and then add the solid fungicide of preparation in embodiment 2, add 1g respectively in each plate, and mix; Process 3(liquid bacterial agent group) for adding ZSW-3-5-1a liquid bacterial agent, specific practice is: by the liquid bacterial agent fermention medium C of preparation in embodiment 2 according to the dilution proportion of 1:1000, in each plate, add 10ml respectively; Above-mentioned plate is put into incubator, 28 DEG C of cultivations, after one week, each plate adds 10ml sterilized water respectively, cultivates after 2 weeks and adopts 0.5mol/l NaHCO 3lixiviate-molybdenum antimony resistance colorimetric method measures content of available phosphorus in soil, adopts the content of ammonium acetate lixiviate-flame spectrophotometric determination soil effective K.
Experimental result as shown in Figure 4, as can be seen from Figure 4, ZSW-3-5-1a liquid bacterial agent and solid fungicide all can significantly improve the content of rapid available phosphorus and available potassium in soil, ZSW-3-5-1a liquid bacterial agent makes rapid available phosphorus and quick-acting potassium content in soil add 82.86% and 154.25% than control group respectively, ZSW-3-5-1a solid fungicide makes rapid available phosphorus and quick-acting potassium content in soil add 61.33% and 156.07% than control group respectively, in addition, blank group (CK) all slightly improves than the rapid available phosphorus of primary soil and quick-acting potassium content.
Embodiment 6
Potted plant soil is identical with embodiment 5, and experiment arranges 3 process altogether, chooses corn seed of uniform size and puts into the little triangular flask of sterilizing, with 95% alcohol leaching 5min, remove alcohol, add 3% NaClO solution surface sterilizing 2 min, remove clorox, with aseptic washing 6 ~ 8 times.Experiment 3 process are set altogether, often organize establish altogether 3 parallel.Process 1(CK) be blank group; Process 2 is for adding ZSW-3-5-1a liquid bacterial agent group, and process 3 is for adding ZSW-3-5-1a solid fungicide group.Select diameter 24 cm, the every basin of flowerpot of high 35cm fills native 15 kg, above-mentioned 150 g solid fungicides used by the every basin of reinforcing body microbial inoculum group, and add liquid bacterial agent that embodiment 3 prepared by the liquid bacterial agent group fermention medium C dilution proportion according to 1:1000, every basin adds 400ml; Each process all do 3 parallel, maize planting, every basin sowing 5-6 grain seed, after emerging, select the corn that growing way is homogeneous, every basin field planting 2, watered the without phosphorus plant nutrition liquid of Holand every 2 weeks once, every basin waters 500 ml.Plant strain growth was gathered in the crops after 45 days, measure respectively corn field upper part height, stem thick, and underground part weight in wet base and dry weight, measure the content of phosphorus in maize leaf on the ground.
Described without phosphorus plant nutrition liquid consists of: Ca (NO 3) H 2o 1.18g, KCl 1.4 g, MgSO 47H 2o 0.49 g, FeCl 30.005 g, KNO 30.51g, Gibson trace element 1 ml, distilled water 1000 ml, pH 6.8 ~ 7.0.
Consisting of of Gibson liquid microelement: H 3bO 32.86 g, ZnSO 47H 2o 0.22 g, CuSO 45H 2o 0.08 g, MnSO 44H 2o 2.03 g, Na 2moO 42H 2o 1.26 g, H 2o 1000 ml.
Shown in experimental result table 2.As can be seen from Table 2: ZSW-3-5-1a significantly can promote the growth of corn, every growth indexes of corn is all better than blank group and wild strain ZSW-3.Add ZSW-3-5-1a microbial inoculum group corn field upper part fresh weight, dry weight and blade phosphorus content and increase by 3.05%, 20.14% and 56.57% respectively than blank group, add 2.69%, 16.28% and 28.1% respectively than wild strain ZSW-3.Therefore, the ZSW-3-5-1a growth-promoting ability that ZSW-3 screens after mutagenesis obviously strengthens, and has the potentiality being developed as microbial fertilizer bacterial strain.
Embodiment 7
Solid fungicide provided by the invention is used to can be used for promoting soybeans they grow.
Potted plant soil and seed treatment are all identical with embodiment 4, experiment arranges 3 process altogether, process 1(CK) be blank group, 2 (ZSW-3) are for adding the solid-state microbial inoculum group of ZSW-3 wild strain in process, process 3(ZSW-3-5-1a) for adding the solid-state microbial inoculum group of ZSW-3-5-1a.Select diameter 14 cm, the every basin of flowerpot of high 15cm fills native 1.5 kg, solid fungicide and soil are added soil according to the consumption of 10g/kg soil also fully mix, manufacturing soybean, after emerging, every basin retains 1 strain, and plant strain growth was gathered in the crops after 45 days, measure plant fresh weight and dry weight respectively, measure soybeans upper part phosphorus content.
Experimental result is as shown in table 3.Result shows: ZSW-3-5-1a significantly can promote the growth of soybean, and every growth indexes of soybean is all better than blank group and wild strain ZSW-3.Add ZSW-3-5-1a microbial inoculum group soybeans upper part fresh weight, dry weight and blade phosphorus content and increase by 17.39%, 15.48% and 21.79% respectively than blank group, add 7.89% and 15.48% respectively than wild strain ZSW-3 inoculation soybean fresh weight and dry weight.
Embodiment 8
Liquid bacterial agent provided by the invention is used to can be used for promoting the growth of leaf mustard.
Potted plant soil and Seeds preprocess are all identical with embodiment 5, and experiment is established blank group (CK1) respectively, added the dead liquid bacterial agent group (CK2) of ZSW-3-5-1a and add ZSW-3-5-1a liquid bacterial agent group.Plantation leaf mustard, after emerging, every basin field planting 3, root (with fermention medium E diluted liquid microbial inoculum) is filled with the liquid bacterial agent of 1:200, every basin adds 50ml microbial inoculum, and watered the without phosphorus plant nutrition liquid of Holand every 2 weeks once, every basin waters 50 ml, plant strain growth was gathered in the crops after 45 days, measured plant plant height, over-ground part fresh weight and dry weight, root fresh weight and dry weight and mustard leaf part phosphorus content respectively.
Experimental result is as shown in table 4.Result shows: result shows: ZSW-3-5-1a significantly can promote the growth of leaf mustard, and every growth indexes of leaf mustard is all better than blank group.Adding ZSW-3-5-1a microbial inoculum group leaf mustard over-ground part fresh weight, over-ground part dry weight, root fresh weight, root dry weight and blade phosphorus content and increase by 29.97%, 10.93%, 0%, 14% and 14.5% respectively than blank group (CK1), adding 5.64%, 2.14%, 10.99%, 32.56% and 8.4 respectively than adding the dead liquid bacterial agent group (CK2) of ZSW-3-5-1a.Therefore, ZSW-3-5-1a microbial inoculum growth promotion ability is comparatively strong, can use as microbial fertilizer, improves crop yield, promotes the Sustainable development of agricultural.
<110> Northwest University
<120> mono-kind has not village pseudomonas and the application thereof of dissolving P capacity
<160> 1
<170> PatentIn Version 2.1
<210> 1
<211> 977
<212> DNA
<213> not village pseudomonas ( pseudomonas vranovensis)
<220>
<400> 1
CTGCAGTCGA GCGGATGAGA AGAGCTTGCT CTTCGATTCA GCGGCGGACG GGTGAGTAAT 60
ACCTAGGAAT CTGCCTGGTA GTGGGGGACA ACGTTTCGAA AGGAACGCTA ATACCGCATA 120
CGCCCTACGG GGGAAAGCAG GGGACCTTCG GGCCTTGCGC TATCAGATGA GCCTAGGTCG 180
GATTAGCTAG TTGGTGAGGT AATGGCTCAC CAAGGCGACG ATCCGTAACT GGTCTGAGAG 240
GATGATCAGT CACACTGGAA CTGAGACACG GTCCAGACTC CTACGGGAGG CAGCAGTGGG 300
GAATATTGGA CAATGGGCGA AAGCCTGATC CAGCCATGCC GCGTGTGTGA AGAAGGTCTT 360
CGGATTGTAA AGCACTTTAA GTTGGGAGGA AGGGTACTTA CCTAATACGT GAGTATTTTG 420
ACGTTACCGA CAGAATAAGC ACCGGCTAAC TCTGTGCCAG CAGCCGCGGT AATACAGAGG 480
GTGCAAGCGT TAATCGGAAT TACTGGGCGT AAAGCGCGCG TAGGTGGTTC GTTAAGTTGG 540
ATGTGAAATC CCCGGGCTCA ACCTGGGAAC TGCATCCAAA ACTGGCGAGC TAGAGTAGGG 600
CAGAGGGTGG TGGAATTTCC TGTGTAGCGG TGAAATGCGT AGATATAGGA AGGAACACCA 660
GTGGCGAAGG CGACCACCTG GGCTCATACT GACACTGAGG TGCGAAAGCG TGGGGAGCAA 720
ACAGGATTAG ATACCCTGGT AGTCCACGCC GTAAACGATG TCAACTAGCC GTTGGAATCC 780
TTGAGATTTT AGTGGCGCAG CTAACGCATT AAGTTGACCG CCTGGGGAGT ACGGCCGCAA 840
GGTTAAAACT CAAATGAATT GACGGGGGCC CGCACAAGCG GTGGAGCATG TGGTTTAATT 900
CGAAGCAACG CGAAAACCTT ACCAGGCCTT GACATGCAGA GAACTTTCCA GAGATGGATT 960
GGTGCCTTCG GGAACTC 977

Claims (5)

1. there is a bacterial strain ZSW-3-5-1a for dissolving P capacity, its Classification And Nomenclature be not village pseudomonas ( pseudomonas vranovensis), be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.8750.
2. contain the microbiobacterial agent of not village pseudomonas ZSW-3-5-1a described in claim 1.
3. the preparation method of microbiobacterial agent described in claim 2, comprises the following steps:
1) strain fermentation
1. by frozen not village pseudomonas ZSW-3-5-1a quick-thawing under 37 DEG C of conditions, be equipped with in the test tube of 10-15ml liquid nutrient medium A according to the inoculum size access of 0.5-1%, 28-32 DEG C, quiescent culture 8-12 hour, obtain the primary seed solution of ZSW-3-5-1a;
Liquid nutrient medium A consists of: Tryptones 5 g, yeast powder 5 g, NaCl 10g, distilled water 1000ml, pH value 7.2;
2. secondary triangular flask liquid culture
Be equipped with in the triangular flask of 50-100ml liquid nutrient medium A by primary seed solution according to the inoculum size access of 3-5%, 28-32 DEG C, 150-200rpm cultivate 8-12 hour;
3. ferment tank
The one-level nutrient solution of ZSW-3-5-1a is accessed in the fermentor tank that fermention medium E is housed according to the inoculum size of 5-10% respectively, carry out fermentation culture, tank temperature 28-32 DEG C, cultivate pH6.8-7.5, within aerated culture 48-72 hour, obtain bacteria suspension, be liquid phosphorus decomposing microbial inoculum, living bacteria count can reach 2.1-5 × 10 9cFU/ml;
Fermention medium E consists of: glucose 10-15g, wheat bran 5-10g, dregs of beans 30-50g, Semen Maydis powder 3-5g, (NH 4) SO 40.2-0.4g, KH 2pO 45-10g, K 2hPO 40.1-0.2g, KCl 0.2-0.5 g, Ca 2(PO 4) 310-13 g, MgSO 47H 2o 0.25-0.5g, MgCl 26H 2o 5-10g, water 1000ml, pH 6.8 ~ 7.5;
2) straw powder, turfy soil, wheat bran and dregs of beans were pulverized 60 mesh sieves, the ratio uniform mixing being 20-30:20-30:5-10:30-50 according to mass ratio adds the fermention medium E of 30-50% again, after mixing, with cloth bag or high-temperature resistance plastice packed, 1-1.5Kg/ bag, 0.1MPa sterilizing 2-3 hour, takes out cooling and is fermentation solid state substrate;
3) by the bacteria suspension obtained in step 1) according to the dosage of 100-200ml/kg and step 2) in the fermentation solid state substrate of gained mix, the 28-32 DEG C of 5-7 days that ferments, namely obtain solid fungicide, cell density reaches 1-2 × 10 9cFU/g.
4. the not application of village pseudomonas in Promoting plant growth described in claim 1.
5. the using method of microbiobacterial agent described in claim 2, it is characterized in that: liquid bacterial agent soaks seed or plant root 2-5 hour when plant seed soaking or leaching root with 100-500 liquid bacterial agent diluent doubly, fill with root in nursery stage or vegetative period and adopt 1000-5000 liquid bacterial agent diluent 10-40ml/Kg soil dosage pouring diluent doubly, whole vegetative period fills with root 1-3 time; Solid-state microbial inoculum is as base manure and use of topdressing, and using dosage is 5-15g/Kg soil.
CN201410237364.1A 2014-05-30 2014-05-30 Pseudomonas vranovensis with phosphorus solubilizing capability and application thereof Pending CN104371946A (en)

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Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105112319A (en) * 2015-07-08 2015-12-02 南京农业大学 Pseudomonas phosphate dissolving bacterium Y2, bioorganic fertilizer prepared through using bacterium, and application of fertilizer
CN105838635A (en) * 2015-04-27 2016-08-10 兰州大学 Method for repairing hexavalent chromium and naphthalene combinedly polluted environment by Pseudomonas fluorescens strain LZ-4
CN106754371A (en) * 2016-12-06 2017-05-31 广州富生源环保工程有限公司 A kind of microbial bacterial agent preparation method and application for improving soil phosphorus utilization
CN109439570A (en) * 2018-10-29 2019-03-08 四川大宇中和农业科技发展有限公司 One plant of phosphorus decomposing pseudomonad and its application
CN109486702A (en) * 2018-10-23 2019-03-19 湖南杂交水稻研究中心 A kind of composite bacteria agent and preparation method thereof enhancing salt tolerance of wheat
CN110004082A (en) * 2019-04-01 2019-07-12 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) A kind of bacterium bacterial strain and application suitable for the regulation of industrialized agriculture nitrogen and phosphorus pollution
CN110106111A (en) * 2019-05-06 2019-08-09 华东理工大学 A kind of preparation method, formulation products and its application of pseudomonad preparation
CN111718874A (en) * 2020-06-19 2020-09-29 中国林业科学研究院林业研究所 Phosphorus-solubilizing bacterium RP22, fermentation product, microbial inoculum and application thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
TVRZOVA L等: "Pseudomonas moraviensis sp. nov. and Pseudomonas vranovensis sp. nov., soil bacteria isolated on nitroaromatic compounds, and emended description of Pseudomonas asplenii", 《INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY》 *

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* Cited by examiner, † Cited by third party
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CN105838635A (en) * 2015-04-27 2016-08-10 兰州大学 Method for repairing hexavalent chromium and naphthalene combinedly polluted environment by Pseudomonas fluorescens strain LZ-4
CN105838635B (en) * 2015-04-27 2019-06-14 兰州大学 Utilize the method for Pseudomonas fluorescens bacterial strain LZ-4 repairing hexavalent chromium and naphthalene combined pollution environment
CN105112319A (en) * 2015-07-08 2015-12-02 南京农业大学 Pseudomonas phosphate dissolving bacterium Y2, bioorganic fertilizer prepared through using bacterium, and application of fertilizer
CN105112319B (en) * 2015-07-08 2018-02-16 南京农业大学 One pseudomonas category phosphate-solubilizing bacteria Y2 and its biological organic fertilizer and the application of preparation
CN106754371A (en) * 2016-12-06 2017-05-31 广州富生源环保工程有限公司 A kind of microbial bacterial agent preparation method and application for improving soil phosphorus utilization
CN109486702A (en) * 2018-10-23 2019-03-19 湖南杂交水稻研究中心 A kind of composite bacteria agent and preparation method thereof enhancing salt tolerance of wheat
CN109439570A (en) * 2018-10-29 2019-03-08 四川大宇中和农业科技发展有限公司 One plant of phosphorus decomposing pseudomonad and its application
CN110004082A (en) * 2019-04-01 2019-07-12 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) A kind of bacterium bacterial strain and application suitable for the regulation of industrialized agriculture nitrogen and phosphorus pollution
CN110004082B (en) * 2019-04-01 2022-04-19 山东省科学院生态研究所(山东省科学院中日友好生物技术研究中心) Bacterial strain suitable for nitrogen and phosphorus pollution regulation and control in facility agriculture and application
CN110106111A (en) * 2019-05-06 2019-08-09 华东理工大学 A kind of preparation method, formulation products and its application of pseudomonad preparation
CN111718874A (en) * 2020-06-19 2020-09-29 中国林业科学研究院林业研究所 Phosphorus-solubilizing bacterium RP22, fermentation product, microbial inoculum and application thereof
CN111718874B (en) * 2020-06-19 2022-02-11 中国林业科学研究院林业研究所 Phosphorus-solubilizing bacterium RP22, fermentation product, microbial inoculum and application thereof

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