CN104232532B - A kind of plant endogenesis bacillus pumilus and its microbial inoculum, preparation method and application - Google Patents
A kind of plant endogenesis bacillus pumilus and its microbial inoculum, preparation method and application Download PDFInfo
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Abstract
The present invention provides a kind of bacillus pumilus and its microbial inoculum of plant endogenesis, preparation method and application.The bacillus pumilus J14 1 of the plant endogenesis（Bacillus pumilus J14‑1）, China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved in, deposit number is CGMCC No.9601.Bacterial strain J14 1 and its microbial inoculum are applied in salt tolerance of crops is improved, bacterial strain can be colonized in crop body, significantly improve the salt resistance ability of seeding corn and other crops, so as to significantly improve the germination percentage of seeding corn and other crops and biomass, solves the problems such as grain drop in production caused by the common crops salt resistance ability such as corn is poor in salinized soil.
Description
Technical field
The present invention relates to a kind of bacillus pumilus of plant endogenesis (Bacillus pumilus), further relate to utilize the bud
The microbial bacterial agent of spore bacillus production, and the plant endogenesis bacillus pumilus and microbial inoculum are in salt tolerance of crops is improved
Application.
Background technology
Saline-alkali soil refers to the soil of soil salt content too high (more than 0.3%), and the salinization of soil of soil can increase agricultural production
Cost, even result in soil and fall into disuse.According to statistics, about 5.27 hundred million mu of the saline alkali land resource area in China, wherein Saline 0.88
Hundred million mu, it is mainly distributed on North China, northwest and the Northeast.The salinization of soil makes China cause substantial amounts of grain drop in production, agriculture every year
Grain drop in production caused by the salinization of soil of field not only causes serious economic loss, and also the grain security of country is constituted sternly
Threaten again.
Administering alkaline land improving measure has physics, chemistry and biology and agricultural tillage measure etc., including water conservancy improvement is arranged
Apply and (irrigation, draining, warp, plant rice, antiseepage etc.)；Agricultural improvement measure (is leveled land, improves farming, apply soil moved in improve the original, apply fertilizer, broadcasting
Kind, crop rotation, kind interplanting etc.)；Biological modification measure (plantation salt-tolerant plant and herbage, green manure, afforestation etc.)；Chemical modifying
Measure (applies upgrade materials, such as gypsum, ardealite, calcium sulfite), and utilizes the microbial strains screened from soil
The fertilizer of formation improves soil property etc..
Substantial amounts of endogenetic bacteria, fungi and actinomyces in the plant of nature be present, these bacterial strains not but not draw
The disease of host plant is played, the energy of the anti-adverse environmental factor of host plant itself by the interaction with plant, can be improved on the contrary
Power.These plant endogenesis microbial resources are developed to have important practical significance to industrial and agricultural production.
The content of the invention
The present invention mainly out of, plant with relatively strong saline and alkaline tolerance, screens itself salt resistance ability by force and can carried
The bacterium of high crop salt tolerant alkali ability, to improve the salt resistance ability of common crops.Alkaline land improving is realized with improved soil
Microbial manure is compared, and endophytic bacterium can enter in root of the crop or stem, mutualism relation is formed with plant, by changing
Become the physiological status of plant, improve the salt resistance ability of crop itself, the use behaviour of endophytic bacterium of the invention and its microbial inoculum
It is low to make simplicity, cost.
The first technical problem that the present invention solves is to overcome the harmful effect of the salinization of soil, there is provided a kind of plant endogenesis bud
Spore bacillus, crop itself salt resistance ability can be improved, improve the common crops salt resistance ability such as corn in salinized soil it is poor and
Caused grain drop in production.
The second technical problem that the present invention solves is to provide a kind of microbial bacteria containing above-mentioned plant endogenesis bacillus
It agent, can be used in salt affected soil, improve the growth of the salt resistance ability and crop of common crops in salt affected soil.The present invention
The bacterial strain and microbial inoculum of offer can use as microbial manure in salinization of soil farmland, improve germination percentage of the crop in, life
Object amount or grain yield.
The 3rd technical problem that the present invention solves is that the short and small gemma bar category bacterium of plant endogenesis and microbial inoculum are resistance in raising crop
The application of salt ability.
Specifically, the present invention proposes following technical scheme.
A kind of the first aspect of the present invention, there is provided the bacillus pumilus J14-1 (Bacillus of plant endogenesis
pumilus J14-1；Hereinafter referred to as bacterial strain J14-1), the deposit number of bacterial strain is CGMCC No.9601.Bacterium provided by the invention
Strain J14-1, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), address：Court of Beijing
Institute of Microorganism, Academia Sinica of the positive institute 3 of area's North Star West Road 1, deposit number are CGMCC No.9601, and preservation date is
On August 26th, 2014.
Bacterial strain J14-1 is applied in salt tolerance of crops is improved.Preferably, the J14-1 bacterial strains are applied to salt affected soil,
The salt affected soil refers to the soil that concentration containing soluble-salt is 0~2.0%.The strain of i (bacillus) pumilus J14-1 can
Colonize in crop body.Preferably, the crop is corn.
Another aspect of the present invention, a kind of microbial bacterial agent, it produces to obtain using above-mentioned bacterial strains J14-1, its activity into
It is divided into above-mentioned plant endogenesis strain of i (bacillus) pumilus J14-1.
The microbial bacterial agent is applied in salt tolerance of crops is improved.Preferably, the microbial bacterial agent is applied to saline and alkaline
Soil, the salt affected soil refer to the soil that concentration containing soluble-salt is 0~2.0%.
Another aspect of the present invention, the microbial bacterial agent are obtained by the preparation method comprised the steps：
1) inclined-plane kind culture：It is oblique by test tube is inoculated in after the strain of i (bacillus) pumilus J14-1 activation described in claim 1
On face；
2) shaking flask kind culture：The obtained test tube strains of step 1) are inoculated in broth bouillon, and isothermal vibration culture is extremely
Exponential phase；
3) seeding tank inoculated and cultured：The shaking flask strain that step 2) obtains is inoculated in kind according to 10% inoculum concentration (volume ratio)
Cultivated in sub- tank to exponential phase；
4) tank culture is produced：The seeding tank strain that step 3) obtains is inoculated in production tank by 10% inoculum concentration (volume ratio)
In, temperature control is cultivated at 30~35 DEG C, obtains microbial inoculum.
Plant endogenesis bacillus pumilus J14-1 and its microbial inoculum provided by the invention can apply to concentration containing soluble-salt
For 0~2.0% soil (salt affected soil), bacterial strain can be colonized in crop body, significantly improve the salt tolerant energy of seeding corn and other crops
Power, so as to significantly improve seeding corn and other crops germination percentage, and significantly improve the biomass of crop.Use plant endogenesis of the present invention
Bacillus J14-1 and its microbial inoculum can expand the planting range of corn, improve the cultivated area of crop.
Embodiment
The present invention mainly out of, plant with relatively strong saline and alkaline tolerance, screens itself salt resistance ability by force and can carried
The bacterium of high crop salt tolerant alkali ability, to improve the salt resistance ability of common crops.The short and small bud of plant endogenesis provided by the invention
Spore bacillus J14-1 and its microbial inoculum can grow in saliferous (NaCl) measures the culture medium such as LB culture mediums for 16% in itself, containing
0.5%th, in the aqueous solution of 1.0% and 2.0% salt (NaCl) concentration, corn seed germination rate can be significantly improved.By seed
It is put into after being soaked 4 hours in microbial inoculum of the present invention, plants in the soil that salt content is 1.0%, the work such as corn can be significantly improved
Thing germination percentage, and significantly improve the biomass of crop.Bacterial strain J14-1 is used in the soil containing 2.0% soluble salt, can be improved
The survival rate of seeding corn and other crops.
Preservation information
Bacillus (Bacillus) the strain of i (bacillus) pumilus J14-1 of plant endogenesis provided by the invention
(Bacillus pumilus J14-1), is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center
(CGMCC), address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica, postcode 100101；
Deposit number is CGMCC No.9601, and preservation date is August in 2014 26.
The J14-1 bacterial strains of the present invention have following property：
1. morphological feature：Thalline is shaft-like.
2. physiological and biochemical property：Gram positive, nitrate reduction negative, arginine hydrolysis are negative；Contact
Enzyme, oxidizing ferment, esterase are positive；Do not produce H2S, indoles, using arabinose, mannose, glucose, rhamnose, cotton are not utilized
Sub- sugar；After LB medium cultures 3 days, diameter 0.5mm, white, opaque, dry tack free, the irregular bacterium colony in edge are formed.
3.16S rDNA Sequence Identifications
Using following primer：8F (5 '-AGAGTTTGATCCTGGCTCAG-3 '), 1492R (5 '-TACCTTGTTA
CGACTT-3 '), enter performing PCR amplification, amplification is to 16S rDNA genetic fragments from bacterial strain J14-1, by obtained 16S rDNA gram
Beijing Bo Aiyuanshang bio tech ltd is sent to be sequenced after the grand T-easy carriers to pGEM.16S rDNA partial sequence is shown in
Sequence table SEQ ID No：Base sequence shown in 1, found with the sequence alignment in GenBank, the 16S rDNA of J14-1 bacterial strains
Sequence and the homology highest of bacillus pumilus (Bacillus pumilus) strain sequence, up to more than 99%.
Specific steps：
One plant of Suaeda heteroptera plant is taken, Suaeda heteroptera root system is cut to several sections, then to plant surface sterilization (preferably using time chlorine
Sour sodium and 70% ethanol), clean with aseptic water washing, and collect flushing water and be coated on LB culture medium flat plates, surface after testing
After sterile, pulverized with sterile mortar, plant tissue residue is collected, in the LB culture mediums containing 2% salinity after being diluted with sterilized water
(dusty yeast 5.0g/L, peptone 10.0g/L, NaCl 20g/L, agar powder 20g/L, 1L, pH 7.0 or so being settled to water) is flat
It is coated with plate, after cultivating 7 days, the line of picking single bacterium colony preserves after purification.
The invention also discloses the microbial bacterial agent using the production of above-mentioned J14-1 bacterial strains, its active component is J14-1 bacterium
Body.
The preparation method of the microbial bacterial agent is as described below：
Production microbial inoculum technique be：Inclined-plane kind-shaking flask kind-seeding tank-production tank-product, the packaging of the product
Formulation is liquid bacterial agent or solid absorption microbial inoculum.
In one preferred embodiment, the preparation of the microbial inoculum can specifically use following steps：
(1) inclined-plane kind culture：By bacterial strain J14-1 (original seed), in high salt LB culture mediums, (composition is as follows：Dusty yeast 5g/L, egg
White peptone 10g/L, NaCl 160g/L, agar powder 20g/L, pH 7.0~7.2) cultivate on flat board, it is standby.
(2) shaking flask kind culture：Test tube kind is inoculated in 200ml broth bouillons (beef extract 3.0g/L, peptone 10.0g/
L, NaCl 5.0g/L, other are water, pH 7.0~7.2) in, constant-temperature shaking culture to exponential phase, prepare inoculation seed
Tank.
(3) seeding tank inoculated and cultured：Fermentation medium is prepared, its component and percentage by weight are：Glucose 0.8%,
(NH4)2SO41%th, K2HPO40.2%th, MgSO40.05%th, NaCl 0.01%, CaCO30.3%th, dusty yeast 0.02%, remaining is
Water, pH value 7.2~7.5,400L fermentation mediums are added into 500L seeding tanks, 121 DEG C of high pressure moist heat sterilizations, are cooled to 30 DEG C
Afterwards, the shaking flask strain that step 2) obtains will be inoculated with into seeding tank by the inoculum concentration of 10% (volumn concentration), cultivated to right
Number growth period, mixing speed are 220 revs/min, and filtrated air intake is 1：0.8 (volume ratio of air and nutrient solution).
(4) tank culture is produced：Produce (the throwing identical with fermentation medium of tank (5 tons of tankage size of production) used medium composition
4.5 tons of doses), the production tank after feeding intake is in 1.1kg/cm2Pressure under, 121 DEG C of high pressure moist heat sterilizations, 30 are cooled to after sterilizing
Below DEG C, lead to filtrated air and keep germ-free condition standby；The seed liquor of logarithmic phase will be reached by 10% (volumn concentration)
Inoculum concentration access production tank, the production tank temperature control after inoculation produce filtrated air in the incubation of tank at 30~35 DEG C
Throughput be 1：(0.6~1.2) (volume ratio of air and nutrient solution), mixing speed are 180~240 revs/min and (are preferably
240 revs/min), whole technological process incubation time is 48~60 hours；After fermentation ends thalline quantity reach 1,000,000,000/ml with
On.
(5) after the completion of fermenting, nutrient solution goes out tank and is directly distributed into liquid dosage form or use with plastic barrel or Packaging Bottle
Adsorption by peat is distributed into solid fungicide formulation with packaging bag.
Selectivity between endophyte and host be present, a kind of specific endophyte genotype needs to find correct host's base
Because type can just be successfully established symbiosis.Bacillus megaterium is related at present and bacillus licheniformis improves the salt tolerant energy of crop
The report of power, but applicable crops are only limitted to wheat, can not be applied to corn.In addition, short and small gemma is not had been reported that at present
Bacillus can improve salt tolerance of crops, particularly, not report that bacillus pumilus can improve the salt resistance ability of corn.
The present invention can improve the plant endogenesis bacillus J14-1 of salt tolerance of crops and microbial bacterial agent to be made with production
With cost it is low, easy to use the advantages of, the salt resistance abilities of seeding corn and other crops can be significantly improved.Bacterial strain provided by the invention and bacterium
Agent can use as microbial manure in salinization of soil farmland, improve germination percentage of the crop in salt affected soil, biomass or
Grain yield.The planting range of corn can be expanded using the microbial inoculum, improve the cultivated area of crop.
Following embodiment facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method, it is conventional method unless otherwise specified.Test material used in following embodiments, it is certainly unless otherwise specified
What routine biochemistry reagent shop was commercially available, specific as shown in table 1, corn seed used is ordinary maize seed.Test below
It is respectively provided with and repeats three times, results averaged.Signified salt content in following examples, it is quality hundred unless otherwise specified
Divide content, signified salt is NaCl.
The commercial source information table of 1 each reagent of table
Embodiment 1：Bacillus pumilus J14-1 acquisition and identification
First, Bacillus pumilus J14-1 acquisition
In June, 2013 is taken from one plant of Inner Mongol Ulansuhai Nur periphery Suaeda heteroptera plant, after Suaeda heteroptera root system is cut into several sections,
Surface sterilization is done with sodium hypochlorite, 70% ethanol, it is clean with aseptic water washing, and collect flushing water and be coated on LB culture medium flat plates
On, after testing after surface sterile, pulverized with sterile mortar, plant tissue residue is collected, dense containing 2% salt after being diluted with sterilized water
It is coated with the LB culture medium flat plates of degree, after cultivating 7 days, the line of picking single bacterium colony preserves after purification.
LB medium components containing 2% salinity：Dusty yeast 5.0g, peptone 10.0g and NaCl 20g, agar powder 20g,
1L, pH 7.0 or so are settled to water.
2nd, Bacillus pumilus J14-1 identification
(1) morphological feature：Thalline is shaft-like.
(2) physiological and biochemical property：Gram positive, nitrate reduction negative, arginine hydrolysis are negative；Contact
Enzyme, oxidizing ferment, esterase are positive；Do not produce H2S, indoles, using arabinose, mannose, glucose, rhamnose, cotton are not utilized
Sub- sugar；After LB medium cultures 3 days, diameter 0.5mm, white, opaque, dry tack free, the irregular bacterium colony in edge are formed.
(3) 16S rDNA Sequence Identifications
Using following primer：8F (5 '-AGA GTT TGA TCC TGG CTC AG-3 '), 1492R (5 '-TAC CTT
GTT ACG ACT T-3 '), enter performing PCR amplification, amplification is to 16S rDNA fragments from bacterial strain J14-1, by obtained 16S
RDNA send Beijing Bo Aiyuanshang bio tech ltd to be sequenced after being cloned into pGEM T-easy carriers.16S rDNA part
Sequence is shown in the SEQ ID No of sequence table：Base sequence shown in 1, found with the sequence alignment in GenBank, bacterial strain J14-1's
16S rDNA sequences and bacillus pumilus (Bacillus pumilus) homology highest, physiology, biochemical indicator and short and small bud
The reference culture of spore bacillus it is consistent, bacterial strain J14-1 is accredited as bacillus pumilus.
3rd, bacterial strain J14-1 preservation
By above qualification result, confirm the new strains that above-mentioned bacterial strains are bacillus pumilus, be named as
Bacillus pumilus J14-1, are preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation
Number it is CGMCC No.9601.
Embodiment 2：Bacterial strain J14-1 improves germination percentage of the corn seed in the aqueous solution of different salinity
Picking bacterial strain J14-1 single bacteriums are fallen within 3ml LB fluid nutrient mediums, 30 DEG C, 165 revs/min of shaken cultivations 24 hours,
Obtain fresh bacterium solution.Being inoculated into 200mL fermentation mediums by the inoculum concentration of 2% (volume ratio), (its component and percentage by weight are：
Glucose 0.8%, (NH4)2SO41%th, K2HPO40.2%th, MgSO40.05%th, NaCl 0.01%, CaCO30.3%th, dusty yeast
0.02%, remaining is water, pH value 7.2-7.5) in expand culture, 120r/min, 30 DEG C of culture 48h.
Corn seed of uniform size is divided into two groups, one group is placed in inoculation immersion 4 hours in bacterium solution, and another group is used culture
Base soaks 4 hours.The corn seed after immersion is respectively placed in the filter paper containing 0.5%, 1.0%, 2.0% salt solution after naturally dry
On, then seed is put in lucifuge quiescent culture in 28 DEG C of incubator respectively, observe its germination at 7 days.
The germination percentage of corn=(germination corn seed quantity/Tests Seeds quantity) × 100%.
As a result (table 2) shows, the corn seed after bacterial strain immersion is in the aqueous solution of 0.5~2.0% salt (NaCl) concentration
Can fast germination, its germination percentage is apparently higher than the corn seed for not being inoculated with bacillus J14-1.
The influence of germination percentage of the J14-1 bacterial strains of table 2 to corn in the salting liquid of various concentrations
Embodiment 3：Bacterial strain J14-1 improves biomass of the corn in salty soil, survival rate
Picking bacterial strain J14-1 single bacteriums are fallen within 3ml LB fluid nutrient mediums, 30 DEG C, 165 revs/min of shaken cultivations 24 hours,
Obtain fresh bacterium solution.Being inoculated into 200mL fermentation mediums by the inoculum concentration of 2% (volume ratio), (its component and percentage by weight are：
Glucose 0.8%, (NH4)2SO41%th, K2HPO40.2%th, MgSO40.05%th, NaCl 0.01%, CaCO30.3%th, dusty yeast
0.02%, remaining is water, pH value 7.2-7.5) in expand culture, 120r/min, 30 DEG C of culture 48h.
Corn seed of the same size is chosen, is put into bacterial strain J14-1 fresh bacterium solution and soaks 4 hours, control seed exists
Soaked 4 hours in LB culture mediums.Then seed is seeded into respectively in the soil that soluble-salt concentration is 0.5%, 1.0%, 2.0%
In earth.Basin alms bowl is put under sunlight and cultivated, regular watering maintains soil moisture content 20%.The growth feelings of corn are observed at 10 days
Condition.After taking part corn root to be cut to several sections simultaneously, surface sterilization is done with sodium hypochlorite, 70% ethanol, it is clean with aseptic water washing,
And collect flushing water and be coated on LB culture medium flat plates, to confirm disinfection situation；Pulverized with sterile mortar, it is residual to collect plant tissue
Slag, it is coated with after being diluted with sterilized water on the LB culture medium flat plates containing 2% salinity, after cultivating 7 days, observation bacterial strain J14-1 exists
Situation is colonized in corn root.
The results are shown in Table 3, the results showed that, bacterial strain immersion after corn seed 1.0~2.0% salt (NaCl) concentration saliferous
The speed of growth in soil is significantly faster than that the corn seed for not being inoculated with bacillus；In the soil of 2% salinity, it is not inoculated with
Corn can not grow in the soil of bacterial strain, and corn can grow after soaking seed.Simultaneously test result indicates that these bacteriums are in corn
Root in successfully colonize.
Influence of the inoculating strain of table 3 to soil maize planting
Embodiment 4 utilizes above-mentioned bacillus pumilus J14-1 production microbial bacterial agents
1st, by bacterial strain J14-1 (original seed), in high salt LB culture mediums, (composition is as follows：Dusty yeast 5g/L, peptone 10g/L,
NaCl 160g/L, agar powder 20g/L, pH 7.0~7.2) cultivate on flat board, it is standby.
2nd, the single bacterium colony on picking high salt LB culture medium flat plates is inoculated in 100ml broth bouillons (0.3 gram of beef extract, egg
White 1.0 grams of peptone, 0.5 gram of NaCl, remaining is water, pH value 7.0~7.2) in, constant-temperature shaking culture to exponential phase, prepare to connect
Kind seeding tank.
3rd, fermentation medium is prepared, its component and percentage by weight are：Glucose 0.8%, (NH4)2SO41%th,
K2HPO40.2%th, MgSO40.05%th, NaCl 0.01%, CaCO30.3%th, dusty yeast 0.02%, remaining is water, pH value 7.2~
7.5,400L fermentation mediums are added into 500L seeding tanks, 121 DEG C of high pressure moist heat sterilizations, after being cooled to 30 DEG C, by shaking flask strain
It is inoculated with into seeding tank, is cultivated to exponential phase by the inoculum concentration of 10% (volumn concentration), mixing speed is 220 revs/min,
Filtrated air intake is 1：0.8 (volume ratio of air and nutrient solution).
4th, it is identical with fermentation medium (4.5 tons of inventory) to produce tank (5 tons of tankage size of production) used medium composition, throws
Production tank after material is in 1.1kg/cm2Pressure under, 121 DEG C of high pressure moist heat sterilizations, less than 30 DEG C are cooled to after sterilizing, lead to it is sterile
Air keeps germ-free condition standby；The seed liquor for reaching logarithmic phase is accessed into production by the inoculum concentration of 10% (volumn concentration)
Tank, for the production tank temperature control after inoculation at 30~35 DEG C, the throughput for producing filtrated air in the incubation of tank is 1：1.0
(volume ratio of air and nutrient solution), mixing speed be 180~240 revs/min (being preferably 240 revs/min), whole technological process
Incubation time is 60 hours；Thalline quantity reaches 1,000,000,000/ml after fermentation ends.
5th, nutrient solution goes out tank and is directly distributed into liquid dosage form or using mud with plastic barrel or Packaging Bottle after the completion of fermenting
Charcoal absorption is distributed into solid fungicide formulation with packaging bag.
From upper embodiment, the bacillus pumilus J14-1 and its microbial inoculum of plant endogenesis provided by the invention can be carried
The survival rate of germination percentage of the crops such as high corn under the conditions of saliferous, the speed of growth or corn.Present invention successfully solves
In the present situation that the common crops such as corn can not grow in the higher soil of salt content or growing way is poor, significantly reduce production and make
Workload during, production is low with use cost, expand the soil types of the common cereal crops such as suitable planting corn with
Area, to ensuring staple food supply, safeguarding that national food security etc. is significant.
The foregoing is only presently preferred embodiments of the present invention, all equivalent changes done according to scope of the invention as claimed with
Modification, it should all belong to the covering scope of the claims in the present invention.
Claims (10)
1. a kind of bacillus pumilus J14-1 (Bacilluspumilus) of plant endogenesis, it is preserved in Chinese microorganism strain guarantor
Hide administration committee common micro-organisms center, deposit number CGMCCNo.9601, the bacillus pumilus J14-1 can be colonized
Into crop body, the crop is corn.
2. applications of the bacillus pumilus J14-1 of the plant endogenesis described in claim 1 in corn salt resistance ability is improved.
3. the bacillus pumilus J14-1 of plant endogenesis according to claim 2 answering in corn salt resistance ability is improved
With, it is characterised in that the bacillus pumilus J14-1 of the plant endogenesis is applied to salt affected soil, and the salt affected soil refers to
Concentration containing soluble-salt is 0~2.0% soil.
4. the bacillus pumilus J14-1 of the plant endogenesis according to Claims 2 or 3 is in corn salt resistance ability is improved
Using, it is characterised in that the bacillus pumilus J14-1 is colonized in Corn.
5. a kind of microbial bacterial agent, it is characterised in that it utilizes the bacillus pumilus of the plant endogenesis described in claim 1
J14-1 produces to obtain, and its active component is the bacillus pumilus J14-1 of the plant endogenesis described in claim 1；It is described short and small
Bacillus J14-1 is 1,000,000,000/more than ml.
6. microbial bacterial agent according to claim 5, the microbial inoculum utilizes the short and small bud of the plant endogenesis described in claim 1
Spore bacillus J14-1 and fermentation medium produce to obtain, and the fermentation medium contains following component in mass：Glucose 0.7
~1.0%, (NH4)2SO40.8~1.2%, K2HPO40.1~0.3%, MgSO40.03~0.07%, NaCl 0.005~
0.015%th, CaCO30.2~0.4%, dusty yeast 0.01~0.03%, remaining is water, pH value 7.2~7.5.
7. application of the microbial bacterial agent in corn salt resistance ability is improved described in claim 5 or 6.
8. application of the microbial bacterial agent in the salt resistance ability for improving corn described in claim 7, it is characterised in that described micro-
Bacteria agent is applied to salt affected soil, and the salt affected soil refers to the soil that concentration containing soluble-salt is 0~2.0%.
9. application of the microbial bacterial agent in the salt resistance ability for improving corn according to claim 7 or 8, its feature exist
In the active component bacillus pumilus J14-1 is colonized in Corn.
10. the preparation method of the microbial bacterial agent described in claim 5 or 6, it is characterised in that it is by comprising the steps
Preparation method obtains：
1) inclined-plane kind culture：Test tube slant will be inoculated in after strain of i (bacillus) pumilus J14-1 activation described in claim 1
On；
2) shaking flask kind culture：The obtained test tube strains of step 1) are inoculated in broth bouillon, isothermal vibration culture to logarithm
Growth period；
3) seeding tank inoculated and cultured：The shaking flask strain that step 2) obtains is inoculated in seeding tank according to 10 volume ratio % inoculum concentration
It is middle to cultivate to exponential phase；
4) tank culture is produced：The seeding tank strain that step 3) obtains is inoculated in production tank by 10 volume ratio % inoculum concentration,
Temperature control is cultivated at 30~35 DEG C, obtains microbial inoculum.
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| CN107099477B (en) * | 2017-06-05 | 2021-02-02 | 江苏师范大学 | Endophytic curvularia lunata with salt tolerance and application thereof |
| CN110892805B (en) * | 2019-05-29 | 2023-01-03 | 青岛农业大学 | Application of biological stimulin for improving salt tolerance of corn seed germination |
| CN110590417B (en) * | 2019-08-01 | 2021-12-17 | 宁夏医科大学 | Bacillus pumilus bacterial fertilizer and application thereof |
| CN110577911B (en) * | 2019-09-17 | 2021-03-26 | 山东农业大学 | Bacillus pumilus and application thereof |
| CN110982753B (en) * | 2019-12-27 | 2021-07-27 | 天津开发区坤禾生物技术有限公司 | Bacillus coagulans MES847, microbial inoculum, chicken feed, preparation method and application |
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| AP2015008329A0 (en) * | 2012-08-31 | 2015-03-31 | Bayer Cropscience Lp | Method of increasing abiotic stress resistance of a plant |
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