CN109897806A - One plant of bacillus megaterium for promoting the growth of crop tolerance to salt alkali and salt-soda soil microbial fertilizer special and application - Google Patents

One plant of bacillus megaterium for promoting the growth of crop tolerance to salt alkali and salt-soda soil microbial fertilizer special and application Download PDF

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CN109897806A
CN109897806A CN201910300563.5A CN201910300563A CN109897806A CN 109897806 A CN109897806 A CN 109897806A CN 201910300563 A CN201910300563 A CN 201910300563A CN 109897806 A CN109897806 A CN 109897806A
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salt
fermentation
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plant growth
plant
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CN109897806B (en
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张瑞福
陈小娟
徐志辉
荀卫兵
邵佳慧
张楠
冯海超
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Nanjing Agricultural University
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Abstract

The present invention discloses the bacillus megaterium that one plant promotes the growth of crop tolerance to salt alkali and salt-soda soil microbial fertilizer special and application, bacterial strain T4-9 belongs to Bacillus megaterium (strain Patent Deposit Designation CGMCC17334), experiments have shown that, after bacterial strain T4-9 microbial manure applies different degrees of salt affected soil, it can be improved crop tolerance to salt stress ability, increasing crop yield and can reducing soil salt content improves the physicochemical property of soil.Application of the bacterial strain T4-9 microbial manure on salt-soda soil can alleviate the injury of salt stress on crop, improve the yield of salt-soda soil crop, provide feasible program for the improvement and agricultural use in salt-soda soil.

Description

One plant of dedicated micro- life of bacillus megaterium and salt-soda soil for promoting crop tolerance to salt alkali to grow Object fertilizer and application
Technical field
The invention belongs to agricultural intensive production technologies, and in particular to one plant of plant growth-promoting rhizobacteria T4-9 and its application, Applying in salt-soda soil has the salt-soda soil microbial fertilizer special for promoting corn salt/alkali-tolerance growing plants plant growth-promoting rhizobacteria T4-9, The yield of corn can be improved, reduce alkaline land soil salt content, improve alkaline land soil physicochemical property, realize salt-soda soil Agricultural use.
Background technique
The salinization of soil is global environmental problem, and China salt-soda soil gross area is 99,130,000 hectares, accounts for about whole world salt Alkali the gross area 10%.Wherein beach saline land is widely distributed, is related to the coastline in 11 province (area) cities, China, accounts for China The 40% of the salt-soda soil gross area.Soil salinization problem is generally existing, and shows the trend got worse, gives agricultural production Bring great obstacle.China's grain demand grows to even greater heights, and cultivated land resource is nervous, solves the problems, such as alkaline land improving to country Have great importance with the demand of the people.
Plant growth-promoting rhizobacteria (PGPR) is that one kind is colonized in plant rhizosphere, and promotes plant growth to a certain extent With a kind of beneficial bacterium for resisting stress.PGPR can improve plant rhizosphere nutrient utilizability by various mechanism, generate volatilization Property gas, adjust hormone levels etc. and promote plant growth, in biotic, PGPR passes through secretion secondary metabolites (such as antibiotic, polyketides, bacteriocin and hydrolase) antagonism pathogen, can also inhibit allergenic by nutrient competition The sprouting of son, inhibits the growth of fungi, and PGPR helps plant that plant is induced to generate own system when resisting biotic sometimes Resistance is to avoid the infringement from pathogenetic bacteria, fungi, virus and nematode etc..PGPR, can under Abiotic stress conditions It is made a response by adjusting ethylene synthase speed to abiotic stress such as saline and alkaline, arid, waterlogging, temperature and pollutants.
Microbial organic fertilizer can increase the utilization rate of chemical fertilizer, improves soil property, increases soil fertility, and promote plant pair The absorption of nutrient enhances crop anti-adversity, and coordinate plant growth increases agricultural product quality and yield, and reduction agriculture is residual, promotes Utilization of waste as resource, and to eco-friendly, researching and developing salt-soda soil dedicated functionality organic microbial fertilizer, (enhancing is made The resistance to salt stress of object improves salt-soda soil crop yield) it is a feasible outlet.
Summary of the invention
The purpose of the present invention is to provide one plant of plant growth-promoting rhizobacteria T4-9, which, which has, produces ammonia, solution Phos, produces Thermophilic iron element produces the growth-promotings abilities such as IAA.
Another object of the present invention is to low, the realities that soil is difficult to be utilized for salt-soda soil crop yield in agricultural production Border problem develops salt-soda soil dedicated functionality microbial organic fertilizer, can promote salt-soda soil crop normal growth, mention The yield of high salinity ground crops and the soil physico-chemical property for improving salt-soda soil to a certain extent.
The purpose of the present invention can be achieved through the following technical solutions:
One plant of plant growth-promoting rhizobacteria T4-9, the bacterial strain belong to Bacillus megaterium, are Gram-positive Bacillus megaterium, on March 15th, 2019 in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation (Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3), deposit number CGMCC17334.
The locality different salt affected soil sample of upper plant grew well but source, extracts soil supension, trains in LB solid Dilution spread on base is supported, is isolated and purified, screening obtains the different bacillus of form.Different bacillus is inoculated in respectively Water planting Rhizosphere of Crops preliminary screening finds well-grown processing, and NaCl concentration is 140mM when corn water planting, and passes through corn It is verified with the earth culture potted plant experiment of wheat, earth culture corn and wheat potted plant experiment NaCl concentration are 200mM, finally prove inoculation bacterium Strain the T4-9 plant height of corn and wheat, overground part fresh weight when corn, wheat rhizosphere are all remarkably higher than not inoculating strain T4-9's Processing.
Salinity be 140mM water planting corn, corn growth after two weeks, Rhizosphere of Crops be inoculated with T4-9 zymocyte liquid, connect Bacteria concentration is about 107A/mL measures the content in maize leaf with environment stress related substances after three days, compared with the control group, Soluble sugar total amount (TSS), peroxidase (POD) activity significantly improve in maize leaf, malonaldehyde (MDA) and hydrogen peroxide (H2O2) content, superoxide dismutase (SOD) and catalase (CAT) activity significantly reduce.
The growth-promoting ability for measuring bacterial strain T4-9 finds that the bacterial strain has and produces ammonia, solution Phos, produces the abilities such as thermophilic iron element, really Determining bacterial strain T4-9 is the raw bacillus of rhizosphere growth-promoting, using sheep blood plate, the hemolytic reaction of bacterial strain T4-9 is measured, experiments have shown that bacterium Strain T4-9 hemolytic reaction is negative, and can be used for the development and utilization of fertilizer.
Above-mentioned plant growth-promoting rhizobacteria T4-9 is preparing the application in microbial manure.The microbial manure is preferred For with the salt-soda soil microbial fertilizer special for improving plant salt tolerance alkaline stress ability.
It is a kind of to be wrapped with the salt-soda soil microbial fertilizer special for improving plant salt tolerance alkaline stress ability, in the microbial manure Containing above-mentioned plant growth-promoting rhizobacteria T4-9.Preferably, the plant growth-promoting rhizobacteria T4-9 is in the microbial manure Content is not less than 1 × 108A spore/g.It is further preferred that nutrient content >=5%, organic matter in the microbial manure contain Amount >=45%.
Still more preferably, which is prepared using following steps:
(1) the above-mentioned plant growth-promoting rhizobacteria T4-9 molasses culture medium for being inoculated into improvement is carried out liquid fermentation, is fermented Condition are as follows: pH 7.3~7.6, cultivation temperature are 34~36 DEG C, and fermentation period is 45~50h；Ferment middle and later periods formation gemma, hair Contain bacterium or gemma amount >=1 × 10 in zymotic fluid10A/mL；
(2) it using the cow dung pig manure stalk hybrid composting after decomposed as the vehicle organics material of the microbial manure, will walk Suddenly (1) preparation plant growth-promoting rhizobacteria T4-9 fermentation liquid and it is decomposed after cow dung pig manure stalk hybrid composting progress it is secondary solid Body ferments, and plant growth-promoting rhizobacteria T4-9 fermentation liquid 30L is added in SOLID ORGANIC material per ton and is uniformly mixed, is turned over halfway Heap makes its fermentation temperature lower than 50 DEG C, and fermentation terminates after 7 days, and the viable bacteria of plant growth-promoting rhizobacteria T4-9 or gemma amount reach 1 ×108A/g or more obtains the cow dung pig manure stalk hybrid composting microbial organic fertilizer of secondary solid fermentation.
The condition for probing into bacillus T4-9 liquid fermentation is learnt through bacillus single factor test and the test of orthogonal liquid fermentation The molasses culture liquid making method of improvement used are as follows: weight percentage, molasses 5%, bean cake powder 0.72%, sodium chloride 0.5%, potassium dihydrogen phosphate 0.1%, calcium chloride 0.1%, magnesium sulfate 0.1%, ferrous sulfate 0.05%, zinc sulfate 0.05%, sulphur Sour manganese 0.05%, water 93.33%, pH value are adjusted to 7.5 ± 0.1,121 DEG C of sterilizing 20min.
It is further preferred that bacillus liquid fermentation condition are as follows: pH 7.3~7.6, cultivation temperature are 35 DEG C, and shaking table turns Speed is 180~200 revs/min, and the liquid fermentation period is 48 hours, and the fermentation middle and later periods forms gemma, viable bacteria in fermentation liquid or Gemma amount >=1 × 1010A/mL.
The vehicle organics material of the microbial manure is purchased from the organic materials in market, mixed for the cow dung pig manure stalk after decomposed Compost is closed, wherein nutrient content is greater than the 5%, content of organic matter and is greater than 45%.
Three pieces of salt are selected in Shandong Binzhou " Bohai Sea silo Wudi template model Project Areas " at the beginning of in late June, 2018 to 10 months The different salt-soda soil of alkali degree height carries out plot experiment, and varieties of plant is the corn of permanent source No.1, applies different fertilizer respectively Processing, pouring water during which is managed collectively with top dressing, consistent with other crop field plot of surrounding, is sampled after three months to corn It surveys and produces the soil physico-chemical property of soil sampling measurement simultaneously, verify the fertilizer efficiency of the microbial organic fertilizer.
The microbial organic fertilizer can be exclusively used in salt-soda soil corn yield obstacle and alkaline land improving.
Beneficial effects of the present invention:
The bacterial strain that corn salt/alkali-tolerance can be promoted to grow the present invention provides one plant, and its liquid fermentation optimum condition is explored, By the bacterium solution after bacteria liquid fermentation, microorganism is made in secondary fermentation after evenly mixing with common cow dung pig manure stalk hybrid composting Fertilizer, the microbial manure have the advantage that
1. microbial manure provided by the invention is exclusively used in improving salt-soda soil corn yield and to a certain extent conditioned soil Earth physicochemical property, the effect for not only acting as volume increase can also improve soil；
2. effective strain in microbial organic fertilizer provided by the invention is originated from salt-soda soil, there is adaptation to saline-alkali environment Property, it has both and produces ammonia, solution Phos, produce the abilities such as thermophilic iron element, and hemolytic reaction is negative, do not have harm to the mankind；
3. culture solution primary raw material needed for liquid fermentation microbial organic fertilizer early period provided by the invention is molasses, dregs of beans Powder, low raw-material cost, and improve the utilization efficiency of these materials.And under optimal conditions of fermentation, shorter fermentation period Effective bacterium number amount or spore content in bacterium solution can be made to reach aequum；
4. vehicle organics material used in microbial organic fertilizer provided by the invention is that common cow dung pig manure stalk mixes Compost, does not need the carrying of specific organic fertilizer, after secondary fermentation can normal use, the raw material for doing carrier common is easy to get into This is low.
Detailed description of the invention
Fig. 1 is that 140mMNaCl difference connects bacterium processing to the influence diagram of corn plant height
Fig. 2 is 140mM NaCl water planting corn (A), wheat (B) figure
Wherein, Mock control:0mM NaCl does not connect bacterium processing；Salt control:140mM NaCl does not connect at bacterium Reason；
T4-9:140mM NaCl is inoculated with T4-9 processing
Fig. 3 is 200mM NaCl earth culture corn (A), wheat (B) figure
Wherein, Mock control:0mM NaCl does not connect bacterium processing；Salt control:200mM NaCl does not connect at bacterium Reason；
T4-9:200mM NaCl is inoculated with T4-9 processing
Fig. 4 is 140mM NaCl maize leaf TSS, POD, MDA, SOD, CAT, H2O2Containing spirogram
Fig. 5 is single-factor fermenting experiment result
Fig. 6 is effective bacterium number amount and gemma amount under bacterial strain T4-9 optimal liquid fermentation condition
Fig. 7 is salt-soda soil field plot corn map
Wherein, T1: not applying fertilizer, T2: common fertilizer, T3: common organic fertilizer, T4:T4-9 organic fertilizer
Specific embodiment
1. soil sample acquisition and bacillus separation screening
Saline-alkali soil in all parts of the country is acquired, and carries out potted plant experiment, it is that capital sweet tea is purple that varieties of plant is distinguished on these soil Glutinous corn is spent, normal to water, harvesting corn plant after one month.The soil supension of Rhizosphere Soil, 90 DEG C of water-baths are extracted respectively Dilution spread, 37 DEG C of overnight incubations respectively isolate and purify the bacterium on plate, filter out gemma altogether on LB plate after 15min 124 plants of bacillus, glycerol tube is stored in -80 DEG C of refrigerators.
2. improving salt tolerance of crops bacterial strain screening
By in the culture dish of sterilizing, the sterilizing filter of sterile water wetting is placed in culture dish bottom after corn seed surface sterilization Paper, be protected from light as 4 DEG C refrigerator vernalization 3 days；By chitting piece as in round plastic basket, 30, seed are placed on each corbie, For plastic crate as on 1.6L keg, adding water about 1.4L to be protected from light film shading with black to contacting with seed, it is primary to change water daily, and 4 Photomask is removed after it, it is primary to change water daily morning and evening later, after a week, changes the 1/4MS nutrient solution processing of the NaCl containing 140mM into Corn, while different bacterium solutions, inoculum density 10 are inoculated in Rhizosphere of Crops respectively73 weights are respectively arranged in a/mL, each bacterial strain It is multiple.Plant height, overground part fresh weight and the root long etc. of corn are measured after 7 days, preliminary screening goes out several plants of preferable bacterial strains of effect (see figure 1)。
Corn and wheat earth culture verification experimental verification are used later, vernalization are protected from light in culture dish after the surface of the seed disinfection, wait plant It after son germination, moves into bottom and pricks in the 700mL plastic cup in hole, each plastics cup soil 1kg plants corn seed 3, wheat seed 10,12 repetitions are arranged in each bacterial strain, every other day pour deionized water 50mL, after 10 days, every other day pour containing 200mM The salt water 50mL of NaCl, and each plastic cup meets bacterium 10mL respectively, and benefit meets bacterium 3mL within the 5th day, and corn and wheat are measured after 10 days The indexs such as plant height and dry weight and fresh weight of plant seedlings.Prove that bacterial strain T4-9 is the plant height that can significantly improve corn and wheat and fresh weight (see Fig. 2 and figure 3).It is analyzed by 16sRDNA, which belongs to Bacillus megaterium, is the huge gemma bar of Gram-positive Bacterium can be creamy white smooth regular bacterium colony in 12h on 30~37 DEG C of LB solid mediums, on blood agar plate, a small number of bacterial strains Generate zone of hemolysis.On March 15th, 2019 in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation, protect Hiding CGMCC17334.
3. the measurement in plant with environment stress related substances content:
Corn seed is subjected to surface sterilization processing, is protected from light vernalization in 4 DEG C, as in corbie after germination, corbie is put in small In bucket, add water to touch corn seed be it is best, it is protected from light with black light-blocking film, it is primary to change water daily, 3-4 Photomask is removed after it, it is primary to change water daily morning and evening later, after a week, changes the 1/4MS nutrient solution processing of the NaCl containing 140mM into Corn, while bacterium solution, inoculum density 10 are inoculated in Rhizosphere of Crops respectively73 repetitions are arranged in a/mL.Corn is measured after 3 days Every content with salt stress related substances, Soluble adhesion molecule are measured with anthrone colorimetry in blade, content of hydrogen peroxide With xylenol orange method measure, remaining and with corresponding reagent box measurement.
(1) anthrone colorimetry:
1. the production of directrix curve
20mL scale test tube 6 are taken, is numbered respectively from 1~6, is separately added into 0,0.2,0.4,0.6,0.8,1.0mL 100 μ g/mL sucrose solutions, and mended with water to 2mL, sugared content is respectively 0,20,40,60,80,100 μ g in each pipe, is then pressed 0.5mL anthrone ethyl acetate reagent and the 5mL concentrated sulfuric acid are added into test tube for sequence, sufficiently vibrate, test tube is put into boiling water immediately In bath, 1min is accurately kept the temperature by pipe, cooled to room temperature after taking-up is compared with blank, its suction is surveyed under 630nm wavelength Luminosity, using sugared content as abscissa, draws standard curve, and find out normal linearity equation using absorbance as ordinate.
2. the extraction of soluble sugar
It weighs and has crushed 0.10~0.30g of sample and be put into 20mL scale test tube, 5~10mL distilled water is added, plastics are thin Film sealing, is extracted 30min (extract 2 times) in boiling water, and extracting solution filters in 25mL volumetric flask, repeated flushing test tube and residual Slag is settled to scale.
3. chromogenic assay
It draws 0.5mL sample liquid (3 repetitions) in 20mL scale test tube, adds distilled water 1.5mL, with production standard curve The step of, it is separately added into anthrone ethyl acetate reagent, concentrated sulfuric acid solution in order, develops the color and measures absorbance.By normal linearity Equation finds out the amount (μ g) of sugar, calculates the sugared content in test sample.
(2) xylenol orange method:
1. the production of standard curve
10mL centrifuge tube six are taken, 0,0.4,0.8,1.2,1.6,2.0mL10 μm of ol/L hydrogen peroxide is separately added into, adds water To 2mL, when colour developing, add 4mL working reagent to each pipe, 30 DEG C of water-baths develop the color 30min, go out to measure absorbance value in 560nm, draw Standard curve.
2. the extraction of sample
Plant tissue samples 0.5g is taken, the pre- cold acetone grinding homogenate of 2mL is added, 10000r/min is centrifuged 10min, and supernatant is fixed Hold to 3mL, as hydrogen peroxide extracting solution, 1mL taken to add 3mL extractant, mix, then plus 5mL distilled water, mix, 5000r/min It is centrifuged 1min, upper strata aqueous phase is sample extracting solution.
3. chromogenic assay
2mL sample extracting solution is taken, 4mL working reagent is added, after equally being handled by standard curve method, is surveyed at 560nm Determine absorbance value, checks in concentration of hydrogen peroxide from standard curve.
The result shows that bacterial strain T4-9 is inoculated in potting wheat and Rhizosphere of Crops, wheat and jade under Salt Strees Condition can be improved The growth indexes (plant height, overground part fresh weight) of rice, and improve soluble sugar total amount (TSS), peroxidase in maize leaf (POD) active, reduce malonaldehyde (MDA), hydrogen peroxide (H in maize leaf2O2) content, reduce superoxide dismutase (SOD), the activity of catalase (CAT) (see Fig. 4).
4. other growth-promoting abilities of bacterial strain measure
⑴NH3The measurement of generation ability
Strain to be tested access is filled in the test tube of peptone water (10g/L) culture solution, 28 DEG C, 170r min-1Oscillation training 72h is supported, 1mL Nessler ' s reagent (0.09mol/L potassium mercuric iodide and 2.5molL is added in each test tube-1Potassium hydroxide Mixed liquor), there is the bacterial strain of tan precipitate to produce ammonia positive reaction.
(2) thermophilic iron element generates the measurement of ability
Each bacterial strain point to be measured is connected in LB solid medium tablets, 28 DEG C are cultivated for 24 hours, then by CAS solid medium (liquid of blue detection containing CAS, 30.24g L-1PIPES, 0.9% agarose) topple over and be laid on LB plate, 15- 30min It observes afterwards and whether occurs yellowish or pink haloing around thallus, its thermophilic iron element ability of production of preliminary judgement.Separately by strains tested Culture solution supernatant cross film degerming after, with CAS detection liquid mix in equal volume, sufficiently react, survey OD630, light absorption value As is obtained, with double Water is steamed as control zeroing；It is mixed in equal volume with nonvaccinated culture medium with CAS detection liquid, surveys OD630, obtain light absorption value Ar.As/ The relative amount of thermophilic iron element in Ar value representative sample, thermophilic iron cellulose content measurement are repeated 3 times, and As/Ar value is lower, produces thermophilic iron element ability It is stronger.General to produce the higher bacterial strain of siderophore ability, As/Ar is lower than 0.5.
(3) IAA generates ability measurement
Strain to be tested (addition 0.5gL in Landy culture medium-1L-Trp, about 2.5mmolL-1)25℃、140r min-1Under the conditions of shaken cultivation 72h, take culture solution 1mL, 12000 × g is centrifuged 5min, takes 500 μ L supernatants to be added isometric Salkowski reagent (10.8molL-1Sulfuric acid ferric trichloride containing 4.5g), room temperature dark place is developed the color after 30min, and light is surveyed at 530nm Density value is compared with blank cultures, and makees standard curve with the corresponding optical density of pure IAA, calculates the quantum of output of IAA (mgL-1)。
(4) dissolving P capacity measures
Each bacterial strain to be measured activates in LB culture medium, takes 1mL culture solution, 6000r min-1It is centrifuged 5min, abandons supernatant, thallus It is resuspended afterwards three times with sterile water washing, strain to be tested access phosphate liquid is supported into base (NBRIP), not connecing bacterium is blank control, 28℃、170r min-1Under the conditions of shaken cultivation 7d, molybdenum antimony resistance colorimetric method quantitative determines Soluble phosphorus content and corresponding pH value.
It the results are shown in Table 1, bacterial strain T4-9 is plant growth-promoting rhizobacteria, and ammonia, solution Phos can be produced, produce thermophilic iron element, produce by having The growth-promotings ability such as IAA.
Table 1 is that bacterial strain T4-9 produces ammonia, phosphorus decomposing, produces thermophilic iron element ability
5. bacterial strain T4-9 bacillus liquid fermentation condition is explored
(1) single-factor fermenting experiment
Table 2 is the fermenting experiment arrangement of bacillus liquid
Table 3 is the carbon nitrogen content table of each substance
(2) orthogonal fermenting experiment
Table 3 is orthogonal fermenting experiment condition
Single-factor fermenting experiment result is shown in Fig. 5, filters out optimal liquid fermentation condition by single-factor fermenting experiment and carries out Orthogonal fermenting experiment, effectively bacterium number amount and gemma amount are shown in Fig. 6 under bacterial strain T4-9 optimal liquid fermentation condition, by orthogonal fermentation reality Test the molasses culture medium formula and fermentation condition of the improvement for designing bacterial strain T4-9.
The molasses culture medium preparation method of improvement are as follows: weight percentage, molasses 5%, bean cake powder 0.72%, chlorination Sodium 0.5%, potassium dihydrogen phosphate 0.1%, calcium chloride 0.1%, magnesium sulfate 0.1%, ferrous sulfate 0.05%, zinc sulfate 0.05%, Manganese sulfate 0.05%, water 93.33%, pH value are adjusted to 7.5 ± 0.1,121 DEG C of sterilizing 20min.
The plant growth-promoting rhizobacteria T4-9 molasses culture medium for being inoculated into improvement is carried out liquid fermentation, fermentation condition are as follows: pH 7.3~7.6, cultivation temperature is 35 DEG C, and shaking speed is 180~200 revs/min, and the liquid fermentation period is 48 hours, in fermentation Later period forms gemma, viable bacteria or gemma amount >=1 × 10 in fermentation liquid10A/mL.
6. prepared by microbial organic fertilizer
Organic materials is purchased from Jiangyin Lianye Biotechology Co., Ltd., for the cow dung pig manure stalk hybrid reactor after decomposed Fertilizer, wherein nutrient content is greater than the 5%, content of organic matter and is greater than 45%.By the T4-9 bacterium solution and organic materials after liquid fermentation Carry out secondary solid fermentation after evenly mixing, T4-9 bacterium solution 30L be added in organic materials per ton and is mixed and stirred for uniformly, halfway into Row turning makes its fermentation temperature lower than 50 DEG C, and fermentation terminates after 7 days, and viable bacteria or gemma amount is made to reach 1 × 108A/g or more is obtained Obtain the microbial organic fertilizer of the cow dung pig manure stalk hybrid composting of secondary solid fermentation.
7. the fertilizer efficiency of field district test verifying microbial organic fertilizer
The plot of 4th area 1 the 2nd, 3,4 furrow, the 6th plot of area 6 are chosen respectively in Bohai Sea silo Wudi template model Project Areas 5th, 6,30th area 47 Qi He plot the 2nd, 3,4 furrow, take pedotheque on each plot at random respectively, take back laboratory survey Determine physiochemical properties of soil.
Table 4 is the dose of each fertilizer
3 repetitions, random distribution is respectively set in plot experiment.
Rotary tillage is carried out after each cell applies fertilizer, is sowed after rotary tillage, is watered every two days.It is applied in the corn typhon phase Top dressing is primary.The plant height, stem thickness and dry weight and fresh weight of plant seedlings etc. of sampled measurements corn when top dressing.Sampling is carried out to corn after three months and surveys production (the results are shown in Table 6), and soil sampling measurement soil physico-chemical property (the results are shown in Table 7).
Table 6 is 3 ‰ (a), 4 ‰ (b), 5 ‰ (c) salinity plot field plot trial corn yield tables
a
b
c
Note: T1: not applying fertilizer, T2: common fertilizer, T3: common organic fertilizer, T4:T4-9 organic fertilizer
Table 7 is 3 ‰ (a), 4 ‰ (b), 5 ‰ (c) salinity plot field plot trial plot soil physico-chemical property tables
a
b
c
Note: T1: not applying fertilizer, T2: common fertilizer, T3: common organic fertilizer, T4:T4-9 organic fertilizer
" Bohai Sea silo Wudi template model Project Areas " field plot trial shows that bacterial strain T4-9 microbial manure applies not After the salt affected soil of degree, the resistance to salt stress ability of corn can be improved, increase corn yield and can reduce soil salt content Improve the physicochemical property (as shown in table 6 and table 7) of soil.Application of the bacterial strain T4-9 microbial manure on salt-soda soil can be alleviated Injury of the salt stress to corn improves the yield (see Fig. 7) of salt-soda soil corn, and providing for the improvement in salt-soda soil and agricultural use can Row scheme.

Claims (9)

  1. It is that Gram-positive is huge 1. one plant of plant growth-promoting rhizobacteria T4-9, the bacterial strain belong to Bacillus megaterium Bacterium anthracoides are protected on March 15th, 2019 in China Committee for Culture Collection of Microorganisms's common micro-organisms center's preservation Hiding CGMCC17334.
  2. 2. plant growth-promoting rhizobacteria T4-9 described in claim 1 is preparing the application in microbial manure.
  3. 3. according to the application in claim 2, it is characterised in that the microbial manure is with the raising plant salt tolerance alkali side of body The salt-soda soil microbial fertilizer special for the ability of compeling.
  4. 4. a kind of with the salt-soda soil microbial fertilizer special for improving plant salt tolerance alkaline stress ability, it is characterised in that the microorganism It include plant growth-promoting rhizobacteria T4-9 described in claim 1 in fertilizer.
  5. 5. microbial manure according to claim 4, it is characterised in that the plant growth-promoting rhizobacteria T4-9 is micro- at this Content in bio-feritlizer is not less than 1 × 108A spore/g.
  6. 6. microbial manure according to claim 4 or 5, it is characterised in that nutrient content in the microbial manure >= 5%, the content of organic matter >=45%.
  7. 7. microbial manure according to claim 4 or 5, it is characterised in that the microbial manure uses following steps system It is standby:
    (1) the plant growth-promoting rhizobacteria T4-9 described in claim 1 molasses culture medium for being inoculated into improvement is carried out liquid fermentation, Fermentation condition are as follows: pH7.3~7.6, cultivation temperature are 34~36 DEG C, and fermentation period is 45~50h；Ferment middle and later periods formation bud Spore contains bacterium or gemma amount >=1 × 10 in fermentation liquid10A/mL；
    (2) using the cow dung pig manure stalk hybrid composting after decomposed as the vehicle organics material of the microbial manure, by step (1) The plant growth-promoting rhizobacteria T4-9 fermentation liquid of preparation and it is decomposed after cow dung pig manure stalk hybrid composting carry out secondary solid fermentation, Plant growth-promoting rhizobacteria T4-9 fermentation liquid 30L is added in SOLID ORGANIC material per ton and is uniformly mixed, midway carries out turning, makes it Fermentation temperature is lower than 50 DEG C, and fermentation terminates after 7 days, and the viable bacteria of plant growth-promoting rhizobacteria T4-9 or gemma amount reach 1 × 108A/ G obtains the cow dung pig manure stalk hybrid composting microbial organic fertilizer of secondary solid fermentation.
  8. 8. microbial manure according to claim 7, which is characterized in that the molasses culture of improvement described in step (1) Base preparation method are as follows: weight percentage, molasses 5%, bean cake powder 0.72%, sodium chloride 0.5%, potassium dihydrogen phosphate 0.1%, calcium chloride 0.1%, magnesium sulfate 0.1%, ferrous sulfate 0.05%, zinc sulfate 0.05%, manganese sulfate 0.05%, water 93.33%, pH value is adjusted to 7.5 ± 0.1,121 DEG C of sterilizing 20min.
  9. 9. a kind of preparation method with the salt-soda soil microbial fertilizer special for improving plant salt tolerance alkaline stress ability, feature exist It is prepared in including the following steps:
    (1) the plant growth-promoting rhizobacteria T4-9 described in claim 1 molasses culture medium for being inoculated into improvement is carried out liquid fermentation, Fermentation condition are as follows: pH7.3~7.6, cultivation temperature are 34~36 DEG C, and fermentation period is 45~50h；Ferment middle and later periods formation bud Spore contains bacterium or gemma amount >=1 × 10 in fermentation liquid10A/mL；
    (2) using the cow dung pig manure stalk hybrid composting after decomposed as the vehicle organics material of the microbial manure, by step (1) The plant growth-promoting rhizobacteria T4-9 fermentation liquid of preparation and it is decomposed after cow dung pig manure stalk hybrid composting carry out secondary solid fermentation, Plant growth-promoting rhizobacteria T4-9 fermentation liquid 30L is added in SOLID ORGANIC material per ton and is uniformly mixed, midway carries out turning, makes it Fermentation temperature is lower than 50 DEG C, and fermentation terminates after 7 days, and the viable bacteria of plant growth-promoting rhizobacteria T4-9 or gemma amount reach 1 × 108A/g More than, obtain the cow dung pig manure stalk hybrid composting microbial organic fertilizer of secondary solid fermentation.
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Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110551656A (en) * 2019-09-10 2019-12-10 北方民族大学 Bacillus strain and preparation method and application thereof
CN110616177A (en) * 2019-10-29 2019-12-27 山东大学 Bacillus with high fermentation density and fermentation production method thereof
CN110904001A (en) * 2019-11-18 2020-03-24 中国科学院东北地理与农业生态研究所 Saline-alkali-tolerant phosphate-solubilizing wild rice rhizosphere bacterium and application thereof
CN112175880A (en) * 2020-10-21 2021-01-05 山西大地生态环境技术研究院有限公司 Saline-alkali-resistant phosphate solubilizing bacterium and application thereof
CN112723922A (en) * 2021-02-24 2021-04-30 黑龙江省农业科学院畜牧研究所 Animal manure leavening agent and bio-organic fertilizer for improving saline-alkali soil
CN113412697A (en) * 2021-07-28 2021-09-21 青岛万慧源环保科技有限公司 Improvement method of saline-alkali soil
CN113416675A (en) * 2021-06-27 2021-09-21 山东农业大学 Saline-alkali-resistant rhizosphere growth-promoting bacterium and application thereof
CN115029278A (en) * 2022-06-23 2022-09-09 河北农业大学 Bacillus megaterium and application thereof in promoting crop growth
CN116731916A (en) * 2023-06-01 2023-09-12 中化农业（临沂）研发中心有限公司 Salt-tolerant bacillus megaterium and application thereof
CN117586914A (en) * 2023-11-22 2024-02-23 上海交通大学 Salt-tolerant phosphate-dissolving nitrogen-fixing bacterium for soil and application thereof

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103409351A (en) * 2013-08-19 2013-11-27 南京农业大学 Growth promoting strain used for promoting banana growth and microbial organic fertilizer produced with same
CN104928212A (en) * 2015-06-03 2015-09-23 华南农业大学 Bacillus megaterium strain X3 and preparation method and application thereof
US20160113288A1 (en) * 2012-08-14 2016-04-28 Marrone Bio Innovations, Inc. Bacillus Megaterium as Drought/Salt Inducer

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20160113288A1 (en) * 2012-08-14 2016-04-28 Marrone Bio Innovations, Inc. Bacillus Megaterium as Drought/Salt Inducer
CN103409351A (en) * 2013-08-19 2013-11-27 南京农业大学 Growth promoting strain used for promoting banana growth and microbial organic fertilizer produced with same
CN104928212A (en) * 2015-06-03 2015-09-23 华南农业大学 Bacillus megaterium strain X3 and preparation method and application thereof

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
高亮等: "生物有机肥在盐碱地上的应用效果及其对玉米的影响", 《山西农业科学》 *

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CN110551656A (en) * 2019-09-10 2019-12-10 北方民族大学 Bacillus strain and preparation method and application thereof
CN110616177A (en) * 2019-10-29 2019-12-27 山东大学 Bacillus with high fermentation density and fermentation production method thereof
CN110616177B (en) * 2019-10-29 2022-06-14 山东大学 Bacillus with high fermentation density and fermentation production method thereof
CN110904001A (en) * 2019-11-18 2020-03-24 中国科学院东北地理与农业生态研究所 Saline-alkali-tolerant phosphate-solubilizing wild rice rhizosphere bacterium and application thereof
CN110904001B (en) * 2019-11-18 2021-06-11 中国科学院东北地理与农业生态研究所 Saline-alkali-tolerant phosphate-solubilizing wild rice rhizosphere bacterium and application thereof
CN112175880B (en) * 2020-10-21 2023-09-12 山西大地生态环境技术研究院有限公司 Salt-tolerant alkaline-hydrolysis phosphorus bacteria and application thereof
CN112175880A (en) * 2020-10-21 2021-01-05 山西大地生态环境技术研究院有限公司 Saline-alkali-resistant phosphate solubilizing bacterium and application thereof
CN112723922A (en) * 2021-02-24 2021-04-30 黑龙江省农业科学院畜牧研究所 Animal manure leavening agent and bio-organic fertilizer for improving saline-alkali soil
CN113416675A (en) * 2021-06-27 2021-09-21 山东农业大学 Saline-alkali-resistant rhizosphere growth-promoting bacterium and application thereof
CN113412697A (en) * 2021-07-28 2021-09-21 青岛万慧源环保科技有限公司 Improvement method of saline-alkali soil
CN115029278A (en) * 2022-06-23 2022-09-09 河北农业大学 Bacillus megaterium and application thereof in promoting crop growth
CN116731916A (en) * 2023-06-01 2023-09-12 中化农业（临沂）研发中心有限公司 Salt-tolerant bacillus megaterium and application thereof
CN116731916B (en) * 2023-06-01 2024-04-12 中化农业（临沂）研发中心有限公司 Salt-tolerant bacillus megaterium and application thereof
CN117586914A (en) * 2023-11-22 2024-02-23 上海交通大学 Salt-tolerant phosphate-dissolving nitrogen-fixing bacterium for soil and application thereof

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