CN107502638A - A kind of preparation method of ecological marine alga biostimulant - Google Patents
A kind of preparation method of ecological marine alga biostimulant Download PDFInfo
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Abstract
The present invention relates to a kind of preparation method of ecological marine alga biostimulant, the marine alga cleaned after crushing is added in agitator tank, adds certain water, at a certain temperature stirring saccharification, marine alga filtrate and marine alga slag are obtained through press filtration, marine alga slag is added in fermentation tank and accesses saccharomycete, certain water after fermentation is added, is mixed after fermentation a period of time is sterilized with marine alga filtrate, obtain mixed liquor, corynebacterium ammoniagenes are accessed into mixed liquor, ferments after certain time, produces ecological marine alga biostimulant.The present invention has conditioning soil, promotes the effect of plant growth.
Description
Technical field
The present invention relates to a kind of preparation method of ecological marine alga biostimulant.
Background technology
Contain the natural stimulin for stimulating plant growth in marine alga.The marine alga of depths is grown on, due to residing ring
Border is cold and high salinity, therefore contains resistance material again in marine alga.By these materials extracting without destruction in marine alga, it is
The direction of current research.
At present, the extracting method of marine alga active component mainly has chemical method, Physical and bioanalysis.Chemical method is mainly
Using acid, alkali and organic solvent processing alginic cell, make cell resolution or endogenous substance solubilising, this method is simple to operate, easily
Realize, and the method that most alga fertilizer manufacturing enterprises use both at home and abroad.But chemical reagent is to the work in alginic cell
Property material composition destruction be sizable, and remain organic reagent be also potentially hazardous to environment.The original of Physical
Reason is the physical environmental condition such as control pressure, temperature, crushes alginic cell using mechanical force, and content release, the method carries
Take and not exclusively cause much to waste.Bioanalysis includes two methods of enzymic degradation and microbial fermentation, and its cardinal principle is to destroy
Alginic cell wall construction and macromolecular substances of degrading are the small-molecule substance that plant easily absorbs, the relative extraction of this method compared with
Completely.
At present using enzymic degradation and microbial fermentation processes extraction marine algae extract, the incomplete problem of extraction be present.
The content of the invention
The technical problem to be solved by the invention is to provide a kind of preparation method of ecological marine alga biostimulant, have
Soil is nursed one's health, promotes the effect of plant growth.
In order to solve the above-mentioned technical problem, the present invention uses following technical scheme：
A kind of preparation method of ecological marine alga biostimulant, it is characterised in that follow the steps below：
1) marine alga fragment, is prepared：Marine alga after cleaning is crushed, obtains marine alga fragment；
2) saccharification marine alga, is prepared：By marine alga fragment and water according to mass ratio 5~30:70~95 are fitted into agitator tank,
1~12h is stirred under the conditions of 50~100 DEG C, must be saccharified marine alga；
3) marine alga filtrate and marine alga slag, are prepared：To be saccharified marine alga press filtration, and gained filtrate is marine alga filtrate, residue obtained to be
Marine alga slag；
4) fermentation of seaweed thing, is prepared：By marine alga slag, urea, magnesium sulfate, sodium chloride, potassium dihydrogen phosphate and glucose according to
Mass ratio 95.5:1:0.5:1:1:1 is added in the fermentation tank to sterilize and mixes, and is adjusted to water content as 50~70% with water, obtains
The mass ratio of mixed culture medium, access candida utili, candida utili and mixed culture medium is 0.2~2:98~
99.8, it is 25~35 DEG C in temperature, under the conditions of oxygen content is 0.1~0.5mg/L, ferments 2~7 days, is 70~150 DEG C in temperature
Under the conditions of sterilize 20~90min, obtain fermentation of seaweed thing；
5) ecological seaweed liquid biostimulant and marine alga residue, are prepared：The marine alga filtrate that step 3) obtains is added to
Stirred and evenly mixed in the fermentation of seaweed thing that step 4) obtains, obtain mixture, corynebacterium ammoniagenes prepared by access thawing method, mixture and
The mass ratio of corynebacterium ammoniagenes prepared by thawing method is 95~99:1~5, it is 25~35 DEG C in temperature, oxygen content is 1~5mg/L
Under the conditions of, ferment 2~7 days, gained filtrate is ecological seaweed liquid biostimulant after press filtration；Filter residue is marine alga residue；
6) ecological marine alga biostimulant, is prepared：Ecological seaweed liquid biostimulant it is concentrated and spray drying after, i.e.,
Obtain ecological marine alga biostimulant；
The marine alga is that one kind in Enteromorpha, sea-tangle, sargassum, opotism and kelp or arbitrary proportion are two or more.
The preparation of thawing method corynebacterium ammoniagenes follows the steps below：
A) expansion of strain is further cultured for：Using the thalline in exponential phase as seed, it is inoculated with 10% inoculum concentration
In fluid nutrient medium, 48h is cultivated in 30 DEG C of shaking table 160r/min, after 8000r/min centrifuges 10min, expansion is obtained and is further cultured for
Strain；
B) thawing method improves the permeability of cell：Thawing method is at least repeated once, and its step is：The expansion that step 1) is obtained
It is further cultured for strain greatly to be put into -20 DEG C of refrigerator, freezes 4h, takes out, dissolve at room temperature；
PH7.2~7.4 of the fluid nutrient medium, and be made into according to following mass ratio：Beef infusion broth 1.0L, peptone
10.0g, NaCl 5.0g, agar 25.0g；
The preparation method of wherein beef infusion broth follows the steps below：Lean beef is cleaned, and chopping, weighs 550 grams of immersions
In 1375ml water, soak a night, filtering, 0.6kg/cm after filtrate packing2Sterilize 40min.
Step 1) the marine alga is Fresh Laminaria Japonica.
The mass ratio 20 of step 2) the marine alga fragment and water:80；Saccharification temperature be 65 DEG C, speed of agitator be 20~
80r/min, stir 6h.
Step 4) the mixed culture medium to water content is 60%, and the mass ratio of candida utili and mixed culture medium is
0.5:99.5, it is 30 DEG C in temperature, under the conditions of oxygen content is 0.2mg/L, ferments 6 days, is sterilized under the conditions of being 120 DEG C in temperature
20min。
The mass ratio of corynebacterium ammoniagenes prepared by step 5) mixture and thawing method is 98:2, it is 28 DEG C in temperature, it is oxygen-containing
Measure under the conditions of 3mg/L, to ferment 5 days.
The candida utili and corynebacterium ammoniagenes are purchased from Chinese industrial Microbiological Culture Collection administrative center, numbering
Respectively CICC 31170 and CICC 10168.
The present invention has following advantageous effects：
1st, the present invention uses new fresh seaweed, and containing in new fresh seaweed largely can be with the stimulin of stimulating growth and resistance thing
Matter, activity are good.
2nd, the present invention is fermented under anaerobic using the candida utili of aerobic and anaerobism, can prevent seaweed nutritive into
The loss divided, and alcohol caused by anaerobic fermentation has booster action to marine alga extraction.
3rd, the present invention is fermented under anoxic conditions using aerobic and anaerobism corynebacterium ammoniagenes, produces ATP, and ATP is a kind of
High energy phosphate compound, in cell, it can realize energy storage and exoergic with ADP mutual conversion, so as to ensure that cell items
The energy supply of vital movement, promote plant growth.It is unreasonable due to long-term fertilization, cause soil acidification, corynebacterium ammoniagenes
Reducible N03--And urea, producing ammonia makes tunning be in alkalescent, can improve acidified soil.
4th, heavy metal of the present invention has certain fixation.Contain polypeptide, alginic acid and other in fermentate of the present invention
Protide, heavy metal can make albumen, alginic acid and polypeptide inactivation, while heavy metal also can be by albumen, alginic acid and polypeptide
It is fixed, so as to prevent crop from absorbing.
5th, current soil hardening is serious, and the microorganism applied in ground is often in anaerobic condition, more can using aerobic and oxygen bacterium
Adapt to present soil environment.
6th, the permeability of corynebacterium ammoniagenes cell is improved by thawing method, ATP caused by corynebacterium ammoniagenes can be made more to hold
Extracellular, product of the product effects significantly better than undressed corynebacterium ammoniagenes is easily penetrated into from cell membrane.
Embodiment
The present invention is further illustrated with reference to instantiation.
Embodiment 1
1) marine alga fragment, is prepared：Fresh Laminaria Japonica after cleaning is crushed, obtains marine alga fragment；
2) saccharification marine alga, is prepared：By marine alga fragment and water according to mass ratio 20:80 are fitted into agitator tank, at 65 DEG C, stir
Under the conditions of mix rotating speed is 45r/min, 6h is stirred, must be saccharified marine alga；
3) marine alga filtrate and marine alga slag, are prepared：Using the filter press of 650 mesh filter clothes to the marine alga press filtration that is saccharified, gained filtrate
For marine alga filtrate, residue obtained is marine alga slag；
4) fermentation of seaweed thing, is prepared：By marine alga slag, urea, magnesium sulfate, sodium chloride, potassium dihydrogen phosphate and glucose according to
Mass ratio 95.5:1:0.5:1:1:1 is added in the fermentation tank to sterilize and mixes, and is adjusted to water content as 60% with water, must mix
The mass ratio of culture medium, access candida utili, candida utili and mixed culture medium is 0.5:99.5, it is 30 in temperature
DEG C, under the conditions of oxygen content is 0.2mg/L, ferment 6 days, sterilize 20min under the conditions of being 120 DEG C in temperature, obtains fermentation of seaweed thing；
5) ecological marine alga biostimulant and marine alga residue, are prepared：The marine alga filtrate that step 3) obtains is added to step
4) stirred and evenly mixed in the fermentation of seaweed thing obtained, obtain mixture, corynebacterium ammoniagenes prepared by access thawing method, mixture and thawing
The mass ratio of corynebacterium ammoniagenes prepared by method is 98:2, it is 28 DEG C in temperature, under the conditions of oxygen content is 3mg/L, ferments 5 days, pressure
Gained filtrate is ecological seaweed liquid biostimulant after filter；Filter residue is marine alga residue；
6) ecological marine alga biostimulant, is prepared：Ecological seaweed liquid biostimulant it is concentrated and spray drying after, i.e.,
Obtain ecological marine alga biostimulant.
The preparation of wherein thawing method corynebacterium ammoniagenes follows the steps below：
A) expansion of strain is further cultured for：Using the thalline in exponential phase as seed, it is inoculated with 10% inoculum concentration
In fluid nutrient medium, 48h is cultivated in 30 DEG C of shaking table 160r/min, after 8000r/min centrifuges 10min, expansion is obtained and is further cultured for
Strain；
B) thawing method improves the permeability of cell：In triplicate, its step is thawing method：The expansion that step 1) is obtained is again
Culture strain is put into -20 DEG C of refrigerator, freezes 4h, is taken out, is dissolved, be placed again into -20 DEG C of refrigerator at room temperature, is freezed
4h, take out, dissolve, be placed again into -20 DEG C of refrigerator at room temperature, freeze 4h, take out, dissolve at room temperature；
PH7.2~7.4 of the fluid nutrient medium, and be made into according to following mass ratio：Beef infusion broth 1.0L, peptone
10.0g, NaCl 5.0g, agar 25.0g；
The preparation method of wherein beef infusion broth follows the steps below：Lean beef is cleaned, and chopping, weighs 550 grams of immersions
In 1375ml water, soak a night, filtering, 0.6kg/cm after filtrate packing2Sterilize 40min.
Embodiment 2
1) marine alga fragment, is prepared：Fresh Laminaria Japonica after cleaning is dried, dry sea-tangle is crushed using pulverizer, obtained
Marine alga fragment；
2) saccharification marine alga, is prepared：By marine alga fragment and water according to mass ratio 20:80 are fitted into agitator tank, at 65 DEG C, stir
Under the conditions of mix rotating speed is 45r/min, 6h is stirred, must be saccharified marine alga；
3) marine alga filtrate and marine alga slag, are prepared：Using the filter press of 650 mesh filter clothes to the marine alga press filtration that is saccharified, gained filtrate
For marine alga filtrate, residue obtained is marine alga slag；
4) fermentation of seaweed thing, is prepared：By marine alga slag, urea, magnesium sulfate, sodium chloride, potassium dihydrogen phosphate and glucose according to
Mass ratio 95.5:1:0.5:1:1:1 is added in the fermentation tank to sterilize and mixes, and is adjusted to water content as 60% with water, must mix
The mass ratio of culture medium, access candida utili, candida utili and mixed culture medium is 0.5:99.5, it is 30 in temperature
DEG C, under the conditions of oxygen content is 0.2mg/L, ferment 6 days, sterilize 20min under the conditions of being 120 DEG C in temperature, obtains fermentation of seaweed thing；
5) ecological marine alga biostimulant and marine alga residue, are prepared：The marine alga filtrate that step 3) obtains is added to step
4) stirred and evenly mixed in the fermentation of seaweed thing obtained, obtain mixture, corynebacterium ammoniagenes prepared by access thawing method, mixture and thawing
The mass ratio of corynebacterium ammoniagenes prepared by method is 98:2, it is 28 DEG C in temperature, under the conditions of oxygen content is 3mg/L, ferments 5 days, pressure
Gained filtrate is ecological seaweed liquid biostimulant after filter；Filter residue is marine alga residue；
6) ecological marine alga biostimulant, is prepared：Ecological seaweed liquid biostimulant it is concentrated and spray drying after, i.e.,
Obtain ecological marine alga biostimulant.
The preparation of wherein thawing method corynebacterium ammoniagenes follows the steps below：
A) expansion of strain is further cultured for：Using the thalline in exponential phase as seed, it is inoculated with 10% inoculum concentration
In fluid nutrient medium, 48h is cultivated in 30 DEG C of shaking table 160r/min, after 8000r/min centrifuges 10min, expansion is obtained and is further cultured for
Strain；
B) thawing method improves the permeability of cell：In triplicate, its step is thawing method：The expansion that step 1) is obtained is again
Culture strain is put into -20 DEG C of refrigerator, freezes 4h, is taken out, is dissolved, be placed again into -20 DEG C of refrigerator at room temperature, is freezed
4h, take out, dissolve, be placed again into -20 DEG C of refrigerator at room temperature, freeze 4h, take out, dissolve at room temperature；
PH7.2~7.4 of the fluid nutrient medium, and be made into according to following mass ratio：Beef infusion broth 1.0L, peptone
10.0g, NaCl 5.0g, agar 25.0g；
The preparation method of wherein beef infusion broth follows the steps below：Lean beef is cleaned, and chopping, weighs 550 grams of immersions
In 1375ml water, soak a night, filtering, 0.6kg/cm after filtrate packing2Sterilize 40min.
Embodiment 3
1) marine alga fragment, is prepared：Enteromorpha after clean packing drying is crushed, obtains marine alga fragment；
2) saccharification marine alga, is prepared：By marine alga fragment and water according to mass ratio 20:80 are fitted into agitator tank, at 65 DEG C, stir
Under the conditions of mix rotating speed is 45r/min, 6h is stirred, must be saccharified marine alga；
3) marine alga filtrate and marine alga slag, are prepared：Using the filter press of 650 mesh filter clothes to the marine alga press filtration that is saccharified, gained filtrate
For marine alga filtrate, residue obtained is marine alga slag；
4) fermentation of seaweed thing, is prepared：By marine alga slag, urea, magnesium sulfate, sodium chloride, potassium dihydrogen phosphate and glucose according to
Mass ratio 95.5:1:0.5:1:1:1 is added in the fermentation tank to sterilize and mixes, and is adjusted to water content as 60% with water, must mix
The mass ratio of culture medium, access candida utili, candida utili and mixed culture medium is 0.5:99.5, it is 30 in temperature
DEG C, under the conditions of oxygen content is 0.2mg/L, ferment 6 days, sterilize 20min under the conditions of being 120 DEG C in temperature, obtains fermentation of seaweed thing；
5) ecological marine alga biostimulant and marine alga residue, are prepared：The marine alga filtrate that step 3) obtains is added to step
4) stirred and evenly mixed in the fermentation of seaweed thing obtained, obtain mixture, corynebacterium ammoniagenes prepared by access thawing method, mixture and thawing
The mass ratio of corynebacterium ammoniagenes prepared by method is 98:2, it is 28 DEG C in temperature, under the conditions of oxygen content is 3mg/L, ferments 5 days, pressure
Gained filtrate is ecological seaweed liquid biostimulant after filter；Filter residue is marine alga residue；
6) ecological marine alga biostimulant, is prepared：Ecological seaweed liquid biostimulant it is concentrated and spray drying after, i.e.,
Obtain ecological marine alga biostimulant.
The preparation of wherein thawing method corynebacterium ammoniagenes follows the steps below：
A) expansion of strain is further cultured for：Using the thalline in exponential phase as seed, it is inoculated with 10% inoculum concentration
In fluid nutrient medium, 48h is cultivated in 30 DEG C of shaking table 160r/min, after 8000r/min centrifuges 10min, expansion is obtained and is further cultured for
Strain；
B) thawing method improves the permeability of cell：In triplicate, its step is thawing method：The expansion that step 1) is obtained is again
Culture strain is put into -20 DEG C of refrigerator, freezes 4h, is taken out, is dissolved, be placed again into -20 DEG C of refrigerator at room temperature, is freezed
4h, take out, dissolve, be placed again into -20 DEG C of refrigerator at room temperature, freeze 4h, take out, dissolve at room temperature；
PH7.2~7.4 of the fluid nutrient medium, and be made into according to following mass ratio：Beef infusion broth 1.0L, peptone
10.0g, NaCl 5.0g, agar 25.0g；
The preparation method of wherein beef infusion broth follows the steps below：Lean beef is cleaned, and chopping, weighs 550 grams of immersions
In 1375ml water, soak a night, filtering, 0.6kg/cm after filtrate packing2Sterilize 40min.
Embodiment 4
1) marine alga fragment, is prepared：New fresh sargassum after cleaning is crushed, obtains marine alga fragment；
2) saccharification marine alga, is prepared：By marine alga fragment and water according to mass ratio 20:80 are fitted into agitator tank, at 65 DEG C, stir
Under the conditions of mix rotating speed is 45r/min, 6h is stirred, must be saccharified marine alga；
3) marine alga filtrate and marine alga slag, are prepared：Using the filter press of 650 mesh filter clothes to the marine alga press filtration that is saccharified, gained filtrate
For marine alga filtrate, residue obtained is marine alga slag；
4) fermentation of seaweed thing, is prepared：By marine alga slag, urea, magnesium sulfate, sodium chloride, potassium dihydrogen phosphate and glucose according to
Mass ratio 95.5:1:0.5:1:1:1 is added in the fermentation tank to sterilize and mixes, and is adjusted to water content as 60% with water, must mix
The mass ratio of culture medium, access candida utili, candida utili and mixed culture medium is 0.5:99.5, it is 30 in temperature
DEG C, under the conditions of oxygen content is 0.2mg/L, ferment 6 days, sterilize 20min under the conditions of being 120 DEG C in temperature, obtains fermentation of seaweed thing；
5) ecological marine alga biostimulant and marine alga residue, are prepared：The marine alga filtrate that step 3) obtains is added to step
4) stirred and evenly mixed in the fermentation of seaweed thing obtained, obtain mixture, corynebacterium ammoniagenes prepared by access thawing method, mixture and thawing
The mass ratio of corynebacterium ammoniagenes prepared by method is 98:2, it is 28 DEG C in temperature, under the conditions of oxygen content is 3mg/L, ferments 5 days, pressure
Gained filtrate is ecological seaweed liquid biostimulant after filter；Filter residue is marine alga residue；
6) ecological marine alga biostimulant, is prepared：Ecological seaweed liquid biostimulant it is concentrated and spray drying after, i.e.,
Obtain ecological marine alga biostimulant.
The preparation of wherein thawing method corynebacterium ammoniagenes follows the steps below：
A) expansion of strain is further cultured for：Using the thalline in exponential phase as seed, it is inoculated with 10% inoculum concentration
In fluid nutrient medium, 48h is cultivated in 30 DEG C of shaking table 160r/min, after 8000r/min centrifuges 10min, expansion is obtained and is further cultured for
Strain；
B) thawing method improves the permeability of cell：The repetition of thawing method is secondary, and its step is：The expansion that step 1) is obtained is again
Culture strain is put into -20 DEG C of refrigerator, freezes 4h, is taken out, is dissolved, be placed again into -20 DEG C of refrigerator at room temperature, is freezed
4h, take out, dissolve at room temperature；
PH7.2~7.4 of the fluid nutrient medium, and be made into according to following mass ratio：Beef infusion broth 1.0L, peptone
10.0g, NaCl 5.0g, agar 25.0g；
The preparation method of wherein beef infusion broth follows the steps below：Lean beef is cleaned, and chopping, weighs 550 grams of immersions
In 1375ml water, soak a night, filtering, 0.6kg/cm after filtrate packing2Sterilize 40min.
Embodiment 5
1) marine alga fragment, is prepared：New fresh sargassum after cleaning is crushed, obtains marine alga fragment；
2) saccharification marine alga, is prepared：By marine alga fragment and water according to mass ratio 10:90 are fitted into agitator tank, at 55 DEG C, stir
Under the conditions of mix rotating speed is 30r/min, 7h is stirred, must be saccharified marine alga；
3) marine alga filtrate and marine alga slag, are prepared：Using the filter press of 800 mesh filter clothes to the marine alga press filtration that is saccharified, gained filtrate
For marine alga filtrate, residue obtained is marine alga slag；
4) fermentation of seaweed thing, is prepared：By marine alga slag, urea, magnesium sulfate, sodium chloride, potassium dihydrogen phosphate and glucose according to
Mass ratio 95.5:1:0.5:1:1:1 is added in the fermentation tank to sterilize and mixes, and is adjusted to water content as 60% with water, must mix
The mass ratio of culture medium, access candida utili, candida utili and mixed culture medium is 0.7:99.3, it is 30 in temperature
DEG C, under the conditions of oxygen content is 0.4mg/L, ferment 6 days, sterilize 30min under the conditions of being 100 DEG C in temperature, obtains fermentation of seaweed thing；
5) ecological marine alga biostimulant and marine alga residue, are prepared：The marine alga filtrate that step 3) obtains is added to step
4) stirred and evenly mixed in the fermentation of seaweed thing obtained, obtain mixture, corynebacterium ammoniagenes prepared by access thawing method, mixture and thawing
The mass ratio of corynebacterium ammoniagenes prepared by method is 97:3, it is 28 DEG C in temperature, under the conditions of oxygen content is 4mg/L, ferments 5 days, pressure
Gained filtrate is ecological seaweed liquid biostimulant after filter；Filter residue is marine alga residue；
6) ecological marine alga biostimulant, is prepared：Ecological seaweed liquid biostimulant it is concentrated and spray drying after, i.e.,
Obtain ecological marine alga biostimulant.
The preparation of wherein thawing method corynebacterium ammoniagenes follows the steps below：
A) expansion of strain is further cultured for：Using the thalline in exponential phase as seed, it is inoculated with 10% inoculum concentration
In fluid nutrient medium, 48h is cultivated in 30 DEG C of shaking table 160r/min, after 8000r/min centrifuges 10min, expansion is obtained and is further cultured for
Strain；
B) thawing method improves the permeability of cell：Thawing method is repeated four times, and its step is：The expansion that step 1) is obtained is again
Culture strain is put into -20 DEG C of refrigerator, freezes 4h, is taken out, is dissolved, be placed again into -20 DEG C of refrigerator at room temperature, is freezed
4h, take out, dissolve, be placed again into -20 DEG C of refrigerator at room temperature, freeze 4h, take out, dissolve at room temperature, be placed again into -
In 20 DEG C of refrigerator, 4h is freezed, takes out, dissolves at room temperature；
PH7.2~7.4 of the fluid nutrient medium, and be made into according to following mass ratio：Beef infusion broth 1.0L, peptone
10.0g, NaCl 5.0g, agar 25.0g；
The preparation method of wherein beef infusion broth follows the steps below：Lean beef is cleaned, and chopping, weighs 550 grams of immersions
In 1375ml water, soak a night, filtering, 0.6kg/cm after filtrate packing2Sterilize 40min.
The beneficial effect of marine alga liquid of the present invention is further illustrated with reference to experimental data：
Experiment one：
1st, material to be tested is tested
1 materials and methods：
1.1 test site and experimental subjects：The yellow sweet large cherry garden in Yantai Fushan.
1.2 experiment detections：Survey width of blade, blade Determination of Chlorophyll a contents, the content of chlorophyll b, the large cherry of large cherry
Yield and large cherry sugariness.
1.3 material to be tested：For common process (only spraying water), contrast 1 (except corynebacterium ammoniagenes are direct without the processing of thawing method
Using outer, other preparation methods are consistent with embodiment 1), compared with embodiment 2 and embodiment 1 do effect.
1.4 experimental design：Experiment sets 4 processing, and 2 repetitions, respectively common process (only spraying water), contrast 1 are (except production ammonia
Bar bacterium directly uses outside without the processing of thawing method, and other preparation methods are consistent with embodiment 1), embodiment 2 and embodiment 1.
Handle in three times, using foliage-spray, diluted concentration per treatment is 200mg/kg, and every mu of spray 50kg, is for the first time large cherry
Before blooming, in processing on March 5th, 2016 once, after second of processing is beared fruit for large cherry, processing time is April 25 in 2016,
Before third time processing is large cherry harvesting, processing time is on May 7th, 2016.
This experiment is consistent with other management in addition to processing difference except testing.
2 results and analysis
Common process (only spray water), contrast 1 (in addition to corynebacterium ammoniagenes are without thawing method processing directly use, other making
Method is consistent with embodiment 1), compared with embodiment 2 and embodiment 1 handle, data are shown in Table 1.
Table 1 respectively handles the influence to width of blade, chlorophyll a, b content, large cherry yield and sugariness
| Common process | Contrast 1 | Embodiment 1 is handled | Embodiment 2 is handled | |
| Mean leaf width (cm) | 4.46 | 4.47 | 4.52 | 4.49 |
| Average Chlorophyll-a Content (mg/g) | 0.58 | 0.62 | 0.78 | 0.63 |
| Average content of chlorophyll b (mg/g) | 0.19 | 0.21 | 0.22 | 0.20 |
| Chlorophyll-a Content/content of chlorophyll b | 3.05 | 3.00 | 3.55 | 3.15 |
| Average large cherry yield (kg/ mus) | 762.5 | 786.7 | 821.4 | 792.7 |
| Average sugariness (%) | 8.4 | 8.6 | 10.5 | 8.8 |
Present invention processing as can be seen from Table 1 is significantly better than common process, the corynebacterium ammoniagenes system handled using thawing method
Standby ecological marine alga biostimulant positive effect be better than contrast 1 (except corynebacterium ammoniagenes are without the processing of thawing method directly in addition to use,
Other preparation methods are consistent with embodiment 1) effect；It is better than to use again using the extract of Fresh Laminaria Japonica extraction and dries sea-tangle
The extract of extraction, the present invention can substantially increase large cherry yield and improve large cherry quality.
Experiment two：
Laboratory experiment
1 experiment material and method
1.1 experiment material：500ml beakers, chromium chloride, caddy, the present invention and deionized water.
1.2 experimental design：Each 0.2g of chromium chloride, caddy will be taken to be dissolved separately in fill 2 of 200ml deionized waters
In 500ml beakers, respectively toward the addition 2ml present invention in two beakers.
2. experimental phenomena：Chromium chloride solvent solution is clear green solution, and solution becomes muddy after adding the present invention,
The a piece of light blue flocculation of beaker bottom after 5 minutes；Caddy solvent solution is clear blue-green solution, after adding the present invention
Solution becomes muddy, a piece of light blue flocculation of beaker bottom after 5 minutes.
It can be seen that heavy metal of the present invention has certain fixation by above-mentioned experimental phenomena, heavy metal quilt can be prevented
Crop absorbs, it is ensured that food security.
Field experiment
1st, material to be tested is tested
1 materials and methods：
1.1 test sites and experimental subjects：Yantai Fushan bar wheat paddock.
1.2 experiment detections：The content of beary metal surveyed in wheat.
1.3 material to be tested：Prepared for common organic fertilizer (dregs of beans is primary raw material) and using marine alga residue of the present invention
Organic fertilizer do effect and compare.
1.4 experimental design：Experiment sets 2 processing, 2 repetitions, respectively common organic fertilizer (dregs of beans is primary raw material)
With the organic fertilizer prepared using marine alga residue of the present invention.Applied as base fertilizer, every mu of application 160kg, 4 mu altogether, 1 mu is one
Handle unit.
This experiment is consistent with other management in addition to processing difference except testing.
2 results and analysis
Common organic fertilizer (dregs of beans is primary raw material) is compared with the organic fertilizer processing prepared using marine alga residue of the present invention,
Data are shown in Table 2
Table 2 respectively handles cadmium in medium and small wheat, chromium, lead, arsenic content
| Cadmium (%) | Chromium (%) | Lead (%) | Arsenic (%) | |
| Common fertilizer | 0.00004 | 0 | 0.00001 | 0 |
| Organic fertilizer of the present invention | 0.00001 | 0 | 0 | 0 |
The present invention can prevent heavy metal to be absorbed by crops it can be seen from above-mentioned experimental data, it is ensured that food security.
Experiment three：
Laboratory experiment
1 experiment material and method
1.1 experiment material：500ml beakers, 1+1 hydrochloric acid, dropper, the present invention and deionized water.
1.2 experimental design：4 500ml beakers are taken to add 200ml deionized waters, one (Duplicate Samples) of experiment are to be used in beaker
Dropper adds 3 to drip, and two (Duplicate Samples) of experiment are that beaker dropper adds the 3 drop present invention (pH is consistent with water), then with dropper point
3 drop 1+1 hydrochloric acid is not instilled into experiment one and experiment two, the pH of liquid in two beakers is detected with acidometer.
Two experimental results are shown in Table 3
Table 3
| Experiment one | Experiment two | |
| Sample 1pH | 1.67 | 4.27 |
| Sample 2pH | 1.73 | 4.31 |
The present invention has buffer pH effect as can be seen from Table 2, can prevent soil acidification.
Field experiment
1st, material to be tested is tested
1 materials and methods：
1.1 test sites and experimental subjects：Yantai Fushan bar Crops ' field.
1.2 experiment detections：PH after pH and tomato harvest is surveyed before tomato field is applied fertilizer.
1.3 material to be tested：For common organic liquid Water soluble fertilizer (solid content is consistent with the present invention) and the present invention.
1.4 experimental design：Experiment sets 2 processing, 2 repetitions, respectively common organic liquid Water soluble fertilizer (solid content and sheet
Invention is consistent) and the present invention.Drip irrigation is applied, every mu of each drip irrigation 5kg, and each handle after drip irrigation is divided into transplanting three times is handled for 15 days
Once, post processing is beared fruit once, once, 4 mu altogether, 1 mu is a processing unit for processing in 15 days before harvesting.
This experiment is consistent with other management in addition to processing difference except testing.
2 results and analysis
Common liq Water soluble fertilizer (solid content is consistent with the present invention) is compared with present invention processing, and data are shown in Table 2
Ph changes during table 3 is respectively handled
| Before processing pH | Contrast 1pH | Contrast 2pH | |
| Common liq organic water-soluble fertilizer | 5.87 | 5.87 | 5.87 |
| The present invention | 5.87 | 5.91 | 5.92 |
As can be seen from Table 3, the conventional present invention can improve soil pH, improve current acidified soil；Reason is production ammonia rod
The ammonia of bacillus output is alkalescent, can improve soil pH.
In view of the foregoing it is apparent that the present invention can improve fruit quality with obvious stimulation plant growth, also having prevents from making
Thing absorbs heavy metal and nurses one's health the effect of acidified soil, has soil remediation and ecological protection function.Using new fresh seaweed system
Standby ecological marine alga biostimulant effect is better than the ecological marine alga biostimulant that its marine alga dries thing preparation.
Claims (7)
1. a kind of preparation method of ecological marine alga biostimulant, it is characterised in that follow the steps below：
1）, prepare marine alga fragment：Marine alga after cleaning is crushed, obtains marine alga fragment；
2）, prepare saccharification marine alga：By marine alga fragment and water according to mass ratio 5~30:70~95 are fitted into agitator tank, 50~
1~12h is stirred under the conditions of 100 DEG C, must be saccharified marine alga；
3）, prepare marine alga filtrate and marine alga slag：To be saccharified marine alga press filtration, and gained filtrate is marine alga filtrate, and residue obtained is marine alga
Slag；
4）, prepare fermentation of seaweed thing：By marine alga slag, urea, magnesium sulfate, sodium chloride, potassium dihydrogen phosphate and glucose according to quality
Than 95.5:1:0.5:1:1:1 is added in the fermentation tank to sterilize and mixes, and is adjusted to water content as 50~70% with water, must mix training
The mass ratio of foster base, access candida utili, candida utili and mixed culture medium is 0.2~2:98~99.8, in temperature
Spend for 25~35 DEG C, under the conditions of oxygen content is 0.1~0.5mg/L, ferment 2~7 days, gone out under the conditions of being 70~150 DEG C in temperature
20~90min of bacterium, obtain fermentation of seaweed thing；
5）, prepare ecological seaweed liquid biostimulant and marine alga residue：By step 3）Obtained marine alga filtrate is added to step
4）Stirred and evenly mixed in obtained fermentation of seaweed thing, obtain mixture, corynebacterium ammoniagenes prepared by access thawing method, mixture and thawing
The mass ratio of corynebacterium ammoniagenes prepared by method is 95~99:1~5, it is 25~35 DEG C in temperature, oxygen content is 1~5mg/L conditions
Under, ferment 2~7 days, gained filtrate is ecological seaweed liquid biostimulant after press filtration；Filter residue is marine alga residue；
6）, prepare ecological marine alga biostimulant：Ecological seaweed liquid biostimulant is concentrated with after spray drying, produces life
State seaweed bio stimulin；
The marine alga is that one kind in Enteromorpha, sea-tangle, sargassum, opotism and kelp or arbitrary proportion are two or more.
2. the preparation method of ecological marine alga biostimulant as claimed in claim 1, it is characterised in that：Thawing method produces ammonia bar
The preparation of bacterium follows the steps below：
a）The expansion of strain is further cultured for：Using the thalline in exponential phase as seed, liquid is inoculated in 10% inoculum concentration
In culture medium, 48h is cultivated in 30 DEG C of shaking table 160r/min, after 8000r/min centrifuges 10min, expansion is obtained and is further cultured for strain；
b）Thawing method improves the permeability of cell：Thawing method is at least repeated once, and its step is：By step 1）Obtained expansion is again
Culture strain is put into -20 DEG C of refrigerator, freezes 4h, is taken out, is dissolved at room temperature；
PH7.2~7.4 of the fluid nutrient medium, and be made into according to following mass ratio：Beef infusion broth 1.0L, peptone
10.0g, NaCl 5.0g, agar 25.0g；
The preparation method of wherein beef infusion broth follows the steps below：Lean beef is cleaned, and chopping, is weighed 550 grams and is soaked in
In 1375ml water, soak a night, filtering, 0.6kg/cm after filtrate packing2Sterilize 40min.
3. the preparation method of ecological marine alga biostimulant as claimed in claim 1, it is characterised in that：Step 1）The marine alga
It is Fresh Laminaria Japonica.
4. the preparation method of ecological marine alga biostimulant as claimed in claim 1, it is characterised in that：Step 2）The marine alga
The mass ratio 20 of fragment and water:80；Saccharification temperature is 65 DEG C, and speed of agitator is 20~80r/min, stirs 6h.
5. the preparation method of ecological marine alga biostimulant as claimed in claim 1, it is characterised in that：Step 4）The mixing
Culture medium to water content is 60%, and the mass ratio of candida utili and mixed culture medium is 0.5:99.5, it is 30 DEG C in temperature,
Under the conditions of oxygen content is 0.2mg/L, ferment 6 days, sterilize 20min under the conditions of being 120 DEG C in temperature.
6. the preparation method of ecological marine alga biostimulant as claimed in claim 1, it is characterised in that：Step 5）Mixture and
The mass ratio of corynebacterium ammoniagenes prepared by thawing method is 98:2, it is 28 DEG C in temperature, under the conditions of oxygen content is 3mg/L, fermentation 5
My god.
7. the preparation method of the ecological marine alga biostimulant as described in claim 1 to 6 any one, it is characterised in that：Institute
State candida utili and corynebacterium ammoniagenes are purchased from Chinese industrial Microbiological Culture Collection administrative center, numbering is respectively CICC
31170 and CICC 10168.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108059535A (en) * | 2017-12-29 | 2018-05-22 | 梁涵 | A kind of production method of ecological fertilizer |
| CN108184520A (en) * | 2017-12-29 | 2018-06-22 | 曲翠平 | A kind of ecological agriculture leisure picking management mode |
| CN108220341A (en) * | 2017-12-29 | 2018-06-29 | 梁涵 | A kind of preparation method of ecology seaweed biostimulant |
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|---|---|---|---|---|
| CN101024847A (en) * | 2007-04-06 | 2007-08-29 | 山东大学 | Method for producing alcohol and feed by utilizing seaweed chemical waste material |
| CN102987077A (en) * | 2012-12-26 | 2013-03-27 | 日照超凡生物技术有限公司 | Preparation method of seaweed fermented feed |
| CN103416581A (en) * | 2013-03-16 | 2013-12-04 | 上海理工大学 | Method for producing feedstuff by using seaweed industrial residues |
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2017
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101024847A (en) * | 2007-04-06 | 2007-08-29 | 山东大学 | Method for producing alcohol and feed by utilizing seaweed chemical waste material |
| CN102987077A (en) * | 2012-12-26 | 2013-03-27 | 日照超凡生物技术有限公司 | Preparation method of seaweed fermented feed |
| CN103416581A (en) * | 2013-03-16 | 2013-12-04 | 上海理工大学 | Method for producing feedstuff by using seaweed industrial residues |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108059535A (en) * | 2017-12-29 | 2018-05-22 | 梁涵 | A kind of production method of ecological fertilizer |
| CN108184520A (en) * | 2017-12-29 | 2018-06-22 | 曲翠平 | A kind of ecological agriculture leisure picking management mode |
| CN108220341A (en) * | 2017-12-29 | 2018-06-29 | 梁涵 | A kind of preparation method of ecology seaweed biostimulant |
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Application publication date: 20171222 |