CN105884415A - Compound alga oligosaccharide microbial fertilizer and preparation method thereof - Google Patents
Compound alga oligosaccharide microbial fertilizer and preparation method thereof Download PDFInfo
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- CN105884415A CN105884415A CN201610249929.7A CN201610249929A CN105884415A CN 105884415 A CN105884415 A CN 105884415A CN 201610249929 A CN201610249929 A CN 201610249929A CN 105884415 A CN105884415 A CN 105884415A
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention provides a compound alga oligosaccharide microbial fertilizer and a preparation method thereof and relates to a microbial fertilizer. The compound alga oligosaccharide microbial fertilizer is prepared from, by volume, an alga oligosaccharide solution 20%-80% and bacillus mucilaginosus fermented liquid 20%-80%, wherein the effective number of living bacteria is greater than or equal to 0.2 billion/mL, the alga oligosaccharide content is greater than or equal to 0.5 g/kg, and a pH value is 6.8-7.2. The preparation method comprises the steps that bacillus mucilaginosus is activated by using a solid nitrogen-free culture medium and then is inoculated to a seed culture medium to obtain primary seed fermentation liquid; the primary seed fermentation liquid is inoculated to a seed fermentation tank, and culture is performed to obtain secondary seed liquid; the second seed liquid is inoculated to a fermentation culture medium, and fermentation culture continues to prepare the bacillus mucilaginosus fermented liquid; the alga oligosaccharide solution and the bacillus mucilaginosus fermented liquid are mixed to obtain the compound alga oligosaccharide microbial fertilizer. The compound alga oligosaccharide microbial fertilizer has the effects of improving soil, improving fertilizer efficiency, adjusting crop growth, improving crop quality and the like.
Description
Technical field
The present invention relates to microbial manure, especially relate to a kind of alga oligosaccharide composite microbe fertilizer and preparation method thereof.
Background technology
On March 18th, 2015, the Ministry of Agriculture issues " to the year two thousand twenty fertilizer application amount zero growth rate action scheme ", along with change
The fertile cancellation producing the preferential policy such as the electricity charge, value-added tax, fertilizer industry is changed to general merchandise by specialty goods；Nitrogenous fertilizer,
Phosphate fertilizer production capacity surplus, fertilizer industry entirety profit level is the highest, is faced with production capacity and carries out a difficult problem for structural adjustment, and this gives
Microbial manure brings chance.Country remains microbial manure production intermediate links implementation and exempts value-added tax preferential policy, this meaning
Taste the application by promoting microbial manure.Within 2015, China's chemical fertilizer average utilization is 35.2%, and microbial manure has raising
The effect of chemical fertilizer utilization ratio, can meet the needs of chemical fertilizer " zero growth rate ", and following microbial manure will account for the 15% of fertilizer total amount
Left and right, application area more than 400,000,000 mu.
Bacillusmusilaginosiengineering (Bacillus Mucilaginosus), also known as silicate bacteria, is bacillus, its bacterium colony
Circular, colourless, transparent, protuberance, bacterium colony is smooth, moistening glossy, neat in edge.Bacillusmusilaginosiengineering can pass through it
Thalline own metabolism produces the materials such as organic acid, aminoacid, polysaccharide, hormone and promotes the growth of plant, and can decompose containing potassium
Mineral, discharge potassium and other application elements, are also equipped with molten phosphorus and fixed nitrogen function (Wu Zuowei etc., 2004), are to be widely used in
The production strain of China's microbial manure, played an important role in agricultural microbial preparation field.But, due to soil ring
The difference in border, especially at some soil such as salt-soda soil or heavy metal pollution, microorganism often cannot be survived because toleration is the highest,
Can not the most surely grow in soil, play its effect.
Alga oligosaccharide is at the straight-chain or branched-chain compound of 2～10 by the Sargassum polysaccharides degree of polymerization that obtains of degraded, is widely present in brown
In the Sargassums such as algae, red algae, green algae and cyanophyceae.Alga oligosaccharide have raising chemical fertilizer utilization ratio (Zhang Chaoxia etc., 2014),
Coordinate plant growth (Idrees et al., 2012), raising stress resistance of plant (Vera et al., 2011), minimizing phytopathy
The effect of insect pest sickness rate, plays an important role in agricultural production as new green environment protection bio-fertilizer.Alga oligosaccharide can also increase
Add soil microbe quantity, promote the change of structure of soil microbial community, it is used in microbial manure, for improving merit
Help can be played by bacterium at the colonization ability of soil.
Chinese patent CN201410359147.X discloses the preparation method of a kind of alga oligosaccharide bio-fertilizer, by Pacific Ocean heat color bacillus
(Flammeovirga Pacifica) H2 inoculates in product sugar culture-medium, centrifugal after fermentation, collects supernatant, by supernatant
After tubular membrane filters, collect filter liquor, add Protease Treatment, obtain alga oligosaccharide bio-fertilizer；This alga oligosaccharide is biological
Fertilizer can significantly improve soil available N and P potassium content and soil organic matter content, promotes the growth of Flavobacterium in soil, but to planting
The growth-promoting effect of thing does not has embodiment.Chinese patent CN201010256317.3 provides a kind of oligo-alginate fertilizer, this oligo-alginate
Can directly be prepared as plant promoting agent, or be aided with substantial amounts of element and/or trace element makes compound alginate fertilizer, Fructus Cucumidis sativi is increased
Product effect is obvious.Visible alga oligosaccharide is improving Soil structure and soil microorganism system, promotes that plant growing aspect has very well
Application effect.
Open a kind of double algae oligosaccharide granular fertilizers of Chinese patent CN201410154473.7 and preparation method thereof, are characterized in: double algaes are few
Sugar granular fertilizer component includes brown alga oligose solution and chlorella oligosaccharide solution mixed liquor, the most also containing carbamide, ammonium sulfate, ammonium chloride,
At least 2 kinds of component combinations in MAP, potassium sulfate, potassium chloride, attapulgite/bentonite, furfural dregs, probiotics；The party
Brown algae degradation solution and chlorella degradation solution, respectively by Brown algae alkaline hydrolysis, chlorella acidolysis, are then neutralized by method, prepare brown alga oligose solution and
Chlorella oligosaccharide solution, crosses colloid mill homogenizer after mixing, prepare double algae oligosaccharide living solution, then by double algae mixed liquors in stirring
Under the conditions of be directly added in granular fertilizer production process；The alga oligosaccharide used in this pair of algae oligosaccharide granular fertilizer is brown alga oligose and chlorella
Oligosaccharide mixed liquor, has degeneration-resistant diseases prevention, promotees root upgrading, the effect of enhancement fertilizer efficiency.
At present, alga oligosaccharide bio-feritlizer is combined a great number of elements and/or middle trace element mostly, the molecular weight phase of alga oligosaccharide used
Difference is very big, have impact on alga oligosaccharide action effect.And, most based on brown alga oligose and chlorella oligosaccharide, and red algae oligosaccharide,
Especially the Thallus Gracilariae oligosaccharide of the degree of polymerization 4～8 and the application being combined in microbial manure of function bacterium yet there are no relevant report and
Research.
Summary of the invention
Present invention aims to the problems referred to above that prior art exists, it is provided that preparation is simple, can directly impose on soil or leaf
Face sprays, and surely grows soil capacity by force, can effectively improve a kind of sea of soil, enhancement fertilizer efficiency, promotion plant growing, improving quality
Algae oligosaccharide composite microbe fertilizer and preparation method thereof.
Described alga oligosaccharide composite microbe fertilizer each component by volume percentage ratio is: alga oligosaccharide solution 20%～80%, gel-shaped
Fermentation of bacillus liquid 20%～80%；Wherein alga oligosaccharide content >=0.5g/kg, living bacteria count >=200,000,000/mL, pH value is
6.8～7.2.
Described bacillusmusilaginosiengineering is No. 01 bacterial strain of bacillusmusilaginosiengineering (Bacillus Mucilaginosus) three torch,
It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, address: Beijing on 07 02nd, 2010
North Star West Road, Chaoyang District 1 No. 3 Institute of Microorganism, Academia Sinica of institute, postcode: 100101, deposit number is CGMCC
No.3995。
The preparation method of described alga oligosaccharide composite microbe fertilizer, comprises the following steps:
1) after bacillusmusilaginosiengineering solid nitrogen-free agar being activated, it is inoculated in seed culture medium, obtains one-level kind
Sub-fermentation liquid；Primary seed solution is seeded in seed fermentation tank, cultivates and obtain secondary seed solution；Described bacillusmusilaginosiengineering
For No. 01 bacterial strain of bacillusmusilaginosiengineering (Bacillus Mucilaginosus) three torch, protected on 07 02nd, 2010
Being hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.3995；
2) secondary seed solution of gained being inoculated in fermentation medium, after continuing fermentation culture, prepared bacillusmusilaginosiengineering is sent out
Ferment liquid；
3) by step 2) the bacillusmusilaginosiengineering fermentation liquid collected mixes with alga oligosaccharide solution, obtains alga oligosaccharide and be combined micro-
Bio-fertilizer.
In step 1) in, described solid nitrogen-free agar consist of sucrose 5～10g/L, dipotassium hydrogen phosphate 0.3～0.8g/L,
Magnesium sulfate 0.1～0.4g/L, sodium chloride 0.15～0.35g/L, calcium carbonate 0.5～1.2g/L, agar 20g/L.PH before sterilizing
Value is 7.0～7.5；The condition of described activation culture is to cultivate 48～72h under the conditions of 28～32 DEG C；
The composition of described seed culture medium can be sucrose 5～10g/L, dipotassium hydrogen phosphate 0.3～0.8g/L, magnesium sulfate 0.1～
0.4g/L, sodium chloride 0.15～0.35g/L, calcium carbonate 0.5～1.2g/L, before sterilizing, pH value is 7.0～7.5.Described one-level
The condition of seed culture can cultivate 24～36h in 30～35 DEG C under the conditions of 150～200rpm；The bar that described secondary seed is cultivated
Part can be in seed fermentation tank, under the conditions of 30～35 DEG C, keeps tank rotating speed 125～150rpm, cultivates 24～48h.
In step 2) in, described fermentation medium consist of alga oligosaccharide solution 0～20mL/L, yeast extract 1.0～1.8g/L,
Starch 5.0～7.5g/L, bean cake powder 5.0～7.0g/L, magnesium sulfate 0.8～1.6g/L, dipotassium hydrogen phosphate 1.0～1.8g/L,
Calcium carbonate 5.0～10.0g/L, sodium chloride 0.1～0.4g/L, before sterilizing, pH value is 7.0～7.5；Described seed liquor is seeded to
In fermentation tank culture medium fermentation inoculum concentration by volume percentage ratio is seed liquor 10%；Described fermentation condition can be: rotating speed control
System 200～250rpm, by regulation rotating speed and ventilation make dissolved oxygen control at 20% (V/V), tank temperature control at 30 DEG C,
Tank pressure is maintained at 0.05～0.06MPa, and pH value controls 7.2 by the ammonia of 10% and the hydrochloric acid of 10%, fermentation time 36～
48h。
In step 3) in, described alga oligosaccharide is to utilize microorganism directly to degrade the oligosaccharide that the degree of polymerization is 4～8 of Thallus Gracilariae gained；
In described alga oligosaccharide solution, the mass percentage concentration of alga oligosaccharide is 4～5g/L；Described alga oligosaccharide solution, step 2) gained
Bacillusmusilaginosiengineering fermentation liquid by volume percentage ratio can be (20%～80%): (20%～80%).
Described alga oligosaccharide composite microbe fertilizer, living bacteria count >=200,000,000/mL, alga oligosaccharide content >=0.5g/kg, pH value is
6.8～7.2.
A kind of alga oligosaccharide composite microbe fertilizer that the present invention provides is compounded by alga oligosaccharide solution and bacillusmusilaginosiengineering fermentation liquid
Forming, preparation technology is simple.The bacterial strain that the present invention uses is bacillusmusilaginosiengineering CGMCC No.3995, has dissolving phosphor and dissolving potassium
Function, can secrete various plants hormone, promotes plant growing.And alga oligosaccharide has induction plant stress-resistance, coordinate plant growth
Metabolism isoreactivity, acts on jointly with bacillusmusilaginosiengineering, it is possible to improvement soil, enhancement fertilizer efficiency, increasing yield and improving quality.
The method have the advantages that
1, according to the regulation of NY 882-2004, the function bacterium used by the present invention being carried out ability of dissolving potassium mensuration, this function bacterium contains potassium
In the medium, available potassium relative increase reaches 25.1% to feldspar powder, has relatively strong solution potassium capability；
2, according to the regulation of NY/T 1847-2010, the function bacterium used by the present invention being carried out dissolving P capacity mensuration, this function bacterium exists
In, organophosphor culture fluid inorganic containing slightly solubility, titanium pigment relative amount increases 440mg/L and 46.9mg/L respectively, this merit
Can have the strongest dissolving P capacity by bacterium.
3, carry out producing promotion plant growth active matter mass-energy to the function bacterium used by the present invention according to the regulation of NY/T 1847-2010
Power measures, and this function bacterium can secrete phytohormone, and total amount can reach 194mg/L, has the strongest plant growth-promoting ability.
4, the function bacterium that the present invention is multiplex has extremely strong salt tolerant, resistance to heavy metal ability, high salt concentration, the consolidating of high concentration cadmium
Well-grown in state culture medium, adapts to multiple soil environment.
5, the alga oligosaccharide composite microbe fertilizer of the present invention has the effect of improvement soil；Alga oligosaccharide is conducive to soil useful micro-
Biological activity, improves the colonization ability of function bacterium, can improve Soil structure simultaneously, promotes that fertilizer is absorbed by plant.
6, the alga oligosaccharide composite microbe fertilizer preparation of the present invention is simple, has obvious increasing yield and improving quality effect, safety collar to plant
Protect, there is good promotional value.
Accompanying drawing explanation
Fig. 1 is available potassium in soils content after different administration amount alga oligosaccharide composite microbe fertilizer foliage-spray.
Fig. 2 is soil available phosphorus content after different administration amount alga oligosaccharide composite microbe fertilizer foliage-spray.
Fig. 3 is soil organic matter content after different administration amount alga oligosaccharide composite microbe fertilizer foliage-spray.
Fig. 4 is Caulis et Folium Lactucae Sativae fresh weight after different administration amount alga oligosaccharide composite microbe fertilizer foliage-spray.
In Fig. 1～4, respectively it is labeled as: matched group, i.e. fresh water spraying；4mL represents experimental group, sprays 4mL dilution 800 every time
Alga oligosaccharide composite microbe fertilizer III again, totally 2 times；6mL represents experimental group, sprays 6mL every time and dilutes the Sargassum of 800 times
Oligosaccharide composite microbe fertilizer III, totally 2 times；8mL represents experimental group, and the alga oligosaccharide every time spraying 8mL dilution 800 times is combined
Microorganism fertilizer III, totally 2 times；10mL represents experimental group, sprays 10mL every time and dilutes the alga oligosaccharide complex microorganism of 800 times
Fertile III, totally 2 times.
Detailed description of the invention
In order to be further appreciated by the present invention, below by embodiment and accompanying drawing, the present invention is elaborated, but the present invention not office
It is limited to following example.
The preparation of embodiment 1 alga oligosaccharide composite microbe fertilizer
1) by bacillusmusilaginosiengineering with solid nitrogen-free agar (sucrose 5g/L, dipotassium hydrogen phosphate 0.5g/L, magnesium sulfate 0.1g/L,
Sodium chloride 0.2g/L, calcium carbonate 0.6g/L, agar 20g/L, distilled water configures, and before sterilizing, pH value is 7.0,121 DEG C, 20min
After sterilizing use) activate after, be inoculated into seed culture medium (10g/L, dipotassium hydrogen phosphate 0.8g/L, magnesium sulfate 0.4g/L,
Sodium chloride 0.35g/L, calcium carbonate 1.0g/L, distilled water configures, and before sterilizing, pH value is 7.2,121 DEG C, makes after 20min sterilizing
With) in, under the conditions of 30 DEG C, 200rpm shaken cultivation 24h, obtain first order seed fermentation liquid；Primary seed solution is seeded to
In seed fermentation tank, at 30 DEG C, keep fermentation tank agitation revolution 150rpm, cultivate 36h cultivation and obtain secondary seed solution.
2) by step 1) secondary seed solution of gained be inoculated into fermentation medium (alga oligosaccharide solution 5mL/L, yeast extract 1.0g/L,
Starch 5.0g/L, bean cake powder 5.0g/L, magnesium sulfate 0.8g/L, dipotassium hydrogen phosphate 1.0g/L, calcium carbonate 5.0g/L, sodium chloride
0.1g/L, distilled water configures, and before sterilizing, pH value is 7.2,121 DEG C, uses after 20min sterilizing) in, inoculum concentration is by volume
Percentage ratio is the 10% of seed liquor；In sweat, fermentation tank rotating speed controls at 250rpm, is made by regulation rotating speed and ventilation
Dissolved oxygen controls at 20% (V/V), and tank temperature control is at 30 DEG C, and tank pressure is maintained at 0.05～0.06MPa, and pH value passes through 10%
Ammonia and 10% hydrochloric acid control 7.2, cultivate 48h after fermentation terminate, prepare bacillusmusilaginosiengineering fermentation liquid.
3) by step 2) the bacillusmusilaginosiengineering fermentation liquid collected mixes with alga oligosaccharide solution by volume percentage ratio 80%: 20%
Close, obtain alga oligosaccharide composite microbe fertilizer I；By step 2) the bacillusmusilaginosiengineering fermentation liquid collected is molten with alga oligosaccharide
Liquid by volume percentage ratio 60% 40% mixing, obtains alga oligosaccharide composite microbe fertilizer II；By step 2) gel-shaped collected
Fermentation of bacillus liquid and alga oligosaccharide solution by volume percentage ratio 40% 60% mixes, and obtains alga oligosaccharide composite microbe fertilizer
Ⅲ；By step 2) the bacillusmusilaginosiengineering fermentation liquid collected mixes with alga oligosaccharide solution by volume percentage ratio 20%: 80%,
Obtain alga oligosaccharide composite microbe fertilizer IV.
Embodiment 2
7 test group of EXPERIMENTAL DESIGN, are respectively as follows:
Negative control group CK: clear water；
Positive controls CK1: alga oligosaccharide solution；
Positive controls CK2: bacillusmusilaginosiengineering fermentation liquid；
Test group A: alga oligosaccharide composite microbe fertilizer I；
Experiment group B: alga oligosaccharide composite microbe fertilizer II；
Test group C: alga oligosaccharide composite microbe fertilizer III；
Test group D: alga oligosaccharide composite microbe fertilizer IV.
Soil sieves, and removes impurity removing, stone etc., mixes thoroughly, makes test soil sample, every basin dress soil 2kg.Conventional fertilizer application, with urine
Element 0.7g/kg, KH2PO4The addition of 0.5g/kg, KCl 0.085g/kg adds azophoska solution to soil sample.Select seed
Semen Brassicae campestris full, of the same size, with the hypochlorite disinfectant 15min of 1%, is rinsed well with deionized water, then spend from
Sub-water soaking 12h.By the Plantula Brassicae chinensis planting seed of dipped water in basin, after germinateing one week, the seedling selecting growing way consistent enters
Row final singling, the strain of every basin 5, sprinkling is watered so that field capacity is about 60%～70%.By molten for above-mentioned test group alga oligosaccharide
Liquid, bacillusmusilaginosiengineering fermentation liquid, alga oligosaccharide composite microbe fertilizer dilute with water 500 times respectively.Take the above-mentioned dilution of 10mL
The alga oligosaccharide solution crossed, bacillusmusilaginosiengineering fermentation liquid, alga oligosaccharide composite microbe fertilizer 7d after Brassica campestris L final singling carry out leaf
Face sprays.Cultivate 40d in the controlled environment chamber, topdress once mid-term.
Result is as follows:
1) comparing comparison CK, different disposal group all can make soil microbe quantity, soil available phosphorus, effective potassium and organic matter contain
Amount has raising in various degree, and content of organic matter maximum improves 22.8% (processing B), and bacterial number maximum improves 4 times of (sun
Property matched group CK1), different disposal is shown in Table 1 to the impact of soil property.
Table 1
2) comparing comparison CK, different disposal group all can make Brassica campestris L fresh weight, plant height, root length, the number of blade have raising in various degree,
Especially processing D can make Brassica campestris L fresh weight increase by 54.8%, and plant height increases by 2.5%, and root length increases by 29.3%.And, alga oligosaccharide
Composite microbe fertilizer can also increase rape leaf number, generally increases by 1～2.Different disposal is shown in Table 2 to the impact of yield of rape.
Table 2
| Process | Brassica campestris L fresh weight (g) | Amplification (%) | Plant height (cm) | Amplification (%) | Root length (cm) | Amplification (%) | The number of blade |
| CK | 4.2 | - | 21.6 | - | 4.6 | - | 7 |
| CK1 | 5.3 | 26.2 | 21.2 | -2.0 | 5.3 | 15.5 | 8 |
| CK2 | 5.1 | 21.4 | 22.2 | 2.5 | 5.5 | 19.6 | 8 |
| A | 5.5 | 31.0 | 21.6 | 0 | 5.6 | 21.7 | 9 |
| B | 5.2 | 23.8 | 22.0 | 1.8 | 5.5 | 19.6 | 8 |
| C | 5.6 | 33.3 | 22.4 | 3.7 | 6.1 | 32.6 | 8 |
| D | 6.5 | 54.8 | 22.2 | 2.5 | 6.0 | 29.3 | 8 |
Embodiment 3
Soil sieves, and removes impurity removing, stone etc., mixes thoroughly, makes test soil sample, every basin dress soil 2kg.Conventional fertilizer application, with urine
Element 0.7g/kg, KH2PO4The addition of 0.5g/kg, KCl 0.085g/kg adds azophoska solution to soil sample.Select seed
Caulis et Folium Lactucae Sativae seed full, of the same size, with the hypochlorite disinfectant 15min of 1%, is rinsed well with deionized water, then spend from
Sub-water soaking 12h.By the Caulis et Folium Lactucae Sativae planting seed of dipped water in basin, after germinateing one week, the seedling selecting growing way consistent is carried out
Final singling, the strain of every basin 5, sprinkling is watered so that field capacity is about 60%～70%.
By above-mentioned test group alga oligosaccharide composite microbe fertilizer dilute with water 800 times.Take respectively on 4mL, 6mL, 8mL, 10mL
State the alga oligosaccharide composite microbe fertilizer III diluted 7d after Brassica campestris L final singling and carry out foliage-spray, enter again every the identical amount of application of 7d
Row sprays once, twice totally, using clear water as comparison.Cultivate 40d in the controlled environment chamber, topdress once mid-term.
Analyze soil constituent and the Growth of Lettuce situation of different experiments group, find soil with organic matter content, effective potassium, available phosphorus
Content and the yield of Caulis et Folium Lactucae Sativae have and improve in various degree, concrete outcome is shown in Fig. 1～4.
Embodiment 4
Test material is TIEGUANYIN, Planting Years 7 years.
Test site is bimodal village, Shu Yang town, Nanjing County, Zhangzhou, Fujian city, and experimental field soil is yellow-red soil, the soil organism
19.4g/kg, full nitrogen 0.96g/kg, alkali-hydrolyzable nitrogen 49.7mg/kg, available phosphorus 21.9mg/kg, available potassium 76mg/kg, pH value
4.9。
Test sets 4 process.Process A alga oligosaccharide composite microbe fertilizer III+conventional fertilizer application；Process B alga oligosaccharide composite microbial
Thing fertilizer III+80% conventional fertilizer application；Process C and spray equivalent clear water+conventional fertilizer application；Process D and spray equivalent clear water (not applying fertilizer).Institute
State alga oligosaccharide >=2g/kg in alga oligosaccharide composite microbe fertilizer III, pH value be 6.8, living bacteria count >=200,000,000/mL.
Test sets 3 repetitions, wherein processes A and processes B two each 30 mu of sheet area of demonstration, and other set est and pilot project area
Each 1 mu, plot area plucked by sample is 30m2, use the group arrangement of random district.Spray on alga oligosaccharide complex microorganism fat guy Folium Camelliae sinensis
Applying method: spray once when summer tea bud is sprouted, spray once after 15d again, execute 2 times altogether, concentration is 400 times, each 666.7m2
It is watered 200kg with alga oligosaccharide composite microbe fertilizer 500mL uniformly to spray.Process C, D and spray equivalent clear water the same period.Trophophase
Between other management methods consistent.
In 666.7m on June 14th, 20152Execute Folium Camelliae sinensis formula fertilizer (N: P2O5∶K2O is 21: 6: 13) 75kg.Process B 666.7
m2Execute Folium Camelliae sinensis formula fertilizer (N: P2O5∶K2O is 21: 6: 13) 60kg.Process A, B on June 14th, 2015 and 30 days
Respectively spray alga oligosaccharide composite microbe fertilizer III 1 times, use 400 diluents.Process C, D and spray equivalent clear water the same period.Summer
Tea was gathered on August 17th, 2015, surveyed yield.
Result is as follows:
1) A Folium Camelliae sinensis blade is processed more dark green than processing B, C, D blade.Process A, B, C, D Camellia sinensis average bud head density (individual
/0.1m2) it is respectively 64.9,63.8,60.2,46.6.Process A, B and add 4.7,3.6/0.1m respectively than processing C2；
Process the average hundred-bud weight of A, B, C, D Camellia sinensis (1 bud 3 leaf) and be respectively 269.3g, 270.0g, 236.7g, 206.0g.Place
Reason A, B add 32.6g and 33.3g respectively than processing C.
2) each community different disposal is shown in Table 3 to the impact of tea yield, processes the every 666.7m of A2Fresh tea yield is divided than processing B, C
Not Zeng Jia 17.8kg and 22.2kg, increase production 8.33%, 10.6% respectively；By 5: 1 give money as a gift raw tea calculate, process A do not have 666.7m2
Dry raw tea yield increases production 3.56kg, 4.44kg respectively than processing B, C, respectively volume increase 8.33% and 10.6%.Process at B ratio
Reason C volume increase 2.13%, processes C ratio and processes D volume increase 42.4%.
Table 3
Test processes cell production variance analysis (being shown in Table 4), F=26.14 > F0.01=9.78, and between explanation processes, volume variance reaches
Pole significant level.
Table 4 different disposal tea yield variance analysis
| Source of variation | Degree of freedom | Quadratic sum | Mean square | F value | F0.05 | F0.01 |
| Between process | 3 | 368.604 | 122.868 | 26.14** | 4.76 | 9.78 |
| Between repetition | 2 | 2.937 | 1.469 | 0.31 | 5.14 | 10.92 |
| Error | 6 | 28.205 | 4.701 | |||
| Total variation | 11 | 399.746 |
Test being processed cell production and carries out LSD method multiple comparisons (being shown in Table 3), result shows: processes A and processes B, C, D
Volume variance big pole significant level, processes B inconspicuous with process C difference, illustrates that Folium Camelliae sinensis sprays alga oligosaccharide bio-fertilizer volume increase effect
Fruit reaches pole significant level.Meanwhile, in the case of conventional fertilizer application reduces 20%, Folium Camelliae sinensis sprays alga oligosaccharide composite microbe fertilizer fertilizer efficiency
The most notable with conventional fertilizer application process volume variance.
3) process polyphenol content 13.2% (in terms of butt), Vc content 25.7mg/100g in A fresh tea, process C tea many
Phenol content 12.7% (in terms of butt), Vc content 14.1mg/100g, processing A increases by 0.5% than polyphenol content in C fresh tea
(in terms of butt), Vc content increase 11.6mg/100g.Folium Camelliae sinensis sprays alga oligosaccharide composite microbe fertilizer can promote fresh tea quality
Improve.
The alga oligosaccharide composite microbe fertilizer of the above verification experimental verification present invention has and the most significantly increases production, promotes fertilizer efficiency, improvement product
The effect of matter.
Additionally, can be in any combination in the middle of the various different embodiments of the present invention.
Claims (10)
1. an alga oligosaccharide composite microbe fertilizer, it is characterised in that each component by volume percentage ratio is: alga oligosaccharide solution 20%～
80%, bacillusmusilaginosiengineering fermentation liquid 20%～80%；Wherein alga oligosaccharide content >=0.5g/kg, living bacteria count >=200,000,000
/ mL, pH value is 6.8～7.2；
Described bacillusmusilaginosiengineering is No. 01 bacterial strain of bacillusmusilaginosiengineering (Bacillus mucilaginosus) three torch,
Being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on 07 02nd, 2010, deposit number is
CGMCC No.3995。
The preparation method of a kind of alga oligosaccharide composite microbe fertilizer the most as claimed in claim 1, it is characterised in that comprise the following steps:
1) after bacillusmusilaginosiengineering solid nitrogen-free agar being activated, it is inoculated in seed culture medium, obtains one-level kind
Sub-fermentation liquid；Primary seed solution is seeded in seed fermentation tank, cultivates and obtain secondary seed solution；Described bacillusmusilaginosiengineering
For No. 01 bacterial strain of bacillusmusilaginosiengineering (Bacillus Mucilaginosus) three torch, protected on 07 02nd, 2010
Being hidden in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number is CGMCC No.3995；
2) secondary seed solution of gained being inoculated in fermentation medium, after continuing fermentation culture, prepared bacillusmusilaginosiengineering is sent out
Ferment liquid；
3) by step 2) the bacillusmusilaginosiengineering fermentation liquid collected mixes with alga oligosaccharide solution, obtains alga oligosaccharide and be combined micro-
Bio-fertilizer.
The preparation method of a kind of alga oligosaccharide composite microbe fertilizer the most as claimed in claim 1, it is characterised in that in step 1) in,
Described solid nitrogen-free agar consist of sucrose 5～10g/L, dipotassium hydrogen phosphate 0.3～0.8g/L, magnesium sulfate 0.1～0.4g/L,
Sodium chloride 0.15～0.35g/L, calcium carbonate 0.5～1.2g/L, agar 20g/L, before sterilizing, pH value is 7.0～7.5.
The preparation method of a kind of alga oligosaccharide composite microbe fertilizer the most as claimed in claim 1, it is characterised in that in step 1) in,
The condition of described activation culture is to cultivate 48～72h under the conditions of 28～32 DEG C.
The preparation method of a kind of alga oligosaccharide composite microbe fertilizer the most as claimed in claim 1, it is characterised in that in step 1) in,
Described seed culture medium consist of sucrose 5～10g/L, dipotassium hydrogen phosphate 0.3～0.8g/L, magnesium sulfate 0.1～0.4g/L,
Sodium chloride 0.15～0.35g/L, calcium carbonate 0.5～1.2g/L, before sterilizing, pH value is 7.0～7.5.
The preparation method of a kind of alga oligosaccharide composite microbe fertilizer the most as claimed in claim 1, it is characterised in that in step 1) in,
The condition that described first order seed is cultivated is in 30～35 DEG C, cultivates 24～36h under the conditions of 150～200rpm；Described secondary seed
The condition cultivated can be in seed fermentation tank, under the conditions of 30～35 DEG C, keeps tank rotating speed 125～150rpm, cultivates 24～48h.
The preparation method of a kind of alga oligosaccharide composite microbe fertilizer the most as claimed in claim 1, it is characterised in that in step 2) in,
Described fermentation medium consist of alga oligosaccharide solution 0～20mL/L, yeast extract 1.0～1.8g/L, starch 5.0～7.5g/L,
Bean cake powder 5.0～7.0g/L, magnesium sulfate 0.8～1.6g/L, dipotassium hydrogen phosphate 1.0～1.8g/L, calcium carbonate 5.0～10.0g/L,
Sodium chloride 0.1～0.4g/L, before sterilizing, pH value is 7.0～7.5.
The preparation method of a kind of alga oligosaccharide composite microbe fertilizer the most as claimed in claim 1, it is characterised in that in step 2) in,
Described seed liquor be seeded in fermentation tank culture medium fermentation inoculum concentration by volume percentage ratio is seed liquor 10%；Described fermentation
Condition can be: rotating speed controls 200～250rpm, makes dissolved oxygen control at 20% (V/V) by regulation rotating speed and ventilation,
Tank temperature control is at 30 DEG C, and tank pressure is maintained at 0.05～0.06MPa, and pH value is controlled by the ammonia of 10% and the hydrochloric acid of 10%
7.2, fermentation time 36～48h.
The preparation method of a kind of alga oligosaccharide composite microbe fertilizer the most as claimed in claim 1, it is characterised in that in step 3) in,
Described alga oligosaccharide is to utilize microorganism directly to degrade the oligosaccharide that the degree of polymerization is 4～8 of Thallus Gracilariae gained；Described alga oligosaccharide solution
The mass percentage concentration of middle alga oligosaccharide is 4～5g/L.
The preparation method of a kind of alga oligosaccharide composite microbe fertilizer the most as claimed in claim 1, it is characterised in that in step 3)
In, described alga oligosaccharide solution, step 2) the bacillusmusilaginosiengineering fermentation liquid by volume percentage ratio of gained is (20%～80%):
(20%～80%).
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