CN103304278B - Seaweed fertilizer comprising seaweed leach liquor and preparation method thereof - Google Patents
Seaweed fertilizer comprising seaweed leach liquor and preparation method thereof Download PDFInfo
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- CN103304278B CN103304278B CN201310165667.2A CN201310165667A CN103304278B CN 103304278 B CN103304278 B CN 103304278B CN 201310165667 A CN201310165667 A CN 201310165667A CN 103304278 B CN103304278 B CN 103304278B
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Abstract
The invention relates to a seaweed fertilizer comprising a seaweed leach liquor. The seaweed fertilizer is prepared by the following steps: (1) seaweed extraction; (2) preparation of a fermentation medium; (3) inoculation and fermentation. The seaweed fertilizer provided by the invention is an active biological fungicide fertilizer of seaweed produced by pure physical and biological methods, and has the activity of seaweed and the living body adjusting function of biological fungicide. The number of effective living bacteria is greater than or equal to 2.0*10 to the power of 9 CFU/ml. Meanwhile, the fertilizer is rich in plant growth-regulating activity substances such as alginic acid, seaweed oligosaccharide, gibberellin (GA) and indoleacetic acid (IAA) biologically fermented, degraded and converted from seaweed gel. The fertilizer is directly absorbed by root systems of crops conveniently and fast in fertilizer efficiency, and can effectively accelerate high yield and early-ripe of plants, enhance the disease-resistance and improve the quality of crops, so that the fertilizer is an excellent special fertilizer for organic food.
Description
Technical field
The present invention relates to a kind of marine alga bacterial manure that contains algae leaching liquor and preparation method thereof, belong to organic biofertilizer field.
Background technology
Marine alga (Alga) is submarine algae, is botanic spore plant.They are generally considered to be simple plant, and principal character is: without vascular bundle tissue, there is no the differentiating phenomenon of real root, stem, leaf; Do not bloom, without fruit and seed; Reproductive organ, without the protective tissue of specialization, often directly produces spore or gamete by single cell; And without embryo's formation.Simple in structure due to algae, thus some botanists by it with mushroom with " the thallose group " that be attributed to lower plant.
Marine alga belongs to lower plant, of a great variety.According to the difference of contained pigment in marine alga, can be divided into green alga (as sea lettuce), brown alga (as sea-tangle, sargassun, marine alga, bulk kelp etc.), blue-green algae (as Shu Maozao), xanthophyta (as halosphaera), chrysophyceae (as calcium plate algae) etc.
Brown alga is a higher monoid in phycophyta.The shape of brown alga and different sizes, from being the algae that grows nonparasitically upon another plant (Ectocarpus (Ectocarpus)) of heterotrichy to the kelp (Laminaria (Laminaria), Macrocystis (Macrocystis) etc.) of 1～100 meter of complicated, huge length.Rock algae is a type for brown alga, grows floating on water (Sargassum (Sargassum)) or grows nonparasitically upon another plant in rock coast, mainly comprises that fucus (Fucus), marine alga belong to (Ascophyllum).
Marine alga fertilizer is using marine plant marine alga as main raw material, the bio-feritlizer of being processed into through science, main component is that the natural bioactivity substance that is conducive to growth and development of plants and the marine alga of from marine alga, extracting absorb also enrichment mineral nutrient element in vivo from ocean, comprises the trace elements such as Sargassum polysaccharides, phenols poly-compounds, N.F,USP MANNITOL, trimethyl-glycine, plant growth regulating substance (phytokinin, Plant hormones regulators,gibberellins, growth hormone and dormin etc.) and nitrogen, phosphorus, potassium, iron, boron, molybdenum, iodine.Therefore, marine alga fertilizer have nutritious, activity by force, be easily absorbed, can improve the feature such as soil, safety nuisance free.Owing to having above-mentioned a series of advantage, marine alga fertilizer becomes the focus that people competitively research and develop just gradually.
Tang Jie, Shi Jingwu discloses a kind of production method of functional marine alga fertilizer in Chinese invention patent CN98101040.It is main raw material that the method adopts marine organisms brown alga, taking potassium hydroxide as digestive pharmaceutical, digestion temperature is 70 DEG C～100 DEG C, can make nutritive substance contained in brown alga, trace element and spontaneous growth Auto-regulator, the soluble status that becomes to be for the benefit of absorbed by plants, be equipped with the hydrolyzed solution of obtaining property fish more, not only regulated the pH value of extracting solution, increased the nutritive substance such as amino acid in extracting solution simultaneously.In the technique of strengthening mineral matter nutritional, can make brown alga extracting solution and positively charged ion more than divalence directly form complex salt, greatly reduce production cost.
Xu Jiachao, the people such as Li Zhaojie disclose a kind of production technique of liquid seaweed manure material in Chinese invention patent CN03138729, comprise and use freshwater soaking sea-tangle, sea-tangle is pulverized or sheared, acid adding makes sea-tangle degraded, the polymerization degree of the Sargassum polysaccharides producing is controlled at below 100, adds degradation solution digestion, filtration, concentrated that alkaline matter makes gained.The liquid seaweed manure material of producing is except containing the nutritive ingredient of marine alga, also contains marine active composition-seaweed gel oligose, plant-growth regulator and a large amount of trace element as phosphorus, calcium, potassium, magnesium, zinc, iron etc.
Although above-mentioned marine alga fertilizer nutritive ingredient is abundant, activeconstituents is poor, function singleness, and Application Areas still has certain limitation.
Summary of the invention
For the deficiencies in the prior art, the object of this invention is to provide a kind of marine alga bacterial manure that contains algae leaching liquor and preparation method thereof.
For achieving the above object, one of technical scheme provided by the invention, is a kind of marine alga bacterial manure that contains algae leaching liquor, and described marine alga bacterial manure is obtained by following preparation method:
(1) marine alga lixiviate
After marine alga is pulverized, add water to stir, lixiviate 2-15 hour; Centrifugation gained supernatant liquor, is algae leaching liquor;
(2) preparation of fermention medium
Get step (1) gained algae leaching liquor 1000 weight parts, add 6-10 weight part peptone, 1-5 weight part yeast extract paste, 5-10 weight part sodium-chlor, 0.5-1 weight part KH
2pO
4, 0.1-0.5 weight part MgSO
4, regulate pH to 7-8, obtain required fermention medium;
(3) inoculate and ferment
Subtilis (preserving number is CGMCC No.6616) is inoculated in step (2) gained substratum, and fermentation, obtains marine alga bacterial manure.
Preferably, in step (1), the add-on of water is by weight marine alga: water=1:10-1:25
Preferably, in step (2), PH conditioning agent used is ammoniacal liquor.
Preferably, in step (3), the inoculum size of subtilis is the 0.1-5% with respect to culture volume ratio.
Preferably, in step (3), subtilis first activates before inoculation, and activation temperature is 35-40 DEG C.
Preferably, zymotechnique described in step (3) is liquid ventilating fermentation technique, and temperature is 35-40 DEG C, and ferment liquid pH value is 7-8, and air flow is 1:0.5-1:1.5, and tank pressure is 0.02-0.03MPa.
Two of technical scheme provided by the invention, is a kind of method of preparing the marine alga bacterial manure that contains algae leaching liquor, comprises the following steps:
(1) marine alga lixiviate
After marine alga is pulverized, add water to stir, lixiviate 2-15 hour; Centrifugation gained supernatant liquor is algae leaching liquor;
(2) preparation of fermention medium
Get step (1) gained algae leaching liquor 1000 weight parts, add 6-10 weight part peptone, 1-5 weight part yeast extract paste, 5-10 weight part sodium-chlor, 0.5-1 weight part KH
2pO
4, 0.1-0.5 weight part MgSO
4, regulate pH to 7-8, obtain required fermention medium;
(3) inoculate and ferment
The subtilis of activation is inoculated in step (2) gained substratum, and fermentation, obtains marine alga bacterial manure.
Bacterial strain used in the present invention is by contriver's separation screening bacillus subtilis BS-1 bacterial strain out, Latin title: Bacillus subtilis, this bacterial strain is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center on September 21st, 2012, deposit number is CGMCC No.6616, depositary institution address: Datun Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101.
Beneficial effect of the present invention:
(1) marine alga bacterial manure of the present invention is that a kind of pure physics and biological method of utilizing produced the marine alga active bio fertilizer bacterial agent obtaining, and has the live body regulatory function of marine alga activity and bacteria agent concurrently, has realized the organic unity of seaweed bio activity with bacteria agent;
(2) the present invention subtilis BS-1 used obtains through long-term screening, is a kind of bio-feritlizer bacterial strain with functions such as high-performance bio control, active growth-promotings, is applied to agriculture production remarkably productive;
(3) conditioning agent of the present invention using ammoniacal liquor as algae leaching liquor substratum, can effectively regulate pH, does not produce precipitation simultaneously, has protected the significant condition of the marine active substance in marine alga;
(4) living bacteria count>=2.0 × 10 of marine alga bacterial manure of the present invention
9cFU/ml, be rich in by seaweed gel through plant growth regulating activity materials such as the Lalgine of biological fermentation Degradation and Transformation, alga oligosaccharide, Plant hormones regulators,gibberellins (GA), indolylacetic acids (IAA) simultaneously, be convenient to directly be absorbed by crop root, fertilizer efficiency is fast, and can effectively promote high-yield plant, precocity, improving disease resistance ability, improve the quality of crop, is a kind of organic food special fertilizer material of high-quality.
Brief description of the drawings
Fig. 1 is preparation method's schematic diagram of a kind of marine alga bacterial manure that contains algae leaching liquor of the present invention.
Embodiment
Embodiment 1
(1) bladder wrack lixiviate
Get 40 kilograms of bladder wracks, be crushed to 80 orders, sieving for standby; The deionized water of pH7.0～7.5 is preheated to 30 DEG C, according to bladder wrack: water=1:10(weight ratio) the ratio 400kg that adds water, fully stir, lixiviate 15 hours, vat liquor centrifugal 20 minutes through 3000rpm, gets supernatant liquor, is bladder wrack vat liquor, for subsequent use;
(2) preparation of fermention medium
Get step (1) gained bladder wrack vat liquor 100kg, add 1kg peptone (1%, W/W), yeast extract paste (0.5%, W/W) 0.5kg; Sodium-chlor (1%, W/W) 0.5kg; KH
2pO
4(0.1%W/W) 0.1kg; MgSO
4(0.01%, W/W) 0.05kg; Utilize ammoniacal liquor to regulate medium pH to 7.0;
(3) subtilis BS-1 seed activation
Be CGMCC No.6616 by subtilis BS-1(preserving number) transfer in fresh test tube slant from the inclined-plane of preserving, cultivate 24h in 35 DEG C of constant incubators.Get cultured inclined-plane, choose a full ring with transfering loop, be inoculated in the 250mL triangular flask that fills 50mL aseptic seed substratum, 35 DEG C of shaking tables, 180r/min shaking culture 18h.Corresponding seed quality criteria: OD600 value reaches 5 times of 0.5(dilutions), bacterial content reaches 8.0 × 108CFU, and microscopy thalline is full, and gemma has just started to form.Inoculum size 0.1%, adopts the inoculation of flame inoculation method;
(4) fermentation
Fermentation parameter control is as follows: leavening temperature: 35 DEG C; In ferment liquid pH:7.0(fermenting process, pH is on a declining curve, by the NaOH auto-feeding control of 0.1M); Mixing speed: 0-4h, 150rpm; 4-20h, 200rpm; 20-24h, 100rpm.Air flow: 0-4h, 1:0.5; 4-10h, 1:0.8; 10-20h, 1:1.2; 20-24h, 1:0.5; Tank pressure: 0.02～0.03MPa, will remain malleation in tank body in fermenting process, and according to how many adjusting tank pressures of fermentor tank inner foam, when foam is too much, can suitably increase tank pressure; Froth breaking: according to produce foam number, the mode adding by stream is added bubble enemy's (sterilizing), or in substratum process for preparation, directly adds 0.05～0.1% bubble enemy.The factor such as the number of foam and air flow size, mixing speed height, broth viscosity, thalline content, defoamer are how many is relevant, can control flexibly during the fermentation;
Every sampling in 2 hours, microscopy thalli morphology, OD600, the parameters such as sporulation ratio and total reducing sugar, total nitrogen.Fermentation time reached about 24 hours, stopped tank when the gemma rate in the visual field reaches 90%.Thalline content can reach 1.2 × 10
9cFU/ml, stops opening cold water after tank and lowers the temperature, and temperature is generally down to 20 DEG C of left and right.Again fermented liquid is transferred in storage tank, detect qualified after, metering packing.
Embodiment 2
(1) blue-green algae lixiviate
Get 40 kilograms of blue-green algaes, be crushed to 120 orders, sieving for standby; Deionized water is preheated to 30 DEG C, according to blue-green algae: water=1:25(weight ratio) the ratio 1000kg that adds water, fully stir, lixiviate 2 hours, vat liquor centrifugal 30 minutes through 2000rpm, gets supernatant liquor, is blue-green algae vat liquor, for subsequent use;
(2) preparation of fermention medium
Get step (1) gained blue-green algae vat liquor 100kg, add 0.6kg peptone (1%, W/W), yeast extract paste (0.5%, W/W) 0.1kg; Sodium-chlor (1%, W/W) 1kg; KH
2pO
4(0.1%W/W) 0.05kg; MgSO
4(0.01%, W/W) 0.01kg; Utilize ammoniacal liquor to regulate medium pH to 8.0;
(3) subtilis BS-1 seed activation
Be CGMCC No.6616 by subtilis BS-1(preserving number) transfer in fresh test tube slant from the inclined-plane of preserving, cultivate 24h in 40 DEG C of constant incubators.Get cultured inclined-plane, choose a full ring with transfering loop, be inoculated in the 250mL triangular flask that fills 50mL aseptic seed substratum 40 DEG C of shaking tables, 180r/min shaking culture 18h, corresponding seed quality criteria: OD600 value reaches 5 times of 0.5(dilutions), bacterial content reaches 8.0 × 108CFU, and microscopy thalline is full, and gemma has just started to form, inoculum size 0.2%, adopts the inoculation of flame inoculation method;
(4) fermentation
Fermentation parameter control is as follows: leavening temperature: 40 DEG C; In ferment liquid pH:8.0(fermenting process, pH is on a declining curve, by the NaOH auto-feeding control of 0.1M); Mixing speed: 0-4h, 150rpm; 4-20h, 200rpm; 20-24h, 100rpm.Air flow: 0-4h, 1:0.5; 4-10h, 1:0.8; 10-20h, 1:1.2; 20-24h, 1:0.5; Tank pressure: 0.02～0.03MPa;
Other step is with embodiment 1.
Embodiment 3
(1) sea-tangle lixiviate
Get 40 kilograms of sea-tangles, be crushed to 100 orders, sieving for standby; Deionized water is preheated to 35 DEG C, according to sea-tangle: water=1:20(weight ratio) the ratio 800kg that adds water, fully stir, lixiviate 8 hours, vat liquor centrifugal 20 minutes through 2500rpm, gets supernatant liquor, is sea-tangle vat liquor, for subsequent use;
(2) preparation of fermention medium
Get step (1) gained sea-tangle vat liquor 100kg, add 0.8kg peptone (1%, W/W), yeast extract paste (0.5%, W/W) 0.3kg; Sodium-chlor (1%, W/W) 0.7kg; KH
2pO
4(0.1%W/W) 0.08kg; MgSO
4(0.01%, W/W) 0.03kg; Utilize ammoniacal liquor to regulate medium pH to 7.2;
(3) subtilis BS-1 seed activation
Be CGMCC No.6616 by subtilis BS-005(preserving number) transfer in fresh test tube slant from the inclined-plane of preserving, cultivate 24h in 38 DEG C of constant incubators.Get cultured inclined-plane, choose a full ring with transfering loop, be inoculated in the 250mL triangular flask that fills 50mL aseptic seed substratum 38 DEG C of shaking tables, 180r/min shaking culture 18h, corresponding seed quality criteria: OD600 value reaches 5 times of 0.5(dilutions), bacterial content reaches 8.0 × 108CFU, and microscopy thalline is full, and gemma has just started to form, inoculum size 1%, adopts the inoculation of flame inoculation method;
(4) fermentation
Fermentation parameter control is as follows: leavening temperature: 38 DEG C; In ferment liquid pH:7.2(fermenting process, pH is on a declining curve, by the NaOH auto-feeding control of 0.1M); Mixing speed: 0-4h, 150rpm; 4-20h, 200rpm; 20-24h, 100rpm.Air flow: 0-4h, 1:0.5; 4-10h, 1:0.8; 10-20h, 1:1.2; 20-24h, 1:0.5; Tank pressure: 0.02～0.03MPa;
Other step is with embodiment 1.
Get respectively embodiment 1-3 gained seaweed fertilizer and certain commercially available seaweed fertilizer A, measure respectively the content of its living bacteria count, Lalgine, alga oligosaccharide, Plant hormones regulators,gibberellins (GA), indolylacetic acid (IAA), the results are shown in Table 1.
Table 1
| Seaweed fertilizer | Living bacteria count (CFU/ml) | Lalgine | Alga oligosaccharide | Plant hormones regulators,gibberellins | Indolylacetic acid |
| Embodiment 1 | 5.2×10 9 | 2.2% | 800ppm | 160ppm | 100ppm |
| Embodiment 2 | 8.1×10 9 | 2.3% | 700ppm | 200ppm | 80ppm |
| Embodiment 3 | 6.7×10 9 | 2.0% | 820ppm | 190ppm | 98ppm |
| Commercially available seaweed fertilizer A | 0 | 1.8% | 20ppm | 10ppm | 16ppm |
Can find out the living bacteria count of marine alga bacterial manure of the present invention and obviously being improved than existing commercially available prod through the content of the plant growth regulating activity materials such as the Lalgine of biological fermentation Degradation and Transformation, alga oligosaccharide, Plant hormones regulators,gibberellins (GA), indolylacetic acid (IAA) by seaweed gel from data shown in table 1.
Application Example
Getting embodiment 1 gained seaweed fertilizer produces for vegetables:
In growing vegetables base, Shouguang City of Shandong Province, choose 100 mu of identical green house of vegetables of condition, be divided into A, B, C, D Si Ge community, each community area is 25 mu, plantation Chinese cabbage, at Chinese cabbage Seedling Stage, its fertilising situation is respectively:
A district does not apply any fertilizer;
B district respectively uses seaweed fertilizer once according to the rate of fertilizer application of 10 liters/mu after Chinese cabbage complete germination He after 3 weeks;
C district respectively uses urea fertilizer once according to the rate of fertilizer application of 12 kgs/acre after Chinese cabbage complete germination He after 3 weeks;
D district respectively uses urea fertilizer once according to the rate of fertilizer application of 12 kgs/acre after Chinese cabbage complete germination He after 3 weeks; And after applying two days of urea fertilizer, then use respectively seaweed fertilizer once according to the rate of fertilizer application of 10 liters/mu at every turn;
Si Ge community adopts identical irrigation and way to manage.
After Chinese cabbage growth 60 days, check the content of its output and nitrate, nitrite, result is as follows:
A district Chinese cabbage output is minimum, only has 5000 kgs/acre;
The Chinese cabbage output in B district and C district is basic identical, is 7200 kgs/acre, exceeds 44% than A district;
D district Chinese cabbage output is 8300 kgs/acre, exceeds 66% than A district, exceeds 15.3% than B district, C district.
Claims (6)
1. a marine alga bacterial manure that contains algae leaching liquor, is characterized in that, described marine alga bacterial manure is obtained by following preparation method:
(1) marine alga lixiviate
After marine alga is pulverized, add water to stir, lixiviate 2-15 hour; Centrifugation gained supernatant liquor, is algae leaching liquor;
(2) preparation of fermention medium
Get step (1) gained algae leaching liquor 1000 weight parts, add 6-10 weight part peptone, 1-5 weight part yeast extract paste, 5-10 weight part sodium-chlor, 0.5-1 weight part KH
2pO
4, 0.1-0.5 weight part MgSO
4, regulate pH to 7-8, and pH adjusting agent used is ammoniacal liquor, obtain required fermention medium;
(3) inoculate and ferment
Be that CGMCC No.6616 is inoculated in step (2) gained substratum by subtilis preserving number, fermentation, obtains marine alga bacterial manure.
2. the method for the marine alga bacterial manure that contains algae leaching liquor according to claim 1, is characterized in that, in step (1), the add-on of water is by weight marine alga: water=1:10-1:25.
3. the method for the marine alga bacterial manure that contains algae leaching liquor according to claim 1, is characterized in that, in step (3), the inoculum size of subtilis is the 0.1-5% with respect to culture volume.
4. the method for the marine alga bacterial manure that contains algae leaching liquor according to claim 1, is characterized in that, in step (3), subtilis first activates before inoculation, and activation temperature is 35-40 DEG C.
5. the method for the marine alga bacterial manure that contains algae leaching liquor according to claim 1, it is characterized in that, zymotechnique described in step (3) is liquid ventilating fermentation technique, temperature is 35-40 DEG C, ferment liquid pH value is 7-8, air flow is 1:0.5-1:1.5, and tank pressure is 0.02-0.03MPa.
6. a method of preparing the marine alga bacterial manure that contains algae leaching liquor described in claim 1, is characterized in that, comprises the following steps:
(1) marine alga lixiviate
After marine alga is pulverized, add water to stir, lixiviate 2-15 hour; Centrifugation gained supernatant liquor is algae leaching liquor;
(2) preparation of fermention medium
Get step (1) gained algae leaching liquor 1000 weight parts, add 6-10 weight part peptone, 1-5 weight part yeast extract paste, 5-10 weight part sodium-chlor, 0.5-1 weight part KH
2pO
4, 0.1-0.5 weight part MgSO
4, regulate pH to 7-8, obtain required fermention medium;
(3) inoculate and ferment
Subtilis is inoculated in step (2) gained substratum, and fermentation, obtains marine alga bacterial manure.
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