CN103304278A - 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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- CN103304278A CN103304278A CN2013101656672A CN201310165667A CN103304278A CN 103304278 A CN103304278 A CN 103304278A CN 2013101656672 A CN2013101656672 A CN 2013101656672A CN 201310165667 A CN201310165667 A CN 201310165667A CN 103304278 A CN103304278 A CN 103304278A
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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 the 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 the vascular bundle tissue, do not have the differentiating phenomenon of real root, stem, leaf; Do not bloom, without fruit and seed; Reproductive organ often directly produces spore or gamete by single cell without the protective tissue of specialization; And without embryo's formation.Because algae is simple in structure, so some botanists follow mushroom with " the thallose group " that be attributed to lower plant it.
Marine alga belongs to lower plant, and is of a great variety.According to the difference of contained pigment in the marine alga, it can be divided into green alga (such as sea lettuce), brown alga (such as sea-tangle, sargassun, marine alga, bulk kelp etc.), blue-green algae (such as Shu Maozao), xanthophyta (such as halosphaera), chrysophyceae (such as calcium plate algae) etc.
Brown alga is a higher monoid in the phycophyta.The shape of brown alga and different sizes, the kelp (Laminaria (Laminaria), Macrocystis (Macrocystis) etc.) from the algae that grows nonparasitically upon another plant (Ectocarpus (Ectocarpus)) that is heterotrichy to 1～100 meter of complicated, huge length.The rock algae is the type of brown alga, grows floating on water (Sargassum (Sargassum)) or grows nonparasitically upon another plant in rock coast, comprises that mainly fucus (Fucus), marine alga belong to (Ascophyllum).
Marine alga fertilizer is as main raw material with the marine plant marine alga, 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 extracting from marine alga absorb also enrichment mineral nutrient element in vivo from the 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, easily be absorbed, can improve the characteristics 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 disclose a kind of production method of functional marine alga fertilizer in Chinese invention patent CN98101040.It is main raw material that the method adopts the marine organisms brown alga, take potassium hydroxide as digestive pharmaceutical, the digestion temperature is 70 ℃～100 ℃, can make nutritive substance contained in the brown alga, trace element and spontaneous growth Auto-regulator, become for the benefit of by the soluble status of plant absorbing, be equipped with the hydrolyzed solution of obtaining property fish more, not only regulated the pH value of extracting solution, increased simultaneously the nutritive substance such as amino acid in the extracting solution.On the technique of strengthening mineral matter nutritional, can make the above positively charged ion of brown alga extracting solution and 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 the freshwater soaking sea-tangle, sea-tangle is pulverized or shears, acid adding makes the sea-tangle degraded, the polymerization degree of the Sargassum polysaccharides that produces is controlled at below 100, adds degradation solution digestion, filtration, concentrated that alkaline matter makes gained.The liquid seaweed manure material of producing also contains marine active composition-seaweed gel oligose, plant-growth regulator and a large amount of trace element such as phosphorus, calcium, potassium, magnesium, zinc, iron etc. except the nutritive ingredient that contains marine alga.
Although above-mentioned marine alga fertilizer nutritive ingredient is abundant, activeconstituents is poor, and function singleness, Application Areas still have certain limitation.
Summary of the invention
For the deficiencies in the prior art, the purpose 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 the marine alga pulverizing, add entry and 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, namely get required fermention medium;
(3) inoculation and fermentation
Subtilis (preserving number is CGMCC No.6616) is inoculated in step (2) the gained substratum, and fermentation namely gets marine alga bacterial manure.
Preferably, the add-on of water is by weight marine alga: water=1:10-1:25 in the step (1)
Preferably, used PH conditioning agent is ammoniacal liquor in the step (2).
Preferably, the inoculum size of the middle subtilis of step (3) is the 0.1-5% with respect to the culture volume ratio.
Preferably, subtilis activates first before inoculation in the step (3), and activation temperature is 35-40 ℃.
Preferably, zymotechnique described in the step (3) is liquid ventilating fermentation technique, and temperature is 35-40 ℃, 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 methods that prepare the marine alga bacterial manure that contains algae leaching liquor, may further comprise the steps:
(1) marine alga lixiviate
After the marine alga pulverizing, add entry and 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, namely get required fermention medium;
(3) inoculation and fermentation
The subtilis of activation is inoculated in step (2) the gained substratum, and fermentation namely gets 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 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 a kind of marine alga active bio fertilizer bacterial agent that utilizes pure physics and biological method production to obtain, 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 used subtilis BS-1 of the present invention obtains through long-term screening, is a kind of have high-performance bio control, active bio-feritlizer bacterial strain of urging functions such as giving birth to, is applied to agriculture production remarkably productive;
(3) the present invention can effectively regulate pH with the conditioning agent of ammoniacal liquor as the algae leaching liquor substratum, does not produce precipitation simultaneously, has protected the significant condition of the marine active substance in the marine alga;
(4) living bacteria count of marine alga bacterial manure of the present invention 〉=2.0 * 10
9CFU/ml, be rich in simultaneously 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), be convenient to directly be absorbed by crop root, fertilizer efficiency is fast, and can effectively promote high-yield plant, precocity, improving the disease resistance ability, improve the quality of crop, is a kind of organic food special fertilizer material of high-quality.
Description of drawings
Fig. 1 is a kind of preparation method's schematic diagram that contains the marine alga bacterial manure of 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 ℃, and according to bladder wrack: ratio water=1:10(weight ratio) adds water 400kg, fully stir, and lixiviate 15 hours, vat liquor centrifugal 20 minutes through 3000rpm is got supernatant liquor, is the bladder wrack vat liquor, and is for subsequent use;
(2) preparation of fermention medium
Get step (1) gained bladder wrack vat liquor 100kg, and interpolation 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 with subtilis BS-1(preserving number) transfer in fresh test tube slant from the inclined-plane of preserving, cultivate 24h in 35 ℃ 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 ℃ of shaking tables, 180r/min shaking culture 18h.Corresponding seed quality criteria: the OD600 value reaches 5 times of 0.5(dilutions), bacterial content reaches 8.0 * 108CFU, and the microscopy thalline is full, and gemma has just begun to form.Inoculum size 0.1% adopts the inoculation of flame inoculation method;
(4) fermentation
Fermentation parameter control is as follows: leavening temperature: 35 ℃; PH is on a declining curve in the ferment liquid pH:7.0(fermenting process, 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 the tank body in the fermenting process, and according to the tank pressure of how much regulating of fermentor tank inner foam, when foam is too much, can suitably increase tank pressure; Froth breaking: according to producing what of foam, add bubble enemy (sterilizing) by the mode that stream adds, perhaps in the substratum process for preparation, directly add 0.05～0.1% bubble and oppose.How much etc. factor is relevant with air flow size, mixing speed height, broth viscosity, thalline content, defoamer for what of foam, 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 behind the tank and lowers the temperature, and temperature generally is down to about 20 ℃.Again fermented liquid is transferred in the storage tank, detect qualified after, metering packing gets final product.
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 ℃, and according to blue-green algae: ratio water=1:25(weight ratio) adds water 1000kg, fully stir, and lixiviate 2 hours, vat liquor centrifugal 30 minutes through 2000rpm is got supernatant liquor, is the blue-green algae vat liquor, and is for subsequent use;
(2) preparation of fermention medium
Get step (1) gained blue-green algae vat liquor 100kg, and interpolation 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 with subtilis BS-1(preserving number) transfer in fresh test tube slant from the inclined-plane of preserving, cultivate 24h in 40 ℃ 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 ℃ of shaking tables, 180r/min shaking culture 18h, corresponding seed quality criteria: the OD600 value reaches 5 times of 0.5(dilutions), bacterial content reaches 8.0 * 108CFU, and the microscopy thalline is full, and gemma has just begun to form, inoculum size 0.2% adopts the inoculation of flame inoculation method;
(4) fermentation
Fermentation parameter control is as follows: leavening temperature: 40 ℃; PH is on a declining curve in the ferment liquid pH:8.0(fermenting process, 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 ℃, and according to sea-tangle: ratio water=1:20(weight ratio) adds water 800kg, fully stir, and lixiviate 8 hours, vat liquor centrifugal 20 minutes through 2500rpm is got supernatant liquor, is the sea-tangle vat liquor, and is for subsequent use;
(2) preparation of fermention medium
Get step (1) gained sea-tangle vat liquor 100kg, and interpolation 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 with subtilis BS-005(preserving number) transfer in fresh test tube slant from the inclined-plane of preserving, cultivate 24h in 38 ℃ 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 ℃ of shaking tables, 180r/min shaking culture 18h, corresponding seed quality criteria: the OD600 value reaches 5 times of 0.5(dilutions), bacterial content reaches 8.0 * 108CFU, and the microscopy thalline is full, and gemma has just begun to form, inoculum size 1% adopts the inoculation of flame inoculation method;
(4) fermentation
Fermentation parameter control is as follows: leavening temperature: 38 ℃; PH is on a declining curve in the ferment liquid pH:7.2(fermenting process, 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 |
[0069]?
| 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 improved than existing commercially available prod by the content of 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) from data shown in the table 1.
Application Example
Get embodiment 1 gained seaweed fertilizer and be used for vegetables production:
In growing vegetables base, Shouguang City, Shandong Province, choose 100 mu of green house of vegetables that conditionally complete is identical, be divided into A, B, C, four residential quarters of D, each residential quarter area is 25 mu, plantation Chinese cabbage, at the Chinese cabbage Seedling Stage, its fertilising situation is respectively:
The A district does not apply any fertilizer;
The B district is respectively using seaweed fertilizer once according to 10 liters/mu rate of fertilizer application behind the Chinese cabbage complete germination He after 3 weeks;
The C district is respectively using urea fertilizer once according to 12 kg/acres rate of fertilizer application behind the Chinese cabbage complete germination He after 3 weeks;
The D district is respectively using urea fertilizer once according to 12 kg/acres rate of fertilizer application behind the Chinese cabbage complete germination He after 3 weeks; And after applying two days of urea fertilizer, use respectively seaweed fertilizer once according to 10 liters/mu rate of fertilizer application more at every turn;
Identical irrigation and way to manage are adopted in four residential quarters.
After Chinese cabbage growth 60 days, check the content of its output and nitrate, nitrite, the result is as follows:
A district Chinese cabbage output is minimum, only has 5000 kg/acres;
The Chinese cabbage output in B district and C district is basic identical, is 7200 kg/acres, exceeds 44% than the A district;
D district Chinese cabbage output is 8300 kg/acres, exceeds 66% than the A district, exceeds 15.3% than B district, C district.
Claims (7)
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 the marine alga pulverizing, add entry and 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, namely get required fermention medium;
(3) inoculation and fermentation
Subtilis (preserving number is CGMCC No.6616) is inoculated in step (2) the gained substratum, and fermentation namely gets marine alga bacterial manure.
2. the method that contains the marine alga bacterial manure of algae leaching liquor according to claim 1 is characterized in that, the add-on of water is by weight marine alga: water=1:10-1:25 in the step (1)
3. the method that contains the marine alga bacterial manure of algae leaching liquor according to claim 1 is characterized in that, used PH conditioning agent is ammoniacal liquor in the step (2).
4. the method that contains the marine alga bacterial manure of algae leaching liquor according to claim 1 is characterized in that, the inoculum size of subtilis is the 0.1-5% with respect to culture volume in the step (3).
5. the method that contains the marine alga bacterial manure of algae leaching liquor according to claim 1 is characterized in that, subtilis activates first before inoculation in the step (3), and activation temperature is 35-40 ℃.
6. the method that contains the marine alga bacterial manure of algae leaching liquor according to claim 1 is characterized in that, zymotechnique described in the step (3) is liquid ventilating fermentation technique, temperature is 35-40 ℃, 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.
7. one kind prepares the described method that contains the marine alga bacterial manure of algae leaching liquor of claim 1, it is characterized in that, may further comprise the steps:
(1) marine alga lixiviate
After the marine alga pulverizing, add entry and 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, namely get required fermention medium;
(3) inoculation and fermentation
Subtilis is inoculated in step (2) the gained substratum, and fermentation namely gets marine alga bacterial manure.
，
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| CN105936881B (en) * | 2016-06-01 | 2019-08-13 | 中国科学院烟台海岸带研究所 | One kind is for alignic thermophilic sugared bacillus and its application method of degrading |
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| CN108484314A (en) * | 2018-03-19 | 2018-09-04 | 辽宁省农业科学院 | It is a kind of promote hickory chick growth microorganism liquid seaweed extract and its application |
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| CN110683870A (en) * | 2019-11-06 | 2020-01-14 | 中国农业科学院烟草研究所 | Preparation method and application of biofertilizer prepared from kelp residues and bacillus |
| CN112209768A (en) * | 2020-10-23 | 2021-01-12 | 内蒙古阜丰生物科技有限公司 | Method for producing alginic acid fertilizer |
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| CN113322217A (en) * | 2021-08-04 | 2021-08-31 | 中国科学院烟台海岸带研究所 | Vibrio freundii and application thereof in preparation of multifunctional small-tank fertilizer |
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