CN101717301B - Capsule type microbial fertilizer and preparation method thereof - Google Patents
Capsule type microbial fertilizer and preparation method thereof Download PDFInfo
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- CN101717301B CN101717301B CN200910237872A CN200910237872A CN101717301B CN 101717301 B CN101717301 B CN 101717301B CN 200910237872 A CN200910237872 A CN 200910237872A CN 200910237872 A CN200910237872 A CN 200910237872A CN 101717301 B CN101717301 B CN 101717301B
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Abstract
The invention relates to a capsule type microbial fertilizer and a preparation method thereof, and belongs to the technical fields of microbiology engineering and plant nutrition. In the capsule type microbial fertilizer, environmentally-friendly and biodegradable substances are taken as a wall material, and useful microorganisms are taken as a core material and are capsulated by a piercing-solidifying incuber method; the environment-friendly and biodegradable wall material is calcium alginate, sodium carboxymethylcellulose and bentonite; and the useful microorganisms are Raoultella planticola, Klebsiella oxytoca, Enterobacter cloacae and Bacillus subtilis. The grain sizes of wet and dry capsules are respectively between 2.40 and 3.50mm and between 0.8 and 1.5mm; the water content is 90 to 98 percent; the diameter diffusion coefficient of dry capsules is 1.3 to 1.5; the quality diffusion coefficient is 1.9 to 2.5; the embedding rate is over 99 percent; the viable count is 1,014CFU/g; and the releasable viable count is 109 to 10CFU/g. After the capsule type microbial fertilizer is stored for 2 months, the effective bacteria amount can reach 108-9CFU/g. The microbial fertilizer has the advantages of long shelf life and stable product, overcomes influence of various factors in the application process, and has the effects of preventing diseases, resisting diseases, relieving salt stress, promoting plant growth and the like in the seedling stage of crops.
Description
Technical field
The present invention relates to develop a kind of capsule type microbial fertilizer and preparation method, microorganism belonging to genus engineering and plant nutrition technical field.The exploitation of this microbial fertilizer formulation can solve in the microbial fertilizer that effectively the thalline preservation period is short, product is unstable, in application process, receive problem such as multiple factor affecting easily, can effectively improve the flexibility etc. of survival time, long-lasting, stability and the environment of thalline.
Background technology
At present, the main still chemical fertilizer of employed fertilizer in the agricultural production process, but because its utilization ratio is low, residual serious, long-term a large amount of use, adverse consequences such as caused anthropogenic soil to harden, organicly descend, soil nutrient is out of proportion.Therefore, need a kind of better product badly and replace chemical fertilizer, promptly microbial fertilizer manufacturing and using.
Some special efficacies of the short living bacterium (PGPR) of plant rhizosphere, promotion plant-growth that can be direct or indirect.The first, the nitrogen of PGPR in can fixed air is organonitrogen, supplies with self and uses with plant materials, has improved the nitrogen nutrition of crop to a great extent, thereby has reached the effect of crop yield.The second, PGPR can produce and secrete variously has the carrier of high-affinity to iron, and dissolving also combines the ferro element in the soil to supply the vegetable cell utilization; In addition, generation have a liking for the iron element, make the pathogenic bacteria of plant rhizosphere can not get enough iron and grow normally and grow surely, also reduced the harm of pathogenic bacteria to crop.The 3rd, PGPR can synthesize different plant hormones, like growth hormone, ethene, phytokinin, VITAMINs etc., can promote plant-growth in different developmental phases, and especially the stimulating plant root system grows; Simultaneously, the synthetic microbiotic has also limited breeding and the growth of pathogenic bacterium.The 4th, PGPR solubilized mineral substance.The mikrobe phosphorus decomposing is because its secretion organic acid can reduce pH in soil, makes some indissolubles or insoluble phosphorus ore thing be transformed into the phosphorus that plant can absorb.
The earliest microbial fertilizer is that Germany in 1895 releases " Nitragin " rhizobium inoculant in the world.All there has been the rhizobium inoculant industry in country such as the U.S., Australia, Britain to 20th century 30, the forties, and development is very fast afterwards.At present, have at least more than 70 country that oneself microbial fertilizer manufacturing enterprise, product technology standard and quality surveillance system are arranged.China more than 500 tame microbial fertilizer enterprise basically forms the microbial fertilizer industry, has more than 300 at the product of the formal registration of the national Ministry of Agriculture, becomes the important member in the fertilizer family gradually.The microbial fertilizer purposes has all kinds such as molten phosphorus, potassium decomposing, short life, diseases prevention, stalk become thoroughly decomposed, and has the multinomial function of nutrition, short life, diseases prevention, degraded soil pollutant.Microbial fertilizer uses area and constantly enlarge, and the minority product begins to get into the world market increasing production, improve product quality, reducing effects such as fertilizer amount, diseases prevention gradually by Agricultural management person, peasant's understanding.
The use of microbial fertilizer overcome utilization rate of fertilizer low with problem such as residual contamination, and obtained significant effect to a great extent.At present, see that the finished product formulation of China's microbial fertilizer mainly is divided into two kinds of liquid and solids from microbial fertilizer finished product proterties.Liquid has plenty of by fermented liquid directly bottles, and also has to use the Dormant oils front cover; Solid dosage mainly is carrier with the peat composed of rotten mosses, divide pulvis, two kinds of formulations of particle, and also useful in recent years vermiculite is a sorbent material; Also useful fermented liquid concentrates the goods of postlyophilization.But mentioned microorganism fertilizer formulation; Mikrobe all directly is exposed in the soil in application process; And receive the influence of multiple adverse environmental factors, weather condition easily; Like factor such as microbial population, illumination in the temperature of soil, humidity, pH, the soil, cause problems such as the microbial fertilizer long-lasting is poor, viable bacteria survival time weak point, product performance instability, and then influenced the promotion and application of microbial fertilizer product greatly.
Therefore, the exploitation of microbial fertilizer formulation also is an importance of studying over nearly 10 years.In microbial inoculum, adding some effect of inoculation that can improve mikrobe and do not suppress nutritive substance, the environmental friendliness material of its survival again, is an aspect of exploratory development in recent years.The invention provides a kind of capsule type microbial fertilizer and preparation method.
Encapsulated technology is a kind of resist technology, is about to some and has reactive behavior, susceptibility or easy volatile material and be encapsulated as fine particle, and this technology is widely used in fields such as food, medicine, makeup, textiles, feed.Because encapsulated technology separates core and environment on every side, has avoided light, O
2, temperature, pH etc. influence, effectively protected core.In recent years, capsule technique was widely used in the immobilization aspect of mikrobe, animal and plant cells, enzyme and other multiple biologically active substances and chemicals.The present invention is wrapped in one or more useful microbes in biodegradable and the environmentally friendly material, is prepared into capsule type microbial fertilizer.This product has overcome the influence of various factors in application process; Improved greatly microbial fertilizer long-lasting, prolonged the beneficial microorganism survival time, strengthened the stability of product; Especially to crops seedling stage have diseases prevention, disease-resistant, alleviate salt stress, promote the effect of plant-growth, and then influenced the promotion and application of this microbial fertilizer product greatly.
Summary of the invention
To the objective of the invention is that effectively the thalline preservation period is short in the existing microbial fertilizer, product is unstable in order solving, in application process, to receive problem such as multiple factor affecting easily, a kind of capsule type microbial fertilizer and preparation method are provided.
The objective of the invention is to realize through following technical proposals.
A kind of capsule type microbial fertilizer of the present invention comprises wall material and core, adopts orifice-precipitation bath method to carry out encapsulated to useful microbe; Wherein, the wall material is environmentally friendly and the biodegradable material, and core is a useful microbe;
Described environmentally friendly and biodegradable wall material are meant alginate calcium, Xylo-Mucine and wilkinite;
Said useful microbe is meant plants living Raoul bacterium (Raoultella planticola), acid-producing Klebsiella bacterium (Klebsiella oxytoca), enterobacter cloacae (Enterobacter cloacae), subtilis (Bacillus subtilis).These mikrobes can reduce the amount of interior ethene (Eth) of plant body and endogenous dormin (ABA); Plant is to Na under the reduction salt stress
+The absolute absorption amount, increase K
+, Ca
2+And Fe
2+Absorption, remove secondary salinization effectively to the coercing of crop, the crops seedling stage disease is had good prevention, resistant function, can promote plant strain growth and to the absorption of nutritive substance.
A kind of preparation method of capsule type microbial fertilizer, concrete performing step is following:
Step 1, useful microbe activation, the liquid culture of cryopreservation is centrifugal, abandon supernatant, with bacterium mud cryopreservation, subsequent use; Comprising:
(1) NA culture medium preparation
With NaCl (5.0g), peptone (10.0g), Carnis Bovis seu Bubali cream (3.0g), H
2O (1000ml) mixes, configuration NA substratum; If solid NA substratum then adds 1.8% agar; Sterilization, subsequent use;
(2) activation of bacterial strain
The useful microbe of cryopreservation is rule on the NA solid medium, 25-32 ℃ following constant temperature culture 2-3 days; Picking list bacterium colony, streak culture on new NA solid medium, so 2-3 time;
(3) beneficial microorganism is cultivated
With the useful microbe after the activation, be linked in the NA liquid nutrient medium, under 25-32 ℃, the condition of 150-220rpm, be cultured to early stage stationary phase, centrifugal, abandon supernatant, preserve bacterium mud, subsequent use.
Step 2; Sodium-alginate, Xylo-Mucine, wilkinite are mixed according to certain ratio; Be that sodium alginate concentration (W/V) is 1.5-5%, sodium-alginate is 5-12 with the ratio (W/W) of Xylo-Mucine: 1, and sodium-alginate is 5-10 with the ratio (W/W) of wilkinite: 1; Sterilization, subsequent use;
Step 3, compound concentration (W/V) is the calcium chloride solution of 1-5%; Sterilization, subsequent use;
Step 4, with the wall material solution of the bacterium mud of step 1 and step 2 according to 1-3: 1 mixed is even, in the mixed solution calcium chloride solution that splashes into step 3 at the uniform velocity with certain volume, fixedly forms capsule after 20-60 minute; Take out capsule,, promptly obtain capsule type microbial fertilizer with aseptic water washing 2-3 time.
Wet, the dried capsular particle diameter that makes through above-mentioned steps is respectively 2.40-3.50mm and 0.80-1.50mm, and water cut is 90-98%, and doing capsular diameter spread coefficient is 1.3-1.5; The mass diffusion coefficient is 1.9-2.5; Embedding rate is higher than 99%, is close to 100%, and contained viable count reaches 10
14CFU/g, releasable viable count reaches 10
9-10CFU/g.At room temperature be dried to constant weight, store after two months, effectively bacteria containing amount can reach 10
8-9CFU/g.
Beneficial effect:
This capsule type microbial fertilizer has overcome the influence of various soil factors and climatic factor; The encapsulated effect that has played slowly-releasing, controlled release; Thereby reached the effect of a long-term fertilizer efficiency; Improved greatly microbial fertilizer long-lasting, prolonged the beneficial microorganism survival time, strengthened the stability of product, especially to crops seedling stage have diseases prevention, disease-resistant, alleviate salt stress, promote the effect of plant-growth, and then influenced the promotion and application of this microbial fertilizer product greatly.
Description of drawings
Fig. 1 is wet capsular picture;
Fig. 2 is for doing capsular picture;
Fig. 3 is the Electronic Speculum picture of capsule type microbial fertilizer (5.0kv 10.3mm * 80)
Fig. 4 is the Electronic Speculum picture of capsule type microbial fertilizer (5.0kv 10.6mm * 300).
Embodiment
Below in conjunction with accompanying drawing and embodiment the present invention is elaborated.
A kind of capsule type microbial fertilizer of the present invention and preparation method; With biodegradable such as alginate calcium, Xylo-Mucine and wilkinite and environmentally friendly material is the wall material; Diseases prevention, one or more beneficial microbe strains disease-resistant, that alleviate salt stress, promotion plant-growth of obtaining with screening from alkaline land soil are core, adopt orifice-precipitation bath method to carry out the exploitation of capsule type microbial fertilizer.
Embodiment 1
Sodium alginate concentration (W/V) 2.5%
Sodium-alginate/Xylo-Mucine (W/W) 5: 1
The ratio (W/W) of sodium-alginate and wilkinite 5: 1
Calcium chloride concentration (W/V) 5%
Bacterium mud/wall material (V/V) 1: 1
Sodium-alginate, Xylo-Mucine, wilkinite are mixed according to aforementioned proportion, and the preparation calcium chloride solution; Moist heat sterilization is 20 minutes under 121 ℃ of conditions, and is subsequent use.With the bacterium mud of prior cultured useful microbe centrifugal preservation under the 10000rpm condition proportionally with wall material solution mixing; And then draw mixed solution splashing in the calcium chloride solution at the uniform velocity with syringe; Fixing 30 minutes; Use the aseptic water washing capsule again 3 times, promptly obtain capsule type microbial fertilizer.
Gained is wet, do capsular particle diameter is respectively 2.65-2.90mm and 1.00-1.25mm, and water cut is 94%, does capsular diameter spread coefficient and be 1.35, the mass diffusion coefficient is 2.13, and embedding rate is 99.99%, and contained viable count reaches 10
14CFU/g, releasable viable count reaches 10
9CFU/g.At room temperature be dried to constant weight, store after two months, effectively bacteria containing amount can reach 10
8CFU/g.
Embodiment 2
Sodium alginate concentration (W/V) 2.0%
Sodium-alginate/Xylo-Mucine (W/W) 10: 1
The ratio (W/W) of sodium-alginate and wilkinite 10: 1
Calcium chloride concentration (W/V) 5%
Bacterium mud/wall material (V/V) 3: 1
The preparation method is identical with embodiment 1.
Gained capsule character is: wet, do capsular particle diameter and be respectively 2.48-2.70mm and 0.90-1.10mm, water cut is 95%, does capsular diameter spread coefficient and be 1.42, the mass diffusion coefficient is 2.0, and embedding rate is 99.99%, and contained viable count reaches 10
14CFU/g, releasable viable count reaches 10
9-10CFU/g.At room temperature be dried to constant weight, store after two months, effectively bacteria containing amount can reach 10
8-9CFU/g, the gained capsule is like accompanying drawing 1,2,3, shown in 4.
Embodiment 3
Sodium alginate concentration (W/V) 1.5%
Sodium-alginate/Xylo-Mucine (W/W) 8: 1
The ratio (W/W) of sodium-alginate and wilkinite 8: 1
Calcium chloride concentration (W/V) 2%
Bacterium mud/wall material (V/V) 2: 1
The preparation method is identical with embodiment 1.
Gained is wet, do capsular particle diameter is respectively 2.38-2.55mm and 0.85-1.00mm, and water cut is 96%, does capsular diameter spread coefficient and be 1.40, the mass diffusion coefficient is 1.9, and embedding rate is 99.99%, and contained viable count reaches 10
14CFU/g, releasable viable count reaches 10
9-10CFU/g.At room temperature be dried to constant weight, store after two months, effectively bacteria containing amount can reach 10
8-9CFU/g.
Embodiment 4
Sodium alginate concentration (W/V) 3.0%
Sodium-alginate/Xylo-Mucine (W/W) 12: 1
The ratio (W/W) of sodium-alginate and wilkinite 8: 1
Calcium chloride concentration (W/V) 2%
Bacterium mud/wall material (V/V) 2: 1
The preparation method is identical with embodiment 1.
Gained is wet, do capsular particle diameter is respectively 2.90-3.20mm and 1.18-1.38mm, and water cut is 92%, does capsular diameter spread coefficient and be 1.38, the mass diffusion coefficient is 2.24, and embedding rate is 99.99%, and contained viable count reaches 10
14CFU/g, releasable viable count reaches 10
9CFU/g.At room temperature be dried to constant weight, store after two months, effectively bacteria containing amount can reach 10
8CFU/g.
Embodiment 5
Sodium alginate concentration (W/V) 5%
Sodium-alginate/Xylo-Mucine (W/W) 8: 1
The ratio (W/W) of sodium-alginate and wilkinite 10: 1
Calcium chloride concentration (W/V) 2%
Bacterium mud/wall material (V/V) 2: 1
The preparation method is identical with embodiment 1.
Gained is wet, do capsular particle diameter is respectively 3.10-3.45mm and 1.20-1.54mm, and water cut is 90%, does capsular diameter spread coefficient and be 1.36, the mass diffusion coefficient is 2.38, and embedding rate is 99.99%, and contained viable count reaches 10
14CFU/g, releasable viable count reaches 10
8-9CFU/g.At room temperature be dried to constant weight, store after two months, effectively bacteria containing amount can reach 10
7-8CFU/g.
Microbial fertilizer long preservative period of the present invention, product be stable, in application process, overcome multiple factor affecting, to crops seedling stage have diseases prevention, disease-resistant, alleviate salt stress, promote effects such as plant-growth.
Claims (1)
1. a capsule type microbial fertilizer is characterized in that: comprise wall material and core, adopt orifice-precipitation bath method to carry out encapsulated to useful microbe; Wherein, the wall material is environmentally friendly and the biodegradable material, and core is a useful microbe;
Described environmentally friendly and biodegradable wall material are meant sodium-alginate, Xylo-Mucine and wilkinite;
Described useful microbe is meant plants living Raoul bacterium, acid-producing Klebsiella bacterium, enterobacter cloacae, subtilis;
The preparation process of said capsule type microbial fertilizer:
Step 1, useful microbe activation, the liquid culture of cryopreservation is centrifugal, abandon supernatant, with bacterium mud cryopreservation, subsequent use; Comprising:
(1) NA culture medium preparation
With 5.0g NaCl, 10.0g peptone, 3.0g Carnis Bovis seu Bubali cream, 1000ml H
2O mixes, configuration NA substratum; If solid NA substratum then adds 1.8% agar; Sterilization, subsequent use;
(2) activation of bacterial strain
The useful microbe of cryopreservation is rule on the NA solid medium, 25-32 ℃ following constant temperature culture 2-3 days; Picking list bacterium colony, streak culture on new NA solid medium, so 2-3 time;
(3) beneficial microorganism is cultivated
With the useful microbe after the activation, be linked in the NA liquid nutrient medium, under 25-32 ℃, the condition of 150-220rpm, be cultured to early stage stationary phase, centrifugal, abandon supernatant, preserve bacterium mud, subsequent use;
Step 2 is mixed sodium-alginate, Xylo-Mucine, wilkinite according to certain ratio, promptly sodium alginate concentration W/V is 1.5-5%, and sodium-alginate is 5-12 with the ratio W/W of Xylo-Mucine:
1, sodium-alginate is 5-10:1 with the ratio W/W of wilkinite; Sterilization, subsequent use;
Step 3, compound concentration W/V is the calcium chloride solution of 1-5%; Sterilization, subsequent use;
Step 4, the wall material solution of the bacterium mud of step 1 and step 2 is even according to the mixed of 1-3:1, in the mixed solution calcium chloride solution that splashes into step 3 at the uniform velocity with certain volume, fixedly form capsule after 20-60 minute; Take out capsule,, promptly obtain capsule type microbial fertilizer with aseptic water washing 2-3 time.
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| CN102199056A (en) * | 2010-12-15 | 2011-09-28 | 沈阳科丰牧业科技有限公司 | Method for prolonging storage life of compound microbial fertilizer |
| CN102199050B (en) * | 2010-12-15 | 2013-06-12 | 沈阳科丰牧业科技有限公司 | Composite microbial fertilizer and preparation method thereof |
| CN102409035A (en) * | 2011-05-06 | 2012-04-11 | 江南大学 | Preparation method and application of microecological bactericide for sustained release in water |
| CN102503663B (en) * | 2011-10-14 | 2013-12-25 | 上海市农业科学院 | Preparation method for microcapsule organic fertilizer |
| EP2773599B1 (en) * | 2011-11-04 | 2019-06-19 | Terragen Holdings Limited | Microbial inoculants and fertilizer compositions comprising the same |
| CN104744108A (en) * | 2013-12-27 | 2015-07-01 | 汪鹤亭 | A novel process of preparing an agricultural-bacterium-containing capsule by a 'carrier water transferring' method |
| CN107032873A (en) * | 2016-12-27 | 2017-08-11 | 青岛苏贝尔作物营养有限公司 | Alleviate the microbial manure and preparation method of vegetables in greenhouse booth salt stress |
| CN106748496A (en) * | 2016-12-29 | 2017-05-31 | 寻甸民生生物工程有限责任公司 | A kind of biological organic fertilizer and preparation method thereof |
| CN107857649A (en) * | 2017-12-02 | 2018-03-30 | 长沙无道工业设计有限公司 | A kind of microbial organic fertilizer and preparation method thereof |
| WO2020021549A1 (en) * | 2018-07-25 | 2020-01-30 | Lavie Bio Ltd. | Encapsulated microorganisms and methods of using same |
| CN109536410B (en) * | 2018-12-17 | 2020-07-17 | 石河子大学 | Salt-tolerant growth-promoting composite microbial inoculum and preparation method and application thereof |
| CN112062632A (en) * | 2020-08-28 | 2020-12-11 | 西安工程大学 | Synthetic method of coated biochar slow-release fertilizer |
| CN114231517A (en) * | 2021-12-06 | 2022-03-25 | 上海纳米技术及应用国家工程研究中心有限公司 | Preparation method of cell immobilization material for large-scale production of exosome, product and application thereof |
| CN116874337B (en) * | 2023-09-08 | 2024-02-02 | 深圳市五谷网络科技有限公司 | Liquid biological slow-release fertilizer and preparation method thereof |
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