CN1535255A

CN1535255A - Biological fertilizer compositions comprising manure, sludge or garbage

Info

Publication number: CN1535255A
Application number: CNA028083571A
Authority: CN
Inventors: 张令玉
Original assignee: Ultra Biotech Ltd
Current assignee: Ultra Biotech Ltd
Priority date: 2001-03-01
Filing date: 2002-03-01
Publication date: 2004-10-06
Also published as: AU2002237398B2; WO2002070436A2; EP1363865A2; WO2002070436A3

Abstract

The present invention provides biological fertilizer compositions that comprise yeast cells and an organic substrate. The yeast cells of the invention have an enhanced ability to fix atmospheric nitrogen, decompose phosphorus minerals and compounds, decompose potassium minerals and compounds, decomposes complex carbon compounds, overproduce growth factors, overproduce ATP, decompose undesirable chemicals, suppress growth of pathogenic microorganisms, or reduces undesirable odor. The organic substrates used can be manure, sludge, or garbage. The biological fertilizer composition of the invention can replace mineral fertilizers in supplying nitrogen, phosphorus, and potassium to crop plants. Methods of manufacture biological fertilizer compositions, and methods of uses are also encompassed.

Description

The Biofertiliser composition that contains muck, mud or rubbish

1. invention field

The present invention relates to contain the bio-feritlizer of yeast and organic substrate.Yeast in the present composition is stimulated to exercise multiple function, comprises organic materials is converted into harmless plant nutrition.The invention still further relates to the method for preparing bio-feritlizer, and use this bio-feritlizer to increase the method for crop yield.

2. background of invention

The use of fertilizer is that to keep the high yield crop growth necessary.In the required basic nutrition of plant health growth, the most of farm crop on the most of soils need a large amount of nitrogen (with NO ₃ ^-Or NH ₄ ⁺Form absorb), phosphorus is (with H ₂PO ₄ ^-Form absorb) and potassium (with K ⁺Form absorb) nutrition (Wichmann, W., et al., IFA World Fertilizer Use Manual).These a large amount of nitrogen, phosphorus and potassium nutrition owner will provide with the form of mineral manure, be natural mineral matter or artificial chemical preparations (K.F.Isherwood through processing, 1998, Mineral Fertilizer Useand the Environment, United Nations Environmental Programme TechnicalReport No.26).

Although mineral manure is providing very important aspect the sufficient agricultural prods to the mankind, it also is familiar with the harm of environment in recent years.Mineral manure can damage soil.For example, most of nitrogenous fertilizer can make soil acidification, therefore are unfavorable for the growth of plant and other soil organisms.Widely-used chemical nitrogen fertilizer also can suppress the activity of natural nitrogen-fixing microorganism, has reduced the natural fertilizer of soil thus.Life-time service mineral manure can also cause the serious environmental pollution.For example, because losing of nitrogenous fertilizer that leaching and soil erosion cause and phosphate fertilizer causes the eutrophication of soil and phreatic pollution and surface water.The soil of purifying contaminated and water are a complexity and difficult task.The cost of this task also is huge.

When seeking the terms of settlement of this problem, some revert to organic fertilizer, for example muck (Wichmann, W., et al., IFA World Fertilizer Use Manual).The ight soil as fertilizer sources is used the initial stage that can trace back to agricultural.Livestock produces a large amount of muck.For example, produce the refuse that surpasses 13,600 ten thousand tonnes (dry weights) on the annual farm of the U.S. (comprising the raising activity of captive animal).Muck keep and improve aspect the soil valuable because it contains plant nutrition, soil ulmin and organism.The nitrogen, phosphorus and the potassium that studies show that the raising cow of high per-cent are drained in muck.

Because muck need carry out careful processing and could therefrom obtain maximum value, some peasants may be unwilling to spend necessary time and efforts.Muck must be stored to reduce losing of nutrition by careful.Must be applied to correct farm crop kind in the suitable time.Usually, muck can not provide all plant nutritions that need, and still will apply very a large amount of organic fertilizer to soil.Therefore, a kind of trend that the muck as fertilizer sources is worth of underestimating is arranged.Muck also may contain the adverse chemical product, for example microbiotic and hormone.Only maybe can not obtain and the relatively cheap under-developed country of labor force in the artificial manure costliness, the muck as fertilizer sources is just attractive.

In addition, muck can contain the nitrogen and the phosphorus of conspicuous level, if incorrect processing can threaten the water source.If muck is not properly stored or removes, can pose a health risk and environment.For example, muck can cause atmospheric pollution, i.e. smell and dust; And superfluous nutrition, organism, salt and pathogen contamination surface water and underground water.For example, muck contains pathogenic microorganism such as intestinal bacteria, Salmonellas, Shigellae and campylobacter jejuni.

The bio-feritlizer of use microorganism has been suggested the substitute as mineral manure.Naturally occurring nitrogen-fixing microorganism comprises bacterium, and for example root nodule bacterium, vinelandii and azospirillum (are seen for example United States Patent (USP) 5,071, No. 462), and fungi, for example flavus/aspergillus oryzae (is seen for example United States Patent (USP) 4,670, No. 037) in bio-feritlizer, use.The naturally occurring microorganism that can to make phosphate rock ore deposit or other insoluble phosphoric acid salt solubilising be soluble phosphate is also used in bio-feritlizer, use respectively with nitrogen-fixing microorganism that (for example United States Patent (USP) 5,912, No. 398) or (for example United States Patent (USP) 5 to unite use with nitrogen-fixing microorganism, 484, No. 464).The bacterial isolates of genetic modification is developed, and uses in bio-feritlizer.Developed and a kind of method based on recombinant DNA technology, produce be used for bio-feritlizer can more effective fixed nitrogen, divide phosphorus decomposing and divide the bacterial isolates of potassium decomposing, for example see United States Patent (USP) 5,578, No. 486; The open WO95/09814 of PCT; Chinese patent is open: CN1081662A; CN 1082016A; CN 1082017A; CN 1103060A; And CN1109595A.

But, can not enough substitute mineral manure effectively usually based on the natural bio-feritlizer of microorganism that exists.Therefore development can substitute mineral manure provides nitrogen, phosphorus and potassium to produce the high quality agricultural prods to farm crop, avoids the better bio-feritlizer of mineral manure associated problem just very important simultaneously.

The invention provides the bio-feritlizer that can substitute mineral manure based on non-recombination yeast, and the effective using method favourable to environment of some organic materials.

Here citing document and do not mean that any document of admitting here to be quoted is relevant prior art, or admit that the document quoted has substantial effect to the patentability of the application's claim.These documents are based on the obtainable information of applicant all about the statement on date with about the statement of content, do not constitute any admitting about these document dates and content exactness.

3. summary of the invention

The present invention relates to Biofertiliser composition.Biofertiliser composition of the present invention comprises the most nearly nine kinds of different yeast cell components, organic substrate component and optional inorganic matrix components.Particularly, each yeast cell component of composition has a kind of in following ten kinds of functions at least, promptly be respectively fixedly atmospheric nitrogen, decompose insoluble phosphorus or potassium inorganics, keep phosphorus compound in the microenvironment balance, decompose complicated carbonaceous material or compound, excessive generation somatomedin, excessive generation ATP, suppress pathogenic microorganism growth, decompose the adverse chemical product, reduce organic smell.The organic substrate component can comprise muck, mud or rubbish.Yeast cell component of the present invention can be used as additive, mixes with organic materials to form bio-feritlizer.

In one embodiment, Biofertiliser composition of the present invention prepares by the organic substrate component is mixed with at least seven to nine primary yeast cellular components, wherein the cell of six primary yeast cellular components is exercised fixedly atmospheric nitrogen, decompose inorganic phosphor-contained thing or keep the balance of its surrounding environment phosphorus compound, decompose and contain the potassium inorganics, decompose complicated carbonaceous material or compound, the basic function of excessive generation somatomedin and excessive generation ATP, wherein the cell of other component is exercised and is suppressed the pathogenic microorganism growth, the subsidiary function of decomposing organic substrate smell in adverse chemical product and the minimizing Ru 2006101161.

In preferred embodiments, the yeast that the present invention uses is that the on sale and/or public can obtain on the market, such as but not limited to yeast saccharomyces cerevisiae (Saccharomyces cerevisiae).Usually, yeast cell component of the present invention is to prepare by cultivate a plurality of yeast cell under activation condition, and the ability that makes these a plurality of cells exercise these functions is activated or strengthens.Therefore, in another embodiment, the present invention includes a kind of method in activation or ten kinds of functions of reinforcement yeast cell enforcement.The invention still further relates to the preparation method of fertilizer, comprise the organic substrate component is mixed with yeast cell of the present invention, dry then and pack finished product.

The invention further relates to the method for using Ru 2006101161 of the present invention.Biofertiliser composition of the present invention is used to support and promote the growth and the maturation of each kind of plant.

4. accompanying drawing summary

Fig. 1: the activation of yeast cell.1 yeast cell culture; 2 containers; 3 electromagnetism field sources.

Fig. 2: the formation of symbiosis sample relation between the yeast strain.4 fixed nitrogen zymic electromagnetism field sources; 5P-decomposes zymic electromagnetism field source; 6K-decomposes zymic electromagnetism field source; 7C-decomposes zymic electromagnetism field source; 8 yeast cell cultures; 9 containers.

Fig. 3: yeast cell adapted soil kind.10 input electrodes; 11 containers; 12 electrodes; 13 yeast cell cultures; 14 electromagnetism field sources; 15 temperature regulators.

Fig. 4: the crushing process of organic substrate.16 organic raw material; 17 crushers; 18 shredders; 19 pulverous organic substrates.

Fig. 5: the crushing process of inorganic matrix.20 inorganic raw materials; 21 crushers; 22 shredders; 23 Powdered inorganic matrixes.

Fig. 6: yeast fermentation process.24 activatory yeast cell; 25 yeast cell culture tank, starch: water (35 ℃)=1: 2.5-3.5, carry out half aerobic fermentation at 28 to 30 ℃; The culture of 26 results.

Fig. 7: mix organic and inorganic raw material.27 inorganic materials; 28 starch; 29 organic materialss; 30 agitators; 31 mixtures; 32 wait to be delivered to the mixture of fertilizer preparatory phase.

Fig. 8: mixed yeast cell.33 fixed nitrogen yeast; 34P-decomposes yeast; 37K-decomposes yeast; 55C-decomposes microorganism; 35ATP-produces yeast; 36GF-produces yeast; 52 suppress the yeast of pathogenic agent; 53 decompose the yeast of adverse chemical product; 54 deodorizing yeast; 38 yeast mixts; 56 wait to be delivered to the mixture of fertilizer preparatory phase.

Fig. 9: fertilizer preparation process.39 yeast mixts; 40 organic and inorganic materials mixtures; 41 tablets presss; 42 fertiliser granulates.

Figure 10: drying process.43 fertiliser granulates; 44 first drying machines; 45 second drying machines; 46 exsiccant fertilizer.

Figure 11: cooling and wrapping process.47 exsiccant fertilizer; 48 water coolers; 49 separators; 50 sack packers; 51 finished products.

5. detailed description of the Invention

The invention provides the Biofertiliser composition that comprises yeast cells and organic substrate. The present invention also provides the method for preparing Biofertiliser composition and the method for using this Biofertiliser composition.

Biofertiliser composition of the present invention can substituted chemistry/inorganic fertilizer, to plant particularly crop plants nitrogen (N), phosphorus (P) and potassium (K) are provided. In Biofertiliser composition of the present invention, contained organic substrate such as muck, mud or rubbish provide a kind of environment acceptable and economic method that reuses these materials.

According to the present invention, Biofertiliser composition comprises brid guano fertilizer and multiple yeast cells component. Every primary yeast cellular component is a yeast cells group who comprises a plurality of yeast cells that can exercise required function. Yeast cells component of the present invention can provide following six kinds of basic functions: (1) is atmospheric nitrogen fixedly; (2) divide phosphorus decomposing inorganic matter or compound, perhaps keep the balance of phosphorus compound; (3) divide potassium decomposing inorganic matter or compound; (4) decompose material with carbon element or compound complicated or HMW; (5) excessive generation growth factor; (6) excessive generation ATP. Yeast cells component of the present invention can provide following miscellaneous function: (7) suppress the growth of pathogen, (8) degraded adverse chemical product, or (9) reduce the smell of organic material.

In one embodiment, Biofertiliser composition of the present invention comprises (I) brid guano fertilizer; (II) a kind of in following at least yeast cells component: (a) the first yeast cells component contains the yeast cells of first group of fixed nitrogen; (b) the second yeast cells component, contain the yeast cells of second component solution phosphorus compound; Or (c) the 3rd yeast cells component, contain the yeast cells of the 3rd component solution potassium compound; A kind of in (III) following at least: (d) the 4th yeast cells component contains the 4th group of yeast cells that suppresses the pathogenic microorganisms growth; (e) the 5th yeast cells component, contain the 5th group of antibiotic yeast cells of degraded; Or (f) the 6th yeast cells component, contain the 6th group of yeast cells that reduces this Biofertiliser composition smell. Therefore, Biofertiliser composition of the present invention comprises at least two primary yeast cellular components, a kind ofly provides a kind of in listed three kinds of basic functions, and another kind provides miscellaneous function. In another embodiment, above-mentioned Biofertiliser composition further comprises a kind of in following at least: (g) the 7th yeast cells component contains the 7th group of yeast cells that the complex carbon compound is converted into simple carbohydrates; (h) the 8th yeast cells component, contain the yeast cells of the 8th group of excessive generation growth factor; Or (i) the 9th yeast cells component, contain the yeast cells of the 9th group of excessive generation atriphos. In preferred embodiments, the yeast cells component that provides all six kinds of basic functions to add at least a miscellaneous function is provided Biofertiliser composition of the present invention. Therefore, preferred Biofertiliser composition comprises seven kinds, eight kinds or nine kinds of different yeast cells components.

Many group yeast cell of the present invention can be added in the muck or existing organic fertilizer of any kind of, to improve their performance.

Organic substrate in the Ru 2006101161 provides the source of nitrogen, phosphorus and potassium.Choose wantonly, Ru 2006101161 can comprise the inorganic component that contains mineral substance, and it provides the source of extra phosphorus and/or potassium, and other mineral substance is such as but not limited to calcium, magnesium and sulphur; And trace nutrient, such as but not limited to boron, copper, iron, manganese, molybdenum and zinc.

Biofertiliser composition of the present invention is compared with organic fertilizer with mineral manure has many advantages.Because the Metabolic activity that bio-feritlizer of the present invention utilizes viable yeast with raw material for example phosphorus and the potassium compound in atmospheric nitrogen, the matrix components be converted into plant nutrition, the regulation and control that yeast cell is subjected to soil nutrient content to the conversion and the release portion of these nutrition.The nutrient contg of soil depends in part on environment and the continuous demand that changes of plant again.Therefore, the plant nutrition of this Biofertiliser composition discharges and can be accepted by edaphic condition, and can be kept for some time.

Biofertiliser composition of the present invention also provides three kinds of subsidiary functions except providing the nutrition to plant, alleviates some bad characteristics that restriction muck, mud and rubbish use as organic fertilizer.Be present in the health that pathogenic bacterium in muck, mud and the rubbish threaten people and domestic animal.Biofertiliser composition of the present invention can comprise a kind of yeast cell component that can suppress pathogenic bacterium propagation, has reduced the danger of infecting thus, and has avoided using chemical to control the needs that these pathogenic agent are sent out.Can comprise the yeast cell component that another kind can reduce muck, mud or waste odours in the said composition, make to comprise in the fertilizer that these materials are easier to be accepted.Can also comprise another kind of yeast cell component with degraded adverse chemical product, for example see the antibiotic feed additive in muck, mud or the rubbish.These subsidiary functions have alleviated the negative impact of using muck, mud or rubbish that environment is caused usually in fertilizer.Provide every kind of the yeast cell component of subsidiary function can be separately to combine with other the six kinds yeast cell components that basic function is provided or they are united to each other with other six kinds of components and combine the various subsidiary functions that provide required.

Though following term is considered to have in this area clear and definite implication, following still they being illustrated is beneficial to explain the present invention.

Phrase used herein " fixed nitrogen " or " fixedly atmospheric nitrogen " comprise with the nitrogen transformation in dinitrogen or the atmosphere being the bioprocess of a kind of and more kinds of nitrogenous (N) compound, described nitrogenous compound comprises, but be not limited to ammonia, ammonium salt, urea, nitrite and nitrate.

Phrase used herein " divides phosphorus decomposing inorganics or compound " and refers to phosphorus (P) compound, be present in water-fast phosphorus compound such as phosphate rock in the mineral substance such as but not limited to those, be converted into that one or more biologies can utilize or easier absorption, promptly can be used to survive and/or the bioprocess of the different phosphate compound of growing by plant and other yeast.For example, the gained phosphorus compound can be more soluble in water or be weak acid, therefore can be absorbed by the root of plant.Biological can the utilization with the limiting examples that can absorb phosphorus compound comprises multiple phosphate radical, for example PO ₄ ^3-, H ₂PO ₄ ^-, HPO ₄ ^2-

The phrase used herein balance of phosphorus compound " keep " refers to and biology can not be utilized or water-fast phosphorus compound is converted into that one or more are easier of the bioprocess of biological utilisation or water-soluble different phosphorus compound, and wherein this process is to the superfluous of phosphorus in the local environment (P) compound or lack responsive.This conversion process downward modulation in (as greater than about 180ppm) local environment when the P-compound level is high, (as less than about 60ppm) this process raises when the P-compound level is low.

Phrase used herein " divide potassium decomposing inorganics or compound " refer to potassium (K) compound such as but not limited to those be present in potassium-bearing mineral matter and the material water-fast potassium compound be converted into one or more can be by biological utilisation or easier by the bioprocess of the different potassium compound of plant and the absorption of other yeast.For example, the gained potassium compound can be more soluble in water, therefore can be absorbed by the root of plant.

Phrase used herein " decompose complicated or high molecular carbon inorganics, material or compound " refers to organic or inorganic carbon molecule (for example, Fu Za carbohydrate such as Mierocrystalline cellulose and xylogen) bio-transformation with complexity and is one or more lower molecular weights (for example simple carbohydrates), is used for the carbon compound of surviving and/or growing by plant and yeast easily.This process comprises those reactions with carbon atom long-chain fracture in the polymerization carbon compound.

Term used herein " somatomedin " refers to yeast growth required molecule usually, include but not limited to VITAMIN, particularly vitamin B complex, for example vitamin B-1, riboflavin (vitamin B-2), vitamin B-12, nicotinic acid (B-3), pyridoxol (B-6), pantothenic acid (B-5); Folic acid; Vitamin H; Para-amino benzoic acid; Choline; And inositol.

In the present invention, above-mentioned five kinds of functions and excessive generation somatomedin and ATP are known as basic function.

Phrase used herein " inhibition pathogenic growth " refers to owing to there is a yeast cell of the present invention in muck, mud or spam samples, the reduced number of the pathogenic microorganism that exists in the sample after after a while or do not increase.Need be appreciated that is not having under the situation of yeast cell, and the number of pathogenic agent can increase by nature in the sample.Many these microorganisms cause the disease of humans and animals, can comprise for example kind of Escherichia, salmonella, Shigella, Mycobacterium, Staphylococcus, bacillus, streptococcus and Diplococcus of bacterium.

Phrase used herein " degraded adverse chemical product " refers to the bad compound in the fertilizer is converted into the biology or the biological process of inactive form, for example is the compound of lower molecular weight with these compound decomposition.Microbiotic is present in the organic materials usually, and these compounds should not occur in fertilizer, because the potentially dangerous of being taken in by the people is arranged, for example uses the vegetables that contain the chemical fertilizer growth of polluting organic materials by edible, and the possible diffusion of antibiotics resistance in the environment.A lot of microbiotic are added in the animal-feed to protect various domestic animals, and for example chicken, turkey and pig avoid bacterium and parasitic disease, and promote growth.A large amount of antibiotic feed additives are drained by animal, thereby gather in muck and mud.Many kinds of microbiotic use in animal surgery, such as but not limited to aminoglycoside, tetracyclines, β-Nei acyl Ammonia, glycopeptide class and Macrolide.The antibiotic example that U.S.'s approval is used on the farm includes but not limited to bacitracin methylene disalicylate, Zinc-bacitracin, Moenomycin. Flavophospholopol, terramycin, duomycin, penicillin, Tylosin/sulphamethazine, roxarsone, nitrasone, monensin, X-537A, Carbadox, tiamulin, hygromycin B, nystatin, Vulkamycin. PA-93, sulfadimethoxine, ormetoprim, lincomycin, fenbendazole and virginiamycin.These antibiotic existence and amount can detect by any means known in the art in the composition, for example high performance liquid chromatography (HPLC).

The method that phrase used herein " reduces the smell of organic materials " instigates one or more scent of compound concentrations in muck, mud or the rubbish to reduce.The scent of compound, such as but not limited to hydrogen sulfide, ammonia, indoles, skatole (being the 3-Methyl-1H-indole), p-cresol and organic acid, known all is the factor of facilitating of muck, mud or rubbish stench character.The concentration of these malodorous compounds in the air sample of brid guano fertilizer or contact muck can detect by any method well known in the art, include but not limited to vapor-phase chromatography.Smell is biological taste by the olfactory organ perception.The minimizing of the odor intensity relevant with muck, mud or rubbish can subjectively detect.Known several different methods and technology can be measured the intensity of smell.A kind of method of subjective measurement odor intensity is that measurement smell concerning human or animal trier can't be discovered or uncertain needed extent of dilution.In addition, can also use recognition threshold, be the higher concentration that the smell characteristics can be identified.The method of any objective or subjective detection odor intensity and technology can be used for monitoring the effect of the present composition and method.

In the present invention, the smell of inhibition pathogenic growth, degraded adverse chemical product and minimizing organic materials is known as subsidiary function or activity.

The present inventor finds, thereby yeast can be induced and shown seven kinds of different basic functions and three kinds of subsidiary functions under multiple culture condition.Culture condition has determined the activity that the yeast of cultivation is activated or strengthens.5.1 describe each concrete culture condition of ten kinds of functions respectively in detail to 5.10 joints.

According to the present invention, the yeast cell component in the Biofertiliser composition prepares by a plurality of yeast cell are cultivated for some time in the suitable culture base, in the presence of one or more successive alternating electromagnetic field.Culturing process makes yeast spore germination, yeast cell growth and division, can carry out in batches or carry out continuously.Term used herein " alternating electromagnetic field ", " electromagnetic field " or " EM field " are synonyms.The used electromagnetic field of the present invention can produce by several different methods well known in the art.Fig. 1 has described the sketch of exemplary device respectively.Electromagnetic wave source (3) produces the electromagnetic field of required frequency and field intensity, and this electromagnetic wave source comprises one or more signal generators that can generate electromagnetic waves, and preferred sinusoidal wave, the optimized frequency scope is 30MHz-3000MHz.Such signal generator is known in the art.Also can use the signal generator that can produce narrower range of frequency signal.If desired, can also use signal amplifier with the enhancing output signal, thereby increase field intensity.

Electromagnetic field can act on culture in several ways, comprises yeast cell near the signal projector device that links to each other with electromagnetic wave source.In one embodiment, apply electromagnetic field by the signal projector that is immersed in the electrode form in the yeast cell culture (1).In preferred embodiments, an electrode is a metal sheet, and another electrode comprises a plurality of metal wires that are installed in the container (2), and electromagnetic field energy can be uniformly distributed in the culture.The number of electrode used therein line depends on the volume of culture and the diameter of electric wire.For example, volume is the culture of 5000ml, and every 100ml culture can use the electrode wires of a diameter between 0.1 to 1.2mm; For the culture of volume above 1000L, every 1000L culture can use the electrode wires of a diameter between 3 to 30mm.

In preferred embodiments, yeast belong (Sacharomyces), Schizosaccharomyces (Schizosaccharomyces), Sporobolomyces (Sporobolomyces), torulopsis (Torulopsis), Trichosporon (Trichosporon), dimension Ke Shi yeast belong (Wickerhamia), Ashbya, Blastomyces (Blastomyces), mycocandida (Candida), Citeromycesbaodingensis belongs to (Citeromyces), Crebrothecium, Cryptococcus (Cryptococcus), Debaryomyces (Debaryomyces), Endomycopsis (Endomycopsis), Geotrichum (Geotrichum), Hansenula (Hansenula), Kloeckera (Kloeckera), saccharomyces oleaginosus belongs to (Lipomyces), Pichia (Pichia), the yeast of Rhodosporidium (Rhodosporidium) and Rhodotorula (Rhodotorula) can be used for the present invention.

Non-limiting line of yeast strains include Saccharomyces cerevisiae (Saccharomyces cerevisiae Hansen) ACCC2034, ACCC2035, ACCC2036, ACCC2037, ACCC2038, ACCC2039, ACCC2040, ACCC2041, ACCC2042, AS2.1, AS2.4, AS2.11, AS2.14, AS2.16, AS2.56, AS2.69, AS2.70, AS2.93, AS2.98, AS2.101, AS2.109, AS2.110, AS2.112, AS2.139, AS2.173, AS2.174, AS2.182, AS2.196, AS2.242, AS2.336, AS2.346, AS2.369, AS2.374, AS2.375, AS2.379, AS2.380, AS2.382, AS2.390, AS2.393, AS2.395, AS2.396, AS2.397, AS2.398, AS2.399, AS2.400, AS2.406, AS2.408, AS2.409, AS2.413, AS2.414, AS2.415, AS2.416, AS2.422, AS2.423, AS2.430, AS2.431, AS2.432, AS2.451, AS2.452, AS2.453, AS2.458, AS2.460, AS2.463, AS2.467, AS2.486, AS2.501, AS2.502, A AS2.516, AS2.535, AS2.536, AS2.558, AS2.560, AS2.561, AS2.562, AS2.576, AS2.593, AS2.594, AS2.614, AS2.620, AS2.628, AS2.631, AS2.666, AS2.982, AS2.1190, AS2.1364, AS2.1396, IFFI1001, IFFI1002, IFFI1005, IFFI1006, IFFI1008, IFFI1009, IFFI1010, IFFI1012, IFFI1021, IFFI1027, IFFI1037, IFFI1042, IFFI1043, IFFI1045, IFFI1048, IFFI1049, IFFI1050, IFFI1052, IFFI1059, IFFI1060, IFFI1063, IFFI1202, IFFI1203, IFFI1206, IFFI1209, IFFI1210, IFFI1211, IFFI1212, IFFI1213, IFFI1215, IFFI1220, IFFI1221, IFFI1224, IFFI1247, IFFI1248, IFFI1251, IFFI1270, IFFI1277, IFFI1287, IFFI1289, IFFI1290, IFFI1291, IFFI1292, IFFI1293, IFFI1297, IFFI1300, IFFI1301, IFFI1302, IFFI1307, IFFI1308, IFFI1309, IFFI1310, IFFI1311, IFFI1331, IFFI1335, IFFI1336, IFFI1337, IFFI1338, IFFI1339, IFFI1340, IFFI1345, IFFI1348, IFFI1396, IFFI1397, IFFI1399, IFFI1411, IFFI1413; Saccharomyces cerevisiae ellipse (Sacharomyces cerevisiae Hansen Var. ellipsoideus (Hansen) Dekker) ACCC2043, AS2.2, AS2.3, AS2.8, AS2.53, AS2.163, AS2.168, AS2.483, AS2.541, AS2.559, AS2.606, AS2.607, AS2.611, AS2.612; Xue watts yeast (Saccharomyces chevalieri Guillermond) AS2.131, AS2.213; Saccharomyces delbrueckii AS2.285; Saccharomyces delbrueckii Lindner var.mongolicus Lodder et van Rij AS2.209, AS2.1157; less spore yeast (Saccharomyces exiguus Hansen) AS2.349, AS2.1158; fermenting yeast (Saccharomyces fermentati (Saito) Lodder et van Rij) AS2.286, AS2.343; Saccharomyces logos van laer et Denamur ex Jorgensen AS2.156, AS2.327, AS2.335; honey and yeast (Saccharomyces mellis Lodder et Kreger Van Rij) AS2.195; Saccharomyces microellipsoides Osterwalder AS2.699; oval yeast (Saccharomyces oviformis Osterwalder) AS2.100; Roche yeast (Saccharomyces rosei (Guilliermond) Lodder et kreger van Rij) AS2.287; Lu's yeast (Saccharomyces rouxii Boutroux) AS2.178, AS2.180, AS2.370, AS2.371; sake yeast (Saccharomyces sake Yabe) ACCC2045; A Baoli yeast (Candida arborea) AS2.566; Candida Krusei (Castellani) Berkhout AS2.1045; rum beer Candida (Candida lambica (Lindner et Genoud) van.Uden et Buckley) AS2.1182; Candida lipolytica yeast Mother (Candida lipolytica (Harrison) Diddens et Lodder) AS2.1207, AS2.1216, AS2.1220, AS2.1379, AS2.1398, AS2.1399, AS2.1400; nearly smooth Candida Yeast (Candida parapsilosis (Ashford) Langeron et Talice) AS2.590; nearly flat Smooth muscle Candida (Candida parapsilosis (Ashford) et Talice Var.intermedia Van Rij et Verona) AS2.491; Maggi Candida (Candida pulcherriman (Lindner) Windisch) AS2.492; Candida rugousa (Anderson) Diddens et Loddeer AS2.511, AS2.1367, AS2.1369, AS2.1372, AS2.1373, AS2.1377, AS2.1378, AS2.1384; Candida tropicalis (Candida tropicalis (Castellani) Berkout) ACCC2004, ACCC2005, ACCC2006, AS2.164, AS2.402, AS2.564, AS2.565, AS2.567, AS2.568, AS2.617, AS2.1387; produce abdominal false Candida (Candida utilis Henneberg Lodder et Kreger Van Rij) AS2.120, AS2.281, AS2.1180; Crebrothecium ashbyii (Guillermond) Routein AS2.481, AS2.482, AS2.1197; Geotrichum (Geotrichum candidum Link) ACCC2016, AS2.361, AS2.498, AS2.616, AS2.1035, AS2.1062, AS2.1080, AS2.1132, AS2.1175, AS2.1183; Hansenula anomala (Hansenula anomala (Hansen) H et P sydow) ACCC2018, AS2.294, AS2.295, AS2.296, AS2.297, AS2.298, AS2.299, AS2.300, AS2.302, AS2.338, AS2.339, AS2.340, AS2.341, AS2.470, AS2.592, AS2.641, AS2.642, AS2.635, AS2.782, AS2.794; Hansenula arabitolgens Fang AS2.877; Hansenula jadinii Wickerham ACCC2019; Saturn Hansenula (Hansenula satumus (Klocker) H et P sydow) ACCC2020; Hansenula schneggii (Weber) Deker AS2.304; Hansenula subpelliculosa Bedford AS2.738, AS2.740, AS2.760, AS2.761, AS2.770, AS2.783, AS2.790, AS2.798, AS2.866; lemon-shaped Klerk yeast (Kloeckera apiculata (Reess emend.Klocker) Janke) ACCC2021, ACCC2022, ACCC2023, AS2.197, AS2.496, AS2.711, AS2.714; Adams Up's fat yeast (Lipomyces starkeyi Lodder et van Rij) ACCC2024, AS2.1390; powdered Pichia (Pichia farinose (Lindner) Hansen) ACCC2025, ACCC2026, AS2.86, AS2.87, AS2.705, AS2.803; film Bu Pichia (Pichia membranaefaciens Hansen) ACCC2027, AS2.89, AS2.661, AS2.1039; Rhodosporidium toruloides Banno ACCC2028; Rhodotorula (Rhodotorula glutinis (Fresenius) Harrison) ACCC2029, AS2.280, ACCC2030, AS2.102, AS2.107, AS2.278, AS2.499, AS2.694, AS2.703, AS2.704, AS2.1146; Red yeast (Rhodotorula minuta (Saito) Harrison) AS2.277; dark red yeast (Rhodotorula rubar (Demme) Lodder) ACCC2031, AS2.21, AS2.22, AS2.103, AS2.105, AS2.108, AS2.140, AS2.166, AS2.167, AS2.272, AS2.279, AS2.282; card's yeast (Saccharomyces carlsbergensis Hansen) ACCC2032, ACCC2033, AS2.113, AS2.116, AS2.118, AS2.121, AS2.132, AS2.162, AS2.189, AS2.200, AS2.216, AS2.265, AS2.377, AS2.417, AS2.420, AS2.440, AS2.441, AS2.443, AS2.444, AS2.459, AS2.595, AS2.605, AS2.638, AS2.742, AS2.745, AS2.748, AS2.1042; grape juice Yeast (Saccharomyces uvarum Beijer) IFFI1023, 1FFI1032, IFFI1036, IFFI1044, IFFI1072, IFFI1205, IFFI1207; Westergren yeast (Saccharomyces willianus Saccardo) AS2.5, AS2.7, AS2.119, AS2.152, AS2.293, AS2.381, AS2.392, AS2.434, AS2.614, AS2.1189; Saccharomyces (Saccharomyce sp.) AS2.311; Lu's yeast (Saccharomyces ludwigii Hansen) ACCC2044, AS2.243, AS2.508; Saccharomyces sinenses Yue AS2.1395; eight fission yeast spores Mother (Schizosaccharomyces octosporus Beijerinck) ACCC2046, AS2.1148; S. pombe (Schizosaccharomyces pombe Linder) ACCC2047, ACCC2048, AS2.248, AS2.249, AS2.255, AS2.257, AS2.259, AS2.260, AS2.274, AS2.994, AS2.1043, AS2.1149, AS2.1178, IFFI1056; Rose Throw yeast spore color (Sporobolomyces roseus Kluyver et van Niel) ACCC2049, ACCC2050, AS2.619, AS2.962, AS2.1036, ACCC2051, AS2.261, AS2.262; white Torulopsis (Torulopsis candida (Saito) Lodder) ACCC2052, AS2.270; Torulopsis famta (Harrison) Lodder et van Rij ACCC2053, AS2.685; Torulopsis globosa (Olson et Hammer) Lodder et van Rij ACCC2054, AS2.202; Torulopsis inconspicua Lodder et van Rij AS2.75; Trichosporon behrendii Lodder et Kroger van Rij ACCC2055, AS2.1193; Capitula Trichosporon yeast (Trichosporon capitatum Diddens et Lodder) ACCC2056, AS2.1385; Trichosporon cutaneum (de Beurm et al.) Ota ACCC2057, AS2.25, AS2.570, AS2.571, AS2.1374; Wickerhamia fluoresens (Soneda) Soneda ACCC2058, AS2.1388. ...

Known some yeast kind that can be activated or induce and be included in according to the present invention in the present invention has pathogenic to people and/or other live organism.Ashbya gossypii for example, Blastomyces dermatitidis (Blastomyces dermatitidis), Candida albicans (Candida albicans), Candidaparakrusei, candida tropicalis (Candida tropicalis), Ma Telitan Citeromycesbaodingensis (Citeromyces matritensis), Crebrothecium ashbyii, Lauren cryptococcus (Cryptococcus laurentii), novel Cryptococcus (Cryptococcus neoformans), the inferior Dbaly yeast of the Chinese (Debaryomyces hansenii), Debaryomyces kloeckeri, Dbaly yeast (Debaryomyces sp.) and Endomycopsis Fibnligera (Endomycopsis fibuligera).In some cases, preferably do not use such disease yeasts in the Biofertiliser composition of the present invention, for example, if this use is in open place, the health of the entail dangers to mankind and/or live organism.

The yeast that common preferred yeast belongs to.Hansen yeast saccharomyces cerevisiae in the bacterial strain of yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) (Saccharomyces cerevisiae Hansen) is a preferred strain.Most preferred yeast strain is yeast saccharomyces cerevisiae (Saccharomyces cerevisiae) bacterial strain: the AS2.504 that is deposited in the following preservation registration number of Chinese common micro-organisms DSMZ (CGMCC), AS2.558, AS2.413, AS2.397, AS2.69, AS2.109, AS2.607, AS2.516, AS2.561, AS2.422, AS2.393, AS2.631, AS2.982, AS2.560, AS2.467, AS2.415, AS2.375, AS2.628, AS2.1190, AS2.562, AS2.463, AS2.409, AS2.379, AS2.666, AS2.631, AS2.182, AS2.431, AS2.606, AS2.53, AS2.611, AS2.414, AS2.576, AS2.483, IFFI1211,1FF11293, IFFI1308, IFFI1210, IFFI1213, IFFI1307, IFFI1206, IFFI1052, IFFI1301, IFFI1291, IFFI1202, IFFI1021, IFFI1059, IFFI1052, IFFI1441, IFFI1008, IFFI1220, IFFI1302 and IFFI1023.

Usually, the yeast strain that the present invention uses can obtain from private or public laboratory culture thing, perhaps obtain from public DSMZ, and as American type culture collection, Manassas, No. 10801, the big ways for education, Virginia 20110-2209; And China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC) that is positioned at Institute of Microorganism, Academia Sinica, Haidian District, BeiJing, China 2714 mailbox, postcode 100080.

Following yeast strain is preferred for preparing P-balance yeast of the present invention: AS2.558, AS2.118, AS2.103, AS2.132, AS2.121, AS2.189, AS2.216, AS2.265, AS2.417, AS2.420, AS2.200, AS2.162, AS2.440, AS2.277, AS2.441, AS2.443, AS2.444, AS2.605, AS2.595, AS2.638, AS2.742, AS2.748, AS2.14, AS2.16, AS2.56, AS2.69, AS2.70, AS2.109, AS2.112, AS2.375, AS2560, AS2.561, AS2.562, AS2.559, AS2.501, AS2.502, AS2.503, AS2.504, IFFI1001, IFFI1002, IFFI1005, IFFI1006, IFFI1008, IFFI1009, IFFI1010, IFFI1012, IFFI1021, IFFI1027, IFFI1037, IFFI1042, IFFI1060, IFFI1063, IFFI1202, IFFI1203, IFFI1206, IFFI1209, IFFI1210, IFFI1211, IFFI1212, IFFI1213, IFFI1215, IFFI1220, IFFI1220, IFFI1221, IFFI1224, IFFI1247, IFFI1248, IFFI1251, IFFI1270, IFFI1277, IFFI1287, IFFI1289, IFFI1290, IFFI1291, IFFI1292, IFFI1293, IFFI1297, IFFI1300, IFFI1301, IFFI1307, IFFI1308, IFFI1309, IFFI1310, IFFI1311, IFFI1331, IFFI1335, IIFFI1336, IFFI1337, IFFI1338, IFFI1340, IFFI1339, IFFI1345, IFFI1396, IFFI1399, IFFI1411, IFFI1413, IFFI1023, IFFI1032, IFFI1036, IFFI1044 and IFFI1207.

Though preferably use pure yeast strain to begin to prepare yeast cell component of the present invention, be not limited to this.Every primary yeast cellular component can be by cultivating not of the same race or bacterial strain the mixture of yeast cell obtain.The composition of yeast cell component can detect with standard yeast authenticate technology well known in the art.

Compare with other yeast, some yeast can more effectively be exercised a kind of in the required function.Following table has been listed kind and the preservation registration number of multiple yeast strain, and after the inventive method stimulates the preferred function of corresponding bacterial strain.

Before and after cultivating, any yeast kind or bacterial strain exercise that any ability and efficient can easily detect by method known in the art in ten kinds of required functions under condition of the present invention.For example, the amount of fixed nitrogen can detect by 5,578, No. 486 described improvement acetylene reduction methods of United States Patent (USP), whole this specification sheets of introducing of the document.Improvement acetylene reduction method detects the amount of fixed nitrogen by the minimizing of measuring nitrogen molecule in a certain amount of air.The amount that can also detect fixed nitrogen by the ammonia and the nitrate of measurement yeast cell generation (see for example Grewling et al., 1965, Cornell Agr Exp StaBull 960:22-25).The standard method that is used to detect total organic nitrogen is Kai Shi (mensuration nitrogen) method.

The plant that is transformed from the insoluble or biological phosphorus compound that can not utilize can utilize the amount of phosphorus to detect by molybdenum blue method (see for example Murphy et al., 1962, Analytica Chimica Acta27:31-36) or UV (ultraviolet ray) absorption process; And the amount of the utilized potassium that is transformed from the insoluble or biological potassium compound that can not utilize can by for example flame atomic absorption spectrometry (see, Puchyr for example, et al., 1986, J.Assoc.Off.Anal.Chem.69:868-870) detect.Biofertiliser composition is added the ability that yeast provides biology can utilize N, P and K behind the soil can be detected by multiple technologies known in the art.For example, the plant that yeast cell produces in soil can utilize ammonia, nitrate, P and K to extract and quantitative analysis by Morgan (Morgan) soil detection system (see for example Lunt et al., 1950, Conn Agr Exp Sta Bull 541).

Can and analyze multiple organic molecule in muck and the soil with method detection well known in the art, comprise HPLC.Equally, can detect and count number of multiple microorganism in the sample and the sum of microorganism with method well known in the art.

Though be not subjected to the constraint of any theory or mechanism, the present inventor thinks that culture condition can activate and/or strengthen one of yeast cell or one group of expression of gene, thereby make this cell more effective, thereby produce corresponding required result at some Metabolic activity of enforcement.

Term used herein " organic substrate " refers to muck, mud or the rubbish of any kind of.In various embodiments, brid guano fertilizer, cattle manure, pig manure have been used.Usually, the used muck of the present invention is the refuse that animal produces in following animal activity, such as but not limited to farm, farmland, slaughterhouse and market.Term used herein " brid guano fertilizer " extensively comprises the ight soil of the birds that contain domestication and the organic materials of urine, and companion or do not accompany is used for the rubbish that fertilizes the land, for example straw, hay or pad grass traditionally.Brid guano fertilizer includes but not limited to the muck that chicken, duck, turkey, goose, quail, young dove, ostrich etc. produce.Brid guano fertilizer comprises movement or the guano that unacclimated birds produce.

Term used herein " cattle manure " extensively comprises and contains domestication the ruminating animal for example movement of cow, beef cattle and the organic materials of urine usually, and companion or do not accompany is used for the rubbish that fertilizes the land, for example straw, hay or pad grass traditionally.Term used herein " cattle manure " is not limited only to ox, also comprises other herbivore, mainly raises for their milk, meat, skin, hair and fur.Cattle manure includes but not limited to the muck that buffalo, wild ox, yak, horse, monkey, mule, sheep, goat, camel etc. produce.Muck also comprises the movement that non-domestication drove produces.

Term used herein " pig manure " extensively comprises the ight soil that contains pig usually and the organic materials of urine, with or do not accompany rubbish such as for example straw that is used for traditionally fertilizing the land, hay, pad grass.Pig manure includes but not limited to the muck that wild boar, dog, tame pig etc. produce.

Term used herein " mud " extensively comprises any solid matter that precipitates from suspension, for example residue in the municipal treatment plant, in the effluent sewage pond in sewage storage and/or treating processes.This term also comprises semi-solid thing, and sewage and sedimentary mixture.Therefore, this term comprise have extensive viscosity, the mud of density and water-content scope, and by partially disposed or stable mud.

Solid waste for example rubbish also can be used for Biofertiliser composition of the present invention.Solid waste typically refers to because realized its purpose or with through useless material.The source of solid waste comprises inhabitation, commerce, agricultural and industrial activity.Non-industrial solid wastes are the refuse from collecting from the urban district for example, the material that uses when mainly containing the foodstuff materials that abandons or making food, and other dry substance for example paper, textiles or plastics.The most preferably solid waste kind that comprises in the Biofertiliser composition is the high organic content rubbish of (as at least 30%).Residential area and commercial zone (restaurant and restaurant are arranged), rubbish mainly comprises decomposable food waste.Preferably, the used solid waste of the present invention is separated from glass, metal and other inorganic or nondegradable article by classification and selection operation.These operations can be carried out with the known method of regeneration/refuse cleaning industry, for example utilize the mechanicalness operation of the physical characteristics of solid waste as size and density.Pulverize or grind the size that can reduce refuse, obtain material of uniform size, can carry out multiple operation, for example drying etc. the gained material.

Biofertiliser composition of the present invention can comprise optional inorganic matrix component.The inorganic matrix component can include but not limited to phosphate rock or rock phosphate, phosphatic rock, phosphorite, sylvite, rock salt, carnallitite and potash mica.

Owing in muck, mud or the rubbish multiple composition is arranged, need from a collection of organic materials, extract the amount that sample is analyzed the plant nutrition that exists with detection.Can measure the amount of N, P, K, calcium, magnesium, zinc, iron, manganese, copper, sodium and sulphur in the muck with soil analysis method well known in the art.

In multiple embodiments, every kind of Biofertiliser composition of the present invention comprises at least can exercise the seven primary yeast cellular components that six kinds of basic functions add at least a subsidiary function.In the most preferred embodiment, Biofertiliser composition of the present invention comprises nine primary yeast cellular components, and six kinds of basic functions and all three kinds of subsidiary functions wherein are provided.Other substitute prescription also among considering.

In specific embodiments of the present invention, when using a collection of biology can utilize the abundant relatively organic substrate of phosphorus, can prepare the Biofertiliser composition that comprises the yeast cell that to keep phosphorus balance, and not comprise the yeast cell that decomposes inorganic phosphor-contained thing or compound.In addition, if desired, Biofertiliser composition can comprise more a spot of one or more above-mentioned yeast cell components that one of six kinds of basic functions are provided.For example, if Biofertiliser composition is to be used for rich potassic soil, this Biofertiliser composition can be formulated into and contain a small amount of yeast cell that contains potassium inorganics or compound that decomposes.

In another embodiment of the invention, when multiple yeast cell component is present in a kind of mixture, the culturing yeast cell makes the yeast cell with difference in functionality provide mutually aspect nutrition and the somatomedin and/or to interdepend under certain condition.As a result, between the multiple yeast cell of Ru 2006101161 of the present invention, set up symbiosis sample relation.This cultural method is chosen wantonly, but can increase the stability and the effectiveness of composition, makes gained fertilizer become preferably in the medium-term and long-term use of natural soils environment.The culture condition of this optional approach is described in 5.11 joints.

In another embodiment of the invention, the culturing yeast cell makes yeast cell adapt to the soil of particular type under certain condition.This cultural method is chosen wantonly, can be applied to every primary yeast cellular component respectively or be applied to the yeast cell component mixture.As a result, yeast cell is grown better and is survived in specific edatope.The culture condition of this optional approach is described in 5.12 joints.

As used herein, if bio-feritlizer of the present invention is present in the soil or imposes on root, this Biofertiliser composition supports or promotes plant-growth that plant obtains viability, size, weight, bud ratio, growth rate or maturing rate.Therefore, Biofertiliser composition of the present invention can be used in any agricultural, gardening or forest practice.This Biofertiliser composition can be used for the economy of large scale farming in the field of spaciousness or greenhouse, even is used for indoor decorative plant.Preferably promote the growth of crop plants, such as but not limited to cereal crop, vegetable crop, fruit crop, flowers crop and fodder crop with bio-feritlizer.For example, Biofertiliser composition can be used for wheat, barley, corn, soybean, rice, oat, potato, apple, orange, tomato, muskmelon, cherry, lemon, lettuce, Radix Dauci Sativae, sugarcane, tobacco, cotton etc.

The method that Biofertiliser composition of the present invention is used can be identical with conventional fertilizers.Known to various equivalent modifications, can use several different methods and instrument.In one embodiment, the mixture of nutrient solution of yeast strain of the present invention and brid guano fertilizer directly is applied to soil or plant roots.In another embodiment, dry powder of yeast strain of the present invention and brid guano fertilizer are imposed on soil or plant root.Biofertiliser composition can be applied to soil by dispenser, atomizer and other mechanical method, and these methods can be automatizations.Biofertiliser composition can directly be applied to plant, for example soaks seed and/or root.Using like this can be periodic, and for example annually, perhaps once perhaps more frequent as required uses each season of growth.Though in most of the cases be not necessary, Biofertiliser composition of the present invention can also or use in turn with the associating of other kind fertilizer.

In a preferred embodiment, Biofertiliser composition of the present invention is that yeast of the present invention mixes with muck, exsiccant mud or granular rubbish, uses as base fertilizer, is applied to the soil of the farm crop main root system degree of depth.Before using, should and weed a garden the soil scarifying.Biofertiliser composition can be sprinkling upon equably on the soil, add near the hole or long ditch dug with a plow in the soil tree crown.For existing fruit tree, dig 5 to 30cm dark holes and Biofertiliser composition is added wherein.Thoroughly will contain soil, hole or the ditch dug with a plow covering of Biofertiliser composition then with soil and water.After 3 to 7 days, just can plant or sow in this zone.For rice, water flooded the soil 3 to 7 days before rice transplanting.If be used for the shallow root system plant at sand, the degree of depth with 5 to 15cm; Be used for dark root system plant, suggestion is with 5 to 25cm.Be used for the shallow root system plant at clayey soil, the degree of depth with 2 to 10cm; Be used for dark root system plant, suggestion is with 2 to 15cm.Required effect is Biofertiliser composition contact or very near plant roots.Preferably, after applying fertilizer and/or the plantation, do not stir soil.Usually, the application temperature of fertilizer is between 5 ℃ to 45 ℃, and is ideal between 16 ℃ to 30 ℃; Preferred pH scope is between 5.5 to 8.5, and is ideal between 6.5 to 7.5.

Recommend consumption

Farm crop	The amount of bio-feritlizer
Farm crop	The amount of bio-feritlizer	Vegetables (term growth)	????600-900kg/ha
Vegetables (long term growth)	????900-1200kg/ha	Vegetables (term growth)	????600-900kg/ha
Vegetables (long term growth)	????900-1200kg/ha	The ground vegetables	????900-1350kg/ha
Solanum fruit	????900-1350kg/ha	The ground vegetables	????900-1350kg/ha
Solanum fruit	????900-1350kg/ha	Piece root and tuberous vegetable	????750-900kg/ha
Bulbous vegetable	????900-1200kg/ha	Piece root and tuberous vegetable	????750-900kg/ha
Bulbous vegetable	????900-1200kg/ha	Leguminous plants	????600-1050kg/ha
Fruit tree	The 2-5kg/ tree	Leguminous plants	????600-1050kg/ha
Fruit tree	The 2-5kg/ tree	Paddy rice	????600-900kg/ha
Wheat and corn	????750-1200kg/ha	Paddy rice	????600-900kg/ha
Wheat and corn	????750-1200kg/ha	Cotton and peanut	????600-1200kg/ha

5.1-5.10 joint has been described the carbon compound that is used for fixed nitrogen, decomposition phosphorus compound, decomposition potassium compound, decomposes complexity, the yeast cell component that produces somatomedin, generation ATP, inhibition pathogenic agent, degraded adverse chemical product and minimizing smell respectively.The preparation method of every primary yeast cellular component has been described.5.11 joint has been described the method for setting up symbiosis sample relation between the yeast strain of Ru 2006101161 of the present invention.5.12 joint has been described the method that makes yeast cell of the present invention adapt to particular type soil.5.13 joint has been described the preparation of Biofertiliser composition of the present invention.Also describe the preparation method of organic substrate and the manufacture method of bio-feritlizer, comprised mixing, drying, cooling and packing.In the various embodiments of the present invention, used the standard technique of yeast operation, transfer and storage.When carrying out preparation process of the present invention,, favourable though aseptic condition or clean environment are not necessarily.

5.1 fixed nitrogen yeast cell component

Fixed nitrogen is the process that atmospheric nitrogen is converted into ammonia and nitrate.Having been found that 800 kinds of naturally occurring microorganisms nearly of surpassing in 70 genus can fixed nitrogen, mainly is bacterium and cyanobacteria.Some nitrogen-fixing microorganisms, for example root nodule bacterium form symbiotic relationship with plant, especially at fabaceous root.Other, for example vinelandii are independent existence, and can be in soil fixed nitrogen.

Among the present invention, the ability of yeast fixed nitrogen is activated or strengthens, and gained fixed nitrogen yeast cell can be used as the component of Biofertiliser composition of the present invention and uses.

According to the present invention, the yeast cell that nitrogen fixing capacity is reinforced is by preparing cell cultivating in the presence of the electromagnetic field in the suitable culture base.The electromagnetic field frequency of activation or reinforcement yeast nitrogen fixing capacity is usually in 800MHz arrives the scope of 1000MHz.Yeast cell growth can detect the nitrogen fixing capacity of yeast cell by method well known in the art after the sufficiently long time.

The method that the present invention prepares the fixed nitrogen yeast cell is to carry out in the liquid medium within.Contain the absorbable nutrition of yeast cell in the substratum.Usually, carbohydrate such as carbohydrate, for example sucrose, glucose, fructose, dextrose, maltose, wood sugar etc. and starch can separately or be united as the source that can absorb carbon in the substratum and use.The accurate consumption of one or more carbohydrate sources that use in the substratum depends in part on other composition in the substratum, but usually the amount of carbohydrate between substratum weight about 0.1% to 5% between, preferably between about 0.5% to 2%, most preferably from about 1%.In substratum, these carbon sources can be used separately, or several such carbon source is united use.

The inorganic salt that can add in the substratum are that sodium, potassium, calcium, phosphate radical, sulfate radical, the isoionic conventional salt of carbonate can be provided.The limiting examples of the inorganic salt of trophicity has CaCO ₃, KH ₂PO ₄, MgSO ₄, NaCl and CaSO ₄

Table 1: fixed nitrogen zymic medium component

Medium component	Content
Medium component	Content	????KH ₂PO ₄	????0.2g
????K ₂HPO ₄	????0.2g	????KH ₂PO ₄	????0.2g
????K ₂HPO ₄	????0.2g	????MgSO ₄·7H ₂O	????0.25g
????CaCO ₃·5H ₂O	????3.5g	????MgSO ₄·7H ₂O	????0.25g
????CaCO ₃·5H ₂O	????3.5g	????CaSO ₄·2H ₂O	????0.5g
????NaCl	????0.25g	????CaSO ₄·2H ₂O	????0.5g
????NaCl	????0.25g	The pasty state yeast extract	????0.3g
Sucrose	????12.0g	The pasty state yeast extract	????0.3g
Sucrose	????12.0g	Distilled water or autoclaving water	????1000ml

It should be noted that the medium component that provides in the table 1 is not restrictive.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.

Culturing process can be at first with cell density 10 ²-10 ⁵The yeast strain inoculum that cell/ml selects 1ml is inoculated in the 100ml substratum, and preferred 3 * 10 ²-10 ⁴Cell/ml.This process can increase and decrease as required in proportion.Yeast culture can be grown in the presence of an electromagnetism (EM) field or one group of electromagnetic field.If apply one group of electromagnetic field, when when an electromagnetic field switches to another electromagnetic field, yeast culture can be in same container, use same set of electromagnetic wave generator and projector.

Electromagnetic field can be used by any means known in the art, and the frequency of each electromagnetic field arrives about 1000MHz scope about 800, preferably arrives in the 916.000MHz scope 840.000.Such as but not limited to these examples, each electromagnetic field frequency can be about 840,845,850,855,860,865,870,875,880,885,890,895,900,905,910,915 or 920MHz.The field intensity of electromagnetic field arrives in the 200mV/cm scope 10.If use one group of electromagnetic field, each electromagnetic field can have different frequency in the above-mentioned scope, or different field intensity in the above-mentioned scope, or interior different frequency of above-mentioned scope and field intensity.In a preferred embodiment, it is lower that the electromagnetic field that begins in a group and electromagnetic field are subsequently compared the EM field intensity, and yeast cell culture just is placed in the electromagnetic field that field intensity increases gradually like this.Though the electromagnetic field number of using in a group can be arbitrarily, preferably yeast culture is placed one group and add up to 2,3,4,5,6,7 or 8 different electromagnetic field.

Even just can be activated though yeast cell is cultivated several hrs in the presence of electromagnetic field, but yeast cell can continue to cultivate for some time (for example several days to 1 week or more weeks) in the presence of electromagnetic field, usually preferred, should make the activatory yeast cell in the presence of one or more electromagnetic field, breed and grew about altogether 140-280 hour.

For example use exemplary device shown in Figure 1, initial electromagnetic field is used about 12.5mV/cm usually in the 10-20mV/cm scope.Period 1 further cultivates yeast cell another cycle under the essentially identical condition the higher level in the electromagnetic field field intensity increases to the 50-200Mv/cm scope (about usually 125mV/cm) after cultivating.Method of the present invention is carried out in about 23 to 30 ℃ of temperature ranges; But, preferably carry out at 25 to 28 ℃.Culturing process is 0.025 to 0.8mol/m at dissolved oxygen concentration preferably ³Condition under carry out preferred 0.4mol/m ³Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.

When culturing process finished, the fixed nitrogen yeast cell can reclaim from culture by several different methods known in the art, and stored being lower than under about 0 ℃ to 4 ℃ temperature.The fixed nitrogen yeast cell also can store with form of powder in the drying processing.

Can detect the ability of activatory yeast cell fixed nitrogen with any method known in the art.For example, the improvement acetylene reduction method of measurement microorganism fixed nitrogen can be used for assessing prepared zymic nitrogen fixing capacity.United States Patent (USP) 5,578 has been described improvement acetylene reduction method No. 486, with its whole content quotation here as a reference.Also can use alternative method based on 15-N.

Zymic nitrogen fixing capacity of the present invention can prove by following two kinds of methods.

With 1ml activatory yeast strain AS2.628 (2-5 * 10 ⁷) in the 1000mlAshby substratum, in the presence of 8 electromagnetic field of 28 ℃, one group following orders, cultivate: 855MHz, 14mV/cm5 hour; 865MHz, 14mV/cm 5 hours; 875MHz, 14mV/cm 5 hours; 885MHz, 14mV/cm 5 hours; 855MHz, 120mV/cm 30 hours; 865MHz, 120mV/cm 30 hours; 875MHz, 120mV/cm 30 hours; 885MHz, 120mV/cm 30 hours.In other container, identical but do not have to cultivate under the condition of electromagnetic field 1ml and do not activate yeast in contrast.After the cultivation, the 1000ml yeast cell is mixed with the aseptic coal dust powder of 3000g, be dried to water content less than 5% being lower than 70 ℃ then.Pulverous finished product (0.1g) is sealed in (every kind is used 5 culturing bottles in the experiment) in the culturing bottle of 100ml with 10ml Ashby substratum.From culturing bottle, take the 10ml air away with syringe, replace 10ml acetylene (purity＞99%).Culturing bottle was cultivated 24-120 hour at 28 ℃, measured the amount that acetylene reduces with vapor-phase chromatography.The acetylene amount has reduced by 120 μ mol after 120 hours above/g dry powder.Contain and do not activate not significantly minimizing of acetylene in the zymic contrast.

Perhaps, can use isotropic substance nitrogen dilution method.Pulverous finished product (the non-activated and 0.1g activatory yeast of 0.1g) was cultivated respectively 96 hours at 28 ℃.The amount of detection and more every primary yeast fixed nitrogen.Activatory yeast amount of nitrogen fixation be 3.5mg above/g dry powder.Contain and do not activate zymic contrast and do not show tangible fixed nitrogen.

5.2 phosphorus decomposability yeast cell component

Phosphorus compound decomposability of the present invention (P-decomposition) yeast with the insoluble or biological phosphorus containg substances that can not utilize for example phosphate rock be converted into the titanium pigment compound, they can be utilized by plant.

In the present invention, the ability that yeast decomposes insoluble phosphorus containg substances is activated or strengthens, and gained P-decomposes the component that yeast cell can be used as Biofertiliser composition of the present invention to be used.

In multiple embodiments, when solubility in organic matrix or the biological level that can utilize phosphorus are low, in the present composition, use P-to decompose yeast cell.When biology can utilize the level of phosphorus high (common in the brid guano fertilizer), preferably do not use P-to decompose yeast.

According to the present invention, can carry out yeast cell that P-decomposes in the suitable culture base by cell is prepared cultivating in the presence of the electromagnetic field.Be used to activate or electromagnetic field frequency that enhancement microbiological P-decomposes usually at 300MHz in the 500MHz scope.Cell cultures can detect the ability that they decompose phosphorus containg substances by method well known in the art after the sufficiently long time.

The method that the present invention prepares P-decomposition yeast cell is to carry out in the liquid medium within.Contain the absorbable nutrition of yeast cell in the substratum.Usually, carbohydrate such as carbohydrate, for example sucrose, glucose, fructose, dextrose, maltose, wood sugar etc. and starch can separately or be united as the source that can absorb carbon in the substratum and use.The accurate consumption of one or more carbohydrate sources depends in part on other composition in the substratum in the substratum, but usually the amount of carbohydrate between substratum weight about 0.1% to 5% between, preferably between about 0.5% to 2%, most preferably from about 1.5%.In substratum, these carbon sources can be used separately, or several such carbon source is united use.

The inorganic salt that can add in the substratum are that sodium, potassium, calcium, sulfate radical, the isoionic conventional salt of carbonate can be provided.The limiting examples of the inorganic salt of trophicity has CaCO ₃, MgSO ₄, NaCl and CaSO ₄The biology of suitable form can not utilize the phosphorus containg substances of form also to comprise in the substratum as the exsiccant organic substrate.The limiting examples of exsiccant organic substrate comprise muck, mud and 〉=150 purpose rubbish.Other insoluble phosphorus containg substances also can be by respectively or unite use.

Table 2:P-decomposes the zymic medium component

Medium component	Content
Medium component	Content	Sucrose	???15g
????NaCl	???1.2g	Sucrose	???15g
????NaCl	???1.2g	????MgSO ₄·7H ₂O	???0.2g
????CaCO ₃·5H ₂O	???3.0g	????MgSO ₄·7H ₂O	???0.2g
????CaCO ₃·5H ₂O	???3.0g	????CaSO ₄·2H ₂O	???0.3g
????KNO ₃	???0.3g	????CaSO ₄·2H ₂O	???0.3g
????KNO ₃	???0.3g	The pasty state yeast extract	???0.5g
Argol fertilizer, mud or rubbish	1.2g to 2.4g; Powder＞150 orders	The pasty state yeast extract	???0.5g
Argol fertilizer, mud or rubbish	1.2g to 2.4g; Powder＞150 orders	Autoclaving water	???1000ml

It should be noted that the medium component that provides in the table 2 is not restrictive.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.

Electromagnetic field can be used by any means known in the art, and the frequency of each electromagnetic field arrives in about 500MHz scope about 300, preferably arrives in the 435.000MHz scope 340.000.Such as but not limited to these examples, each electromagnetic field frequency can be about 340,345,350,355,360,365,370,375,375,380,385,390,395,400,405,410,415,420,425,430 or 435MHz.The field intensity of electromagnetic field arrives in the 200mV/cm scope 10.If use one group of electromagnetic field, each electromagnetic field can have different frequency in the above-mentioned scope, or different field intensity in the above-mentioned scope, or different frequency and field intensity in the above-mentioned scope.In a preferred embodiment, it is lower that the electromagnetic field that begins in a group and electromagnetic field are subsequently compared the EM field intensity, and yeast cell culture just is placed in the electromagnetic field that field intensity increases gradually like this.Though the electromagnetic field number of using in a group can be arbitrarily, preferably yeast culture is placed one group and add up to 2,3,4,5,6,7 or 8 different electromagnetic field.

Even just can be activated though yeast cell is cultivated several hrs in the presence of electromagnetic field, but yeast cell can continue to cultivate for some time (for example 1 week or more weeks) in the presence of electromagnetic field, usually preferred, should make the activatory yeast cell in the presence of one or more electromagnetic field, breed and grew about altogether 140-280 hour.

For example, use exemplary device shown in Figure 1, initial field intensity is used about 12.5mV/cm usually in the 10-20mV/cm scope.Period 1 further cultivates yeast cell another cycle under the essentially identical condition higher level in the electromagnetic field field intensity increases to the 50-200vM/cm scope (about usually 125mV) after cultivating.Method of the present invention is carried out in about 23 to 30 ℃ of temperature ranges; But, preferably carry out at 25 to 28 ℃.Culturing process is 0.025 to 0.8mol/m at dissolved oxygen concentration preferably ³Condition under carry out preferred 0.4mol/m ³Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.

When culturing process finished, P-decomposed yeast cell and can reclaim from culture by several different methods known in the art, and stored being lower than under about 0 ℃ to 4 ℃ temperature.P-decomposes yeast cell and also can the drying processing store with form of powder.

Biology can utilize for example H of phosphorus in the culture ₃PO ₄, H ₂PO ₄ ^-And HPO ₄ ^2-Amount can detect by any means known in the art, include but not limited to the UV absorption spectroscopy.Can utilize the total amount of phosphorus and not activate the amount that biology can utilize the difference calculating of the amount of phosphorus to increase in the same substratum of yeast by biological in the activatory yeast culture.For example, with 1ml Wine brewing yeast strain AS2.399 (2 * 10 ⁷To 5 * 10 ⁷Cell/ml) is inoculated in the 1000ml substratum according to table 2.Culture is cultivated in the presence of 8 electromagnetic field of 28 ℃, one group following orders: 360MHz, 14mV/cm5 hour; 365MHz, 14mV/cm 5 hours; 370MHz, 14mV/cm 5 hours; 380MHz, 14mV/cm 5 hours; 360MHz, 130mV/cm 30 hours; 365MHz, 130mV/cm 30 hours; 370MHz, 130mV/cm 30 hours; 375MHz, 130mV/cm 30 hours.After testing, biology can utilize the amount of phosphorus increased 330mg above/the ml yeast culture.Do not activate the biological not obviously change of amount that can utilize phosphorus in the zymic contrast.

5.3 phosphorus balance yeast cell component

Phosphorus balance of the present invention (P-balance) yeast also can be converted into the insoluble or biological phosphorus containg substances that can not utilize the biological available phosphorus compound of solubility.But, when the level of phosphorus in the local environment is high, preferably use P-balance yeast.Insoluble or the biological phosphorus containg substances that can not utilize can utilize the level sensitivity of the conversion of phosphorus to phosphorus to the solubility biology; In about 180ppm or higher level, transform and reduce, and at about 60ppm or more low-level, transforming increases.

Among the present invention, P-balance yeast cell preferably uses in comprising solubility or the biological Biofertiliser composition that can utilize the significant relatively organic substrate of phosphorus level.For example, compare with other kind muck, brid guano fertilizer contains high-caliber relatively titanium pigment.

According to the present invention, can P-equilibrated yeast cell by cell is prepared cultivating in the suitable culture base in the presence of the electromagnetic field.The electromagnetic field frequency that is used for activating or strengthens yeast P-equilibrium function usually at 300MHz in the 500MHz scope.Cell cultures can detect the ability that cell decomposes phosphorus containg substances by method well known in the art after the sufficiently long time.

The method that the present invention prepares P-balance yeast cell is to carry out in the liquid medium within.Contain the absorbable nutrition source of yeast cell in the substratum.Usually, carbohydrate such as carbohydrate, for example sucrose, glucose, fructose, dextrose, maltose, wood sugar etc. and starch can separately or be united as the source that can absorb carbon in the substratum and use.The accurate consumption of one or more carbohydrate sources depends in part on other composition in the substratum in the substratum, but usually the amount of carbohydrate between substratum weight about 0.1% to 5% between, preferably between about 0.5% to 2%, most preferably from about 1.5%.In substratum, these carbon sources can be used separately, or several such carbon source is united use.

The inorganic salt that can add in the substratum are that sodium, potassium, calcium, sulfate radical, the isoionic conventional salt of carbonate can be provided.The limiting examples of the inorganic salt of trophicity has CaCO ₃, MgSO ₄, NaCl and CaSO ₄The insoluble phosphorus containg substances of suitable form is also contained in the substratum, and limiting examples comprises＞150 purpose sludge-dryings.Can also respectively or unite and use other insoluble phosphorus containg substances.

Table 3:P-balance zymic medium component

Medium component	Content
Medium component	Content	Sucrose	????15g
????NaCl	????1.2g	Sucrose	????15g
????NaCl	????1.2g	????MgSO ₄·7H ₂O	????0.2g
????CaCO ₃·5H ₂O	????3.0g	????MgSO ₄·7H ₂O	????0.2g
????CaCO ₃·5H ₂O	????3.0g	????CaSO ₄·2H ₂O	????0.3g
????KNO ₃	????0.3g	????CaSO ₄·2H ₂O	????0.3g
????KNO ₃	????0.3g	The pasty state yeast extract	????0.5g
Argol fertilizer, mud or rubbish	1.2g; Powder＞150 orders	The pasty state yeast extract	????0.5g
Argol fertilizer, mud or rubbish	1.2g; Powder＞150 orders	Autoclaving water	????1000ml

It should be noted that the medium component that provides in the table 3 is not restrictive.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.

Electromagnetic field can be used by any means known in the art, and the frequency of each electromagnetic field can arrive in about 500MHz scope about 300, or preferably arrives in the 485.000MHz scope 380.000.Such as but not limited to these examples, each electromagnetic field frequency can be about 380,385,390,395,400,402,405,410,415,420,422,425,430,432,435,440,445,450,455,460,465,470,480 or 485MHz.The field intensity of electromagnetic field arrives in the 300mV/cm scope 90.If use one group of electromagnetic field, each electromagnetic field can have different frequency in the above-mentioned scope, or different field intensity in the above-mentioned scope, or interior different frequency of above-mentioned scope and field intensity.In a preferred embodiment, it is lower that the electromagnetic field that begins in a group and electromagnetic field are subsequently compared the EM field intensity, and yeast cell culture just is placed in the electromagnetic field that field intensity increases gradually like this.Though the electromagnetic field number of using in a group can be arbitrarily, preferably yeast culture is placed one group and add up to 2,3,4,5,6,7 or 8 different electromagnetic field.

Even just can be activated though yeast cell is cultivated several hrs in the presence of electromagnetic field, but yeast cell can continue to cultivate for some time (for example 2 week or more weeks) in the presence of electromagnetic field, usually preferred, should make the activatory yeast cell in the presence of one or more electromagnetic field, breed and grew about altogether 230-480 hour.

For example, use exemplary device shown in Figure 1, initial field intensity is used about 100mV usually in the 50-150mV/cm scope.Period 1 further cultivates yeast cell another cycle under the essentially identical condition increase to higher level in the 200-300mV/cm scope (about usually 250mV/cm) except electromagnetic field intensity after cultivating.Method of the present invention is carried out in about 23 to 30 ℃ of temperature ranges; But, preferably carry out at 25 to 28 ℃.Culturing process is 0.025 to 0.8mol/m at dissolved oxygen concentration preferably ³Condition under carry out preferred 0.4mol/m ³Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.

When culturing process finished, P-balance yeast cell can reclaim from culture by several different methods known in the art, and stored being lower than under about 0 ℃ to 4 ℃ temperature.P-balance yeast cell also can store with form of powder in the drying processing.

Biology in the culture can utilize for example H of phosphorus ₃PO ₄, H ₂PO ₄ ^-And HPO ₄ ^2-Amount can detect by any means known in the art, include but not limited to the UV absorption spectroscopy.Can utilize the total amount of phosphorus and not activate the amount that biology can utilize the difference calculating of the amount of phosphorus to increase in the same substratum of yeast by biological in the activatory yeast culture.For example, with 1ml Wine brewing yeast strain AS2.628 (2 * 10 ⁷To 5 * 10 ⁷Cell/ml) be inoculated in 1000ml contains 200mg/l H ₃PO ₄, H ₂PO ₄ ^-And HPO ₄ ^2-Substratum in.Culture is cultivated in the presence of 8 electromagnetic field of 28 ℃, one group following orders: 385MHz, 99mV/cm 12 hours; 415MHz, 99mV/cm 12 hours; 440MHz, 99mV/cm 12 hours; 460MHz, 99mV/cm 12 hours; 385MHz, 250mV/cm 48 hours; 415MHz, 250mV/cm 48 hours; 440MHz, 250mV/cm24 hour; 460MHz, 250mV/cm 24 hours.After measured, biology can utilize the amount of phosphorus to increase more than 24%.Biology can utilize the amount of phosphorus not show any obvious change in the control group.

5.4 potassium decomposability yeast cell component

Potassium compound decomposability of the present invention (K-decomposition) yeast with insoluble contain the potassium material for example potash mica be converted into soluble potassium, make them can be by plant utilization.

In the present invention, the insoluble ability that contains the potassium material of a plurality of yeast cell decomposition is activated or strengthens, and gained K-decomposes the component use that yeast cell can be used as Biofertiliser composition of the present invention.

According to the present invention, can carry out yeast cell that K-decomposes in the suitable culture base by cell is prepared cultivating in the presence of the electromagnetic field.Be used to activate or strengthen electromagnetic field frequency that yeast K-decomposes usually in the 100MHz-300MHz scope.After yeast cell is cultivated the sufficiently long time, can detect cell by method well known in the art and decompose the ability that contains the potassium material.

The method that the present invention prepares K-decomposition yeast cell is to carry out in the liquid medium within.Contain the absorbable nutrition source of yeast cell in the substratum.Usually, carbohydrate such as carbohydrate, for example sucrose, glucose, fructose, dextrose, maltose, wood sugar etc. and starch can separately or be united as the source that can absorb carbon in the substratum and use.The accurate consumption of one or more carbohydrate sources depends in part on other composition in the substratum in the substratum, but usually the amount of carbohydrate between substratum weight about 0.1% to 5% between, preferably between about 0.5% to 2%, most preferably from about 1.5%.In substratum, these carbon sources can be used separately, or several such carbon source is united use.

Can add inorganic salt in the substratum comprises sodium, calcium, phosphate radical, sulfate radical, the isoionic conventional salt of carbonate can be provided.The limiting examples of the inorganic salt of trophicity has (NH ₄) ₂HPO ₄, CaCO ₃, MgSO ₄, NaCl and CaSO ₄The insoluble potassium material that contains of suitable form is also contained in the substratum, and limiting examples comprises 〉=200 purpose potash micas.Can also respectively or unite and use other insoluble potassium material that contains.

Table 4:K-decomposes the zymic medium component

Medium component	Content
Medium component	Content	Sucrose	????15g
????NaCl	????1.2g	Sucrose	????15g
????NaCl	????1.2g	????MgSO ₄·7H ₂O	????0.2g
????CaCO ₃·5H ₂O	????3.0g	????MgSO ₄·7H ₂O	????0.2g
????CaCO ₃·5H ₂O	????3.0g	????CaSO ₄·2H ₂O	????0.3g
????(NH ₄) ₂HPO ₄	????0.3g	????CaSO ₄·2H ₂O	????0.3g
????(NH ₄) ₂HPO ₄	????0.3g	The pasty state yeast extract	????0.5g
Potash mica	1.0g, powder＞200 orders	The pasty state yeast extract	????0.5g
Potash mica	1.0g, powder＞200 orders	Argol fertilizer, mud or rubbish	1.2-3g; Powder＞150 orders
Autoclaving water	????1000ml	Argol fertilizer, mud or rubbish	1.2-3g; Powder＞150 orders

It should be noted that the medium component that provides in the table 4 is not restrictive.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.

Electromagnetic field can be used by any means known in the art, and the frequency of each electromagnetic field arrives in about 300MHz scope about 100, and preferred 190.000 in the 285.000MHz scope.Such as but not limited to these examples, each electromagnetic field frequency can be about 190,195,200,205,210,215,220,225,230,235,240,245,250,255,260,265,270,275,280 or 285MHz.The field intensity of electromagnetic field arrives in the 200mV/cm scope 10.If use one group of electromagnetic field, each electromagnetic field can have different frequency in the above-mentioned scope, or different field intensity in the above-mentioned scope, or interior different frequency of above-mentioned scope and field intensity.In a preferred embodiment, it is lower that the electromagnetic field that begins in a group and electromagnetic field are subsequently compared the EM field intensity, and yeast cell culture just is placed in the electromagnetic field that field intensity increases gradually like this.Though the electromagnetic field number of using in a group can be arbitrarily, preferably yeast culture is placed one group and add up to 2,3,4,5,6,7 or 8 different electromagnetic field.

For example, use exemplary device shown in Figure 1, initial field intensity is used about 12.5mV/cm usually in the 10-20mV/cm scope.Period 1 further cultivates yeast cell another cycle under the essentially identical condition increase to higher level in the 50-200Mv/cm scope (about usually 125mV/cm) except electromagnetic field intensity after cultivating.Method of the present invention is carried out in about 23 to 30 ℃ of temperature ranges; But, preferably carry out at 25 to 28 ℃.Culturing process is 0.025 to 0.8mol/m at dissolved oxygen concentration preferably ³Condition under carry out preferred 0.4mol/m ³Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.

When culturing process finished, K-decomposed yeast cell and can reclaim from culture by several different methods known in the art, and stored being lower than under about 0-4 ℃ the temperature.K-decomposes yeast cell and also can the drying processing store with form of powder.

The yeast cell of cultivating decomposes the insoluble ability that contains the potassium material and can detect by any means known in the art.For example, with 1ml Wine brewing yeast strain AS2.631 (2 * 10 ⁷To 5 * 10 ⁷Cell/ml) is inoculated in the 1000ml substratum according to table 4.Culture is cultivated in the presence of 8 electromagnetic field of 28 ℃, one group following orders: 210MHz, 14mV/cm 5 hours; 235MHz, 14mV/cm 5 hours; 245MHz, 14mV/cm 5 hours; 255MHz, 14mV/cm5 hour; 210MHz, 120mV/cm 30 hours; 235MHz, 120mV/cm 30 hours; 245MHz, 120mV/cm 30 hours; 255MHz, 120mV/cm 30 hours.Foundation contains the not contrast of active cells of same yeast strain.Can utilize potassium K by biology in any method detection culture known in the art ⁺Amount, include but not limited to flame spectrum and/or atomic absorption spectrometry.The increase of potassium is to calculate by the difference between the basal level of potassium in the substratum before amount of cultivating potassium in table 4 substratum of back and the cultivation.After measured, the biology of the yeast cell of cultivation can utilize the amount of potassium to increase more than the 120mg/ml.Can utilize the amount of potassium obviously not change in the control group.

5.5 complex carbon decomposability yeast cell component

Carbon decomposability of the present invention (C-decomposition) yeast with complexity, high-molecular weight carbon compound and material, particularly Fu Za carbohydrate such as Mierocrystalline cellulose and lignin conversion be simple carbohydrate, for example pentose and hexose.These simple carbohydrate can be utilized by other yeast cell in the local environment, support their growth and activity.

In the present invention, the ability that yeast effectively decomposes the complex carbon compound is activated or strengthens, and gained C-decomposability yeast cell can be used as the component of Biofertiliser composition of the present invention and uses.

According to the present invention, can carry out yeast cell that C-decomposes in the suitable culture base by cell is prepared cultivating in the presence of the electromagnetic field.The electromagnetic field frequency that activation yeast C-decomposes is usually in the 1000MHz-1200MHz scope.Cell cultures can detect the ability that cell decomposes the complex carbon compound by method well known in the art after the sufficiently long time.

The method that the present invention prepares C-decomposition yeast cell is to carry out in the liquid medium within.Contain the absorbable nutrition of yeast cell in the substratum.The complicated carbonaceous material such as the Mierocrystalline cellulose of suitable form, xylogen, coal dust etc. can be as the carbon sources in the substratum.The accurate consumption of one or more carbon sources depends in part on other composition in the substratum in the substratum, but usually the amount of simple carbohydrates between substratum weight about 0.1% to 5% between, preferably between about 0.1% to 1%, most preferably from about 0.5%.In substratum, these carbon sources can be used separately, or several such carbon source is united use.

Can add inorganic salt in the substratum comprises sodium, calcium, phosphate radical, sulfate radical, the isoionic conventional salt of carbonate can be provided.The limiting examples of the inorganic salt of trophicity has (NH ₄) ₂HPO ₄, CaCO ₃, MgSO ₄, NaCl and CaSO ₄

Table 5:C-decomposes the zymic medium component

Medium component	Content
Medium component	Content	Mierocrystalline cellulose	3.0g; Powder＞100 orders
Argol fertilizer, mud or rubbish	5g; Powder＞150 orders	Mierocrystalline cellulose	3.0g; Powder＞100 orders
Argol fertilizer, mud or rubbish	5g; Powder＞150 orders	????NaCl	????0.6g
????MgSO ₄·7H ₂O	????0.3g	????NaCl	????0.6g
????MgSO ₄·7H ₂O	????0.3g	????CaCO ₃·5H ₂O	????1.5g
????CaSO ₄·2H ₂O	????0.4g	????CaCO ₃·5H ₂O	????1.5g
????CaSO ₄·2H ₂O	????0.4g	????(NH ₄) ₂HPO ₄	????0.3g
The pasty state yeast extract	????0.5g	????(NH ₄) ₂HPO ₄	????0.3g
The pasty state yeast extract	????0.5g	????K ₂HPO ₄	????0.5g
Autoclaving water	????1000ml	????K ₂HPO ₄	????0.5g

It should be noted that the medium component that provides in the table 5 is not restrictive.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.

Electromagnetic field can be used by any means known in the art, and the frequency of each electromagnetic field arrives in about 1200MHz scope preferred 1050.000 to 1160.000MHz about 1000.Such as but not limited to these examples, the frequency of each electromagnetic field can be about 1050,1055,1060,1065,1070,1075,1080,1085,1090,1095,1100,1105,1110,1115,1120,1125,1130,1135,1140,1145,1150,1155 or 1160MHz.The field intensity of electromagnetic field arrives in the 200mV/cm scope 10.If use one group of electromagnetic field, each electromagnetic field can have different frequency in the above-mentioned scope, or different field intensity in the above-mentioned scope, or interior different frequency of above-mentioned scope and field intensity.In a preferred embodiment, it is lower that the electromagnetic field that begins in a group and electromagnetic field are subsequently compared the EM field intensity, and yeast cell culture just is placed in the electromagnetic field that field intensity increases gradually like this.Though the electromagnetic field number of using in a group can be arbitrarily, preferably yeast culture is placed one group and add up to 2,3,4,5,6,7 or 8 different electromagnetic field.

For example, use exemplary device shown in Figure 1, initial field intensity is used about 12.5mV/cm usually in the 10-20mV/cm scope.Period 1 further cultivates yeast cell another cycle under the essentially identical condition higher level in the electromagnetic field field intensity increases to the 100-200Mv/cm scope (about usually 125mV/cm) after cultivating.Method of the present invention is carried out in about 23 to 30 ℃ of temperature ranges; But, preferably carry out at 25 to 28 ℃.Culturing process is 0.025 to 0.8mol/m at dissolved oxygen concentration preferably ³Condition under carry out preferred 0.4mol/m ³Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.

When culturing process finished, C-decomposed yeast cell and can reclaim from culture by several different methods known in the art, and stored being lower than under about 0-4 ℃ the temperature.C-decomposes yeast cell and also can the drying processing store with form of powder.

The ability that the yeast cell of cultivating decomposes complicated carbonaceous material can detect by any means known in the art.For example, the index that can change as carbonaceous material concentration complicated in the sample with the change of sample chemical oxygen demand (COD) (COD).For example, with 1ml Wine brewing yeast strain AS2.982 (2 * 10 ⁷To 5 * 10 ⁷Cell/ml) is inoculated in the 30ml substratum according to table 5.Culture is cultivated in 20-28 ℃ of temperature range, in the presence of 8 electromagnetic field of one group of following order: 1050MHz, 16mV/cm 5 hours; 1070MHz, 16mV/cm 5 hours; 1090MHz, 16mV/cm 5 hours; 1110MHz, 16mV/cm 5 hours; 1050MHz, 125mV/cm 30 hours; 1070MHz, 125mV/cm 30 hours; 1090MHz, 125mV/cm 30 hours; 1110MHz, 125mV/cm 30 hours.After the activation, measure based on the change of COD, the amount of carbohydrate is greater than the 330mg/ml yeast culture in the culture.COD does not obviously change in the control cultures.

In addition, the amount of simple carbohydrates can detect by any means known in the art in the culture, includes but not limited to biochemical reaction, chromatography and molecular fluorescence spectrum.

5.6 produce the yeast cell component of somatomedin

The yeast that the present invention produces somatomedin (GF-generation) produces multivitamin and other nutrition, such as but not limited to vitamin B-1, riboflavin (vitamin B-2), vitamin B-12, nicotinic acid (B-3), pyridoxol (B-6), pantothenic acid (B-5), folic acid, vitamin H, para-amino benzoic acid, choline, inositol, its amount can be supported the growth of other yeast strain.

The ability of the excessive generation somatomedin of yeast is activated by method of the present invention or strengthens, and gained GF-generation property yeast cell can be used as the component of Biofertiliser composition of the present invention and uses.

According to the present invention, yeast cell that can excessive generation somatomedin is by preparing yeast cell cultivating in the presence of the electromagnetic field in the suitable culture base.Be used to activate or strengthen electromagnetic field frequency that yeast GF produces usually in the 1300MHz-1500MHz scope.Cell cultures can detect the ability that they produce somatomedin by method well known in the art after the sufficiently long time.

The method that the present invention prepares GF generation property yeast cell is to carry out in the liquid medium within.Contain the absorbable nutrition of yeast cell in the substratum.Usually, carbohydrate such as carbohydrate, for example sucrose, glucose, fructose, dextrose, maltose, wood sugar etc. and starch can separately or be united as the source that can absorb carbon in the substratum and use.The accurate consumption of one or more carbohydrate sources depends in part on other composition in the substratum in the substratum, but usually the amount of carbohydrate between substratum weight about 0.1% to 5% between, preferably between about 0.5% to 2%, most preferably from about 0.8%.In substratum, these carbon sources can be used separately, or several such carbon source is united use.

Can add inorganic salt in the substratum comprises sodium, calcium, phosphate radical, sulfate radical, the isoionic conventional salt of carbonate can be provided.The limiting examples of the inorganic salt of trophicity has NH ₄NO ₃, K ₂HPO ₄, CaCO ₃, MgSO ₄, NaCl and CaSO ₄

Table 6:GF generation property zymic medium component

Medium component	Content
Medium component	Content	Starch	8.0g; Powder＞120 orders
????NaCl	????0.3g	Starch	8.0g; Powder＞120 orders
????NaCl	????0.3g	????MgSO ₄·7H ₂O	????0.2g
????CaCO ₃·5H ₂O	????0.5g	????MgSO ₄·7H ₂O	????0.2g
????CaCO ₃·5H ₂O	????0.5g	????CaSO ₄·2H ₂O	????0.2g
????NH ₄NO ₃	????0.3g	????CaSO ₄·2H ₂O	????0.2g
????NH ₄NO ₃	????0.3g	????K ₂HPO ₄	????0.8g
Autoclaving water	????1000ml	????K ₂HPO ₄	????0.8g

It should be noted that the medium component that provides in the table 6 is not restrictive.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.

Electromagnetic field can be used by any means known in the art, and the frequency of each electromagnetic field arrives in about 1500MHz scope about 1300, and preferred 1340.000 in the 1440.000MHz scope.Such as but not limited to these examples, each electromagnetic field frequency can be about 1340,1345,1350,1355,1360,1365,1370,1375,1380,1385,1390,1395,1400,1405,1410,1415,1420,1425,1430,1435 or 1440MHz.The field intensity of electromagnetic field arrives in the 200mV/cm scope 20.If use one group of electromagnetic field, each electromagnetic field can have different frequency in the above-mentioned scope, or different field intensity in the above-mentioned scope, or different frequency and field intensity in the above-mentioned scope.In a preferred embodiment, it is lower that the electromagnetic field that begins in a group and electromagnetic field are subsequently compared the EM field intensity, and yeast cell culture just is placed in the electromagnetic field that field intensity increases gradually like this.Though the electromagnetic field number of using in a group can be arbitrarily, preferably yeast culture is placed one group and add up to 2,3,4,5,6,7 or 8 different electromagnetic field.

For example, use exemplary device shown in Figure 1, initial field intensity is used about 25mV/cm usually in the 20-40mV/cm scope.Period 1 further cultivates yeast cell another cycle under the essentially identical condition the higher level (about usually 125mV/cm) that increases to except electromagnetic field intensity in the 100-200mV/cm scope after cultivating.Method of the present invention is carried out in about 23 to 30 ℃ of temperature ranges; But, preferably carry out at 25 to 28 ℃.Culturing process is 0.025 to 0.8mol/m at dissolved oxygen concentration preferably ³Condition under carry out preferred 0.4mol/m ³Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.

When culturing process finished, GF produced the property yeast cell and can reclaim from culture by several different methods known in the art, and stored being lower than under about 0-4 ℃ the temperature.GF generation property yeast cell also can store with form of powder in the drying processing.

The ability of the excessive generation somatomedin of yeast cell of cultivating can detect by any means known in the art, includes but not limited to high performance liquid chromatography (HPLC).For example, with 1ml activatory or non-activated Wine brewing yeast strain AS2.413 (2 * 10 ⁷To 5 * 10 ⁷Cell/ml) is inoculated in the 1000ml substratum according to table 6.Culture is cultivated in the presence of 8 electromagnetic field of 28 ℃, one group following orders: 1340MHz, 28mV/cm 5 hours; 1350MHz, 28mV/cm5 hour; 1380MHz, 28mV/cm 5 hours; 1390MHz, 28mV/cm 5 hours; 1340MHz, 135mV/cm 30 hours; 1350MHz, 135mV/cm 30 hours; 1380MHz, 135mV/cm 30 hours; 1390MHz, 135mV/cm 30 hours.The amount that produces somatomedin can or not activate the total amount of VITMAIN B1 in the yeast culture, B2, B6 and B12 and not have by activation to be calculated with the difference between the total amount of like growth factor in the same substratum of zymic.After measured, the amount of somatomedin has increased 350mg above/ml activatory yeast culture.The total amount of somatomedin does not have noticeable change in the control cultures.

5.7 ATP generation property yeast cell component

ATP of the present invention produces the property yeast can excessive generations ATP, its amount can the biological support Ru 2006101161 in other zymic grow.

In the present invention, the ability of the excessive generation of yeast ATP is activated or strengthens, and gained ATP generation property yeast cell can be used as the component of Biofertiliser composition of the present invention and uses.

According to the present invention, the yeast cell that the ATP-with reinforcement produces ability is by preparing cell cultivating in the presence of the electromagnetic field in the suitable culture base.Be used to activate or strengthen electromagnetic field frequency that yeast ATP-produces usually in the 1600MHz-1800MHz scope.Cell cultures can detect the ability that their enhanced produce ATP by method well known in the art after the sufficiently long time.

The method that the present invention prepares ATP generation property yeast cell is to carry out in the liquid medium within.Substratum contains the absorbable nutrition of yeast cell.Usually, carbohydrate such as carbohydrate, for example sucrose, glucose, fructose, dextrose, maltose, wood sugar etc. and starch can separately or be united as the source that can absorb carbon in the substratum and use.The accurate consumption of one or more carbohydrate sources that use in the substratum depends in part on other composition of substratum, but usually the amount of carbohydrate between substratum weight about 0.1% to 5% between, preferably between about 0.5% to 2%, most preferably from about 0.8%.In substratum, these carbon sources can be used separately, or several such carbon source is united use.

Can add inorganic salt in the substratum comprises sodium, calcium, phosphate radical, sulfate radical, the isoionic conventional salt of carbonate can be provided.The limiting examples of the inorganic salt of trophicity has (NH ₄) ₂HPO ₄, K ₂HPO ₄, CaCO ₃, MgSO ₄, NaCl and CaSO ₄

Table 7:ATP generation property zymic medium component

Medium component	Content
Medium component	Content	Starch	10.0g,＞120 orders
??NaCl	??0.2g	Starch	10.0g,＞120 orders
??NaCl	??0.2g	??MgSO ₄·7H ₂O	??0.2g
??CaCO ₃·5H ₂O	??0.8g	??MgSO ₄·7H ₂O	??0.2g
??CaCO ₃·5H ₂O	??0.8g	??CaSO ₄·2H ₂O	??0.2g
??NH ₄NO ₃	??0.2g	??CaSO ₄·2H ₂O	??0.2g
??NH ₄NO ₃	??0.2g	??K ₂HPO ₄	??0.5g
Autoclaving water	??1000ml	??K ₂HPO ₄	??0.5g

It should be noted that the medium component that provides in the table 7 is not restrictive.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.

Electromagnetic field can be used by any means known in the art, and the frequency of each electromagnetic field arrives about 1800MHz scope about 1600, preferably arrives in the 1730.000MHz scope 1630.000.Such as but not limited to these examples, each electromagnetic field frequency can be about 1630,1635,1640,1645,1650,1655,1660,1665,1670,1675,1680,1685,1690,1695,1700,1705,1710,1715,1720,1725 or 1730MHz.The field intensity of electromagnetic field arrives in the 200mV/cm scope 20.If use one group of electromagnetic field, each electromagnetic field can have different frequency in the above-mentioned scope, or different field intensity in the above-mentioned scope, or different frequency and field intensity in the above-mentioned scope.In a preferred embodiment, it is lower that the electromagnetic field that begins in a group and electromagnetic field are subsequently compared the EM field intensity, and yeast cell culture just is placed in the electromagnetic field that field intensity increases gradually like this.Though the electromagnetic field number of using in a group can be arbitrarily, preferably yeast culture is placed one group and add up to 2,3,4,5,6,7 or 8 different electromagnetic field.

Even just can be activated though yeast cell is cultivated several hrs in the presence of electromagnetic field, but yeast cell can continue to cultivate for some time (for example 1 week or more weeks) in the presence of electromagnetic field, usually preferred, should make the activatory yeast cell in the presence of one or more electromagnetic field, breed and grew about altogether 160-300 hour.

For example, use exemplary device shown in Figure 1, initial field intensity is used about 30mV/cm usually in the 20-40mV/cm scope.Period 1 further cultivates yeast cell another cycle under the essentially identical condition the higher level (about usually 150mV/cm) that increases to except electromagnetic field intensity in the 100-200mV/cm scope after cultivating.Method of the present invention is carried out in about 23 to 30 ℃ of temperature ranges; But, preferably carry out at 25 to 28 ℃.Culturing process is 0.025 to 0.8mol/m at dissolved oxygen concentration preferably ³Condition under carry out preferred 0.4mol/m ³Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.

When culturing process finished, ATP produced the property yeast cell and can reclaim from culture by several different methods known in the art, and stored being lower than under about 0-4 ℃ the temperature.ATP generation property yeast cell also can store with form of powder in the drying processing.

The ability of the excessive generation of the yeast cell of cultivating ATP can detect by any means known in the art, includes but not limited to HPLC.For example, with 1ml activatory yeast culture (2 * 10 ⁷To 5 * 10 ⁷Cell/ml) is inoculated in the 1000ml substratum according to table 7.Culture is cultivated in the presence of 8 electromagnetic field of 28 ℃, one group following orders: 1635MHz, 29mV/cm10 hour; 1655MHz, 29mV/cm 10 hours; 1675MHz, 29mV/cm 10 hours; 1695MHz, 29mV/cm 10 hours; 1635MHz, 150mV/cm 30 hours; 1655MHz, 150mV/cm 30 hours; 1675MHz, 150mV/cm 30 hours; 1695MHz, 150mV/cm30 hour.Produce the total amount that the amount of ATP can be by ATP in the yeast culture and do not have in the same substratum of zymic the difference between the ATP total amount calculate.Use activatory Wine brewing yeast strain AS2.536, record that ATP must measure the yeast culture into 170mg/ml in the culture.

5.8 pathogenic agent inhibition yeast cell component

The present invention also provides the yeast cell of the pathogenic microorganism propagation that can suppress to be present in the bio-feritlizer organic substrate constituent materials.Usually, owing in the organic substrate material the utilizable abundant nutrition of these pathogenic microorganisms is arranged, the number of pathogenic agent increases fast after after a while.But in the presence of pathogenic agent inhibition zymic of the present invention, through after a while, the number of organic substrate pathogens in materials remains unchanged or reduces.Though be not subjected to the constraint of any theory or mechanism, the present inventor thinks that the existence of organic substrate pathogens in materials inhibition yeast cell has formed the environment that is unfavorable for the pathogenic microorganism growth.

According to the present invention, the ability of yeast influence/control pathogenic agent number is activated or strengthens by yeast is cultivated in the presence of electromagnetic field.Gained pathogenic agent inhibition yeast cell uses as the component of Biofertiliser composition of the present invention.

The electromagnetic field frequency of activation or reinforcement yeast control pathogenic microorganism number ability is usually in 30MHz arrives the 50MHz scope.Yeast cell growth can detect the ability of impact cell/control pathogenic agent number by method well known in the art after the sufficiently long time.

The method that the present invention prepares pathogenic agent inhibition yeast cell is to carry out in the liquid medium within.Contain the absorbable nutrition source of yeast cell in the substratum.Usually, carbohydrate such as carbohydrate, for example sucrose, glucose, fructose, dextrose, maltose, wood sugar etc. and starch can separately or be united as the source that can absorb carbon in the substratum and use.The accurate consumption of one or more carbohydrate sources depends in part on other composition in the substratum in the substratum, but usually the amount of carbohydrate between substratum weight about 0.1% to 5% between, preferred about 0.5% to 2%, most preferably from about 0.8%.In substratum, these carbon sources can be used separately, or several such carbon source is united use.

Table 8: pathogenic agent inhibition zymic medium component

Medium component	Content
Medium component	Content	Zulkovsky starch	????8.0g
Sucrose	????5g	Zulkovsky starch	????8.0g
Sucrose	????5g	????NaCl	????0.2g
????MgSO ₄·7H ₂O	????0.2g	????NaCl	????0.2g
????MgSO ₄·7H ₂O	????0.2g	????CaCO ₃·5H ₂O	????0.5g
????CaSO ₄·2H ₂O	????0.2g	????CaCO ₃·5H ₂O	????0.5g
????CaSO ₄·2H ₂O	????0.2g	Peptone	????1.5g
????K ₂HPO ₄	????0.5g	Peptone	????1.5g
????K ₂HPO ₄	????0.5g	Autoclaving water	????400ml
Muck, mud or rubbish extract	????600ml	Autoclaving water	????400ml

Used muck, mud or the rubbish extract of substratum is to remove particulate matter by brid guano fertilizer, cattle manure, pig manure, mud or rubbish that 500g is fresh 30-37 ℃ of cultivation 24 hours down, filter liquide in about 600ml warm water (35 ℃ to 40 ℃) to prepare.It should be noted that the medium component that provides in the table 8 is not restrictive.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.

Electromagnetic field can be used by any means known in the art, and the frequency of each electromagnetic field arrives in about 50.000MHz scope about 30.000.Such as but not limited to these examples, each electromagnetic field frequency can be about 30,31,32,33,34,35,36,37,38,39,40,41,42,43,44,45,46,47,48,49 or 50MHz.The field intensity of electromagnetic field arrives in the 200mV/cm scope preferred 10 to 180mV/cm 0.5.If use one group of electromagnetic field, each electromagnetic field can have different frequency in the above-mentioned scope, or different field intensity in the above-mentioned scope, or interior different frequency of above-mentioned scope and field intensity.In a preferred embodiment, it is lower that the electromagnetic field that begins in a group and electromagnetic field are subsequently compared the EM field intensity, and yeast cell culture just is placed in the electromagnetic field that field intensity increases gradually like this.Though the electromagnetic field number of using in a group can be arbitrarily, preferably yeast culture is placed one group and add up to 2,3,4,5,6,7 or 8 different electromagnetic field.

Even just can be activated though yeast cell is cultivated several hrs in the presence of electromagnetic field, but yeast cell can continue to cultivate for some time (for example 1 week or more weeks) in the presence of electromagnetic field, usually preferred, should make the activatory yeast cell in the presence of one or more electromagnetic field, breed and grew about altogether 144-272 hour.

For example, use exemplary device shown in Figure 1, initial field intensity is used about 25mV/cm usually in the 10-30mV/cm scope.Period 1 further cultivates yeast cell another cycle under the essentially identical condition the higher level (about usually 150mV/cm) that increases to except amplitude in the 100-200mV/cm scope after cultivating.Method of the present invention is carried out in about 23 to 30 ℃ of temperature ranges; But, preferably carry out at 25 to 28 ℃.Culturing process is 0.025 to 0.8mol/m at dissolved oxygen concentration preferably ³Condition under carry out preferred 0.4mol/m ³Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.

When culturing process finished, pathogenic agent-inhibition yeast cell can reclaim from culture by several different methods known in the art, and stores under about 0 to 4 ℃ temperature.Pathogenic agent-inhibition yeast cell also can store with form of powder in the drying processing.

The ability of pathogenic agent-inhibition yeast cell control pathogenic agent number can detect by any microorganism count method known in the art, for example optical density(OD), the solid medium flat board dilution method of counting or in microscopically individual cells counting process.Can use that dyeing is distinguished or definite sample in different microorganism strains or kinds, or detect their viability.When estimating pathogenic agent-when the inhibition yeast influences a series of pathogenic microorganism, can monitor the number of more than one representative pathogenic microorganism kinds and assess pathogenic agent-inhibition zymic performance.

For example, in the presence of different concns pathogenic agent inhibition zymic, the organic substrate sample of material that contains the concentration known pathogenic microorganism is cultivated the identical time under similarity condition, and the identical yeast strain that cultural method is handled according to the present invention is not as negative control.Do not add any zymic sample and can be used for detecting the growth of pathogenic agent under the normal condition yet.The number of pathogenic agent before and after detecting and relatively cultivating.

Prepare 1 liter of culture, every milliliter contains 10 at least ¹⁰The pathogenic microorganism of cell.(every milliliter contains 2 * 10 to add 1ml activatory yeast cell in 1 liter of pathogenic microorganism culture ⁷To 5 * 10 ⁷Individual yeast), cultivated 24 hours at 30 ℃.Comprise and contain not activated yeast cell or do not contain the zymic contrast.Detect and compare the number of microorganism in each culture.Be several examples below, wherein studied the malignant bacteria of particular types.

Use Wine brewing yeast strain IFFI1037, in the presence of 8 electromagnetic field of one group of following order, cultivate: 30MHz, 26mV/cm 12 hours; 36MHz, 26mV/cm 12 hours; 43MHz, 26mV/cm 12 hours; 47MHz, 26mV/cm 12 hours; 30MHz, 150mV/cm 24 hours; 36MHz, 150mV/cm 24 hours; 43MHz, 150mV/cm 24 hours; 47MHz, 150mV/cm 24 hours.With respect to not containing zymic contrast, in the sample reduced number of streptococcus aureus more than 2.7%.The number that contains pathogenic agent in the contrast of active cells does not obviously change.

Use Wine brewing yeast strain IFFI1021, in the presence of 8 electromagnetic field of one group of following order, cultivate: 30MHz, 26mV/cm 12 hours; 36MHz, 26mV/cm 12 hours; 42MHz, 26mV/cm 12 hours; 49MHz, 26mV/cm 12 hours; 30MHz, 150mV/cm 24 hours; 36MHz, 150mV/cm 24 hours; 42MHz, 150mV/cm 24 hours; 49MHz, 150mV/cm 24 hours.With respect to not containing zymic contrast, in the sample reduced number of pneumococcus more than 2.8%.The number that contains pathogenic agent in the contrast of active cells does not obviously change.

Use Wine brewing yeast strain IFFI1051, in the presence of 8 electromagnetic field of one group of following order, cultivate: 35MHz, 26mW/cm 12 hours; 39MHz, 26mV/cm 12 hours; 43MHz, 26mV/cm 12 hours; 47MHz, 26mV/cm 12 hours; 35MHz, 150mV/cm 24 hours; 39MHz, 150mV/cm 24 hours; 43MHz, 150mV/cm 24 hours; 47MHz, 150mV/cm 24 hours.With respect to not containing zymic contrast, in the sample reduced number of anthrax bacillus more than 3.1%.The number that contains pathogenic agent in the contrast of active cells does not obviously change.

Use Wine brewing yeast strain IFFI1331, in the presence of 8 electromagnetic field of one group of following order, cultivate: 33MHz, 26mV/cm 12 hours; 36MHz, 26mV/cm 12 hours; 45MHz, 26mV/cm 12 hours; 47MHz, 26mV/cm 12 hours; 33MHz, 150mV/cm 24 hours; 36MHz, 150mV/cm 24 hours; 45MHz, 150mV/cm 24 hours; 47MHz, 150mV/cm 24 hours.With respect to not containing zymic contrast, in the sample Mycobacterium tuberculosis reduced number more than 2.9%.The number that contains pathogenic agent in the contrast of active cells does not obviously change.

Use Wine brewing yeast strain IFFI1345, in the presence of 8 electromagnetic field of one group of following order, cultivate: 30MHz, 26mV/cm 12 hours; 34MHz, 26mV/cm 12 hours; 38MHz, 26mV/cm 12 hours; 49MHz, 26mV/cm 12 hours; 30MHz, 150mV/cm 24 hours; 34MHz, 150mV/cm 24 hours; 38MHz, 150mV/cm 24 hours; 49MHz, 150mV/cm 24 hours.With respect to not containing zymic contrast, in the sample colibacillary reduced number more than 48%.The number that contains pathogenic agent in the contrast of active cells does not obviously change.

Use Wine brewing yeast strain IFFI1211, in the presence of 8 electromagnetic field of one group of following order, cultivate: 30MHz, 26mV/cm 12 hours; 33MHz, 26mV/cm 12 hours; 36MHz, 26mV/cm 12 hours; 38MHz, 26mV/cm 12 hours; 30MHz, 150mV/cm 24 hours; 33MHz, 150mV/cm 24 hours; 36MHz, 150mV/cm 24 hours; 38MHz, 150mV/cm 24 hours.With respect to not containing zymic contrast, in the sample reduced number of salmonella bacterium more than 66%.The number that contains pathogenic agent in the contrast of active cells does not obviously change.

5.9 decompose the yeast cell component of adverse chemical product

The present invention further provides adverse chemical product common in to degrade muck or the mud such as antibiotic yeast cell.

According to the present invention, the yeast cell antibiotic ability of degrading is to activate or strengthen by yeast is cultivated in the presence of electromagnetic field.The gained yeast cell can be used as the component of Biofertiliser composition of the present invention and uses.

Be used for activating or strengthen yeast degradation adverse chemical product particularly the microbiotic ability electromagnetic field frequency usually at about 70MHz in the scope of about 100MHz.Yeast cell growth can detect the antibiotic ability of decomposition that yeast cell is strengthened by method well known in the art after the sufficiently long time.Can be included but not limited to the molecule of β-Nei acyl Ammonia, tetracyclines, polypeptide class, glycopeptide class, aminoglycoside and Macrolide family by the microbiotic of yeast degradation of the present invention.

It is to carry out in the liquid medium within that the present invention prepares microbiotic-degradation property yeast method.Contain the absorbable nutrition source of yeast cell in the substratum.Usually, carbohydrate such as carbohydrate, for example sucrose, glucose, fructose, dextrose, maltose, wood sugar etc. and starch can separately or be united as the source that can absorb carbon in the substratum and use.The accurate consumption of one or more carbohydrate sources depends in part on other composition in the substratum in the substratum, but usually the amount of carbohydrate between substratum weight about 0.1% to 5% between, preferred about 0.5% to 2%, most preferably from about 0.8%.In substratum, these carbon sources can be used separately, or several such carbon source is united use.

Table 9: the zymic medium component of degraded adverse chemical product

Medium component	Content
Medium component	Content	Zulkovsky starch	8.0g,＞120 orders
Sucrose	????5g	Zulkovsky starch	8.0g,＞120 orders
Sucrose	????5g	????NaCl	????0.2g
????MgSO ₄·7H ₂O	????0.2g	????NaCl	????0.2g
????MgSO ₄·7H ₂O	????0.2g	????CaCO ₃·5H ₂O	????0.5g
????CaSO ₄·2H ₂O	????0.2g	????CaCO ₃·5H ₂O	????0.5g
????CaSO ₄·2H ₂O	????0.2g	Peptone	????1.5g
????K ₂HPO ₄	????0.5g	Peptone	????1.5g
????K ₂HPO ₄	????0.5g	Autoclaving water	????1000ml
Muck, mud or rubbish extract	????600ml	Autoclaving water	????1000ml

Used muck, mud or the rubbish extract of substratum is to remove particulate matter by brid guano fertilizer, cattle manure, pig manure, mud or rubbish that 500g is fresh 30-37 ℃ of cultivation 24 hours down, filter liquide in about 600ml warm water (35-40 ℃) to prepare.It should be noted that the medium component that provides in the table 9 is not restrictive.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.

Electromagnetic field can be used by any means known in the art, and the frequency of each electromagnetic field arrives the 100.000MHz scope 70.000.Such as but not limited to these examples, each electromagnetic field frequency can be about 70,71,72,73,74,75,76,77,78,79,80,81,82,83,84,85,86,87,88,89,90,91,92,93,94,95,96,97,98 or 100MHz.The field intensity of electromagnetic field arrives in the 250mV/cm scope 40.If use one group of electromagnetic field, each electromagnetic field can have different frequency in the above-mentioned scope, or different field intensity in the above-mentioned scope, or interior different frequency of above-mentioned scope and field intensity.In a preferred embodiment, it is lower that the electromagnetic field that begins in a group and electromagnetic field are subsequently compared the EM field intensity, and yeast cell culture just is placed in the electromagnetic field that field intensity increases gradually like this.Though the electromagnetic field number of using in a group can be arbitrarily, preferably yeast culture is placed one group and add up to 2,3,4,5,6,7 or 8 different electromagnetic field.

Even just can be activated though yeast cell is cultivated several hrs in the presence of electromagnetic field, but yeast cell can continue to cultivate for some time (for example 1 week or more weeks) in the presence of electromagnetic field, usually preferred, should make the activatory yeast cell in the presence of one or more electromagnetic field, breed and grew about altogether 180-328 hour.

For example, use exemplary device shown in Figure 1, initial field intensity is used about 50mV/cm usually in the 40-60mV/cm scope.Period 1 further cultivates yeast cell another cycle under the essentially identical condition the higher level (about usually 200mV/cm) that increases to except amplitude in the 100-250mV/cm scope after cultivating.Method of the present invention is carried out in about 23 ℃ to 30 ℃ temperature range; But, preferably carry out at 25 to 28 ℃.Culturing process is 0.025 to 0.8mol/m at dissolved oxygen concentration preferably ³Condition under carry out preferred 0.4mol/m ³Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.

When culturing process finished, yeast cell can reclaim from culture by several different methods known in the art, and stored being lower than under about 0 ℃ to 4 ℃ temperature.The yeast cell that reclaims also can store with form of powder in the drying processing.

In order to detect the activity of activatory yeast cell to Antibiotique composition, can with method well known in the art for example HPLC measure the amount that different time points and different culture condition detect Antibiotique composition in the sample down.For example, the microbiotic (can reach every liter of 100mg) with known quantity adds in the aqueous extract of 10 liters of muck.Then, with 0.1ml activatory and non-activated yeast (at least 10 ⁷Cell/ml) add 10 liters to contain in the antibiotic sample was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, by usefulness HPLC Detection and Extraction thing sample, residual antibiotic amount in detection and the comparison extract.

Use Wine brewing yeast strain AS2.293, in the presence of 8 electromagnetic field of one group of following order, cultivate: 77MHz, 48mV/cm 15 hours; 83MHz, 48mV/cm 15 hours; 90MHz, 48mV/cm 15 hours; 96MHz, 48mV/cm 15 hours; 77MHz, 200mV/cm 30 hours; 83MHz, 200mV/cm 30 hours; 90MHz, 200mV/cm 30 hours; 96MHz, 200mV/cm 30 hours.With respect to not containing the zymic contrast, the amount of penicillin G has reduced more than 23% in the sample.Contain that antibiotic concentration does not obviously change in the contrast of active cells not.

Use Wine brewing yeast strain IFFI1063, in the presence of 8 electromagnetic field of one group of following order, cultivate: 70MHz, 48mV/cm 15 hours; 73MHz, 48mV/cm 15 hours; 88MHz, 48mV/cm 15 hours; 98MHz, 48mV/cm 15 hours; 70MHz, 200mV/cm 30 hours; 73MHz, 200mV/cm 30 hours; 88MHz, 200mV/cm 30 hours; 98MHz, 200mV/cm 30 hours.With respect to not containing the zymic contrast, the amount of duomycin has reduced more than 31% in the sample.Contain that antibiotic concentration does not obviously change in the contrast of active cells not.

Use Wine brewing yeast strain IFFI1221, in the presence of 8 electromagnetic field of one group of following order, cultivate: 70MHz, 48mV/cm 15 hours; 74MHz, 48mV/cm 15 hours; 88MHz, 48mV/cm 15 hours; 98MHz, 48mV/cm 15 hours; 70MHz, 200mV/cm 30 hours; 74MHz, 200mV/cm 30 hours; 88MHz, 200mV/cm 30 hours; 98MHz, 200mV/cm 30 hours.With respect to not containing the zymic contrast, the amount of terramycin has reduced more than 28% in the sample.Contain that antibiotic concentration does not obviously change in the contrast of active cells not.

Use Wine brewing yeast strain IFFI1340, in the presence of 8 electromagnetic field of one group of following order, cultivate: 71MHz, 48mV/cm 15 hours; 73MHz, 48mV/cm 15 hours; 77MHz, 48mV/cm 15 hours; 88MHz, 48mV/cm 15 hours; 71MHz, 200mV/cm 30 hours; 73MHz, 200mV/cm 30 hours; 77MHz, 200mV/cm 30 hours; 88MHz, 200mV/cm 30 hours.With respect to not containing the zymic contrast, the amount of doxycycline has reduced more than 33% in the sample.Contain that antibiotic concentration does not obviously change in the contrast of active cells not.

Use Wine brewing yeast strain IFFI1215, in the presence of 8 electromagnetic field of one group of following order, cultivate: 70MHz, 48mV/cm 15 hours; 75MHz, 48mV/cm 15 hours; 82MHz, 48mV/cm 15 hours; 85MHz, 48mV/cm 15 hours; 70MHz, 200mV/cm 30 hours; 75MHz, 200mV/cm 30 hours; 82MHz, 200mV/cm 30 hours; 85MHz, 200mV/cm 30 hours.With respect to not containing the zymic contrast, the amount of tsiklomitsin has reduced more than 26% in the sample.Contain that antibiotic concentration does not obviously change in the contrast of active cells not.

Use Wine brewing yeast strain IFFI1213, in the presence of 8 electromagnetic field of one group of following order, cultivate: 70MHz, 48mV/cm 15 hours; 73MHz, 48mV/cm 15 hours; 80MHz, 48mV/cm 15 hours; 96MHz, 48mV/cm 15 hours; 70MHz, 200mV/cm 30 hours; 73MHz, 200mV/cm 30 hours; 80MHz, 200mV/cm 30 hours; 96MHz, 200mV/cm 30 hours.With respect to not containing the zymic contrast, the amount of Streptomycin sulphate has reduced more than 31% in the sample.Contain that antibiotic concentration does not obviously change in the contrast of active cells not.

Use Wine brewing yeast strain IFFI1206, in the presence of 8 electromagnetic field of one group of following order, cultivate: 71MHz, 48mV/cm 15 hours; 78MHz, 48mV/cm 15 hours; 86MHz, 48mV/cm 15 hours; 98MHz, 48mV/cm 15 hours; 71MHz, 200mV/cm 30 hours; 78MHz, 200mV/cm 30 hours; 86MHz, 200mV/cm 30 hours; 98MHz, 200mV/cm 30 hours.With respect to not containing the zymic contrast, the amount of kantlex has reduced more than 25% in the sample.Contain that antibiotic concentration does not obviously change in the contrast of active cells not.

Use Wine brewing yeast strain IFFI1211, in the presence of 8 electromagnetic field of one group of following order, cultivate: 73MHz, 48mV/cm 15 hours; 79MHz, 48mV/cm 15 hours; 88MHz, 48mV/cm 15 hours; 98MHz, 48mV/cm 15 hours; 73MHz, 200mV/cm 30 hours; 79MHz, 200mV/cm 30 hours; 88MHz, 200mV/cm 30 hours; 98MHz, 200mV/cm 30 hours.With respect to not containing the zymic contrast, the amount of erythromycin has reduced more than 27% in the sample.Contain that antibiotic concentration does not obviously change in the contrast of active cells not.

Use Wine brewing yeast strain IFFI1210, in the presence of 8 electromagnetic field of one group of following order, cultivate: 70MHz, 48mV/cm 15 hours; 77MHz, 48mV/cm 15 hours; 84MHz, 48mV/cm 15 hours; 93MHz, 48mV/cm 15 hours; 70MHz, 200mV/cm 30 hours; 77MHz, 200mV/cm 30 hours; 84MHz, 200mV/cm 30 hours; 93MHz, 200mV/cm 30 hours.With respect to not containing the zymic contrast, the amount of Spiramycin Base has reduced more than 22% in the sample.Contain that antibiotic concentration does not obviously change in the contrast of active cells not.

Use Wine brewing yeast strain IFFI1260, in the presence of 8 electromagnetic field of one group of following order, cultivate: 75MHz, 48mV/cm 15 hours; 78MHz, 48mV/cm 15 hours; 81MHz, 48mV/cm 15 hours; 95MHz, 48mV/cm 15 hours; 75MHz, 200mV/cm 30 hours; 78MHz, 200mV/cm 30 hours; 81MHz, 200mV/cm 30 hours; 95MHz, 200mV/cm 30 hours.With respect to not containing the zymic contrast, the amount of bacitracin has reduced more than 17% in the sample.Contain that antibiotic concentration does not obviously change in the contrast of active cells not.

5.10 reduce the yeast cell component of smell

The present invention also provides the yeast cell of the smell that can reduce muck, mud or rubbish.Though be not subjected to the constraint of any theory or mechanism, the present inventor thinks that yeast cell of the present invention can reduce the smell of muck, mud or rubbish by changing or decomposing malodorous compound known and unknown in these organic materialss.But, there is no need to confirm that these compounds are decomposed.As long as after using yeast cell of the present invention, one group of subjective smell that detects of main body is reduced just passable.

According to the present invention, the yeast cell that can reduce the organic materials smell is by preparing cell cultivating in the presence of the electromagnetic field, in the suitable culture base.The electromagnetic field frequency of activation or this function of reinforcement yeast arrives in about 2380MHz scope about 2160 usually.Yeast cell growth can detect the ability that yeast cell reduces the organic materials smell by method well known in the art after the sufficiently long time.

The method that the present invention prepares the yeast cell that reduces smell is to carry out in the liquid medium within.Substratum contains the absorbable nutrition source of yeast cell.Usually, carbohydrate such as carbohydrate, for example sucrose, glucose, fructose, dextrose, maltose, wood sugar etc. and starch can separately or be united as the source that can absorb carbon in the substratum and use.The accurate consumption of one or more carbohydrate sources depends in part on other composition in the substratum in the substratum, but usually between substratum weight about 0.1% to 5% between, preferred about 0.5% to 2%.Most preferably from about 0.8%.In substratum, these carbon sources can be used separately, or several such carbon source is united use.

Table 10: the zymic medium component that reduces smell

Medium component	Content
Medium component	Content	Muck, mud, rubbish	????100g
????NaCl	????0.2g	Muck, mud, rubbish	????100g
????NaCl	????0.2g	????MgSO ₄·7H ₂O	????0.2g
????CaCO ₃·5H ₂O	????0.5g	????MgSO ₄·7H ₂O	????0.2g
????CaCO ₃·5H ₂O	????0.5g	????CaSO ₄·2H ₂O	????0.2g
????K ₂HPO ₄	????0.5g	????CaSO ₄·2H ₂O	????0.2g
????K ₂HPO ₄	????0.5g	Autoclaving water	????900ml

It should be noted that the medium component that provides in the table 10 is not restrictive.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.

Electromagnetic field can be used by any means known in the art, and the frequency of each electromagnetic field arrives in about 2380.000MHz scope 2160.000, preferably arrives in the 2380MHz scope 2160 to 2250MHz or 2280.Such as but not limited to these examples, each electromagnetic field frequency can be about 2160,2165,2170,2175,2180,2185,2190,2195,2200,2205,2210,2215,2220,2225,2230,2235,2240,2245,2250,2280,2285,2290,2295,2300,2305,2315,2320,2325,2330,2335,2340,2345,2350,2355,2360,2365,2370,2375 or 2380MHz.The field intensity of electromagnetic field arrives in the 320mV/cm scope preferred 30 to 300mV/cm 0.5.If use one group of electromagnetic field, each electromagnetic field can have different frequency in the above-mentioned scope, or different field intensity in the above-mentioned scope, or interior different frequency of above-mentioned scope and field intensity.In a preferred embodiment, it is lower that the electromagnetic field that begins in a group and electromagnetic field are subsequently compared the EM field intensity, and yeast cell culture just is placed in the electromagnetic field that field intensity increases gradually like this.Though the electromagnetic field number of using in a group can be arbitrarily, preferably yeast culture is placed one group and add up to 2,3,4,5,6,7 or 8 different electromagnetic field.

Even just can be activated though yeast cell is cultivated several hrs in the presence of electromagnetic field, but yeast cell can continue to cultivate for some time (for example 2 week or more weeks) in the presence of electromagnetic field, usually preferred, should make the activatory yeast cell in the presence of one or more electromagnetic field, breed and grew about altogether 80-320 hour.

Method of the present invention is carried out in about 23 ℃ to 30 ℃ temperature range; But, preferably carry out to 28 ℃ at 25 ℃.Culturing process is 0.025 to 0.8mol/m at dissolved oxygen concentration preferably ³Condition under carry out preferred 0.4mol/m ³Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.

When culturing process finished, yeast cell can reclaim from culture by several different methods known in the art, and stored being lower than under about 0-4 ℃ the temperature.The yeast cell that reclaims also can store with form of powder in the drying processing.

Can detect the ability that the yeast cell of cultivating reduces the organic materials smell with any means known in the art.Stench chemical such as hydrogen sulfide, ammonia, indoles, p-cresol, skatole and organic acid amount that organic materials detects in the sample can detect with any means known in the art, include but not limited to vapor-phase chromatography, olfactometry, mass spectroscopy or use smell test panel.

In order to detect the activity of activatory yeast cell, can use method well known in the art such as HPLC or mass spectroscopy (as VG micromass) measurement different time points and different breeding conditions to detect the amount of malodorous compound in the sample down to malodorous compound.For example, the malodorous compound (up to every liter of 100mg) with known quantity adds in the aqueous extract of 10 liters of muck.Then, with 0.1ml activation and non-activated yeast (at least 10 ⁷Cell/ml) add 10 liters to contain in the antibiotic sample was cultivated 24 hours at 28 ℃.Comprise the contrast that does not contain any yeast cell.After 24 hours, detect and compare the amount of malodorous compound remaining in the extract.

Therefore, the sulfur-bearing that hydrogen sulfide is relevant with other or contain the caused smell of sulfydryl (SH-) molecule and can be reduced by the yeast of cultivating in the presence of the electromagnetic field in 2160.000 to 2250.000 scopes.Use Wine brewing yeast strain AS2.559 cell, in the presence of 4 electromagnetic field of one group of following order, cultivate: 2165MHz, 240mV/cm 20 hours; 2175MHz, 240mV/cm 20 hours; 2200MHz, 240mV/cm 20 hours; With 2235MHz, 240mV/cm 20 hours.With respect to not containing the zymic contrast, the amount of hydrogen sulfide has reduced more than 13% in the sample.Contain and do not activate not obviously minimizing of malodorous compound in the zymic sample.

Ammonia can be reduced by the yeast of cultivating in the presence of the electromagnetic field in 2160.000 to 2250.000 scopes with the relevant caused smell of NH-compound that contains.Use Wine brewing yeast strain AS2.423 cell, in the presence of 4 electromagnetic field of one group of following order, cultivate: 2160MHz, 250mV/cm 20 hours; 2175MHz, 250mV/cm 20 hours; 2210MHz, 250mV/cm 20 hours; With 2245MHz, 250mV/cm 10 hours.With respect to not containing the zymic contrast, the amount of ammonia has reduced more than 11% in the sample.Contain and do not activate not obviously minimizing of malodorous compound in the zymic sample.

Indoles and other associated molecule such as the caused smell of skatole can be reduced by the yeast of cultivating in the presence of the electromagnetic field in 2160.000 to 2250.000 scopes.Use Wine brewing yeast strain AS2.612 cell, in the presence of 4 electromagnetic field of one group of following order, cultivate: 2165MHz, 240mV/cm 40 hours; 2180MHz, 240mV/cm 20 hours; 2200MHz, 240mV/cm40 hour; With 2220MHz, 240mV/cm 20 hours.With respect to not containing the zymic contrast, the amount of indoles has reduced more than 15% in the sample.Contain and do not activate not obviously minimizing of malodorous compound in the zymic sample.

The caused smell of organic acid (for example formic acid, acetate, propionic acid, butyric acid and other voltaile fatty acid) can be reduced by the yeast of cultivating in the presence of the electromagnetic field in 2280.000 to 2380.000 scopes.Use Wine brewing yeast strain AS2.53 cell, in the presence of 4 electromagnetic field of one group of following order, cultivate: 2315MHz, 290mV/cm 30 hours; 2335MHz, 290mV/cm10 hour; 2355MHz, 290mV/cm 20 hours; With 2375MHz, 290mV/cm 10 hours.With respect to not containing the zymic contrast, the amount of acetate has reduced more than 19% in the sample.Contain and do not activate not obviously minimizing of malodorous compound in the zymic sample.

Methylamine, dimethylamine or Trimethylamine 99 and other aliphatics replace the caused smell of amine and can be reduced by the yeast of cultivating in the presence of the electromagnetic field in 2160.000 to 2250.000 scopes.Use Wine brewing yeast strain AS2.541 cell, in the presence of 4 electromagnetic field of one group of following order, cultivate: 2160MHz, 250mV/cm 20 hours; 2190MHz, 250mV/cm 10 hours; 2210MHz, 250mV/cm 40 hours; 2250MHz, 250mV/cm 40 hours.With respect to not containing the zymic contrast, the amount of methyl substituted amine has reduced more than 23% in the sample.Contain and do not activate not obviously minimizing of malodorous compound in the zymic sample.

The caused smell of p-cresol and related compound can be reduced by the yeast of cultivating in the presence of the electromagnetic field in 2280.000 to 2380.000 scopes.Use Wine brewing yeast strain AS2.163 cell, in the presence of 4 electromagnetic field of one group of following order, cultivate: 2300MHz, 98mV/cm 20 hours; 2370MHz, 98mV/cm 15 hours; 2300MHz, 250mV/cm 20 hours; 2370MHz, 250mV/cm 30 hours.With respect to not containing the zymic contrast, the amount of p-cresol has reduced more than 23% in the sample.Contain and do not activate not obviously minimizing of malodorous compound in the zymic sample.

5.11 the formation of symbiosis sample relation

In another embodiment of the invention, 5.1-5.5 the yeast cell that inorganic phosphor-contained thing or compound, (3) balance phosphorus compound are decomposed in (1) fixed nitrogen of describing in the joint, (2), the insoluble ability that contains the complicated carbon compound of potassium inorganics or compound and (5) decomposition of (4) decomposition just has been activated or strengthen is merged cultivation, make their form symbiosis sample relation, they can be grown and the biology that needn't mainly depend on the outside and provide can utilize nitrogen, phosphorus, potassium and carbon nutrition together thus.The required nutrition of growing produces the property yeast strain by corresponding nutrition in the Ru 2006101161 and can not utilize nutrition to be converted into by the biology with multiple source can to utilize nutrition to provide.Each yeast strain produces the part correlation that needs of the activity of corresponding kind nutrition and other yeast and plant.As a result, solubility, biological available nutrition will be transformed when needs, have therefore avoided the extra losses that causes owing to for example leaching.

The optional approach that can be used for improving the bio-feritlizer performance is described below.At least four mixed being incorporated under the electromagnetic field existence of yeast strains that are equipped with according to the 5.1-5.5 restraining are cultivated in the appropriate liquid substratum.Substratum contains nitrogen, phosphorus, potassium and the carbon nutrition that biology can not utilize form.As limiting examples, atmospheric nitrogen uses as the carbon nutrition source of complexity as use of potassium nutrition source and cellulose powder as nitrogen nutrient source use, potash mica powder as the use of nitrogen nutrition source, phosphate rock powder.The insoluble phosphorous and carbon compound that contains potassium material and complexity of other form also can replace or unite as phosphorus, potassium and the use of carbon nutrition source with any above-mentioned inorganics.Can add inorganic salt in the substratum comprises sodium, calcium, sulfate radical, the isoionic conventional salt of carbonate can be provided.The limiting examples of the inorganic salt of trophicity has CaCO ₃, MgSO ₄, NaCl and CaSO ₄

Table 11: the medium component that is used to form symbiosis sample relation

Medium component	Content
Medium component	Content	????NaCl	????0.5g
????MgSO ₄·7H ₂O	????0.4g	????NaCl	????0.5g
????MgSO ₄·7H ₂O	????0.4g	????CaCO ₃·5H ₂O	????3.0g
????CaSO ₄·2H ₂O	????0.3g	????CaCO ₃·5H ₂O	????3.0g
????CaSO ₄·2H ₂O	????0.3g	The pasty state yeast extract	????0.3g
Potash mica	1.2g; Powder＞200 orders	The pasty state yeast extract	????0.3g
Potash mica	1.2g; Powder＞200 orders	Phosphate rock	1.2g; Powder＞200 orders
Mierocrystalline cellulose	5.0g; Powder＞200 orders	Phosphate rock	1.2g; Powder＞200 orders
Mierocrystalline cellulose	5.0g; Powder＞200 orders	Autoclaving water	????1000ml

It should be noted that the medium component that provides in the table 11 is not restrictive.Those skilled in the art can carry out multiple improvement to substratum according to practice and economic consideration, for example the local provisioning of scale of Pei Yanging and nutrient media components.

Culturing process is 0.025 to 0.8mol/m at dissolved oxygen concentration preferably ³Condition under carry out preferred 0.4mol/m ³Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.This method of the present invention is carried out in about 25 ℃ to 30 ℃ temperature ranges; But, preferably carry out at 28 ℃.When this method begins usually with every kind of about 20ml inoculum of four primary yeast cell bacterial strains with about 10 ⁸Cell/ml cell density is inoculated in the aseptic culture medium.This process for selective can increase or reduce as required in proportion.

Yeast culture growth 12-72 hour, preferably in the presence of four independent electromagnetic field about 48 hours.Can apply electromagnetic field with several different methods, each has following frequency respectively: (1) fixed nitrogen, arrive in about 916MHz scope about 840; (2) phosphorus-decomposition or phosphorus balance arrive in about 500MHz scope about 300; (3) potassium-decomposition is arrived in about 300MHz scope about 100; (4) complex carbon-decomposition is arrived in about 1200MHz scope about 1000.Usually, in each recycle to extinction yeast cell placed intensity in the electromagnetic field of 5mV/cm in the 160mV/cm scope.Use exemplary device shown in Figure 2, used electromagnetic output amplitude in the 0-3000mV scope, preferred 20-1800mV.The amplitude of each electromagnetic field at 0mV to recirculation between the 3000mV, preferably at 20mV between the 1800mV, step-length is 1mV, speed about 2 is to about 10 minutes/one recycle to extinction.

5.12 soil adapts to

Yeast cell of the present invention also must be grown in polytype soil and be exercised their functions separately.The ability of yeast cell survival and growth can be strengthened by making yeast cell of the present invention adapt to specific edaphic condition.

In another embodiment of the invention, according to 5.1-5.10 save yeast cell that any restraining is equipped with can be in containing from the solid of one or more soil source soil or semisolid medium respectively or mixed culture.Following case description this be used for improving the process for selective of bio-feritlizer performance.

Containing 10ml density is 10 ⁶Cell/ml zymic suspension and 1000cm ³Solid medium mixes.This method can increase and decrease as required in proportion.Yeast and soil mixture were cultivated about 48-96 hour preferred about 48 hours in the presence of electromagnetic field.Electromagnetic field can apply by several different methods, and according to the zymic function, its frequency meets 5.1-5.10 and saves one of described frequency.Usually, in this method yeast cell placed intensity in the electromagnetic field of 60mV/cm in the 250mV/cm scope.

This culture is being cultivated under the round-robin temperature between about 4 ℃ to about 48 ℃.For example, in a typical circulation, the temperature of culture can and keep the about 1-2 of this temperature hour from 35-48 ℃ of beginning, transfer to 42-45 ℃ and kept this temperature 1-2 hour then, transfer to 26-30 ℃ and kept the about 2-4 of this temperature hour then, reduce to 5-10 ℃ and kept the about 1-2 of this temperature hour then, temperature is increased to 35-45 ℃ of another circulation of beginning more then.Recirculation is up to end of processing.After last temperature cycle finished, the temperature of culture was reduced to 3-4 ℃ and kept the about 5-6 of this temperature hour.After the adaptation, for example filter with traditional method yeast cell is separated from substratum, reclaims.The yeast cell that adapts to is stored in 4 ℃.Fig. 3 has described an exemplary device of this cultural method.

5.13 the separation of yeast cell and enrichment

Separation or enrichment have been adjusted the yeast cell that forms symbiosis sample relation according to 5.11 joints, make each yeast cell bacterial strain keep function that they obtained or strengthened.According to United States Patent (USP) 5,578, carry out separating of yeast cell with the open described method of CN1110317A of Chinese patent No. 486, this patent integral body is quoted here as a reference.In sepn process, can use the same frequency that is used to activate this yeast cell.Then with isolating yeast cell drying, storage.

5.14 the preparation of bio-feritlizer

Except the yeast cell component, also comprise organic matrix components and optional inorganic materials in the Biofertiliser composition of the present invention.The preparation of muck, mud, rubbish and analogous material is described below, and the step of preparation Biofertiliser composition.

5.14.1 the organic and preparation inorganic matrix component

Can use muck, mud or the rubbish of any kind of in the Biofertiliser composition of the present invention.Can also use the mixture of different sorts muck (fowl, livestock, pig).Organic compound in muck, mud or the rubbish is decomposed by yeast of the present invention.According to the kind of muck, mud, rubbish, except nitrogen, it can contain the phosphorus (P for example of significant quantity ₂O ₅) and potassium (K for example ₂O).Because the difference of kind, feed difference different with strain, storage method and water content, the nutrient contg in fowl, livestock or the pig manure can be different.Also can use the mixture of different sources mud.Because the method and the water content of source, presumable processing, storage, the nutrient contg in the mud can be different.The difference and the storage method of the geographical position in (that for example live, commercial), source, processing are different with water content because the source, and the nutrient contg in the rubbish can be different.The rubbish that contains high organic content (promptly more than 30%) is for preferred.Before every batch of organic substrate is used to prepare Biofertiliser composition, can detect its nutritive value with methods known in the art.

Inorganic materials such as but not limited to phosphate rock and potash mica, can be respectively optionally be included in the present composition as the additional source of phosphorus and potassium.Also can use other material and mineral substance phosphorous or potassium.These mineral compound are decomposed by K-and P-decomposition yeast cell be decomposed into can be grown plant and fertilizer in the biology of yeast cell utilization can utilize potassium and biology can utilize phosphorus.

Any inorganic materials can be united use with organic substrate of the present invention.In addition, in application-specific, if think needs, inorganic components can be removed or be substituted by other composition.For example, contain the brid guano fertilizer that high-level relatively biology can utilize phosphorus if use, phosphoric acid rock salt can be removed.

Muck, mud or rubbish preferably are dried to water content≤5%.Exsiccant muck, mud or rubbish and optional inorganic matrix component are milled to suitable form and size among the present invention before mixing fertilizer.Usually, organic materials and/or inorganic materials are sent to the fragment that crusher is broken into diameter≤5cm.Can use any traditional crusher for this purpose or be equal to machinery.These fragments are delivered to pulverizer by any transportation means and are ground to form 〉=150 purpose powder then.Can use any pulverizer that can fine grinding for this purpose.Then powder being transported to suitable storage vessel storage uses up to other component with fertilizer.Fig. 4 and Fig. 5 have shown the synoptic diagram of this Ginding process.

5.14.2 use somatomedin generation property zymic fermenting process

Among the present invention, the preparation of GF generation property zymic is carried out as seed during the fermentation with the described activation yeast strain of 5.6 joints.Fig. 6 has shown the synoptic diagram of this fermenting process.

Ratio according to every kilogram of starch of 2.5 premium on currency prepares fermention medium.Use cleaning water, preferably, prepare fermention medium without any the water of microorganism.Fermentation the temperature between 20-30 ℃, preferably between 25-28 ℃, clean environment and do not have the strong-electromagnetic field source such as the place of power line and generator is carried out.The device of any contact fermented liquid comprises reactor, pipeline and agitator, must thoroughly clean before each the use.When the fermented substrate when at least 90% was fermented, fermenting process continued about 48-72 hour at 28-30 ℃ usually.Fermentation is preferably carried out under half aerobic condition or oxygen level are the condition of the about 20-60% of maximum dissolved oxygen.Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.After the fermentation, cell counting should reach about 2 * 10 ¹⁰Cell/ml.Fermented liquid is remained in the 15-28 ℃ of temperature range, and must in 24 hours, use.In addition, GF-produce the property yeast can draining, drying and store with powder type.

5.14.3 use ATP generation property zymic fermenting process

Among the present invention, the preparation of ATP generation property zymic is carried out as seed during the fermentation with the described activation yeast strain of 5.7 joints.Fig. 6 has shown the synoptic diagram of this fermenting process.

Ratio according to every kilogram of starch of 2.5 premium on currency prepares fermention medium.Use cleaning water, preferably without any the water of microorganism, most preferably autoclaving water prepares fermention medium.Fermentation the temperature between 20-30 ℃, preferably between 25-28 ℃, clean environment and do not have the strong-electromagnetic field source such as the place of power line and generator is carried out.The device of any contact fermented liquid comprises reactor, pipeline and agitator, must thoroughly clean before each the use.According to leavening temperature, fermenting process continues about 48-72 hour usually.Preferably when this end of processing, at least 90% fermented substrate is fermented.Fermentation is preferably carried out under half aerobic condition or oxygen level are the condition of the about 20-60% of maximum dissolved oxygen.Oxygen level can be controlled by any ordinary method known to those skilled in the art, includes but not limited to stir and/or foaming.After the fermentation, cell counting should reach about 2 * 10 ¹⁰Cell/ml.Fermented liquid is remained in the 15-28 ℃ of temperature range, and must in 24 hours, use.In addition, ATP produce the property yeast can draining, drying and store with powder type.

5.14.4 the preparation of raw mix

With organic substrate (muck, mud or rubbish) and optional inorganic raw material according to the exemplary mixed shown in the table 12.Organic and the inorganic materials and the starch of the sufficient quantity that restraining is equipped with according to 5.10.1 are sent to mixing machine.Can use any traditional mixing machine, such as but not limited to rotary drum mixer.Steel basin is continued rotation, make the powder and the starch uniform mixing of muck, mud or rubbish.Then mixture is delivered to hold-up vessel.Fig. 7 shows the operation of mixing muck, mud or rubbish and inorganic matrix material.

The ratio of table 12 raw material

Material	Per-cent	Requirement
Material	Per-cent	Requirement	The powder of muck, mud or rubbish	??58-63％	〉=150 orders, water-content≤5%
The powder of inorganic materials	??20％	〉=150 orders, water-content≤3%	The powder of muck, mud or rubbish	??58-63％	〉=150 orders, water-content≤5%
The powder of inorganic materials	??20％	〉=150 orders, water-content≤3%	Starch	??10-15％	Conventional starch powder, water-content≤8%

5.14.5. the preparation of yeast mixt

If do not use inorganic materials, the ratio of muck, mud or rubbish can increase to 80%.Prepare yeast mixt in the exemplary ratio shown in the table 13.The nine primary yeast bacterial strains according to 5.1-5.10 restraining dry powdered form fully of sufficient quantity are sent to steel basin.Yeast was mixed about 10-20 minute.Then mixture is delivered to hold-up vessel.Any equipment that is used for mixed yeast comprises steel basin and hold-up vessel, must thoroughly be cleaned before each the use, and is preferably aseptic.Yeast mixt is stored under the temperature that is lower than 20 ℃, must use in 24 hours.Fig. 8 has shown the operation of mixed yeast.Perhaps, the mixture of nine primary yeasts can be dried and store with powder type.

The ratio of table 13 microorganism

Yeast	Amount	Per-cent (dry weight)	Remarks
Yeast	Amount	Per-cent (dry weight)	Remarks	The fixed nitrogen yeast	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Phosphorus-decomposition yeast	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder	The fixed nitrogen yeast	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Phosphorus-decomposition yeast	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder	Potassium-decomposition yeast	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Carbon-decomposition yeast	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder	Potassium-decomposition yeast	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Carbon-decomposition yeast	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder	Pathogenic agent inhibition yeast	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Chemical decomposability yeast	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder	Pathogenic agent inhibition yeast	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Chemical decomposability yeast	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder	Smell minimizing property yeast	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Somatomedin generation property yeast	??25L	??1％	Yeast fermentation broth	Smell minimizing property yeast	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Somatomedin generation property yeast	??25L	??1％	Yeast fermentation broth	ATP generation property yeast	??75L	??3％	Yeast fermentation broth

5.14.6. the preparation of bio-feritlizer

Bio-feritlizer of the present invention is by the organic and inorganic materials mixture of the yeast mixt of 5.14.5 joint and the 5.14.1 joint mixed according to table 14 is prepared.For example, yeast and organic substrate (brid guano fertilizer, pig manure, cattle manure, mud or rubbish) and inorganic materials are sent to tablets press and form particle.Then can fertiliser granulates is dry in one two stage drying process.In first drying stage, fertilizer temperature in first drying machine is no more than for some time that 65 ℃ of following dryings are no more than 10 minutes, make the rapid dormancy of yeast cell.Then fertilizer is delivered to second drying machine, be no more than for some time that 70 ℃ of following dryings are no more than 30 minutes, further remove moisture in temperature.After two stages, water-content should be less than 5%.Preferably, should observe temperature and time of drying in two drying stages, yeast cell just can not lose activity and function like this.Then fertilizer is cooled to room temperature.Can also screen fertilizer with separator, select the fertiliser granulates of preferred size.Can use any separator, such as but not limited to, the adjustable turbo separator of speed and screen sizes.Then the fertilizer of selected size being delivered to the cargo in bulk sack packer packs.

Fig. 9-11 has shown this production method.Fig. 9 is an operation synoptic diagram of producing fertilizer from its component.Figure 10 is the synoptic diagram of drying process.Figure 11 is the synoptic diagram of cooling and wrapping process.

The composition of table 14 bio-feritlizer (1 tonne of fertilizer)

	Amount	Per-cent (dry weight)	Remarks
	Amount	Per-cent (dry weight)	Remarks	Raw mix	??952-956kg	??95.2-95.4％	Dry weight
Yeast mixt	??100L	??4.4-4.8％	Dry weight	Raw mix	??952-956kg	??95.2-95.4％	Dry weight

6. embodiment

Following embodiment has shown the preparation of the exemplary Biofertiliser composition of the present invention.These embodiment have represented the preferred embodiments of the invention.

6.1 contain the Biofertiliser composition of brid guano fertilizer

Preserving number is the yeast cell component that the Wine brewing yeast strain of AS2.628, AS2.631, AS2.982, AS2.413 and AS2.536 can be used to prepare Biofertiliser composition.All these bacterial strains are deposited in Chinese common micro-organisms DSMZ (CGMCC), China Committee for Culture Collection of Microorganisms.With yeast strain AS2.628, for fixed nitrogen is cultivated according to the described method of 5.1 joints; For the P-balance is cultivated according to the described method of 5.3 joints.For K-decomposes, according to the described method culturing yeast strains A S2.631 of 5.4 joints.For C-decomposes, according to the described method culturing yeast strains A S2.982 of 5.5 joints.In order to produce somatomedin, according to the described method culturing yeast strains A S2.413 of 5.6 joints.In order to produce ATP, according to the described method culturing yeast strains A S2.536 of 5.7 joints.In order to suppress the growth of pathogenic agent, according to the described method culturing yeast bacterial strain IFFI1221 of 5.8 joints.For the adverse chemical product of degrading, according to the described method culturing yeast bacterial strain IFFI1293 of 5.9 joints.In order to reduce smell, according to the described method culturing yeast strains A S2.607 of 5.10 joints.

The brid guano fertilizer that is equipped with powder type according to the 5.14.1 restraining.

Somatomedin produces the preparation of property zymic and carries out in as the fermenting process of seed saving described activatory yeast strain AS2.413 with 5.6.Fig. 6 has shown the sketch of this fermenting process.Fermention medium is the ratio preparation that rises every kilogram of starch of clean water and 10 kilograms of starch PMT (Per metric ton) bio-feritlizers according to 2.5-3.5.Fermention medium is according to the ratio inoculation of every liter of substratum of 10ml seed solution.Fermentation is at 28 ± 1 ℃ temperature, 0.4mol/m ³Oxygen concentration, do not have to carry out in the clean environment of electromagnetism field source.After fermenting about 48 hours, barm cell concentration reaches about 2 * 10 ¹⁰Cell/ml.

It is to carry out in as the fermenting process of seed saving described activatory yeast strain AS2.536 with 5.7 that ATP produces the preparation of property zymic.Fig. 6 has shown the sketch of this fermenting process.Fermention medium is the ratio preparation that rises every kilogram of starch of clean water and 10 kilograms of starch PMT (Per metric ton) bio-feritlizers according to 2.5-3.5.Fermention medium is according to the ratio inoculation of every liter of substratum of 10ml seed solution.Fermentation is at 28 ± 1 ℃ temperature, 0.4mol/m ³Oxygen concentration, do not have to carry out about 56 hours in the clean environment of electromagnetism field source.After the fermentation, cell counting reaches about 2 * 10 ¹⁰Cell/ml.

Raw mix is the method preparation according to table 15 and 5.14.4 joint.

The ratio of table 15 raw material

Material	Per-cent	Requirement
Material	Per-cent	Requirement	The powder of dry brid guano fertilizer	??80.3％	〉=150 orders, water-content≤5%
Starch	??15％	Conventional starch powder, water-content≤8%	The powder of dry brid guano fertilizer	??80.3％	〉=150 orders, water-content≤5%

Yeast mixt is to save described method preparation according to table 16 and 5.14.5.

Table 16 zymic ratio (1 tonne of fertilizer)

Yeast	Amount	Per-cent (dry weight)	Remarks
Yeast	Amount	Per-cent (dry weight)	Remarks	Fixed nitrogen yeast AS2.628	??2.0kg	??0.2％	The dry yeast powder
Phosphorus-balance yeast AS2.628	??2.0kg	??0.2％	The dry yeast powder	Fixed nitrogen yeast AS2.628	??2.0kg	??0.2％	The dry yeast powder
Phosphorus-balance yeast AS2.628	??2.0kg	??0.2％	The dry yeast powder	Potassium-decomposition yeast AS2.631	??2.0kg	??0.2％	The dry yeast powder
Carbon-decomposition yeast AS2.982	??2.0kg	??0.2％	The dry yeast powder	Potassium-decomposition yeast AS2.631	??2.0kg	??0.2％	The dry yeast powder
Carbon-decomposition yeast AS2.982	??2.0kg	??0.2％	The dry yeast powder	Pathogenic agent inhibition yeast IFFI1221	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Chemical-degradation property yeast IFFI1293	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder	Pathogenic agent inhibition yeast IFFI1221	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Chemical-degradation property yeast IFFI1293	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder	Smell minimizing property yeast AS2.607	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Produce the yeast AS2.413 of somatomedin	??25L	??1％	Yeast fermentation broth	Smell minimizing property yeast AS2.607	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Produce the yeast AS2.413 of somatomedin	??25L	??1％	Yeast fermentation broth	Produce the yeast AS2.536 of ATP	??75L	??3％	Yeast fermentation broth

Bio-feritlizer is by yeast mixt, organic materials are prepared by the mixed of table 17.Blended yeast and organic materials are sent to tablets press and form particle.Then that fertiliser granulates is dry in one two stage drying process.In first drying stage, fertilizer temperature in first drying machine is no more than 60 ± 2 ℃ of following dryings 5 minutes, make the rapid dormancy of yeast cell.Then fertilizer is delivered to second drying machine, drying is 8 minutes under temperature is no more than 65 ± 2 ℃, further removes moisture.Then fertilizer is cooled to room temperature.Then fertilizer being delivered to the cargo in bulk sack packer packs.

Table 17 Ru 2006101161 (1 tonne of fertilizer)

	Amount	Per-cent (dry weight)	Remarks
	Amount	Per-cent (dry weight)	Remarks	Raw mix	????949kg	????94.9％	Dry weight
Yeast mixt	????100L	????5.1％	Dry weight	Raw mix	????949kg	????94.9％	Dry weight

6.2 contain the Biofertiliser composition of cattle manure

Preserving number is the yeast cell component that the Wine brewing yeast strain of AS2.628, AS2.399, AS2.631, AS2.982, AS2.413 and AS2.536 can be used to prepare Biofertiliser composition.All these bacterial strains are deposited in Chinese common micro-organisms DSMZ (CGMCC), China Committee for Culture Collection of Microorganisms.For fixed nitrogen, according to the described method culturing yeast strains A S2.628 of 5.1 joints; For P-decomposes, according to the described method culturing yeast strains A S2.399 of 5.2 joints.For K-decomposes, according to the described method culturing yeast strains A S2.631 of 5.4 joints.For C-decomposes, according to the described method culturing yeast strains A S2.982 of 5.5 joints.In order to produce somatomedin, according to the described method culturing yeast strains A S2.413 of 5.6 joints.In order to produce ATP, according to the described method culturing yeast strains A S2.536 of 5.7 joints.In order to suppress the growth of pathogenic agent, according to the described method culturing yeast bacterial strain IFFI1308 of 5.8 joints.For the adverse chemical product of degrading, according to the described method culturing yeast bacterial strain IFFI1210 of 5.9 joints.In order to reduce smell, according to the described method culturing yeast bacterial strain IFFI1213 of 5.10 joints.

The cattle manure humidity of using in the present embodiment is equipped with according to the 5.14.1 restraining less than 5%.

It is to carry out in as the fermenting process of seed saving described activatory yeast strain AS2.413 with 5.6 that somatomedin produces the preparation of property zymic.Fig. 6 has shown the sketch of fermenting process.Fermention medium is the ratio preparation according to every kilogram of starch of 2.5 liters of clean waters and 10 kilograms of starch PMT (Per metric ton) bio-feritlizers.Fermention medium is according to the ratio inoculation of every liter of substratum of 10ml seed solution.Fermentation is at 28 ± 1 ℃ temperature, 0.4mol/m ³Oxygen concentration, do not have to carry out in the clean environment of electromagnetism field source.After fermenting about 48 hours, the concentration of yeast cell reaches about 2 * 10 ¹⁰Cell/ml.

It is to carry out in as the fermenting process of seed saving described activatory yeast strain AS2.536 with 5.7 that ATP produces the preparation of property zymic.Fig. 6 has shown the sketch of fermenting process.Fermention medium is the ratio preparation that rises every kilogram of starch of clean water and 10 kilograms of starch PMT (Per metric ton) bio-feritlizers according to 2.5-3.5.Fermention medium is according to the ratio inoculation of every liter of substratum of 10ml seed solution.Fermentation is at 28 ± 1 ℃ temperature, 0.4mol/m ³Oxygen concentration, do not have to carry out about 56 hours in the clean environment of electromagnetism field source.After the fermentation, cell counting reaches about 2 * 10 ¹⁰Cell/ml.

Raw mix is the method preparation according to table 18 and 5.14.4 joint.

The ratio of table 18 raw material

Material	Per-cent	Requirement
Material	Per-cent	Requirement	Dry cattle manure powder	??80.3％	〉=150 orders, water-content≤5%
Starch	??15％	Conventional starch powder, water-content≤8%	Dry cattle manure powder	??80.3％	〉=150 orders, water-content≤5%

Yeast mixt is to save described method preparation according to table 19 and 5.14.5.

Table 19 zymic ratio (1 tonne of fertilizer)

Yeast	Amount	Per-cent (dry weight)	Remarks
Yeast	Amount	Per-cent (dry weight)	Remarks	Fixed nitrogen yeast AS2.628	??2.0kg	??0.2％	The dry yeast powder
Phosphorus-decomposition yeast AS2.399	??2.0kg	??0.2％	The dry yeast powder	Fixed nitrogen yeast AS2.628	??2.0kg	??0.2％	The dry yeast powder
Phosphorus-decomposition yeast AS2.399	??2.0kg	??0.2％	The dry yeast powder	Potassium-decomposition yeast AS2.631	??2.0kg	??0.2％	The dry yeast powder
Carbon-decomposition yeast AS2.982	??2.0kg	??0.2％	The dry yeast powder	Potassium-decomposition yeast AS2.631	??2.0kg	??0.2％	The dry yeast powder
Carbon-decomposition yeast AS2.982	??2.0kg	??0.2％	The dry yeast powder	Pathogenic agent inhibition yeast IFFI1308	?1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Chemical degradation property yeast IFFI1210	?1.0-2.0kg	??0.1-0.2％	The dry yeast powder	Pathogenic agent inhibition yeast IFFI1308	?1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Chemical degradation property yeast IFFI1210	?1.0-2.0kg	??0.1-0.2％	The dry yeast powder	Smell minimizing property yeast IFFI1213	?1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Produce the yeast AS2.413 of somatomedin	?25L	??1％	Yeast fermentation broth	Smell minimizing property yeast IFFI1213	?1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Produce the yeast AS2.413 of somatomedin	?25L	??1％	Yeast fermentation broth	Produce the yeast AS2.536 of ATP	?75L	??3％	Yeast fermentation broth

Bio-feritlizer is by yeast mixt, organic materials are prepared by the mixed of table 20.Blended yeast and organic materials are sent to tablets press and form particle.Then that fertiliser granulates is dry in one two stage drying process.In first drying stage, fertilizer temperature in first drying machine is no more than 60 ± 2 ℃ of following dryings 5 minutes, make the rapid dormancy of yeast cell.Then fertilizer is delivered to second drying machine, drying is 8 minutes under temperature is no more than 65 ± 2 ℃, further removes moisture.Then fertilizer is cooled to room temperature, delivers to the cargo in bulk sack packer then and pack.

Table 20 Ru 2006101161 (1 tonne of fertilizer)

6.3 contain the Biofertiliser composition of pig manure

Preserving number is the yeast cell component that the Wine brewing yeast strain of AS2.628, AS2.399, AS2.631, AS2.982, AS2.413 and AS2.536 can be used to prepare Biofertiliser composition.All these bacterial strains are deposited in Chinese common micro-organisms DSMZ (CGMCC), China Committee for Culture Collection of Microorganisms.For fixed nitrogen, according to the described method culturing yeast strains A S2.628 of 5.1 joints; For P-decomposes, according to the described method culturing yeast strains A S2.399 of 5.2 joints.For K-decomposes, according to the described method culturing yeast strains A S2.631 of 5.4 joints.For C-decomposes, according to the described method culturing yeast strains A S2.982 of 5.5 joints.In order to produce somatomedin, according to the described method culturing yeast strains A S2.413 of 5.6 joints.In order to produce ATP, according to the described method culturing yeast strains A S2.536 of 5.7 joints.In order to suppress the growth of pathogenic agent, according to the described method culturing yeast bacterial strain IFFI1307 of 5.8 joints.For the adverse chemical product of degrading, according to the described method culturing yeast bacterial strain IFFI1206 of 5.9 joints.In order to reduce smell, according to the described method culturing yeast bacterial strain IFFI1052 of 5.10 joints.

The pig manure that use is equipped with according to the 5.14.1 restraining.

It is to carry out in as the fermenting process of seed saving described activatory yeast strain AS2.413 with 5.6 that somatomedin produces the preparation of property zymic.Fig. 6 has shown the sketch of fermenting process.Fermention medium is the ratio preparation that rises every kilogram of starch of clean water and 10 kilograms of starch PMT (Per metric ton) bio-feritlizers according to 2.5-3.5.Fermention medium is according to the ratio inoculation of every liter of substratum of 10ml seed solution.Fermentation is at 28 ± 1 ℃ temperature, 0.4mol/m ³Oxygen concentration, do not have to carry out in the clean environment of electromagnetism field source.After fermenting about 48 hours, barm cell concentration reaches about 2 * 10 ¹⁰Cell/ml.

It is to carry out in as the fermenting process of seed saving described activatory yeast strain AS2.1190 with 5.7 that ATP produces the preparation of property zymic.Fig. 6 has shown the sketch of fermenting process.Fermention medium is the ratio preparation that rises every kilogram of starch of clean water and 10 kilograms of starch PMT (Per metric ton) bio-feritlizers according to 2.5-3.5.Fermention medium is according to the ratio inoculation of every liter of substratum of 10ml seed solution.Fermentation is at 28 ± 1 ℃ temperature, 0.4mol/m ³Oxygen concentration, do not have to carry out about 56 hours in the clean environment of electromagnetism field source.After the fermentation, cell counting reaches about 2 * 10 ¹⁰Cell/ml.

Raw mix is the method preparation according to table 21 and 5.14.5 joint.

The ratio of table 21 raw material

Material	Per-cent	Requirement
Material	Per-cent	Requirement	The dry pig manure of powder type	????80％	〉=150 orders, water-content≤5%
Starch	????15％	Conventional starch powder, water-content≤8%	The dry pig manure of powder type	????80％	〉=150 orders, water-content≤5%

Yeast mixt is to save described method preparation according to table 22 and 5.14.5.

Table 22 zymic ratio (1 tonne of fertilizer)

Yeast	Amount	Per-cent (dry weight)	Remarks
Yeast	Amount	Per-cent (dry weight)	Remarks	Fixed nitrogen yeast AS2.628	?2.0kg	?0.2％	The dry yeast powder
Phosphorus-decomposition yeast AS2.399	?2.0kg	?0.2％	The dry yeast powder	Fixed nitrogen yeast AS2.628	?2.0kg	?0.2％	The dry yeast powder
Phosphorus-decomposition yeast AS2.399	?2.0kg	?0.2％	The dry yeast powder	Potassium-decomposition yeast AS2.631	?2.0kg	?0.2％	The dry yeast powder
Carbon-decomposition yeast AS2.982	?2.0kg	?0.2％	The dry yeast powder	Potassium-decomposition yeast AS2.631	?2.0kg	?0.2％	The dry yeast powder
Carbon-decomposition yeast AS2.982	?2.0kg	?0.2％	The dry yeast powder	Pathogenic agent inhibition yeast IFFI1307	?1.0-2.0kg	?0.1-0.2％	The dry yeast powder
Chemical degradation property yeast IFFI1206	?1.0-2.0kg	?0.1-0.2％	The dry yeast powder	Pathogenic agent inhibition yeast IFFI1307	?1.0-2.0kg	?0.1-0.2％	The dry yeast powder
Chemical degradation property yeast IFFI1206	?1.0-2.0kg	?0.1-0.2％	The dry yeast powder	Smell minimizing property yeast IFFI1052	?1.0-2.0kg	?0.1-0.2％	The dry yeast powder
Produce the yeast AS2.413 of somatomedin	?25L	?1％	Yeast fermentation broth	Smell minimizing property yeast IFFI1052	?1.0-2.0kg	?0.1-0.2％	The dry yeast powder
Produce the yeast AS2.413 of somatomedin	?25L	?1％	Yeast fermentation broth	Produce the yeast AS2.536 of ATP	?75L	?3％	Yeast fermentation broth

Bio-feritlizer is by yeast mixt, organic materials are prepared by the mixed of table 23.Blended yeast and organic materials are sent to tablets press and form particle.Then that fertiliser granulates is dry in one two stage drying process.In first drying stage, fertilizer temperature in first drying machine is no more than 60 ± 2 ℃ of following dryings 5 minutes, make the rapid dormancy of yeast cell.Then fertilizer is delivered to second drying machine, drying is 8 minutes under temperature is no more than 65 ± 2 ℃, further removes moisture.Then fertilizer is cooled to room temperature.Then fertilizer being delivered to the cargo in bulk sack packer packs.

Table 23 Ru 2006101161 (1 tonne of fertilizer)

	Amount	Per-cent (dry weight)	Remarks
	Amount	Per-cent (dry weight)	Remarks	Raw mix	????949kg	????94.9％	Dry weight
Yeast mixt	????100L	????5.1％	Dry weight (51kg/ ton)	Raw mix	????949kg	????94.9％	Dry weight

6.4 contain the Biofertiliser composition of mud

Preserving number is the yeast cell component that the Wine brewing yeast strain of AS2.628, AS2.399, AS2.631, AS2.982, AS2.413 and AS2.536 can be used to prepare Biofertiliser composition.All these bacterial strains are deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC).For fixed nitrogen, according to the described method culturing yeast strains A S2.628 of 5.1 joints; For P-decomposes, according to the described method culturing yeast strains A S2.399 of 5.2 joints.For K-decomposes, according to the described method culturing yeast strains A S2.631 of 5.4 joints.For C-decomposes, according to the described method culturing yeast strains A S2.982 of 5.5 joints.In order to produce somatomedin, according to the described method culturing yeast strains A S2.413 of 5.6 joints.In order to produce ATP, according to the described method culturing yeast strains A S2.536 of 5.7 joints.In order to suppress the growth of pathogenic agent, according to the described method culturing yeast bacterial strain IFFI1301 of 5.8 joints.For the adverse chemical product of degrading, according to the described method culturing yeast bacterial strain IFFI1291 of 5.9 joints.In order to reduce smell, according to the described method culturing yeast bacterial strain IFFI1202 of 5.10 joints.

Powdered exsiccant mud saves described preparation according to 5.14.5.

Raw mix is the method preparation according to table 24 and 5.14.5 joint.

The ratio of table 24 raw material

Material	Per-cent	Requirement
Material	Per-cent	Requirement	Powdered exsiccant mud	??80.3％	〉=150 orders, water-content≤5%
Starch	??15％	Conventional starch powder, water-content≤8%	Powdered exsiccant mud	??80.3％	〉=150 orders, water-content≤5%

Yeast mixt is to save described method preparation according to table 25 and 5.14.5.

Table 25 zymic ratio (1 tonne of fertilizer)

Yeast	Amount	Per-cent (dry weight)	Remarks
Yeast	Amount	Per-cent (dry weight)	Remarks	Fixed nitrogen yeast AS2.628	????2.0kg	????0.2％	The dry yeast powder
Phosphorus-decomposition yeast AS2.399	????2.0kg	????0.2％	The dry yeast powder	Fixed nitrogen yeast AS2.628	????2.0kg	????0.2％	The dry yeast powder
Phosphorus-decomposition yeast AS2.399	????2.0kg	????0.2％	The dry yeast powder	Potassium-decomposition yeast AS2.631	????2.0kg	????0.2％	The dry yeast powder
Carbon-decomposition yeast AS2.982	????2.0kg	????0.2％	The dry yeast powder	Potassium-decomposition yeast AS2.631	????2.0kg	????0.2％	The dry yeast powder
Carbon-decomposition yeast AS2.982	????2.0kg	????0.2％	The dry yeast powder	Pathogenic agent inhibition yeast IFFI1301	????1.0-2.0kg	????0.1-0.2％	The dry yeast powder
Chemical degradation property yeast IFFI1291	????1.0-2.0kg	????0.1-0.2％	The dry yeast powder	Pathogenic agent inhibition yeast IFFI1301	????1.0-2.0kg	????0.1-0.2％	The dry yeast powder
Chemical degradation property yeast IFFI1291	????1.0-2.0kg	????0.1-0.2％	The dry yeast powder	Smell minimizing property yeast IFFI1202	????1.0-2.0kg	????0.1-0.2％	The dry yeast powder
Produce the yeast AS2.413 of somatomedin	????25L	????1％	Yeast fermentation broth	Smell minimizing property yeast IFFI1202	????1.0-2.0kg	????0.1-0.2％	The dry yeast powder
Produce the yeast AS2.413 of somatomedin	????25L	????1％	Yeast fermentation broth	Produce the yeast AS2.536 of ATP	????75L	????3％	Yeast fermentation broth

Bio-feritlizer is by yeast mixt, mud and the optional inorganic materials mixed by table 26 is prepared.Blended yeast and mud are sent to tablets press and form particle.Then that fertiliser granulates is dry in one two stage drying process.In first drying stage, fertilizer temperature in first drying machine is no more than 60 ± 2 ℃ of following dryings 5 minutes, make the rapid dormancy of yeast cell.Then fertilizer is delivered to second drying machine, drying is 8 minutes under temperature is no more than 65 ± 2 ℃, further removes moisture.Then fertilizer is cooled to room temperature.Then fertilizer being delivered to the cargo in bulk sack packer packs.

Table 26 Ru 2006101161 (1 tonne of fertilizer)

	Amount	Per-cent (dry weight)	Remarks
	Amount	Per-cent (dry weight)	Remarks	Raw mix	949kg	?94.9％	Dry weight
Yeast mixt	100L	?4.8％	Dry weight	Raw mix	949kg	?94.9％	Dry weight

6.5 contain the Biofertiliser composition of rubbish

Preserving number is the yeast cell component that the Wine brewing yeast strain of AS2.628, AS2.399, AS2.631, AS2.982, AS2.413 and AS2.536 can be used to prepare Biofertiliser composition.All these bacterial strains are deposited in Chinese common micro-organisms DSMZ (CGMCC), China Committee for Culture Collection of Microorganisms.For fixed nitrogen, according to the described method culturing yeast strains A S2.628 of 5.1 joints; For P-decomposes, according to the described method culturing yeast strains A S2.399 of 5.2 joints.For K-decomposes, according to the described method culturing yeast strains A S2.631 of 5.4 joints.For C-decomposes, according to the described method culturing yeast strains A S2.982 of 5.5 joints.In order to produce somatomedin, according to the described method culturing yeast strains A S2.413 of 5.6 joints.In order to produce ATP, according to the described method culturing yeast strains A S2.536 of 5.7 joints.In order to suppress the growth of pathogenic agent, according to the described method culturing yeast bacterial strain IFFI120 of 5.8 joints.For the adverse chemical product of degrading, according to the described method culturing yeast bacterial strain IFFI1059 of 5.9 joints.In order to reduce smell, according to the described method culturing yeast bacterial strain IFFI1247 of 5.10 joints.

The rubbish that is equipped with according to the 5.14.1 restraining uses as organic substrate.The rubbish that uses in the present embodiment contains at least 30% organic materials.

It is to carry out in as the fermenting process of seed saving described activatory yeast strain AS2.413 with 5.6 that somatomedin produces the preparation of property zymic.Fig. 6 has shown the sketch of fermenting process.Fermention medium is the ratio preparation according to every kilogram of starch of 2.5 liters of clean waters and 10 kilograms of starch PMT (Per metric ton) bio-feritlizers.Fermention medium is according to the ratio inoculation of every liter of substratum of 10ml seed solution.Fermentation is at 28 ± 1 ℃ temperature, 0.4mol/m ³Oxygen concentration, do not have to carry out in the clean environment of electromagnetism field source.After fermenting about 48 hours, barm cell concentration reaches about 2 * 10 ¹⁰Cell/ml.

It is to carry out in as the fermenting process of seed saving described activatory yeast strain AS2.536 with 5.7 that ATP produces the preparation of property zymic.Fig. 6 has shown the sketch of fermenting process.Fermention medium is the ratio preparation according to every kilogram of starch of 2.5 liters of clean waters and 10 kilograms of starch PMT (Per metric ton) bio-feritlizers.Fermention medium is according to the ratio inoculation of every liter of substratum of 10ml seed solution.Fermentation is at 28 ± 1 ℃ temperature, 0.4mol/m ³Oxygen concentration, do not have to carry out about 56 hours in the clean environment of electromagnetism field source.After the fermentation, cell counting reaches about 2 * 10 ¹⁰Cell/ml.

Raw mix is the method preparation according to table 27 and 5.14.5 joint.

The ratio of table 27 raw material

Material	Per-cent	Requirement
Material	Per-cent	Requirement	Powdered exsiccant rubbish	??80.3％	〉=150 orders, water-content≤5%
Starch	??15％	Conventional starch powder, water-content≤8%	Powdered exsiccant rubbish	??80.3％	〉=150 orders, water-content≤5%

Yeast mixt is to save described method preparation according to table 28 and 5.14.5.

Table 28 zymic ratio (1 tonne of fertilizer)

Yeast	Amount	Per-cent (dry weight)	Remarks
Yeast	Amount	Per-cent (dry weight)	Remarks	Fixed nitrogen yeast AS2.628	??2.0kg	??0.2％	The dry yeast powder
Phosphorus-decomposition yeast AS2.399	??2.0kg	??0.2％	The dry yeast powder	Fixed nitrogen yeast AS2.628	??2.0kg	??0.2％	The dry yeast powder
Phosphorus-decomposition yeast AS2.399	??2.0kg	??0.2％	The dry yeast powder	Potassium-decomposition yeast AS2.631	??2.0kg	??0.2％	The dry yeast powder
Carbon-decomposition yeast AS2.982	??2.0kg	??0.2％	The dry yeast powder	Potassium-decomposition yeast AS2.631	??2.0kg	??0.2％	The dry yeast powder
Carbon-decomposition yeast AS2.982	??2.0kg	??0.2％	The dry yeast powder	Pathogenic agent inhibition yeast IFFI1201	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Chemical degradation property yeast IFFI1059	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder	Pathogenic agent inhibition yeast IFFI1201	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Chemical degradation property yeast IFFI1059	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder	Smell minimizing property yeast IFFI1247	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Produce the yeast AS2.413 of somatomedin	??25L	??1％	Yeast fermentation broth	Smell minimizing property yeast IFFI1247	??1.0-2.0kg	??0.1-0.2％	The dry yeast powder
Produce the yeast AS2.413 of somatomedin	??25L	??1％	Yeast fermentation broth	Produce the yeast AS2.536 of ATP	??75L	??3％	Yeast fermentation broth

Bio-feritlizer is by yeast mixt, organic materials are prepared by the mixed of table 29.Blended yeast and organic materials are sent to tablets press and form particle.Then that fertiliser granulates is dry in one two stage drying process.In first drying stage, fertilizer temperature in first drying machine is no more than 60 ± 2 ℃ of following dryings 5 minutes, make the rapid dormancy of yeast cell.Then fertilizer is delivered to second drying machine, drying is 8 minutes under temperature is no more than 65 ± 2 ℃, further removes moisture.Then fertilizer is cooled to room temperature.Then fertilizer being delivered to the cargo in bulk sack packer packs.

Table 29 Ru 2006101161 (1 tonne of fertilizer)

	Amount	Per-cent (dry weight)	Remarks
	Amount	Per-cent (dry weight)	Remarks	Raw mix	??949kg	94.9％	Dry weight
Yeast mixt	??100L	5.1％	Dry weight (51kg/ ton)	Raw mix	??949kg	94.9％	Dry weight

The invention is not restricted to the category of described specific embodiments, they are indivedual examples of all respects of the present invention, and method that function is suitable and component are also in category of the present invention.Undoubtedly, except show here and describe, will be conspicuous for a person skilled in the art according to the description and the accompanying drawing multiple improvement of the present invention of front.These improve also within additional claim scope.

Claims

1. Biofertiliser composition, it contains:

(I) organic substrate;

(II) at least a in following:

(a) the first yeast cell component contains the yeast cell of first group of fixed nitrogen;

(b) the second yeast cell component contains the yeast cell that second component is separated phosphorus compound; Or

(c) the 3rd yeast cell component contains the yeast cell that the 3rd component is separated potassium compound; With

(III) at least a in following:

(d) the 4th yeast cell component contains the 4th group of yeast cell that suppresses the pathogenic microorganism growth;

(e) the 5th yeast cell component contains the 5th group of antibiotic yeast cell of degrading; Or

(f) the 6th yeast cell component contains the 6th group of yeast cell that reduces the Biofertiliser composition smell.

2. the Biofertiliser composition of claim 1, it further comprises at least a in following:

(g) the 7th yeast cell component contains the yeast cell that the 7th group of carbon compound with complexity is converted into simple carbohydrates;

(h) the 8th yeast cell component contains the yeast cell of the 8th group of excessive generation somatomedin; Or

(i) the 9th yeast cell component contains the yeast cell of the 9th group of excessive generation Triphosaden.

3. Biofertiliser composition, it contains:

(I) organic substrate;

(II) at least a in following:

(a) the first yeast cell component prepares by cultivating first group of yeast cell in frequency 840 to 916MHz, in first electromagnetic field of field intensity in 10 to 200mV/cm scopes;

(b) the second yeast cell component prepares by cultivating second group of yeast cell in frequency 300 to 500MHz, in second electromagnetic field of field intensity in 10 to 300mV/cm scopes;

(c) the 3rd yeast cell component prepares by cultivating the 3rd group of yeast cell in frequency 190 to 285MHz, in the 3rd electromagnetic field of field intensity in 10 to 200mV/cm scopes; With

(III) at least a in following:

(d) the 4th yeast cell component prepares by cultivating the 4th group of yeast cell in frequency 30 to 50MHz, in the 4th electromagnetic field of field intensity in 20 to 200mV/cm scopes;

(e) the 5th yeast cell component prepares by cultivating the 5th group of yeast cell in frequency 70 to 100MHz, in the 5th electromagnetic field of field intensity in 40 to 250mV/cm scopes; With

(f) the 6th yeast cell component prepares by cultivating the 6th group of yeast cell in frequency 2160 to 2250MHz, in the 6th electromagnetic field of field intensity in 2280 to 2380mV/cm scopes.

4. the Biofertiliser composition of claim 3 further comprises at least a in following:

(g) the 7th yeast cell component prepares by cultivating the 7th group of yeast cell in frequency 1050 to 1160MHz, in the 7th electromagnetic field of field intensity in 10 to 200mV/cm scopes;

(h) the 8th yeast cell component prepares by cultivating the 8th group of yeast cell in frequency 1340 to 1440MHz, in the 8th electromagnetic field of field intensity in 20 to 200mV/cm scopes;

(i) the 9th yeast cell component prepares by cultivating the 9th group of yeast cell in frequency 1630 to 1730MHz, in the 9th electromagnetic field of field intensity in 20 to 200mV/cm scopes.

5. claim 2 or 4 Biofertiliser composition, wherein each yeast cell component comprises the yeast cell from yeast belong.

6. claim 2 or 4 Biofertiliser composition, wherein each yeast cell component comprises the cell that is selected from following yeast kind respectively: yeast saccharomyces cerevisiae, Xue's watt yeast, Saccharomycesdelbrueckii, saccharomyces exiguus, saccharomyces fermentati, Saccharomyces logos, saccharomyces mellis, Saccharomyces microellipsoides, ellipsoideus yeast, Luo Shi yeast, Lu Shi yeast, saccharomyces sake, saccharomyces uvarum, Wei Shi yeast, saccharomyces ludwigii, Saccharomyces sinenses and saccharomyces carlsbergensis.

7. claim 2 or 4 Biofertiliser composition, wherein each yeast cell component comprises the cell that is selected from following yeast strain respectively: yeast saccharomyces cerevisiae (Saccharomyces cerevisiaeHansen) ACCC2034, ACCC2035, ACCC2036, ACCC2037, ACCC2038, ACCC2039, ACCC2040, ACCC2041, ACCC2042, AS2.1, AS2.4, AS2.11, AS2.14, AS2.16, AS2.56, AS2.69, AS2.70, AS2.93, AS2.98, AS2.101, AS2.109, AS2.110, AS2.112, AS2.139, AS2.173, AS2.174, AS2.182, AS2.196, AS2.242, AS2.336, AS2.346, AS2.369, AS2.374, AS2.375, AS2.379, AS2.380, AS2.382, AS2.390, AS2.393, AS2.395, AS2.396, AS2.397, AS2.398, AS2.399, AS2.400, AS2.406, AS2.408, AS2.409, AS2.413, AS2.414, AS2.415, AS2.416, AS2.422, AS2.423, AS2.430, AS2.431, AS2.432, AS2.451, AS2.452, AS2.453, AS2.458, AS2.460, AS2.463, AS2.467, AS2.486, AS2.501, AS2.502, AS2.503, AS2.504, AS2.516, AS2.535, AS2.536, AS2.558, AS2.560, AS2.561, AS2.562, AS2.576, AS2.593, AS2.594, AS2.614, AS2.620, AS2.628, AS2.631, AS2.666, AS2.982, AS2.1190, AS2.1364, AS2.1396, IFFI1001, IFFI1002, IFFI1005, IFFI1006, IFFI1008, IFFI1009, IFFI1010, IFFI1012, IFFI1021, IFFI1027, IFFI1037, IFFI1042, IFFI1043, IFFI1045, IFFI1048, IFFI1049, IFFI1050, IFFI1052, IFFI1059, IFFI1060, IFFI1063, IFFI1202, IFFI1203, IFFI1206, IFFI1209, IFFI1210, IFFI1211, IFFI1212, IFFI1213, IFFI1215, IFFI1220, IFFI1221, IFFI1224, IFFI1247, IFFI1248, IFFI1251, IFFI1270, IFFI1277, IFFI1287, IFFI1289, IFFI1290, IFFI1291, IFFI1291, IFFI1292, IFFI1293, IFFI1297, IFFI1300, IFFI1301, IFFI1302, IFFI1307, IFFI1308, IFFI1309, IFFI1310, IFFI1311, IFFI1331, IFFI1335, IFFI1336, IFFI1337, IFFI1338, IFFI1339, IFFI1340, IFFI1345, IFFI1348, IFFI1396, IFFI1397, IFFI1399, IFFI1411, IFFI1413; Wine brewing saccharomyces ellipsoideus (Sacharomyces cerevisiaeHansen Var.ellipsoideus (Hansen) Dekker) ACCC2043, AS2.2, AS2.3, AS2.8, AS2.53, AS2.163, AS2.168, AS2.483, AS2.541, AS2.559, AS2.606, AS2.607, AS2.611, AS2.612; Xue's watt yeast (Saccharomyces chevalieriGuillermond) AS2.131, AS2.213; Saccharomyces delbrueckii AS2.285; Saccharomyces delbrueckii Lindner var.mongolicus Lodder et van RijAS2.209, AS2.1157; Saccharomyces exiguus (Saccharomyces exiguus Hansen) AS2.349, AS2.1158; Saccharomyces fermentati (Saccharomyces fermentati (Saito) Lodder et vanRij) AS2.286, AS2.343; Saccharomyces logos van laer et Denamur exJorgensen AS2.156, AS2.327, AS2.335; Saccharomyces mellis (Saccharomyces mellisLodder et Kreger Van Rij) AS2.195; Saccharomyces microellipsoidesOsterwalder AS2.699; Ellipsoideus yeast (Saccharomyces oviformis Osterwalder) AS2.100; Luo Shi yeast (Saccharomyces rosei (Guilliermond) Lodder et kregervan Rij) AS2.287; Lu Shi yeast (Saccharomyces rouxii Boutroux) AS2.178, AS2.180, AS2.370, AS2.371; Saccharomyces sake (Saccharomyces sake Yabe) ACCC2045; Saccharomyces carlsbergensis (Saccharomyces carlsbergensis Hansen) ACCC2032, ACCC2033, AS2.113, AS2.116, AS2.118, AS2.121, AS2.132, AS2.162, AS2.189, AS2.200, AS2.216, AS2.265, AS2.377, AS2.417, AS2.420, AS2.440, AS2.441, AS2.443, AS2.444, AS2.459, AS2.595, AS2.605, AS2.638, AS2.742, AS2.745, AS2.748, AS2.1042; Saccharomyces uvarum (Saccharomyces uvatum Beijer) IFFI1023, IFFI1032, IFFI1036, IFFI1044, IFFI1072, IFFI1205, IFFI1207; Wei Shi yeast (Saccharonzyces willianusSaccardo) AS2.5, AS2.7, AS2.119, AS2.152, AS2.293, AS2.381, AS2.392, AS2.434, AS2.614, AS2.1189; Yeast AS2.311; Lu Shi Saccharomyces ludwigiiHansen) ACCC2044, AS2.243, AS2.508 and Saccharomyces sinenses YueAS2.1395.

8. claim 2 or 4 Biofertiliser composition, wherein each yeast cell component comprises yeast saccharomyces cerevisiae respectively.

9. claim 2 or 4 Biofertiliser composition, it further comprises the inorganic matrix component.

10. claim 2 or 4 Biofertiliser composition, wherein the inorganic matrix component comprises one or more phosphate rocks, phosphatic rock, phosphorite, sylvite, rock salt, carnallitite or potash mica.

11. the Biofertiliser composition of claim 2, it comprises the yeast cell component of claim 1 from (a) to (f), and the yeast cell component of claim 2 from (g) to (i).

12. the Biofertiliser composition of claim 4, it comprises the yeast cell component of claim 3 from (a) to (f), and the yeast cell component of claim 4 from (g) to (i).

13. the Biofertiliser composition of claim 2 or 4, wherein yeast cell component (a) comprises the cell of yeast saccharomyces cerevisiae AS2.628 yeast strain; Yeast cell component (b) comprises the cell of yeast saccharomyces cerevisiae AS2.628 yeast strain; Yeast cell component (c) comprises the cell of yeast saccharomyces cerevisiae AS2.631 yeast strain; Yeast cell component (d) comprises the cell of one or more following yeast strains: yeast saccharomyces cerevisiae IFFI1037, IFFI1021, IFFI1051, IFFI1331, IFFI1345 or IFFI1211; Yeast cell component (e) comprises the cell of one or more following yeast strains: yeast saccharomyces cerevisiae IFFI1063, IFFI1211, IFFI1340, IFFI1215, IFFI1213, IFFI1206, IFFI1211, IFFI1210 or IFFI1260; Yeast cell component (f) comprises the cell of one or more following yeast strains: yeast saccharomyces cerevisiae AS2.559, AS2.423, AS2.612, AS2.53, AS2.541 or AS2.163; Yeast cell component (g) comprises the cell of yeast saccharomyces cerevisiae AS2.982 yeast strain; Yeast cell component (h) comprises the cell of yeast saccharomyces cerevisiae AS2.413 yeast strain; And yeast cell component (i) comprises the cell of yeast saccharomyces cerevisiae AS2.536 yeast strain.

14. the Biofertiliser composition of claim 11 or 12, wherein yeast cell component (a) comprises the cell of yeast saccharomyces cerevisiae AS2.628 yeast strain; Yeast cell component (b) comprises the cell of yeast saccharomyces cerevisiae AS2.628 yeast strain; Yeast cell component (c) comprises the cell of yeast saccharomyces cerevisiae AS2.631 yeast strain; Yeast cell component (d) comprises the cell of one or more following yeast strains: yeast saccharomyces cerevisiae IFFI1037, IFFI1021, IFFI1051, IFFI1331, IFFI1345 or IFFI1211; Yeast cell component (e) comprises the cell of one or more following yeast strains: yeast saccharomyces cerevisiae IFFI1063, IFFI1211, IFFI1340, IFFI1215, IFFI1213, IFFI1206, IFFI1211, IFFI1210 or IFFI1260; Yeast cell component (f) comprises the cell of one or more following yeast strains: yeast saccharomyces cerevisiae AS2.559, AS2.423, AS2.612, AS2.53, AS2.541 or AS2.163; Yeast cell component (g) comprises the cell of yeast saccharomyces cerevisiae AS2.982 yeast strain; Yeast cell component (h) comprises the cell of yeast saccharomyces cerevisiae AS2.413 yeast strain; And yeast cell component (i) comprises the cell of yeast saccharomyces cerevisiae AS2.536 yeast strain.

15. the Biofertiliser composition of claim 2 or 4, wherein said yeast cell is an exsiccant.

16. the preparation method of the Biofertiliser composition of claim 1 or 3 comprises by described order:

(I) by or (c) and at least a yeast cell component (d), (e) or (f) be mixed with the yeast cell mixture with at least a yeast cell component (a) and (b); With

(II) in described yeast cell mixture, add organic substrate and make Biofertiliser composition.

17. the preparation method of the Biofertiliser composition of claim 2 or 4 comprises by described order:

(I) by with at least a yeast cell component (a) and (b) or (c), at least a yeast cell component (d), (e) or (f) with at least a yeast cell component (g), (h) or (i) are mixed with the yeast cell mixture; With

18. the method for claim 16, wherein:

The first yeast cell component (a) be by with first group of yeast cell frequency 840 to 916MHz and field intensity in 10 to 200mV/cm scopes first electromagnetic field or first group of electromagnetic field in cultivate and prepare;

The second yeast cell component (b) be by with second group of yeast cell frequency 300 to 500MHz and field intensity in 10 to 300mV/cm scopes second electromagnetic field or second group of electromagnetic field in cultivate and prepare;

The 3rd yeast cell component (c) be by with the 3rd group of yeast cell frequency 190 to 285MHz and field intensity in 10 to 200mV/cm scopes the 3rd electromagnetic field or the 3rd group of electromagnetic field in cultivate and prepare;

The 4th yeast cell component (d) be by with the 4th group of yeast cell frequency 30 to 50MHz and field intensity in 20 to 200mV/cm scopes the 4th electromagnetic field or the 4th group of electromagnetic field in cultivate and prepare;

The 5th yeast cell component (e) be by with the 5th group of yeast cell frequency 70 to 100MHz and field intensity in 40 to 250mV/cm scopes the 5th electromagnetic field or the 5th group of electromagnetic field in cultivate and prepare;

The 6th yeast cell component (f) be by with the 6th group of yeast cell frequency 2160 to 2250MHz and 2280 to 2380MHz and field intensity in 100 to 300mV/cm scopes the 6th electromagnetic field or the 6th group of electromagnetic field in cultivate and prepare.

19. the method for claim 17, wherein:

The 7th yeast cell component (g) be by with the 7th group of yeast cell frequency 1050 to 1160MHz and field intensity in 10 to 200mV/cm scopes the 7th electromagnetic field or the 7th group of electromagnetic field in cultivate and prepare;

The 8th yeast cell component (h) be by with the 8th group of yeast cell frequency 1340 to 1440MHz and field intensity in 20 to 200mV/cm scopes the 8th electromagnetic field or the 8th group of electromagnetic field in cultivate and prepare;

The 9th yeast cell component (i) be by with the 9th group of yeast cell frequency 1630 to 1730MHz and field intensity in 20 to 200mV/cm scopes the 9th electromagnetic field or the 9th group of electromagnetic field in cultivate and prepare.

20. the method for claim 16 wherein in step (II), adds described yeast cell mixture in organic substrate, starch and the inorganic matrix component.

21. the method for claim 17 wherein in step (II), adds described yeast cell mixture in organic substrate, starch and the inorganic matrix component.

22. the method for claim 16, it further comprises by described order:

(III) with described Biofertiliser composition not being higher than dry for some time under 65 ℃ the temperature, make the yeast cell dormancy;

(IV) with described Biofertiliser composition not being higher than dry for some time under 70 ℃ the temperature, make water-content less than 5%;

(V) described Biofertiliser composition is cooled to envrionment temperature; With

(VI) make described Biofertiliser composition form particle.

23. the method for claim 17, it further comprises by described order:

(IH) with described Biofertiliser composition not being higher than dry for some time under 65 ℃ the temperature, make the yeast cell dormancy;

(VI) make described Biofertiliser composition form particle.

24. a method that promotes plant-growth comprises plant is grown in the presence of the Biofertiliser composition of claim 2 or 4.

25. the method for claim 23 wherein imposes on Biofertiliser composition in the soil of the plant main root system degree of depth.

26. the method for claim 23 is wherein used about 600 to 1350kg/ha Biofertiliser composition.

27. the method for claim 23, wherein plant is cereal crop, vegetable crop, fruit crop, flowers crop or fodder crop.

28. the method for claim 23, wherein plant is wheat, barley, corn, soybean, rice, oat, potato, apple, orange, tomato, muskmelon, cherry, lemon, lettuce, Radix Dauci Sativae, sugarcane, tobacco or cotton.

29. the Biofertiliser composition of claim 1 or 2, wherein the yeast cell of yeast cell component (b) can keep the balance of phosphorus compound.

30. the Biofertiliser composition of claim 3 or 4, wherein yeast cell component (b) is to prepare by second group of yeast cell cultivated in 90 second electromagnetic field in the 300mV/cm scope to 465MHz, field intensity 380 in frequency.

31. the method for claim 18, the electromagnetic field frequency of wherein cultivating the second yeast component (b) arrives in the 465MHz scope 380.

32. the method for claim 19, the electromagnetic field frequency of wherein cultivating the second yeast component (b) arrives in the 465MHz scope 380.

33. claim 1,2,3 or 4 Biofertiliser composition, wherein organic substrate is brid guano fertilizer, cattle manure, pig manure, mud or rubbish.

34. the Biofertiliser composition of claim 8, wherein organic substrate is brid guano fertilizer, cattle manure, pig manure, mud or rubbish.

35. the Biofertiliser composition of claim 9, wherein organic substrate is brid guano fertilizer, cattle manure, pig manure, mud or rubbish.

36. the Biofertiliser composition of claim 11 or 12, wherein organic substrate is brid guano fertilizer, cattle manure, pig manure, mud or rubbish.

37. the method for claim 16, wherein organic substrate is brid guano fertilizer, cattle manure, pig manure, mud or rubbish.

38. the method for claim 17, wherein organic substrate is brid guano fertilizer, cattle manure, pig manure, mud or rubbish.

39. the method for claim 18, wherein organic substrate is brid guano fertilizer, cattle manure, pig manure, mud or rubbish.

40. the method for claim 19, wherein organic substrate is brid guano fertilizer, cattle manure, pig manure, mud or rubbish.