CN1436787A - Antibiotic and its prepn and application - Google Patents
Antibiotic and its prepn and application Download PDFInfo
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- CN1436787A CN1436787A CN 02100874 CN02100874A CN1436787A CN 1436787 A CN1436787 A CN 1436787A CN 02100874 CN02100874 CN 02100874 CN 02100874 A CN02100874 A CN 02100874A CN 1436787 A CN1436787 A CN 1436787A
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Abstract
The present invention relates to one kind of antibiotic and features that through culture of Streptomiyces ahygroscopicus and separating and purifying extractive from its fermented liquid, one new kind of antibiotic is obtained, which has cytidine with one-C4H9O9 side chain and molecular expression C13H21N3O14 and is dissimilar to any of available nucleoside antibiotic. The antibiotic has no toxicity and no environmental pollution and is one kind of pollution-less biologic pesticide for preventing and treating crop's fungus disease with the effect of 60-90%.
Description
Technical field
The invention belongs to biological technical field, more specifically saying so has the not streptomyces hygroscopicus that preserving number is CGMCC NO.0703 by cultivation, obtains a kind of new microbiotic and application thereof.
Background technology
Microbiotic is a low-molecular-weight meta-bolites, is widely used in aspects such as treatment human diseases, livestock industry, agricultural.Can be divided into beta-lactam, tetracyclines, aminoglycoside (aminocyclitol class), Macrolide, ansamycins, peptide class, glycopeptide class (dipropyl peptide in heptan class), ucleosides and several big classes of antitumor antibiotics by antibiotic structure.Nucleoside antibiotics is the important microbiotic of a class, they are big class compounds to nucleosides and nucleotide structure modification, with nucleosides is its common parent nucleus, have antibacterium, antimycotic, anti-trypanosome, antitumor, hoe up weeds, the biological activity (Isono of wider range such as desinsection, strengthening immunity, J Antibot 41:1711-1739,1988):
The difference of different nucleoside antibioticss is the side-chain radical beyond the parent nucleus.The nucleoside antibiotics of using on the agricultural mainly contains blasticidin S (Blasticidin S) and Polyoxin (Polyoxins), and their structural formula is seen accompanying drawing 1 (b) and (c) respectively; Blasticidin S (Takeuchi et al, J Antibiot11; 1-5,1958) be used to prevent and treat rice blast, once be widely used the sixties, substitutes the mercurial of high poison; Polyoxin is used to prevent and treat various fungal diseases (Mitani ﹠amp; Inoue, J Antibiot 21: 492-6,1968).The research of China microbiotic is to follow the tracks of and imitated, and ability of developing voluntarily, formulating and level are very low, have independent intellectual property right brand-new compound report seldom, the agricultural antibiotic of brand new is very little especially.Therefore, have the research of independent intellectual property right compound, not only directly for human disease treatment and agricultural application in new variety are provided, and provide lead compound for the chemist develops new medicine, further strengthen the medicine innovation ability.
Summary of the invention
The purpose of this invention is to provide a kind of new microbiotic and application method thereof that is applied to prevent and treat corps diseases and is different from known microbiotic structure.
Main technical schemes of the present invention: by cultivating not streptomyces hygroscopicus (Streptomycesahygroscopicus).From its fermented liquid, separate again, purified extract, obtain having side chain and be-C
4H
9O
9Nitrogen replace the microbiotic of cytidine.
Microbiotic of the present invention has side chain
4H
9O
9Nitrogen replace cytidine, molecular formula is C
13H
21N
3O
14Molecular weight is 443 compound, and its structural formula is:
Record concrete side chain and be by infrared spectra, carbon spectrum, hydrogen spectrum
Molecular structural formula:
Antibiotic preparation method of the present invention comprises following process:
(1) the microbiotic biological fermentation is cultivated: with organic carbon source, organic nitrogen source, quick-acting carbon sources, inorganic nitrogen-sourced, inorganic salt and water and not streptomyces hygroscopicus join in the container, cultivate not streptomyces hygroscopicus, under 20-34 ℃, carry out one grade fermemtation, two-stage fermentation or three grade fermemtation, 45-100 hour one grade fermemtation time, the two-stage fermentation time was respectively 16-45 hour, 30-80 hour, the three grade fermemtation time was respectively 16-45 hour, 16-45 hour, 30-80 hour, obtain the culture that antibiotic content is the 1000-3500 mcg/ml: cultural method employing rotating speed is that 90-300 rev/min shaking table concussion is cultivated or the employing liquid submerged fermentation is cultivated: tank pressure is kept 0.02-0.10MPa, the air quantity ratio is 1: 0.15-1: 1, mixing speed is 50-300 rev/min: inoculum size is 2-15% (V/V), described organic carbon source is 1-5%, organic nitrogen source is 0.2-5%, quick-acting carbon source 0.5-4%, inorganic nitrogen-sourced 0.2-1.5%, inorganic salt 0.01-1.0%, above percentage ratio is 100 in the weight of water.
(2) separation of antibiotics: with the antibiotic culture that contains of step (1) collection; being acidified to the pH value with oxalic acid is 2.0; leave standstill 12-24 hour after-filtration after the stirring; filtrate is decoloured through the seed activity carbon granule; the activated carbon column effluent liquid is again with the absorption of 732# resin cation (R.C.); elder generation's water takes off earlier; use 0.5N ammoniacal liquor wash-out again, flow velocity 0.5-3ml/ minute, after effluent liquid pH value is greater than 7.0, begin to receive; concentrate and receive liquid extremely; through the decolouring of neutral alumina post, first water wash-out is used the ammoniacal liquor wash-out to the 1/10-1/20 of original volume more again; collecting active high effluent liquid is concentrated into dried; dry powder is gone up the LH-20 sephadex column with 1-5 ml water dissolving back, uses the distilled water wash-out, flow velocity 0.5-1.5ml/ minute; the active high effluent liquid of collection is concentrated into dried, promptly obtains the raw product that antibiotic content is 30-45%.
(3) antibiotic purifying: the raw product that step (2) is obtained is through sephadex G-25 column chromatography, water elution, flow velocity is 0.5-1.5ml/ minute, after high pressure liquid chromatography (HPLC) is checked, the major ingredient amount is merged with top 75%, lyophilize then, be prepared through the HPLC semipreparative column again, obtain antibiotic content and reach the 70-86% highly finished product.
The described not streptomyces hygroscopicus of step (1) comprises not streptomyces hygroscopicus Mount Huang mutation.The mutation of streptomyces hygroscopicus Mount Huang is not to separate the streptomycete that can produce biologically active substance that a strain that obtains is numbered 2-16 from the soil of Mount Huang, Anhui Province, through isolation identification, this streptomycete named be not streptomyces hygroscopicus Mount Huang mutation (Streptomyces ahygroscopicus var.huangsanensis) (Shi Yiping etc., the microorganism journal, 39:84-86,1999).This bacterial strain has been handed over the preservation of DSMZ of Microbe Inst., Chinese Academy of Sciences, and preserving number is CGMCC NO.0703.The synthetic nutrient agar of employing Gao Shi (Chinese common micro-organisms culture presevation administrative center (CGMCC) compiles: bacterial classification catalogue, the third edition, Chinese agriculture science and technology press, 1997) is cultivated in going down to posterity of this bacterial strain
The described organic carbon source of step (1) comprises Semen Maydis powder, dextrin, sweet potato starch, and solubility is sunk powder, wheat-flour; Described organic nitrogen source comprises analysis for soybean powder, groundnut meal, cottonseed meal, coarse colza meal, soybean cake powder, dried silkworm chrysalis meal, yeast powder, yeast extract paste, fish meal, peptone, described quick-acting carbon source comprises maltose, glucose, sucrose, described inorganic nitrogen-sourced ammonium nitrate, sulfuric acid amine, ammonium chloride, SODIUMNITRATE, the saltpetre of comprising, described inorganic salt comprise sal epsom, dipotassium hydrogen phosphate, potassium primary phosphate, lime carbonate, sodium-chlor.
Antibiotic biological activity assay of the present invention: (Chinese common micro-organisms culture presevation management committee compiles at agriculture microorganism center: Chinese agriculture bacterial classification catalogue with bacillus megaterium ACCC10008 (Bacillus megatherium), Chinese agriculture science and technology press, 1991) for detecting bacterium, potato dextrose agar (PDA substratum: 20% murphy juice, 2% glucose, 0.8% agar) for detecting substratum, adopt the biological activity of cup-plate method detect antibiotics.
Antibiotic application method of the present invention: contain antibiotic fermenting culture, raw product or highly finished product and all can be used for preventing and treating the farm crop fungus disease, can directly use or add auxiliary agent or mix use, be diluted with water to working concentration when directly using usually and be the 25-200 mcg/ml with other agricultural chemicals.For containing the fermenting culture that microbiotic is the not streptomyces hygroscopicus of 1000-3500 mcg/ml, when using in the field, the 1/8-1/3 of filtrate through being evaporated to stoste that fermenting culture is obtained after filtration doubly, thickening temperature 40-70 ℃, make concentrated solution, perhaps make pulvis by spray drying process, concentrated solution or pulvis can directly use, also can add auxiliary agent or mix use with suitable chemical pesticide or biological pesticide, for different farm crop, the different sites site of pathological change adopts different methods and different concns: to leaf, stem's disease, often the concentrated solution thin up is used, with dilution 100-400 medicine liquid spray doubly, vegetable disease generally sprays 2-3 time, 7-10 days interval; Seed-borne disease is often carried out seed disinfection, generally with the soup seed soaking after 50-100 times of water dilution 1-24 hour; To seedbed, nutrition pot, can adopt the soup after the dilution of 100-250 times of water to carry out soil disinfection, to soil-borne disease with the medicine liquid irrigation root; Can directly smear affected part fruit tree stem disease with concentrated solution.It is better that microbiotic of the present invention is used to prevent and treat the fungal disease effect, various crop fungal disease prevention effect had certain difference, generally between 60-99%.
Microbiotic of the present invention be must guard against with strong acid, highly basic and is used with.Spraying the antibiotic time was advisable with fine, cloudy day, not before and after heavy rain or dew is not dried and the sunlight intensive sprays medicine noon.Store the place and should be chosen in the not place of direct irradiation of ventilation, drying, sunlight, under cold condition, store, can prolong storage period greatly.
Advantage of the present invention and effect:
Microbiotic of the present invention is that a kind of structure is different from a kind of new microbiotic of known any nucleoside antibiotics, and it is better to be used to prevent and treat farm crop fungus disease effect, and general prevention effect is 60-99%.For example prevent and treat powdery mildew of cucumber, prevention effect is more than 91%, prevents and treats leaf muld of tomato and can reach 86.93%, and compare with chemical pesticide, and product is nontoxic, and is free from environmental pollution, is a kind of non-harmful biological pesticide.In sum, the invention provides that having of a kind of development research voluntarily is efficient, the new microbiotic that is used to prevent and treat corps diseases of wide spectrum, low toxicity characteristic,, promote that agricultural sustainable development has significance for preserving the ecological environment.
Embodiment
Further specify characteristics of the present invention below by specific examples.
Brief Description Of Drawings in the example:
Fig. 1 is the chemical structural formula of microbiotic of the present invention (a) and blasticidin S (b), Polyoxin (c);
Fig. 2 is the antibiotic mass spectrum of the present invention;
Fig. 3 is the antibiotic infrared absorption spectrum of the present invention;
Fig. 4 be the antibiotic carbon spectrogram of the present invention (
13C-NMR);
Fig. 5 is the antibiotic carbon spectrogram of the present invention (DEPT);
Fig. 6 is the antibiotic hydrogen spectrogram of the present invention (DMSO);
Fig. 7 is the antibiotic hydrogen spectrogram of the present invention (D
2The O exchange).
Example 1
This example adopts shaking table concussion method to carry out the antibiotic biofermentation and cultivates.
Containing analysis for soybean powder 2%, glucose 3%, Zulkovsky starch 1%, yeast extract paste 0.2%, MgSO
47H
2O0.05%, CaCO
30.7% inoculation of medium preserving number is the not streptomyces hygroscopicus of CGMCC NO.0703, and the concussion of 24 ℃ of shaking tables is cultivated, and rotating speed is 220 rev/mins, cultivates 45 hours earlier, is inoculated into by 10% inoculum size and cultivates in the identical substratum 80 hours again.
Example 2
This example adopts liquid submerged fermentation method to carry out the microbiotic biological fermentation and cultivates.
Containing soybean cake powder 5%, glucose 1%, Zulkovsky starch 2%, peptone 0.2%, NaCl0.2%, (NH
4)
2SO
47H
2O 0.25%, MgSO
47H
2O 0.05%, K
2HPO
40.02%, CaCO
30.8% inoculation of medium preserving number is the not streptomyces hygroscopicus of CGMCC NO.0703, and 34 ℃ of liquid submerged fermentations are cultivated, and mixing speed is 220 rev/mins, ventilates 1: 0.9, and tank pressure 0.10Mpa cultivated 45 hours.
Example 3
This example is to antibiotic biological activity assay: 3ml PDA solid medium is placed 15 * 150mm test tube, put into the long inclined-plane of 3.5cm after the sterilization; Inoculation bacillus megaterium ACCC10008 on the inclined-plane, cultivated 1 day for 28 ℃, with the sterilized water of 50ml band granulated glass sphere will be tiltedly and on thalline wash, and thalline broken up mixing, make bacteria suspension, dosage by 1-5% adds to bacteria suspension in the PDA substratum, in the culture dish of 15 centimetres of diameters, pour this PDA bacterium liquid 60-100 milliliter into, simultaneously, nutrient solution with filter paper filtering example 1 or example 2, collect filtrate and be used for measuring, wait to contain the PDA of ACCC10008 dull and stereotyped coagulate with after, with the bacteriostatic activity of cup-plate method working sample.
Example 4
This example is separation of antibiotics and purifying.
Fermenting culture from example 1 and example 2 collections is acidified to pH2.5 with 0.5-1% oxalic acid respectively, stirs the back standing over night, filters; Filtrate is through the decolouring of granulated active carbon post, and the activated carbon effluent liquid adsorbs with the 732# cationic resin column, first water wash-out, use 0.5N ammoniacal liquor wash-out again, flow velocity 2ml/ minute, after effluent liquid pH value is greater than 7.0, begin to receive, concentrate and receive the 1/10-1/20 (30-50ml) of liquid to original volume; After the decolouring of neutral alumina post, first water wash-out is used 3% ammoniacal liquor wash-out again, collects active high effluent liquid and contracts deeply to doing; Dry powder dissolves with less water (1-5 milliliter), and last LH-20 sephadex column is used the distilled water wash-out, and flow velocity 1ml/ minute, the active high effluent liquid of collection was concentrated to dried, promptly gets crude product; Crude product coagulates post G-25 column chromatography through dextran, water elution elution speed 1ml/ minute, merges 75% main component content after HPLC detects with top, be prepared through the HPLC semipreparative column again after the lyophilize, can obtain containing the highly finished product of microbiotic 86% after the preparation.Through the routine analysis, antibiotic physico-chemical property of the present invention: cold dry is a micro-yellow powder, the easy moisture absorption, and 239 ℃ of fusing points (carbonization), very easily water-soluble, be slightly soluble in methyl alcohol, be insoluble to organic solvents such as acetone, chloroform, pyridine.
Example 5
This example is that antibiotic structure of the present invention is identified.
According to high resolution mass spectrum, determine that antibiotic molecular weight is 443 (see figure 2)s, ultimate analysis shows, contains C in the antibiotic molecule of the present invention, H, (C31.94%, H4.73% N11.7%), according to the nitrogen rule, calculate that its fraction is C for N and O atom
13H
21N
3O
14Ultra-violet absorption spectrum demonstrates charateristic avsorption band (unimodal) λ max (H of typical nucleoside antibiotics
2O) 269.4nm, λ max (0.1NNaOH) 265.8nm, λ max (0.1N HCL) 275.0nm.Through various spectrographic analysis-by-synthesis, after microbiotic more of the present invention and the chemical shift of cytidine carbon atom, determine that the antibiotic chemical structure of the present invention is to have a nitrogen that contains the peroxide bridge side chain to replace cytidine, microbiotic of the present invention and cytidine carbon atom chemical shift contrast are as follows:
Microbiotic cytidine of the present invention
| Carbon atom NO. | Chemical shift (ppm) | |
| Microbiotic of the present invention | Cytidine | |
| ??2 | ????174.5 | ????166.6 |
| ??4 | ????169.1 | ????156.9 |
| ??5 | ????99.0 | ????95.6 |
| ??6 | ????146.7 | ????142.8 |
| ??1′ | ????85.5 | ????90.1 |
| ??2′ | ????74.1 | ????70.6 |
| ??3′ | ????75.7 | ????75.1 |
| ??4′ | ????78.5 | ????85.2 |
| ??5′ | ????64.0 | ????61.8 |
Infrared spectra 3350cm
-1Locate among the non-level and smooth broad peak explanation microbiotic A of the present invention except that existences-NH-, there be (Fig. 3) in hydroxyl in addition.The antibiotic molecular formula of the present invention is C
13H
21N
3O
14, degree of unsaturation is 5, so-on the NH-connect-C
4H
9O
9Be saturated side chains.Can draw the carbon atom information on this side chain from carbon spectrum (Fig. 4,5), its chemical shift (ppm) is respectively: 58.3 (CH-), 55.9 (CH-), 52.1 (CH2-), 35.5 (CH3-), compose (Fig. 6: DMSO, Fig. 7 D according to hydrogen
2O exchange) can draw the information of reactive hydrogen, low 9ppm place by on the peroxide bridge the chemical shift of company's hydrogen (C-O-O-H), and 839cm in the infrared spectra
-1, 1134cm
-1Two absorption peaks are also pointed out the existence of peroxide bridge.Comprehensive above information, the structure that draws side chain is:
Comprehensive above conclusion, the antibiotic structural formula of the present invention such as accompanying drawing 1 (a).Fig. 1 (b) is oryzacidin S (Blasticidin S) structure.Fig. 1 (c) is Polyoxin (Polyoxins) structure iron.Wherein R1, the R2 in the Polyoxin structure iron, R3 numerical value see the following form:
Polyoxin????R
1??????R
2????????????????R
3
A???????????CH
2OH?
???????OH
B???????????CH
2OH???OH??????????????????OH
D???????????COOH?????OH??????????????????OH
E???????????COOH?????OH??????????????????H
F???????????COOH????
???????OH
G???????????CH
2OH???OH??????????????????H
H???????????CH
3????
???????OH
J???????????CH
3?????OH??????????????????OH
K???????????H??????
????????OH
L???????????H????????OH??????????????????OH
M???????????H????????OH??????????????????H
Example 6
This example is the antibiotic toxicity test of the present invention.
With content is that the antibiotic concentrated solution of the present invention of 10000 mcg/ml is used to carry out toxicity test, according to Tianjin chemical toxicological detection test-results, microbiotic of the present invention is to rat acute per os toxicity maximum tolerated dose, LD
50＞5000mg/kg belongs to low toxicity; To rat acute percutaneous toxicity maximum tolerated dose LD
50＞2000mg/kg belongs to low toxicity; To the rabbit eye irritant test, its stimulus intensity nonirritant level, to the test of rabbit acute skin, its skin irritation intensity belongs to does not have irritating property level.Safe and reliable.
Example 7
This example is a field of the present invention farm crop Application Example.
The fermenting culture that example 1 is collected obtains filtrate after filtration, be evaporated to 1 ' 5 times of stoste, thickening temperature is 65 ℃, and concentrated solution microbiotic of the present invention is content 10000 mcg/ml, is used to prevent and treat powdery mildew of cucumber, leaf muld of tomato, sclerotinia rot of colza and careless mould botrytis cinerea.
(1) Powdery Mildew of control cucumber: powdery mildew of cucumber (Spharotheca fuliginea) is one of main disease of cucumber, generally causes underproduction 10-20%, and serious reaches more than 50%.Concentrated solution is sprayed with 150 times water dilution, 200 jin of water of every mu of spray, and prevention effect is 91.46%, 500 times of the chemical pesticide triadimefon dilutions of using simultaneously, prevention effect 96.36%.
(2) prevent and treat leaf muld of tomato: leaf muld of tomato (Cladosporium fulvum) is one of main disease of tomato.With 200 times, 250 times water dilution back spraying, every mu sprays 200 jin of water to concentrated solution respectively, and prevention effect is respectively 86.93%, 83.07%, and the contrast agricultural chemicals of use is that 75% zinc manganese ethylenebisdithiocarbamate dilutes prevention effect 63.41% with 300 times of water.
(3) control sclerotinia rot of colza: sclerotinia rot of colza (Sclerotinia sclerotiorum) is the main disease of rape, generally causes underproduction 11-73%, and oleaginousness reduces 1-5%.With 100 times of concentrated solution dilute with waters, 150 times, 200 times, at rape flowering spraying control sclerotinia rot of colza, spray medicine 1 time, 150 jin of water of every mu of spray, prevention effect is respectively 82.6%, 78.1%, 69.8%; The mould clever suspension agent 100ml/667m of chemical pesticide 36% powder of Shi Yonging simultaneously
2, 40% control the clever wettable powder 60g/667m that withers
2, prevention effect is respectively 94.4% and 66.1%.
(4) control grey mould fruit rot of strawberry: grey mould fruit rot of strawberry is one of main disease of strawberry.With 100 times of controls of concentrated solution dilution grey mould fruit rot of strawberry, at strawberry their early stage spray medicine, 100 jin of water of every mu of spray, prevention effect is 65.37%.
Claims (7)
1. a microbiotic is characterized in that having-C
4H
9O
9The nitrogen of side chain replaces cytidine, molecular formula is C
13H
21N
3O
14Microbiotic, its molecular structural formula is:
2. according to the described microbiotic of claim 1, it is characterized in that molecular structure is
3. the described antibiotic preparation method of claim 1 may further comprise the steps:
(1) the microbiotic biological fermentation is cultivated: with organic carbon source, organic nitrogen source, quick-acting carbon source, inorganic nitrogen-sourced, inorganic salt and water and not streptomyces hygroscopicus join in the container, cultivate not streptomyces hygroscopicus, under 20-34 ℃, carry out one grade fermemtation, two-stage fermentation or three grade fermemtation, 45-100 hour one grade fermemtation time, the two-stage fermentation time is respectively 16-45 hour, 30-80 hour, the three grade fermemtation time was respectively 16-45 hour, 16-45 hour, 30-80 hour, obtained the culture that antibiotic content is the 1000-3500 mcg/ml; Cultural method employing rotating speed is that 90-300 rev/min shaking table concussion is cultivated or the employing liquid submerged fermentation is cultivated: tank pressure is kept 0.02-0.10MPa, the air quantity ratio is 1: 0.15-1: 1, and mixing speed is 50-300 rev/min; Inoculum size is 2-15% (V/V), and described organic carbon source is that 1-5%, organic nitrogen source are 0.2-5%, quick-acting carbon source 0.5-4%, inorganic nitrogen-sourced 0.2-1.5%, inorganic salt 0.01-1.0%, and above percentage ratio is 100 in the weight of water.
(2) separation of antibiotics: with the antibiotic culture that contains of step (i) collection; being acidified to the pH value with oxalic acid is 2.0; leave standstill 12-24 hour after-filtration after the stirring; filtrate is decoloured through the seed activity carbon granule; the activated carbon column effluent liquid is again with the absorption of 732# resin cation (R.C.); elder generation's water takes off earlier; use 0.5N ammoniacal liquor wash-out again, flow velocity 0.5-3ml/ minute, after effluent liquid pH value is greater than 7.0, begin to receive; concentrate and receive the 1/10-1/20 of liquid to original volume; through the decolouring of neutral alumina post, first water wash-out is used the ammoniacal liquor wash-out more again; collecting active high effluent liquid is concentrated into dried; dry powder is gone up the LH-20 sephadex column with 1-5 ml water dissolving back, uses the distilled water wash-out, flow velocity 0.5-1.5ml/ minute; the active high effluent liquid of collection is concentrated into dried, promptly obtains the raw product that antibiotic content is 30-45%.
(3) antibiotic purifying: the raw product that step (2) is obtained is through sephadex G-25 column chromatography, water elution, flow velocity is 0.5-1.5ml/ minute, after high pressure liquid chromatography (HPLC) is checked, the major ingredient amount is merged with top 75%, lyophilize then, be prepared through the HPLC semipreparative column again, obtain antibiotic content and reach the 70-86% highly finished product.
4. in accordance with the method for claim 3, it is characterized in that the described not streptomyces hygroscopicus of step (1) comprises not streptomyces hygroscopicus Mount Huang mutation (Streptomyces ahygroscopicus var.huangsanensis).
5. in accordance with the method for claim 3, it is characterized in that the described organic carbon source of step (1) comprises Semen Maydis powder, dextrin, sweet potato starch, Zulkovsky starch, wheat-flour; Described organic nitrogen source comprises analysis for soybean powder, groundnut meal, cottonseed meal, coarse colza meal, soybean cake powder, dried silkworm chrysalis meal, yeast powder, yeast extract paste, fish meal, peptone, described quick-acting carbon source comprises maltose, glucose, sucrose, described inorganic nitrogen-sourced ammonium nitrate, ammonium sulfate, ammonium chloride, SODIUMNITRATE, the saltpetre of comprising, described inorganic salt comprise sal epsom, dipotassium hydrogen phosphate, potassium primary phosphate, lime carbonate, sodium-chlor.
6. the described microbiotic application method of claim 1, it is characterized in that containing fermenting culture, raw product or the highly finished product of antibiotic not streptomyces hygroscopicus, be used to prevent and treat the farm crop fungus disease and can directly use or add auxiliary agent or mix use, be diluted with water to working concentration when using and be the 25-200 mcg/ml with other agricultural chemicals.
7. according to the described application method of claim 6, it is characterized in that containing the fermenting culture that microbiotic is the mould that do not absorb water of 1000-3500 mcg/ml, when use in the field, fermenting culture will be after filtration and is obtained concentrated solution or spraying drying is made dry powder at 40-70 ℃ of following concentrating under reduced pressure, to leaf, stem disease evil, with the 100-400 water dilution concentrated solution doubly of concentrated solution weight, the soup that obtains is sprayed, vegetable disease spray 2-3 time, 7-10 days interval; To seed-borne disease, with the soup seed soaking after 50-100 times of water dilution 1-24 hour; To seedbed, nutrition pot, carry out soil disinfection with the soup after the 100-250 times of water dilution; To the soil-borne disease poison, with the doubly water-reducible medicine liquid irrigation root of 100-250; Fruit tree stem disease is directly smeared affected part with concentrated solution.
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| CN1296370C (en) * | 2005-03-07 | 2007-01-24 | 吴元华 | Anti plant virus agricultural cytosin peptidechycin, and its preparing process |
| CN101891508B (en) * | 2009-05-22 | 2012-06-06 | 辽宁微科生物工程有限公司 | Living body biological organic fertilizer and preparation method thereof |
| CN101831396A (en) * | 2010-04-29 | 2010-09-15 | 赤峰制药股份有限公司 | Process for preparing oxytetracycline single colony frozen bacteria |
| CN102946727B (en) * | 2010-05-06 | 2015-08-19 | 诺华有限公司 | The organic peroxide compounds of bacteria inactivation rate |
| CN102946727A (en) * | 2010-05-06 | 2013-02-27 | 诺华有限公司 | Organic peroxide compounds for microorganism inactivation |
| JP2013528586A (en) * | 2010-05-06 | 2013-07-11 | ノバルティス アーゲー | Organic peroxide compounds for inactivation of microorganisms |
| US9426989B2 (en) | 2010-05-06 | 2016-08-30 | Novartis Ag | Organic peroxide compounds for microorganism inactivation |
| CN103416437A (en) * | 2012-05-23 | 2013-12-04 | 徐州诺特化工有限公司 | Preparation method for modified cytidine antibiotic oil suspension |
| CN103421865A (en) * | 2012-05-23 | 2013-12-04 | 徐州诺特化工有限公司 | Preparation method for modified cytidine antibiotic |
| CN102688280A (en) * | 2012-06-13 | 2012-09-26 | 吉林农业大学 | Preparation method for radiation-resistant Chinese medicine fermentation liquor |
| CN103232525A (en) * | 2013-04-17 | 2013-08-07 | 南京工业大学 | Method for extracting polyoxin from streptomyces fermentation broth |
| CN103232525B (en) * | 2013-04-17 | 2015-04-22 | 南京工业大学 | Method for extracting polyoxin from streptomyces fermentation broth |
| CN104604946A (en) * | 2015-01-12 | 2015-05-13 | 邱德文 | Biological protein preparation for preventing and controlling citrus huanglongbing and preparation method for biological protein preparation |
| CN104604946B (en) * | 2015-01-12 | 2017-10-13 | 邱德文 | It is a kind of for biologic protein agents of prevention and control Citrus Huanglongbing pathogen and preparation method thereof |
| WO2018059263A1 (en) * | 2016-09-28 | 2018-04-05 | The University Of Hong Kong | Bismuth (iii) compounds and methods thereof |
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