CN105660704A - Bacillus subtilis and oligosaccharins compound composition, compound preparation and applications of compound composition and compound preparation - Google Patents
Bacillus subtilis and oligosaccharins compound composition, compound preparation and applications of compound composition and compound preparation Download PDFInfo
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- CN105660704A CN105660704A CN201610005812.4A CN201610005812A CN105660704A CN 105660704 A CN105660704 A CN 105660704A CN 201610005812 A CN201610005812 A CN 201610005812A CN 105660704 A CN105660704 A CN 105660704A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
- A01N25/12—Powders or granules
- A01N25/14—Powders or granules wettable
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
Abstract
The invention discloses a bacillus subtilis and oligosaccharins compound composition, a compound preparation and applications of the compound composition and the compound preparation. The bacillus subtilis and oligosaccharins compound composition includes bacillus subtilis and oligosaccharins, and the mass ratio of the bacillus subtilis to the oligosaccharins is 2000 billion to (1.5 g-3.5 g). The compound preparation comprises the compound composition, a dispersing agent, a high-water-absorption polymer and a filling material. The compound composition and the compound preparation can be used for sterilization of plants, especially for prevention and treatment of fusarium wilt. With adopting of compounding of the bacillus subtilis and the oligosaccharins, synergistic interaction can be achieved, so that the bacteriostasis action is better played, at the same time, the amount of the oligosaccharins is reduced, and the production cost is reduced.
Description
Technical field
The present invention relates to a kind of complex preparation for crops sterilization, be specifically related to the complex composition of a kind of bacillus subtilis and amino-oligosaccharide and the wettable powder of preparation thereof.
Background technology
Along with the development of agricultural, the healthy growth of crops is increasingly paid attention to by the mankind, and the droop of crops annoyings peasant always. Droop is a kind of soil-borne disease, and pathogen mainly accumulates in soil, and along with the growth of the continuous cropping time limit, morbidity also increases the weight of year by year, and the sheet that can cause crops is withered, dead. How to prevent and treat the droop harm to crops, have become as the important topic that agricultural industry faces.
Amino-oligosaccharide, chemical name: [(1,4)-2-amino-2-deoxidation-β-D-Glucose], plant self can be induced to produce multiple resistance material, as PAL, superoxide dismutase, peroxidase, polyphenol oxidase, protective plant protecting agent etc. improve the defence capability of self. And the cell of fungus all contains chitin, chitinase can effectively decompose chitin, when fungus, bacterial invasion plant time, under the effect of chitinase fungus, antibacterial cell be decomposed and dead. Additionally the resistance enzyme such as chitinase can activate plant cell, opens vegeto-animal immune system, greatly strengthens the resist ability of plant poor environment to external world. Amino-oligosaccharide also promotes the mitogenetic of root cell when resisting the infringement of pathogen, improves the resistance against diseases of crop root, beneficial microorganism amount reproduction in soil, root rot, droop, verticillium wilt, nematicide etc. are had control effect. And it is sturdy that well developed root system, stipes can be promoted to shorten, alleviate the problem that growing way is too prosperous or too weak, regulate the growth potential of crop.
But at present, the use cost of amino-oligosaccharide is higher, it is difficult to large scale application. How when lower cost, amino-oligosaccharide can be utilized to reach to promote plant growing, antibacterial effect, becomes a difficult problem of research at present.
Summary of the invention
The invention aims to the defect solving exist in prior art, it is provided that a kind of cost is low and has bacillus subtilis and amino-oligosaccharide complex composition and the complex preparation of synergistic function.
In order to achieve the above object, the invention provides a kind of bacillus subtilis and amino-oligosaccharide complex composition, including bacillus subtilis and amino-oligosaccharide;The mass ratio of bacillus subtilis living spores bacterium number and amino-oligosaccharide is 2,000,000,000,000: 1.5g～3.5g (being preferably in a proportion of 2,000,000,000,000: 2.5g).
Present invention also offers a kind of bacillus subtilis and amino-oligosaccharide complex preparation, including bacillus subtilis, amino-oligosaccharide, dispersant, high water absorbency polymer and inserts; In complex preparation, the weight percentage of amino-oligosaccharide is 0.1%～10% (preferred content is 2.5%), the mass ratio of bacillus subtilis living spores bacterium number and amino-oligosaccharide is 2,000,000,000,000: 1.5g～3.5g (being preferably in a proportion of 2,000,000,000,000: 2.5g), the weight percentage of dispersant is 5%～30% (preferred content is 5%), the weight percentage of high water absorbency polymer is 8%～30% (preferred content is 8%), and all the other are inserts.
Wherein, dispersant is sodium lignin sulfonate. High water absorbency polymer is high water absorbency polymer is polyacrylate, isobutene ./maleate (isobutene., maleate copolymer), starch/polyacrylate (starch graft polyacrylate copolymer), polyvinyl alcohol/polyacrylate (polyvinyl alcohol graft copolymerized polyacrylate polymers), acrylic fiber hydrate, how sulfonic formaldehyde condensation compound sodium salt or polymerization propylhomoserin.
Inserts is clay, potter's clay, kieselguhr, Pulvis Talci, Kaolin, corn starch, glucose or white carbon.
Present invention also offers the application in plant sterilization of above-mentioned complex composition and complex preparation, particularly in the application preventing and treating droop aspect. And add water and prepare medicinal liquid when using, in medicinal liquid, bacillus subtilis is 64mg/L with the total content of amino-oligosaccharide.
The present invention has the advantage that the present invention adopts amino-oligosaccharide and bacillus subtilis to carry out composite compared to existing technology, amino-oligosaccharide can make crops produce the disease-resistance substances such as phenolic compound, lignin, protective plant protecting agent, pathogenesis-related proteins, improve the defensive ferment relevant with disease-resistant metabolism and the activity of Active oxygen eliminating enzyme system system, thus blocking the breeding of Pathogen of Fusarium Wilt; Utilize abundant C, N that amino-oligosaccharide itself contains as the raw material of bacillus subtilis bacteria growing simultaneously, the materials such as antibiotic that bacillus subtilis secretion goes out, antibacterial peptide, big ring grease, cyclic peptide, similar phage, can play better bacteriostasis by Synergistic. The present invention adopts composite component bacillus subtilis, the consumption of amino-oligosaccharide can not only be reduced, reduce production cost, and as a kind of microbial bactericide, noresidue is also without resistance problems, its metabolite is less to the negatively influencing of amino-oligosaccharide, has high affinity, and that preventive effect to pathogenic bacteria is used in combination is more excellent with amino-oligosaccharide. The present invention utilizes dispersant, high water absorbency polymer to increase composite material dispersibility in water simultaneously, is adapted at field and uses on a large scale.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention is described in detail.
Amino-oligosaccharide (oligosaccharins) former medicine: Weifang, Shandong Tian Da plant protection company limited (LS20120353) product, amino-oligosacchride cellulose content 90%.
Bacillus subtilis (bacillussubtilis) former medicine: Wuhan City, Hubei Province Tian Hui biological engineering company limited (PD20096823) product, content 500,000,000,000 living spores/gram.
Complex composition embodiment one
Taking the former medicine of 10g amino-oligosaccharide and the former medicine of 24g bacillus subtilis is mixed with complex composition, in prepared compositions, bacillus subtilis living spores bacterium number is 2,000,000,000,000: 1.5g with the mass ratio of amino-oligosaccharide.
Complex composition embodiment two
Taking the former medicine of 10g amino-oligosaccharide and the former medicine of 18g bacillus subtilis is mixed with complex composition, in prepared compositions, bacillus subtilis living spores bacterium number is 2,000,000,000,000: 2.0g with the mass ratio of amino-oligosaccharide.
Complex composition embodiment three
Taking the former medicine of 10g amino-oligosaccharide and the former medicine of 14.4g bacillus subtilis is mixed with complex composition, in prepared compositions, bacillus subtilis living spores bacterium number is 2,000,000,000,000: 2.5g with the mass ratio of amino-oligosaccharide.
Complex composition embodiment four
Taking the former medicine of 10g amino-oligosaccharide and the former medicine of 12g bacillus subtilis is mixed with complex composition, in prepared compositions, bacillus subtilis living spores bacterium number is 2,000,000,000,000: 3.0g with the mass ratio of amino-oligosaccharide.
Complex composition embodiment five
Taking the former medicine of 10g amino-oligosaccharide and the former medicine of 10.3g bacillus subtilis is mixed with complex composition, in prepared compositions, bacillus subtilis living spores bacterium number is 2,000,000,000,000: 3.5g with the mass ratio of amino-oligosaccharide.
Complex preparation embodiment six
Taking the former medicine of amino-oligosaccharide 0.1 part, the former medicine of bacillus subtilis 0.1 part, sodium lignin sulfonate 30 parts, acrylic fiber hydrate 30 parts, Kaolin 39.8 parts, after pulverizing, mix homogeneously, 320 mesh sieves divide, and obtain complex preparation. Wherein, amino-oligosacchride cellulose content is 0.1%, and the mass ratio of bacillus subtilis living spores bacterium number and amino-oligosaccharide is 2,000,000,000,000: 3.5g, and the content of sodium lignin sulfonate is 30%, the content of acrylic fiber hydrate is 30%, and kaolinic content is 39.8%.
Complex preparation embodiment seven
Taking the former medicine of amino-oligosaccharide 11 parts, the former medicine of bacillus subtilis 27 parts, sodium lignin sulfonate 5 parts, starch/polyacrylate 8 parts, clay 49 parts, after pulverizing, mix homogeneously, 320 mesh sieves divide, and obtain complex preparation. Wherein, amino-oligosacchride cellulose content is 10%, and the mass ratio of bacillus subtilis living spores bacterium number and amino-oligosaccharide is 2,000,000,000,000: 1.5g, and the content of sodium lignin sulfonate is 5%, and the content of starch/polyacrylate is 8%, and the content of clay is 49%.
Complex preparation embodiment eight
Taking the former medicine of amino-oligosaccharide 2.8 parts, the former medicine of bacillus subtilis 4 parts, sodium lignin sulfonate 5 parts, polyacrylate 8 parts, white carbon 80.2 parts, after pulverizing, mix homogeneously, 320 mesh sieves divide, and obtain complex preparation. Wherein, amino-oligosacchride cellulose content is 2.5%, and the mass ratio of bacillus subtilis living spores bacterium number and amino-oligosaccharide is 2,000,000,000,000: 2.5g, and the content of sodium lignin sulfonate is 5%, and the content of polyacrylate is 8%, and the content of white carbon is 80.2%.
Effect example one
Toxicity test to withered germ of water-melon
With reference to the Plating (NY/T1156.2-2006) that People's Republic of China's agricultural industry criteria farm-chemical indoor determination test rule is recommended. Step is as follows:
(1) (Fusariumoxys-porumf.sp.niveum, by Hunan agricultural university plant pathology research department separation purification storage and provide for indoor culture withered germ of water-melon. For inoculation.
(2) medicament configuration: first former for amino-oligosaccharide medicine and bacillus subtilis are made into contrast mother solution with sterilized water for solvent, then the product of complex composition embodiment one to five is prepared mother solution with sterilized water for solvent.
(3) chemicals treatment: aseptically, processing according to test is quantitatively adding in aseptic conical flask by the sterilizing PDA culture medium melted in advance, from low concentration to high concentration, quantitatively draw medicinal liquid be successively separately added in above-mentioned conical flask, fully shaking up, then equivalent is poured in the culture dish that four diameters are 9cm and is made pastille flat board.Wherein the drug concentration of amino-oligosaccharide is 4mg/L, 2mg/L, 1mg/L, 0.5mg/L, 0.25mg/L; The drug concentration of bacillus subtilis is 160mg/L, 80mg/L, 40mg/L, 20mg/L, 10mg/L; The drug concentration of the compound product of complex composition embodiment one to five is 64mg/L, 32mg/L, 16mg/L, 8mg/L, 4mg/L; Set aseptic aqueous solution comparison simultaneously.
(4) inoculation and cultivation: by cultured bacterium cake, aseptically with the sterilizing card punch that diameter is 5mm, bacterium cake is cut from colony edge, seeing that pure culture biscuits involvng inoculation is in pastille flat board central authorities with inoculator, mycelia faces up, and covers ware lid, bio-incubator is cultivated (temperature 26 DEG C～28 DEG C, relative humidity 85%～95%), each process repeats for 4 times, if aseptic aqueous solution comparison.
(5) investigation: according to the growing state investigation bacterium pathogenic bacteria silk growing state of bacterium in blank culture dish, with kind of calliper colony diameter (mm), each bacterium colony decussation method vertical survey diameter is each once, takes its meansigma methods. Correlation computations formula is:
D=D1-D2
D is that bacterium colony increases diameter; D1For colony diameter; D2For bacterium cake diameter.
I=100 × (D0-Dt)/D0
I: mycelial growth inhibition rate
D0: blank bacterium colony increases diameter
Dt: chemicals treatment bacterium colony increases diameter
Result is in Table 1, from table 1 it follows that the rejection ability of withered germ of water-melon is above bacillus subtilis by the complex composition of amino-oligosaccharide and bacillus subtilis, and along with increasing of concentration is more high to the concordance rate of withered germ of water-melon.
Table 1 amino-oligosaccharide and bacillus subtilis and complex composition are to withered germ of water-melon toxicity test result
Data in his-and-hers watches 1 carry out statistical analysis, result is in Table 2, from data in table, in complex composition embodiment one to five, the co-toxicity coefficient of withered germ of water-melon is all higher than 120 by the product of preparation, amino-oligosaccharide is described and bacillus subtilis is composite that droop is had potentiation. Wherein, complex composition embodiment three co-toxicity coefficient is the highest, and when namely the mass ratio of bacillus subtilis living spores bacterium number and amino-oligosaccharide is 2,000,000,000,000: 2.5g, synergistic function is the strongest.
Withered germ of water-melon toxicity test result is added up by table 2 amino-oligosaccharide, bacillus subtilis and complex composition
Effect example two
Complex preparation embodiment six to eight index of correlation is detected, and testing result is as shown in the table:
Table 3 complex preparation item controlled index
| Living spores bacillus number (hundred million/gram) | 5～1350 |
| Amino-oligosacchride cellulose content (%) | 0.1～10 |
| Moisture (%) | 3～5 |
| PH value | 6～8 |
| Suspensibility (%) | > 90% |
The complex preparation of complex preparation embodiment eight preparation is carried out Security test:
3% amino-oligosaccharide: Beijing Multigrass Formulation Co., Ltd. (LS20110178).
1, preparation of reagents (being effective ingredient amount):
(1) the former medicine 4g/hm of amino-oligosaccharide2;
(2) the former medicine 8g/hm of amino-oligosaccharide2;
(3) the former medicine 12g/hm of amino-oligosaccharide2;
(4) the former medicine 16g/hm of amino-oligosaccharide2;
(5) the former medicine 20g/hm of amino-oligosaccharide2;
(6) the former medicine 20g/hm of bacillus subtilis2;
(7) the former medicine 30g/hm of bacillus subtilis2;
(8) the former medicine 40g/hm of bacillus subtilis2;
(9) the former medicine 50g/hm of bacillus subtilis2;
(10) the former medicine 60g/hm of bacillus subtilis2;
(11) complex preparation embodiment eight prepares product 150g/hm2(effective ingredient amount is amino-oligosaccharide and bacillus subtilis total amount);
(12) complex preparation embodiment eight prepares product 300g/hm2;
(13) complex preparation embodiment eight prepares product 450g/hm2;
(14) complex preparation embodiment eight prepares product 600g/hm2;
(15) complex preparation embodiment eight prepares product 750g/hm2;
(16) 3% amino-oligosaccharide 4g/hm2;
(17) 3% amino-oligosaccharide 8g/hm2;
(18) 3% amino-oligosaccharide 12g/hm2;
(19) 3% amino-oligosaccharide 16g/hm2;
(20) 3% amino-oligosaccharide 32g/hm2;
(21) blank.
2, the inhibitory action of the growth of Citrullus vulgaris Seedling is measured
Above-mentioned medicament is configured to required series concentration with appropriate amounts of sterilized water respectively stand-by.
Test arrangement carries out in Agricultural University Of Hunan's pesticide biological and ecological research department phjytotron, Citrullus vulgaris Seedling phjytotron condition of culture temperature is 25-28 degree Celsius, illumination every day 16 hours, humidity 92-97%, experimental field soil is more fertile, pH6.5, the content of organic matter 5.4%, other are consistent with greenhouse management.
Treating that the Citrullus vulgaris Seedling young leaves 4-6 leaf phase processes, take the medicament of above-mentioned preparation, spray medicinal liquid with POTTER precision laboratory spray tower (BS00282) respectively, spouting liquid, for often processing 50ml, keeps ground moistening after medicine. Often process and repeat 4 times, within after medicine 10 days, observe each process by ocular estimate and test basin watermelon growing situation. Calculate each medicament variable concentrations growth inhibited effect (poisoning grade scale (point Pyatyi): represent asymptomatic to Citrullus vulgaris Seedling; + represent that young leaves has a small amount of medicine speckle; ++ represent that young leaves has relatively multiple medicines speckle, but grow normal; +++ represent the distortion of young leaves deformity; ++++the represent whole strain of Citrullus vulgaris is withered even dead).
Suppression ratio (%)=(check plot Citrullus vulgaris height of seedling degree treatment region Citrullus vulgaris height of seedling degree) * 100/ check plot Citrullus vulgaris height of seedling degree
Then virulence regression equation is asked to calculate the IC of each medicament by DPS software statistics10And the confidence limit of 80%.
Within after medicine 10 days, observing the Citrullus vulgaris condition of production by ocular estimate, its result is in Table 4. Complex preparation prepared by the visible present invention produces without poisoning under recommending test dose.
The poisoning condition survey result table of 10 days Citrullus vulgaris Seedlings after table 4 amino-oligosaccharide and bacillus subtilis and complex preparation medicine thereof
As can be seen from the above table, amino-oligosaccharide and bacillus subtilis and complex preparation thereof are essentially the same to the growth effect of three Citrullus vulgaris different cultivars, therefore our plant height measurement to variety of watermelon early good (8424), and its measurement result is shown in following table.
Table 5 amino-oligosaccharide and bacillus subtilis and the complex preparation inhibitory action measurement result to Citrullus vulgaris Seedling early good (8424) plant height thereof
Within 30 days, measuring the plant height of the variety of watermelon early good (8424) of each process after medicine, measurement result is shown in following table simultaneously.
Table 6 amino-oligosaccharide is to Citrullus vulgaris Seedling early good (8424) plant height inhibitory action result
As can be seen from the above table, is there is not poisoning by the complex preparation of amino-oligosaccharide and bacillus subtilis within recommendation test dose in Citrullus vulgaris Seedling, is adapted at preventing and treating large-scale popularization application on watermelon blight.
Claims (10)
1. a bacillus subtilis and amino-oligosaccharide complex composition, it is characterised in that: described complex composition includes bacillus subtilis and amino-oligosaccharide; The mass ratio of described bacillus subtilis living spores bacterium number and amino-oligosaccharide is 2,000,000,000,000: 1.5g～3.5g.
2. bacillus subtilis according to claim 1 and amino-oligosaccharide complex composition, it is characterised in that: the mass ratio of described bacillus subtilis living spores bacterium number and amino-oligosaccharide is 2,000,000,000,000: 2.5g.
3. the application in preparing bactericide of the complex composition described in claim 1 or 2.
4. the application in preventing and treating droop of the complex composition described in claim 1 or 2.
5. a bacillus subtilis and amino-oligosaccharide complex preparation, it is characterised in that: described complex preparation includes bacillus subtilis, amino-oligosaccharide, dispersant, high water absorbency polymer and inserts; In described complex preparation, the weight percentage of amino-oligosaccharide is 0.1%～10%, the mass ratio of bacillus subtilis living spores bacterium number and amino-oligosaccharide is 2,000,000,000,000: 1.5g～3.5g, the weight percentage of dispersant is 5%～30%, the weight percentage of high water absorbency polymer is 8%～30%, remaining as inserts.
6. complex preparation according to claim 5, it is characterised in that: the mass ratio of described bacillus subtilis living spores bacterium number and amino-oligosaccharide is 2,000,000,000,000: 2.5g.
7. complex preparation according to claim 5, it is characterised in that: the weight percentage of described amino-oligosaccharide is 2.5%; The weight percentage of described dispersant is 5%; The weight percentage of described high water absorbency polymer is 8%.
8. according to the arbitrary described complex preparation of claim 5 to 7, it is characterised in that: described dispersant is sodium lignin sulfonate; Described high water absorbency polymer be polyacrylate, isobutene ./maleate, starch/polyacrylate, polyvinyl alcohol/polyacrylate, acrylic fiber hydrate, how sulfonic formaldehyde condensation compound sodium salt or polymerization propylhomoserin; Described inserts is clay, potter's clay, kieselguhr, Pulvis Talci, Kaolin, corn starch, glucose or white carbon.
9. claim 5 to 7 arbitrary described complex preparation application in plant sterilization.
10. claim 5 to 7 arbitrary described complex preparation application in preventing and treating plant droop.
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| CN106135291A (en) * | 2016-07-14 | 2016-11-23 | 解晓燕 | A kind of prevent and treat soil-borne disease, improve crop cold resistance compositions |
| CN106717245A (en) * | 2016-11-15 | 2017-05-31 | 云南良禾种业有限公司 | A kind of processing method of disease resistance corn seed high |
| CN107821439A (en) * | 2017-10-27 | 2018-03-23 | 沈阳化工研究院有限公司 | A kind of composition for preventing and treating rice sheath blight disease and preparation method thereof |
| CN109452284A (en) * | 2018-11-19 | 2019-03-12 | 南京工业大学 | A kind of preparation process of amino-oligosaccharide and bacillus compound preparation |
| CN112674115A (en) * | 2021-01-21 | 2021-04-20 | 山东亿嘉农化有限公司 | Compound preparation for preventing and treating ginger stem rot and application thereof |
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Application publication date: 20160615 |