CN101167480A - Composite preparation made from microorganism fermented liquid - Google Patents
Composite preparation made from microorganism fermented liquid Download PDFInfo
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Abstract
A multiple-effect medicament compounded by microbe fermentation liquor has a multiple-effect medicament formulation compounded by microbe fermentation liquor and inorganic acid or organic acid, and inorganic salt or organic salt. The multiple-effect medicament of the compounded medicament comprises very obvious characteristic component of aliphatic acid. The multiple-effect medicament realizes multiple effects of promoting plants growing, increasing plants volume of output, preventing and curing plants diseases, and is a multiple-effect medicament which can promote plants to grow, increase plants volume of output, has preventing and curing effects for plant diseases.
Description
One. technical field
The present invention relates to the new purposes of microbial fermentation solution aspect plant protection and promotion plant growing, belong to microbiology, chemistry and agricultural technology field relate to microbiology, chemistry, agronomy and biochemistry etc.
Two. technical background
Along with the development of chemical industry and the expansion of the agrochemical fertilizer scope of application, the quantity of chemical pesticide and kind are all in continuous increase, and World chemical agricultural chemicals gross yield (in active ingredient) is above 3,000,000 tons now.Because using in a large number for a long time of chemical pesticide and chemical fertilizer brought serious environmental to pollute and the drug-fast continuous increase of sick worm.Many countries have all dropped into substantial contribution and have researched and developed the novel formulation that can substitute or reduce chemical pesticide and chemical fertilizer, to keep the sustainable development of agricultural production.
Environmental pollution and residue of pesticide that agriculture chemical causes are even more serious in China.There are 1.2 hundred million hectares of arable lands in China, and the total amount of application of chemical pesticide reaches 131.2 ten thousand tons, uses 931.3 grams for average every mu, and vegetables and melon and fruit more reach 6.67 kilograms/mu.The fertilizer amount of China has then accounted for 1/3 of global consumption, has brought serious rivers and lakes and underground water eutrophication pollution thus.Causing the output because the grower covets of using in a large number on the one hand of China's fertilizer and pesticide, is owing to lack the use that can effectively reduce fertilizer and agricultural chemicals but the method and the product that can guarantee crop yield and quality on the other hand.
Early stage discover the plant hormone that some microorganisms can the regulation of secretion growth and development of plants, as draw indolylbutyric acid, abscisic acid, gibberellin etc., also have the metabolite of some microorganisms can inducing plant resistance (CN1456059A), as chitin, chitosan etc.And the fatty acid that is present in occurring in nature in a large number also is noted to have certain bacterium and Fungicidally active extremely, and the n-nonanoic acid of high concentration has very strong phytotoxicity also to be used as weed killer herbicide.S.D. people such as savart lattice has obtained good effect (CN1069391A) with direct antifungal in disease position and bacterium that fatty acid is administered to plant, the result of treatment of fatty acid by antifungal and bacterize plant disease emphasized in their research, just there is not an activity but the concentration of making statement fatty acid is very low, too highly then plant is damaged, and fatty acid and its salt or derivative do not have prophylactic activity to plant disease.In the experimental result that they provide, fatty acid and its salt or derivative are that therapeutic action is just played at the disease position that is applied directly to plant, and the minimal effective concentration that the salt of fatty acid and it or derivative are administered on the plant is 0.125%a.i. (1250 mg/litre), and concentration is low to moderate 0.063% o'clock used fatty acid or its salt or derivative and has not just controlled the effect of infection process.
The present invention relates to the multiple-effect preparation that is used for coordinate plant growth growth and controlling plant diseases with the microbial fermentation solution preparation.By the combination of microbial fermentation solution and inorganic acid or organic acid and mineral salt or organic salt, obtained promoting plant growing, increase the outstanding effect on the plant products, and this composition can also effectively prevent plurality of plant diseases.The existence of the characteristic component fatty acid that contains in this multiple-effect preparation is very obvious, although content be lower than that people such as savart lattice proposes can sterilization minimum effective fatty acid concentration 1250 mg/litre.The multiple-effect preparation has been realized the promotion plant growing, increases plant products, the multiple effect of controlling plant diseases, and this uses chemical fertilizer in a large number for minimizing and the pollution of agricultural chemicals in agricultural product and environment has very positive meaning.
Three. summary of the invention
The technical problem that solves
Purpose one of the present invention provides a kind of multiple-effect pharmaceutical formulation made from the combination of microbial fermentation solution and inorganic acid or organic acid and mineral salt or organic salt, this multiple-effect preparation has been realized the promotion plant growing, increase plant products, the multiple effect of controlling plant diseases, be one and can promote plant growing, improve plant products, the existing preventive effect of plant disease is had the multiple-effect preparation of result of treatment again.
Purpose two of the present invention is seeds that the preparation made can be applied to plant, leaf, and root, stem or fruit, and the soil of plant attaching and culture fluid etc., the application process of preparation is for spraying, and pouring is soaked seed and is soaked fruit.
Technical scheme
Those skilled in the art know that microorganism has many useful compositions in the metabolite that fermentation process produces, comprise plant hormone, induction of resistance element etc.These compositions usually need complicated program to extract collection, make preparation again and are used for production practices.Main method of the present invention is with microbial fermentation solution and inorganic acid or organic acid and mineral salt or the direct a kind of pharmaceutical formulation made that makes up of organic salt, synergy by zymotic fluid and inorganic acid or organic acid and mineral salt or organic salt, make preparation obtain the promotion plant growing, improve plant products, the existing preventive effect of plant disease is had the multi-functional of result of treatment again.
The main component of described zymotic fluid including, but not limited to: the saturated or undersaturated fatty acid from C5 to C24, fatty acid is including, but not limited to palmitic acid, palmitoleic acid, stearic acid, linoleic acid, oleic acid, certain herbaceous plants with big flowers acid, lauric acid, myristic acid, n-nonanoic acid, arachidonic acid etc., or the salt of fatty acid or derivative, including, but not limited to: by potassium, sodium, soap or derivative that ammonium plasma or group of containing amido etc. are combined into.Described inorganic acid or organic acid are including, but not limited to sulfuric acid, acetic acid, phosphoric acid, nitric acid, hydrochloric acid, phosphorous acid, sulfurous acid, boric acid, formic acid, acetate, glacial acetic acid, butyric acid, phthalic acid, terephthalic acid (TPA), succinic acid, humic acid, salicylic acid, p-aminobenzoic acid, ortho-aminobenzoic acid, benzoic acid etc.Described mineral salt or organic salt are including, but not limited to calcium chloride, sodium chloride, potassium chloride, superphosphate, sodium tripolyphosphate, sodium polyphosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, sodium sulphate, potassium sulphate, calcium carbonate, calcium bicarbonate, calcium sulphate, nitrate of lime, calisanin etc.
Implementation step of the present invention is:
1) get glucose and Fructus Hordei Germinatus and extract each 20 gram of powder, add 5 gram yeast extract powders, 5 gram trehalose ammonia calcium, agar 15 grams add water to 1 liter.After the sterilization cooling, pour into and make medium (GMYAA) in the culture dish.On medium, rule to cultivate to cultivate in 48 hours with the relevant bacterial classification Pseudomonas glathei TP06 that is selected from Pseudomonas and obtain bacterial classification.
2) bacterial classification is forwarded to from medium contain GMYAB culture fluid (glucose and Fructus Hordei Germinatus extract each 20 gram of powder, add 5 gram yeast extract powders, and 5 gram trehalose ammonia calcium add water to 1 liter),, on shaking table, cultivated 72 hours under the condition of rotating speed 125rpm 30 ℃ of temperature.
3) with 10% inoculum concentration the bacterial classification of cultivating is transferred to fermentation tank and cultivated 72 hours at 30 ℃ of aerobic fementations, make zymotic fluid with identical culture fluid with seeding tank.
4) get in the hydrochloric acid solution that 100 milligrams of nitrate of lime join 120 milliliter 10%, fully stir, add 10 milliliters and contain 20% sodium tripolyphosphate solution, join in 1 liter of zymotic fluid of above-mentioned cultivation after fully stirring, fully stir, promptly make the multiple-effect preparation that contains zymotic fluid.
Embodiment:
Example 1.
1) glucose and Fructus Hordei Germinatus extract each 20 gram of powder, add 5 gram yeast extract powders, 5 gram trehalose ammonia calcium, and agar 15 grams add water to 1 liter.After the sterilization cooling, pour into and make medium (GMYAA) in the culture dish.On medium, rule to cultivate to cultivate in 48 hours with the relevant bacterial classification Pseudomonas glathei TP06 that is selected from Pseudomonas and obtain bacterial classification.
2) bacterial classification forwards to from medium and contains GMYAB culture fluid (glucose and Fructus Hordei Germinatus extract each 20 gram of powder, add 5 gram yeast extract powders, and 5 gram trehalose ammonia calcium add water to 1 liter), 30 ℃ of temperature, cultivates 72 hours on shaking table under the condition of rotating speed 125rpm.
3) with 10% inoculum concentration the bacterial classification of cultivating is transferred to fermentation tank and cultivated 72 hours at 30 ℃ of aerobic fementations, make zymotic fluid with identical culture fluid with seeding tank.
4) get in the hydrochloric acid solution that 100 milligrams of nitrate of lime join 120 milliliter 10%, fully stir, add 10 milliliters and contain 20% sodium tripolyphosphate solution, join in 1 liter of zymotic fluid of above-mentioned cultivation after fully stirring, fully stir, promptly make the multiple-effect preparation that contains zymotic fluid.
The content of fatty acid is measured by following condition with gas chromatography-mass spectrography in the example 2. multiple-effect preparations.
1. instrument Agilent Technologies GC/MS 5975i
2. chromatographic column: HP-5ms (30m * 250 μ m * 0.25 μ m) 5%PhenyMethylSiloxane
3. chromatogram column temperature program: 100 ℃ keep 1min, then with 30 ℃/min temperature programming to 130 ℃, again with 5 ℃/min temperature programming to 250 ℃, with 10 ℃/min temperature programming to 300 ℃, keep 5min again;
4. carrier gas: helium, purity 〉=99.999%, flow velocity are 1.2mL/min;
5. injector temperature: 290 ℃;
6. sample size: 2 μ L;
7. input mode: no split sampling;
8. electron bombardment ionization source: 70eV;
9. quadrupole rod temperature: 150 ℃;
10. ion source temperature: 230 ℃;
11.GC-MS interface temperature: 280 ℃;
Full spectrum scanning; M/z:50 is to 550u for the mass scanning scope; Solvent delay: 3min.
The result such as the table 1 that record.
The content of the main fatty acid of part in the table 1 multiple-effect preparation
| Title | Content (mg/L) |
| Palmitic acid | 3.56 |
| Palmitoleic acid | 1.11 |
| Stearic acid | 0.45 |
| Oleic acid | 3.00 |
| Linoleic acid | 3.12 |
Example 3. multiple-effect preparations increase the effect on the output promoting plant growing
Get 100 milliliters in multiple-effect preparation, 100 times of thin ups evenly are sprayed onto on the new eggplant leaf of transplanting in field, spray once every 15 days, do not use any chemical bactericide and topdress, and adjacent plot compares with the Routine Management that sprays chemical pesticide and topdress.Result such as table 2, the multiple-effect preparation significantly promotes plant growth, and has obtained the effect of increasing production up to 42%.
Table 2 multiple-effect preparation promotes the comparison of eggplant growth and effect of increasing production
Annotate 1: with the significant difference of different letter representations processing and contrast behind the column of figure.
The plot of peach yellows took place by the randomised block design field trial in example 4. at known continuous 3 years.The reason that causes the peach yellows is not made final conclusion as yet, but preliminary investigation thinks it may is because the physiological disease that causes of iron deficiency or infect the pathologic yellow that causes by a kind of viroids, or the peach yellow that causes of the immixture of these two kinds of reasons.Test is divided into two groups, and one group is that peach occurs promptly beginning to handle before the yellow, and another group just begins to handle after the yellow symptom has appearred in peach blade.
100 milliliters in multiple-effect preparation is got in processing, and 100 times of thin ups evenly are sprayed onto on the blade of peach, sprays once every 15 days, does not use any chemical pesticide and topdresses, and compares with spray chemical pesticide and the Routine Management that topdresses.Handle and each 5 strain of contrast, repeat 3 times.Investigation result such as table 3.
The effect of table 3 multiple-effect preparation on control peach yellows
| Handle | The yellows incidence (%) of beginning spray preparation before yellows take place | Yellows begin spray preparation after taking place |
| The multiple-effect preparation | 0.00 | Yellow leaf vein changes green after spraying multiple-effect preparation, and it is green spray 3 rear blades commentaries on classics |
| Contrast | 75.5 | The yellow of complete stool blade, serious blade is little and bleach |
The white seedling of the new transplanting lotus flower plot that root rot harm will take place example 5. is divided into processing and contrast by randomised block design, handles and gets 100 milliliters in multiple-effect preparation, and 100 times of thin ups evenly are sprayed onto on the white seedling of lotus flower of new transplanting, spray once every 7 days.The spirit of chemical pesticide root-rot is sprayed in contrast.Every processing 30 young plants repeat 3 times.Spray 3 " Invest, Then Investigate " results such as table 4, the seedling in contrast plot almost all is subjected to infection process death, and the multiple-effect preparation has significantly reduced the harm of root rot, and the control efficiency of the white seedling root rot of lotus flower is reached 95%.
The effect of table 4, multiple-effect preparation control vegetable root rot
| Handle | Seedling death rate of the onset % |
| The multiple-effect preparation | 12.3a 1 |
| Contrast | 95.7b |
Annotate
1: with the significant difference of different letter representations processing and contrast behind the column of figure.
Example 6. is got 100 milliliters in multiple-effect preparation, and 100 times of thin ups evenly are sprayed onto kind and are subjected in the greenhouse on the Lantana camara of powdery mildew serious harm, handles and each 10 strain of contrast, repeats 3 times.Sprayed once every 7 days.The spray clear water compares.Spray 2 " Invest, Then Investigate " results such as table 5, the multiple-effect preparation has the control efficiency of highly significant to plant powdery mildew.
The effect of table 5, multiple-effect preparation control plant powdery mildew
| Handle | Incidence of disease % |
| The multiple-effect preparation | 21.3a |
| Contrast | 98.2b |
Annotate
1: with the significant difference of different letter representations processing and contrast behind the column of figure.
Example 7. grape seedling kinds after being kept at the Pseudoperonospora cubensis inoculation on the live body plant, place high humidity environment to cultivate 24 hours in the greenhouse.Get 100 milliliters in multiple-effect preparation, 100 times of thin ups, whole strain is evenly sprayed on the grape seedlings of having cultivated 24 hours after the inoculation, and the spray clear water compares.Handle and each 5 strain of contrast, repeat 3 times.7 days " Invest, Then Investigate " results such as table 6, the multiple-effect preparation has the control efficiency of highly significant to downy mildew of garpe.
Table 6, multiple-effect preparation are prevented and treated the effect of downy mildew of garpe
| Handle | Incidence of disease % | Disease index |
| The multiple-effect preparation | 17.2a | 0.8a |
| Contrast | 91.1b | 4.3b |
Annotate
1: with the significant difference of different letter representations processing and contrast behind the column of figure.
Claims (16)
1. a coordinate plant growth made from microbial fermentation solution is grown and the multiple-effect preparation of controlling plant diseases.It is characterized in that: be selected from a kind of microbial fermentation solution and be selected from one or more inorganic acids or organic acid and be selected from the coordinate plant growth growth that the combination of one or more mineral salt or organic salt is made, increase plant products, the existing preventive effect of plant disease is had the multiple-effect preparation of result of treatment again.
2. the active ingredient of microbial fermentation solution as claimed in claim 1 is including, but not limited to the saturated or undersaturated fatty acid from C5 to C24 or the salt or the derivative of fatty acid, the compound of one or more amine or amide-type, the compound of one or more amino acidses, or the compound of one or more polypeptide classes.
3. inorganic acid or organic acid are including, but not limited to sulfuric acid, acetic acid, phosphoric acid, nitric acid as described in wanting 1 as right, hydrochloric acid, phosphorous acid, sulfurous acid, boric acid, formic acid, acetate, glacial acetic acid, butyric acid, phthalic acid, terephthalic acid (TPA), succinic acid, humic acid, salicylic acid, p-aminobenzoic acid, ortho-aminobenzoic acid, benzoic acid etc.
According to claim 1 mineral salt or organic salt including, but not limited to calcium chloride, sodium chloride, potassium chloride, superphosphate, sodium tripolyphosphate, sodium polyphosphate, potassium dihydrogen phosphate, dipotassium hydrogen phosphate, sodium sulphate, potassium sulphate, calcium carbonate, calcium bicarbonate, calcium sulphate, nitrate of lime, calisanin etc.
5. preparation according to claim 1 is characterized in that: described preparation can only contain the salt of a kind of fatty acid or fatty acid or derivative as characteristic component.
6. preparation according to claim 1 is characterized in that: more preferably described preparation contains the composition of the salt of two or more fatty acid and fatty acid or fatty acid or derivative as characteristic component.
7. preparation according to claim 1 is characterized in that: the target site that described preparation can be applied to is including, but not limited to the seed of plant, leaf, root, stem or fruit, or any solid-state or liquid object of plant attaching.
8. preparation according to claim 1, it is characterized in that: the mass concentration of institute's fatty acids is 0.001 mg/litre-20 grams per liter in the liquid when being administered on the target site behind the described preparation dilute with water.
9. preparation according to claim 1, it is characterized in that: the mass concentration of institute's fatty acids is 0.01 mg/litre-1249.9999 mg/litre in the liquid when being administered on the target site behind the preferred described preparation dilute with water.
10. according to claim 1, it is characterized in that described inorganic acid of preparation or organic acid mass concentration are 0.00001-50%, the mass concentration of described mineral salt or organic salt is 0.00001-50%.
11. microbial fermentation solution according to claim 1, it is characterized in that: the preparation method obtains bacterial classification by the relevant bacterial classification Pseudomonas glathei TP06 that is selected from Pseudomonas through the GMYAA medium culture, bacterial classification forwarded in the triangular flask from medium (contain the GMYAB culture fluid) at temperature 30C, on shaking table, cultivated 72 hours under the condition of rotating speed 125rpm, inoculum concentration with 10% is transferred to fermentation tank with the bacterial classification of cultivating and cultivated 72 hours at 30 ℃ of aerobic fementations with identical culture fluid, makes microbial fermentation solution.
12. preparation according to claim 1 is characterized in that: the preparation method directly adds inorganic acid or organic acid and mineral salt or organic salt in microbial fermentation solution, fully stir, and promptly makes described multiple-effect preparation.
13. plant disease according to claim 1 is characterized in that: including, but not limited to by fungi, bacterium, virus, viroids, nematode etc. are infected the pathologic plant disease that causes, or because nitrogen stress, phosphorus, potassium, calcium, magnesium, sulphur, iron, zinc, manganese, the physiological plant disease that nutritive elements such as boron cause.
14., it is characterized in that as fatty acid as described in the claim 2: including, but not limited to palmitic acid, palmitoleic acid, stearic acid, linoleic acid, oleic acid, certain herbaceous plants with big flowers acid, lauric acid, myristic acid, n-nonanoic acid, arachidonic acid etc.
15. as the salt of fatty acid as described in the claim 2 or derivative including, but not limited to: by potassium, sodium, soap or derivative that ammonium plasma or group of containing amido etc. are combined into.
16. as described in claim 6, it is characterized in that the ratio of described fatty acid in total grease of preparation is: palmitic acid 0-95%, stearic acid 0-20%, linoleic acid 0-85%, oleic acid 0-90%, certain herbaceous plants with big flowers acid 0-20%, lauric acid 0-25%, myristic acid 0-20%, n-nonanoic acid 0-20%.
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| CN102325872A (en) * | 2009-02-20 | 2012-01-18 | 丹尼斯科美国公司 | Fermentation broth formulations |
| CN104510729A (en) * | 2013-09-26 | 2015-04-15 | 黄永 | Animal immune resistance-activating composition of fatty acid and derivatives thereof |
| WO2018047918A1 (en) * | 2016-09-08 | 2018-03-15 | イビデン株式会社 | Plant activator and process for producing plant activator |
| WO2018168583A1 (en) * | 2017-03-14 | 2018-09-20 | イビデン株式会社 | Plant functional component content improver, and method for producing plant functional component content improver |
| CN108990983A (en) * | 2018-08-31 | 2018-12-14 | 成都特普生物科技股份有限公司 | A kind of sustained release microbial bacterial agent |
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| JP2019151607A (en) * | 2018-03-06 | 2019-09-12 | イビデン株式会社 | Improver of functional component content in licorice, and manufacturing method of improver of functional component content in licorice |
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| CN102325872A (en) * | 2009-02-20 | 2012-01-18 | 丹尼斯科美国公司 | Fermentation broth formulations |
| CN104510729A (en) * | 2013-09-26 | 2015-04-15 | 黄永 | Animal immune resistance-activating composition of fatty acid and derivatives thereof |
| WO2018047918A1 (en) * | 2016-09-08 | 2018-03-15 | イビデン株式会社 | Plant activator and process for producing plant activator |
| CN109688821A (en) * | 2016-09-08 | 2019-04-26 | 揖斐电株式会社 | The manufacturing method of plant activator and plant activator |
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| WO2018168583A1 (en) * | 2017-03-14 | 2018-09-20 | イビデン株式会社 | Plant functional component content improver, and method for producing plant functional component content improver |
| US11963492B2 (en) | 2017-03-14 | 2024-04-23 | Ibiden Co., Ltd. | Agent for increasing a plant functional component content and a method for manufacturing the same |
| CN110418845A (en) * | 2017-03-14 | 2019-11-05 | 揖斐电株式会社 | The manufacturing method of Plant functional component content enhancer and Plant functional component content enhancer |
| JP2021035362A (en) * | 2017-03-14 | 2021-03-04 | イビデン株式会社 | Plant functional component content improver and manufacturing method of plant functional component content improver |
| JP7088694B2 (en) | 2018-03-06 | 2022-06-21 | イビデン株式会社 | A method for producing a carrot growth promoter and a functional ingredient content improver in carrots, and a carrot growth promoter and a functional ingredient content improver in carrots. |
| JP2019151607A (en) * | 2018-03-06 | 2019-09-12 | イビデン株式会社 | Improver of functional component content in licorice, and manufacturing method of improver of functional component content in licorice |
| JP7088695B2 (en) | 2018-03-06 | 2022-06-21 | イビデン株式会社 | Method for producing functional ingredient content improving agent in licorice and functional ingredient content improving agent in licorice |
| JP2019151606A (en) * | 2018-03-06 | 2019-09-12 | イビデン株式会社 | Carrot growth promoter, improver of functional component content in carrot, and manufacturing method of carrot growth promoter and improver of functional component content in carrot |
| CN108990983A (en) * | 2018-08-31 | 2018-12-14 | 成都特普生物科技股份有限公司 | A kind of sustained release microbial bacterial agent |
| CN115997766A (en) * | 2022-11-21 | 2023-04-25 | 云南农业大学 | Mixtures of phthalic acid, palmitic acid and azelaic acid and their use |
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