CN109362790A - It is a kind of for preventing and treating the complex microbial inoculum and preparation method thereof of pear tree root knot - Google Patents
It is a kind of for preventing and treating the complex microbial inoculum and preparation method thereof of pear tree root knot Download PDFInfo
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Abstract
The invention discloses a kind of for preventing and treating the complex microbial inoculum of pear tree root knot, the weight percent composition content of each component in the complex microbial inoculum includes: composite humic acid 10%-25%, soil supension 5%-20%, copper sulphate 5%-15%, Fungicidal active substance crude extract 3%-10%, azone 3%-8%, jinggangmycin 7%-8%, glycerol 5%-10%, penicillin 1%-3%, planting spirit 1%-3%, ABT root-inducing powder 0.01%-0.1%, and surplus is water.Complex microbial inoculum made of the present invention, rationally, production method is simple for proportion, by the interaction of various ingredients and root nodule germ, pear tree root nodule germ can be killed, improves soil texture, nutrient needed for improving plant growth, therapeutic effect is significant, can effectively promote Growth of Pear Trees development, improve the resistance of pear tree, reduce the use of agriculture chemical, the yield and quality of agricultural product is improved, there is very high economic benefit, ecological benefits and social benefit, using having a bright future.
Description
Technical field
It is specially a kind of for preventing and treating the microorganism of pear tree root knot the present invention relates to the preparation technical field of biological pesticide
Composite bacteria agent and preparation method thereof.
Background technique
The root knot of pear tree is also referred to as root knot, is a kind of bacterial disease, and the rootstock position of main harm pear tree is also endangered
Root, branch, mainly overwintering in invalid body or soil with invalid body and gemma, route of transmission is extensive, rainwater, wind, insect, artificial biography
It has been broadcast that, be the most important trunk disease of pear tree, fallen ill in recent years increasingly heavier, and not only affected the normal growth of pear tree,
And the quality of product, yield are also affected, or even will cause pear tree death, the attention of numerous orchard workers should be introduced.
Therefore, it for the current demand of pear tree root knot prevention and treatment, develops a kind of application convenience, highly effective and safe, have both prevention
It is of great significance with the complex microbial inoculum for the treatment of pear tree root knot.
Summary of the invention
The complex microbial inoculum and its preparation side that the purpose of the present invention is to provide a kind of for preventing and treating pear tree root knot
Method, to solve the problems mentioned in the above background technology.
In order to solve the above technical problem, the present invention provides following technical solutions: a kind of for preventing and treating pear tree root knot
The weight percent composition content of complex microbial inoculum, each component in the complex microbial inoculum is as follows:
Composite humic acid 10%-25%, soil supension 5%-20%, copper sulphate 5%-15%, Fungicidal active substance slightly mention
Liquid 3%-10%, azone 3%-8%, jinggangmycin 7%-8%, glycerol 5%-10%, penicillin 1%-3%, planting spirit 1%-
3%, ABT root-inducing powder 0.01%-0.1%, surplus are water.
Further, the compound humic acid is the stem using a variety of green crops such as corn stover, wheat straw, rice straw
Bar is that raw material is crushed, after mixing treatment, is sterilized, is fermented, carrying out solid fermentation in the big tank of rotation, then being separated
It extracts, finished product is made.
Further, the soil supension is to scalp topsoil by selecting fertile soil, take soil, spare, will
The soil for scalping topsoil is put into sterile water and has in the triangular flask of bead, and setting shaking table oscillation 3-8min divides soil sample uniformly
Being dispersed in dilution becomes soil supension.
It further, is HSAF containing Fungicidal active substance in the Fungicidal active substance crude extract.
Further, the Fungicidal active substance crude extract preparation method the following steps are included:
1) Lysobacter enzymogenes strain OH11, is taken, in streak inoculation to LB solid plate, activation culture under the conditions of 28 DEG C
24-36h；
2) it, is inoculated into the triangular flask containing LB liquid medium using one single colonie of sterilizing toothpick picking, 28
Shake culture 16-24h under the conditions of DEG C, oscillation frequency 150r/min；
3), by above-mentioned steps 2) resulting seed liquor with the ratio that volume ratio is 2% be inoculated into 10%TSB culture medium into
Row fermentation, fermentation temperature are 28 DEG C, per minute by the ratio between the volume of air of fermentation liquid and fermentating liquid volume for 0.5-2.0:1
Mode, carry out fermentation ventilation, mixing speed 80-160r/min, fermentation time 72h；
4) fermentation liquid is filtered using double gauze, boils 10min, then 6000r/min is centrifuged 15 minutes and takes supernatant,
HSAF content in fermentation supernatant is measured with HPLC, and the concentration of HSAF is adjusted to 80mg/L using pure water, obtains bactericidal activity
Substance crude extract.
The invention also discloses a kind of preparation methods of above-mentioned complex microbial inoculum, comprising the following steps:
By composite humic acid, soil supension, copper sulphate, Fungicidal active substance crude extract, azone, jinggangmycin, glycerol, blueness
Mycin, planting spirit, ABT root-inducing powder are dissolved with water indoors in proportion respectively, are then poured into container in order and are set shaking table
Oscillation 5-10min mixes each component uniformly, and the sequence is successively: composite humic acid, soil supension, azone, plant
Withered spirit, ABT root-inducing powder, Fungicidal active substance crude extract, glycerol, penicillin, jinggangmycin, copper sulphate, room temperature are controlled at 16 DEG C
More than.
Compared with prior art, the beneficial effects obtained by the present invention are as follows being:
1), Fungicidal active substance crude extract of the present invention is a kind of new bio sterilization of prevention and control pear tree root knot
Agent, principle active component HSAF, HSAF are a kind of Macrocyclic lactams of structure containing tetra-atomic acid, molecular formula C29H39O6N2, tool
There is efficient, wide spectrum bactericidal activity, there is protection and therapeutic effect, by interfering the polar growth of hypha,hyphae, inhibits production spore
And spore germination, cause eucaryotic cell structure that plasmolysis occurs, its resistivity to environment-stress is reduced, to influence germ
It survives, infect and propagates.
2), the present invention is compound humic acid made from raw material using green plantss caulomes such as corn stover, wheat straw, rice straws,
There is nutrition, healthcare function to pear tree, and pear tree physiological function, strengthen immunity can be activated.
3), contain azone in the present invention, azone can promote fruit tree absorption of drugs as a kind of bleeding agent, improve medicine
Effect.
4), planting spirit can further improve pear tree immunity in the present invention, and ABT root-inducing powder mainly promotes Growth of Pear Trees
New root, substitution are corroded dead old root by root nodule.
To sum up, the present invention made of complex microbial inoculum, proportion rationally, production method is simple, by various ingredients with
And the interaction of root nodule germ, pear tree root nodule germ can be killed, soil texture is improved, nutrient needed for improving plant growth,
Recurrence rate is low, and therapeutic effect is significant, can effectively promote Growth of Pear Trees development, improve the resistance of pear tree, reduce agriculture chemical
Use, improve the yield and quality of agricultural product, reduce environmental pollution, have very high economic benefit, ecological benefits and society
Benefit, using having a bright future.
Specific embodiment
It is clearly and completely described below in conjunction with the technical solution in the embodiment of the present invention, it is clear that described reality
Applying example is only a part of the embodiment of the present invention, instead of all the embodiments.Based on the embodiments of the present invention, this field is general
Logical technical staff every other embodiment obtained without making creative work belongs to what the present invention protected
Range.
Embodiment 1:
It is a kind of for preventing and treating the complex microbial inoculum of pear tree root knot, each component in the complex microbial inoculum
Weight percent composition content is as follows:
Composite humic acid 15%, soil supension 10%, copper sulphate 5%%, Fungicidal active substance slightly mention 7%, azone 5%,
Jinggangmycin 8%, glycerol 10%, penicillin 2%, planting spirit 2%, ABT root-inducing powder 0.05%, surplus are water.
The preparation method of above-mentioned complex microbial inoculum, comprising the following steps:
By composite humic acid, soil supension, copper sulphate, Fungicidal active substance crude extract, azone, jinggangmycin, glycerol, blueness
Mycin, planting spirit, ABT root-inducing powder are dissolved with water indoors in proportion respectively, are then poured into container in order and are set shaking table
Oscillation 5min mixes each component uniformly, and the sequence is successively: composite humic acid, soil supension, azone, plant are withered
Spirit, ABT root-inducing powder, Fungicidal active substance crude extract, glycerol, penicillin, jinggangmycin, copper sulphate, room temperature control 16 DEG C with
On.
In the present embodiment, the compound humic acid is to utilize a variety of green crops such as corn stover, wheat straw, rice straw
Stem be that raw material is crushed, after mixing treatment, sterilized, fermented, carrying out solid fermentation rotating in big tank, then being carried out
Finished product is made in separation and Extraction.
In the present embodiment, the soil supension is to scalp topsoil by selecting fertile soil, take soil, standby
With, by the soil for scalping topsoil be put into sterile water and with bead triangular flask in, set shaking table oscillation 3min keep soil sample equal
Even be dispersed in dilution becomes soil supension.
It in the present embodiment, is HSAF containing Fungicidal active substance in the Fungicidal active substance crude extract.
In the present embodiment, the Fungicidal active substance crude extract preparation method the following steps are included:
1) Lysobacter enzymogenes strain OH11, is taken, in streak inoculation to LB solid plate, activation culture under the conditions of 28 DEG C
24-36h；
2) it, is inoculated into the triangular flask containing LB liquid medium using one single colonie of sterilizing toothpick picking, 28
Shake culture 16h under the conditions of DEG C, oscillation frequency 150r/min；
3), by above-mentioned steps 2) resulting seed liquor with the ratio that volume ratio is 2% be inoculated into 10%TSB culture medium into
Row fermentation, fermentation temperature are 28 DEG C, to pass through the ratio between volume of air and fermentating liquid volume of fermentation liquid per minute as the side of 0.9:1
Formula carries out fermentation ventilation, mixing speed 120r/min, fermentation time 72h；
4) fermentation liquid is filtered using double gauze, boils 10min, then 6000r/min is centrifuged 15 minutes and takes supernatant,
HSAF content in fermentation supernatant is measured with HPLC, and the concentration of HSAF is adjusted to 80mg/L using pure water, obtains bactericidal activity
Substance crude extract.
The present embodiment method of administration: by the present embodiment drug dilution according to drug: water is 1:200 times of liquid and pours into bretschneider pear root
Portion can spray Pears assisting in preventing and treating on a small quantity, 2 kilograms every plant, monthly be administered 1 time, 15 days first meeting effects when disease is serious,
Being administered 1-3 times depending on the state of an illness can cure.
Embodiment 2:
It is a kind of for preventing and treating the complex microbial inoculum of pear tree root knot, each component in the complex microbial inoculum
Weight percent composition content is as follows:
Composite humic acid 25%, soil supension 10%, copper sulphate 15%, Fungicidal active substance crude extract 3%, azone 8%,
Jinggangmycin 7%, glycerol 10%, penicillin 3%, planting spirit 3%, ABT root-inducing powder 0.01%, surplus are water.
The preparation method of above-mentioned complex microbial inoculum, comprising the following steps:
By composite humic acid, soil supension, copper sulphate, Fungicidal active substance crude extract, azone, jinggangmycin, glycerol, blueness
Mycin, planting spirit, ABT root-inducing powder are dissolved with water indoors in proportion respectively, are then poured into container in order and are set shaking table
Oscillation 7min mixes each component uniformly, and the sequence is successively: composite humic acid, soil supension, azone, plant are withered
Spirit, ABT root-inducing powder, Fungicidal active substance crude extract, glycerol, penicillin, jinggangmycin, copper sulphate, room temperature control 16 DEG C with
On.
In the present embodiment, the compound humic acid is to utilize a variety of green crops such as corn stover, wheat straw, rice straw
Stem be that raw material is crushed, after mixing treatment, sterilized, fermented, carrying out solid fermentation rotating in big tank, then being carried out
Finished product is made in separation and Extraction.
In the present embodiment, the soil supension is to scalp topsoil by selecting fertile soil, take soil, standby
With, by the soil for scalping topsoil be put into sterile water and with bead triangular flask in, set shaking table oscillation 4min keep soil sample equal
Even be dispersed in dilution becomes soil supension.
It in the present embodiment, is HSAF containing Fungicidal active substance in the Fungicidal active substance crude extract.
In the present embodiment, the Fungicidal active substance crude extract preparation method the following steps are included:
1) Lysobacter enzymogenes strain OH11, is taken, in streak inoculation to LB solid plate, activation culture under the conditions of 28 DEG C
24h；
2) it, is inoculated into the triangular flask containing LB liquid medium using one single colonie of sterilizing toothpick picking, 28
Shake culture 20h under the conditions of DEG C, oscillation frequency 150r/min；
3), by above-mentioned steps 2) resulting seed liquor with the ratio that volume ratio is 2% be inoculated into 10%TSB culture medium into
Row fermentation, fermentation temperature are 28 DEG C, to pass through the ratio between volume of air and fermentating liquid volume of fermentation liquid per minute as the side of 2.0:1
Formula carries out fermentation ventilation, mixing speed 100r/min, fermentation time 72h；
4) fermentation liquid is filtered using double gauze, boils 10min, then 6000r/min is centrifuged 15 minutes and takes supernatant,
HSAF content in fermentation supernatant is measured with HPLC, and the concentration of HSAF is adjusted to 80mg/L using pure water, obtains bactericidal activity
Substance crude extract.
The present embodiment applies method: by the present embodiment drug dilution according to drug: water is 1:300 times of liquid and pours into pear tree
Root can spray Pears assisting in preventing and treating on a small quantity, 1.5 kilograms every plant, monthly be administered 1 time, first meeting in 12 days when disease is serious
Effect, being administered 1-3 times depending on the state of an illness can cure.
Embodiment 3:
It is a kind of for preventing and treating the complex microbial inoculum of pear tree root knot, each component in the complex microbial inoculum
Weight percent composition content is as follows:
Composite humic acid 10%, soil supension 20%, copper sulphate 15%, Fungicidal active substance crude extract 10%, azone
3%, jinggangmycin 7%, glycerol 5%, penicillin 3%, planting spirit 1%, ABT root-inducing powder 0.1%, surplus are water.
The preparation method of above-mentioned complex microbial inoculum, comprising the following steps:
By composite humic acid, soil supension, copper sulphate, Fungicidal active substance crude extract, azone, jinggangmycin, glycerol, blueness
Mycin, planting spirit, ABT root-inducing powder are dissolved with water indoors in proportion respectively, are then poured into container in order and are set shaking table
Oscillation 7min mixes each component uniformly, and the sequence is successively: composite humic acid, soil supension, azone, plant are withered
Spirit, ABT root-inducing powder, Fungicidal active substance crude extract, glycerol, penicillin, jinggangmycin, copper sulphate, room temperature control 16 DEG C with
On.
In the present embodiment, the compound humic acid is to utilize a variety of green crops such as corn stover, wheat straw, rice straw
Stem be that raw material is crushed, after mixing treatment, sterilized, fermented, carrying out solid fermentation rotating in big tank, then being carried out
Finished product is made in separation and Extraction.
In the present embodiment, the soil supension is to scalp topsoil by selecting fertile soil, take soil, standby
With, by the soil for scalping topsoil be put into sterile water and with bead triangular flask in, set shaking table oscillation 8min keep soil sample equal
Even be dispersed in dilution becomes soil supension.
It in the present embodiment, is HSAF containing Fungicidal active substance in the Fungicidal active substance crude extract.
In the present embodiment, the Fungicidal active substance crude extract preparation method the following steps are included:
1) Lysobacter enzymogenes strain OH11, is taken, in streak inoculation to LB solid plate, activation culture under the conditions of 28 DEG C
24-36h；
2) it, is inoculated into the triangular flask containing LB liquid medium using one single colonie of sterilizing toothpick picking, 28
Under the conditions of DEG C shake culture for 24 hours, oscillation frequency 150r/min；
3), by above-mentioned steps 2) resulting seed liquor with the ratio that volume ratio is 2% be inoculated into 10%TSB culture medium into
Row fermentation, fermentation temperature are 28 DEG C, to pass through the ratio between volume of air and fermentating liquid volume of fermentation liquid per minute as the side of 0.5:1
Formula carries out fermentation ventilation, mixing speed 160r/min, fermentation time 72h；
4) fermentation liquid is filtered using double gauze, boils 10min, then 6000r/min is centrifuged 15 minutes and takes supernatant,
HSAF content in fermentation supernatant is measured with HPLC, and the concentration of HSAF is adjusted to 80mg/L using pure water, obtains bactericidal activity
Substance crude extract.
The method of administration of the present embodiment: by the present embodiment drug dilution according to drug: water is 1:400 times of liquid and pours into pear tree
Root can spray Pears assisting in preventing and treating on a small quantity, 1 kilogram every plant, take effect at the beginning of being monthly administered 1 time, 10 days when disease is serious
Fruit, being administered depending on the state of an illness can cure for 1-3 time, if root nodule in 5cm or more, erect year again irrigation liquid three times after can fully recover.
Complex microbial inoculum made of the present invention, rationally, production method is simple, passes through various ingredients and root nodule for proportion
The interaction of germ can kill pear tree root nodule germ, improve soil texture, nutrient needed for improving plant growth, recurrence rate
Low, therapeutic effect is significant, can effectively promote Growth of Pear Trees development, improve the resistance of pear tree, reduce the use of agriculture chemical,
The yield and quality for improving agricultural product, reduces environmental pollution, and has very high economic benefit, ecological benefits and social benefit, answers
With having a bright future.
The complex microbial inoculum through present inventor for many years repeatedly planting experiment, test, adjust raw material each component
Ratio achieves effect of the present invention.
Finally, it should be noted that the foregoing is only a preferred embodiment of the present invention, it is not intended to restrict the invention,
Although the present invention is described in detail referring to the foregoing embodiments, for those skilled in the art, still may be used
To modify the technical solutions described in the foregoing embodiments or equivalent replacement of some of the technical features.
All within the spirits and principles of the present invention, any modification, equivalent replacement, improvement and so on should be included in of the invention
Within protection scope.
Claims (6)
1. a kind of for preventing and treating the complex microbial inoculum of pear tree root knot, it is characterised in that: in the complex microbial inoculum
Each component weight percent composition content it is as follows:
Composite humic acid 10%-25%, soil supension 5%-20%, copper sulphate 5%-15%, Fungicidal active substance crude extract
3%-10%, azone 3%-8%, jinggangmycin 7%-8%, glycerol 5%-10%, penicillin 1%-3%, planting spirit 1%-
3%, ABT root-inducing powder 0.01%-0.1%, surplus are water.
2. according to claim 1 a kind of for preventing and treating the complex microbial inoculum of pear tree root knot, it is characterised in that: institute
It is that raw material is crushed, mixed that the compound humic acid stated, which is using the stem of a variety of green crops such as corn stover, wheat straw, rice straw,
It after material processing, sterilized, fermented, carrying out solid fermentation in the big tank of rotation, then carrying out separation and Extraction, finished product is made.
3. according to claim 1 a kind of for preventing and treating the complex microbial inoculum of pear tree root knot, it is characterised in that: institute
The soil supension stated is to scalp topsoil by selecting fertile soil, take soil, spare, and the soil for scalping topsoil is put
Enter sterile water and have in the triangular flask of bead, setting shaking table oscillation 3-8min is dispersed in soil sample in dilution as soil
Earth suspension.
4. according to claim 1 a kind of for preventing and treating the complex microbial inoculum of pear tree root knot, it is characterised in that: institute
Containing Fungicidal active substance in the Fungicidal active substance crude extract stated is HSAF.
5. according to claim 4 a kind of for preventing and treating the complex microbial inoculum of pear tree root knot, it is characterised in that: institute
State the preparation method of Fungicidal active substance crude extract the following steps are included:
1) Lysobacter enzymogenes strain OH11, is taken, in streak inoculation to LB solid plate, activation culture 24- under the conditions of 28 DEG C
36h；
2) it, is inoculated into the triangular flask containing LB liquid medium using one single colonie of sterilizing toothpick picking, 28 DEG C of items
Shake culture 16-24h under part, oscillation frequency 150r/min；
3), by above-mentioned steps 2) resulting seed liquor is inoculated into 10%TSB culture medium with the ratio that volume ratio is 2% and sent out
Ferment, fermentation temperature are 28 DEG C, to pass through the ratio between volume of air and fermentating liquid volume of fermentation liquid per minute as the side of 0.5-2.0:1
Formula carries out fermentation ventilation, mixing speed 80-160r/min, fermentation time 72h；
4) fermentation liquid is filtered using double gauze, boils 10min, then 6000r/min is centrifuged 15 minutes and takes supernatant, uses
HPLC measures HSAF content in fermentation supernatant, and the concentration of HSAF is adjusted to 80mg/L using pure water, obtains bactericidal activity object
Matter crude extract.
6. it is according to claim 1-5 a kind of for preventing and treating the complex microbial inoculum of pear tree root knot, it is special
Sign is: the preparation method of the complex microbial inoculum, comprising the following steps:
By composite humic acid, soil supension, copper sulphate, Fungicidal active substance crude extract, azone, jinggangmycin, glycerol, mould
Element, planting spirit, ABT root-inducing powder are dissolved with water indoors in proportion respectively, are then poured into container in order and are set shaking table vibration
Swinging 5-10min mixes each component uniformly, and the sequence is successively: composite humic acid, soil supension, azone, plant are withered
Spirit, ABT root-inducing powder, Fungicidal active substance crude extract, glycerol, penicillin, jinggangmycin, copper sulphate, room temperature control 16 DEG C with
On.
Priority Applications (1)
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Cited By (1)
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| CN113994837A (en) * | 2021-09-07 | 2022-02-01 | 重庆三峡农业科学院 | Efficient prevention and control method for prunus salicina linn root tumor disease |
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