CN108485990A - One kind having prevention and control apple tree root rot microbial bacterial agent and preparation method thereof - Google Patents
One kind having prevention and control apple tree root rot microbial bacterial agent and preparation method thereof Download PDFInfo
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- CN108485990A CN108485990A CN201810287516.7A CN201810287516A CN108485990A CN 108485990 A CN108485990 A CN 108485990A CN 201810287516 A CN201810287516 A CN 201810287516A CN 108485990 A CN108485990 A CN 108485990A
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- 230000002265 prevention Effects 0.000 title claims abstract description 46
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- ZQIYJHBQRBBBRZ-UHFFFAOYSA-N nimbin Natural products COC(=O)CC1C2C(C(OC(=O)C)C3OC4CC(C(=C4C13C)C)c5cocc5)C(C)(C=CC2=O)C(=O)OC ZQIYJHBQRBBBRZ-UHFFFAOYSA-N 0.000 description 1
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- 235000010374 vitamin B1 Nutrition 0.000 description 1
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- 238000005303 weighing Methods 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- 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
-
- 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
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
- C12N11/02—Enzymes or microbial cells immobilised on or in an organic carrier
- C12N11/10—Enzymes or microbial cells immobilised on or in an organic carrier the carrier being a carbohydrate
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- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- Biochemistry (AREA)
- Mycology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Dentistry (AREA)
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- Pest Control & Pesticides (AREA)
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Abstract
The invention discloses one kind having prevention and control apple tree root rot microbial bacterial agent and preparation method thereof, is made from the raw material by following parts by weight：20 30 parts of trichoderma harzianum, 10 20 parts of bafillus natto, 15 25 parts of bacillus subtilis, 20 30 parts of Hansenula anomala, 5 15 parts of azotobacter chroococcum, 5 10 parts of porous magnetic chitosan microball.The present invention has prevention and control apple tree root rot microbial bacterial agent, completely not caused by the use because of chemical agent the problems such as chemical drug resistance, thus is conducive to the No-harmful apple orchard of apple.Complexity, stable structure, a function widely biological flora with varied bacterial micro-organism group are formed by mutual synergistic effect, Relationship With Proliferation, there is good prevention and control effect to apple tree root rot.
Description
Technical field
The invention belongs to biological fields, and in particular to one kind having prevention and control apple tree root rot microbial bacterial agent and its preparation
Method.
Background technology
Apple belongs to the rose family, is deciduous tree, there is stronger polarity, usually grows vigorous, and tree crown is tall and big, and tree is high reachable
It is 15 meters, general 3-5 meters or so high under cultivation condition.Trunk taupe, Lao Pi have irregular lobe or scale off, sprig light
It is sliding.According to surveying and determination, per 6.5-11.2 containing fructose grams of hectogram apple, 2.5-3.5 grams of glucose, 1.0-5.2 grams of sucrose also contains micro
Element zinc, calcium, phosphorus, iron, potassium and vitamin B1, vitamin B2, vitamin C and carrotene etc..Apple enjoys light, and happiness subacidity arrives
Neutral soil, be most suitable for soil layer it is deep, be the well-drained sand of ventilating rich in organic matter, cubsoil.
Armillariella root rot of apple also known as round spot root rot are a kind of root rots distributed more widely, that harm is heavier, and root rot is apple
In the case that root system is weak, caused by the sickle-like bacteria infringement of habitat in by soil.According to investigations, the generation of apple tree root rot and fruit
Tree root system power is closely related.Arid, fertilizer deficiency, the salinization of soil, soil erosion, soil hardening poor aeration, result are excessive, big
Off year is serious and pest and disease damage is all seriously the essential condition for inducing this disease.Apple tree root rot is cause the apple underproduction main
Reason.After main root is susceptible, early stage plant does not show symptom, after with butt rot degree aggravation, plant absorb moisture and support
The function of dividing gradually weakens, and supply falls short of demand for aerial part nutrient, and illumination is strong at noon, when evaporation capacity is big, plant upper leaf
Piece is wilted, but night can restore again.When being in a bad way, wilting situation can not be restored at night.At this point, root skin browning,
And detached with marrow, last complete stool is dead.Occurrent time：It generally mostly falls ill in late March to early April, enters morbidity May and contain
Phase.No matter children, in, veteran may all send out this disease.Existing control method usually has following several：Comprehensive preventive health measures：1, agriculture
Industry measure：1. selecting disease-resistant variety.2. crop rotation.3. blending sand in clay.4. fine site preparation, ditch cleaning draining, rainy days
The not upper furrow of water, ditch is without ponding.The low-lying ponding field of doubles, answers high ridge zanjon, reduces soil moisture in time.5. chemical fertilizer is used less, it is more
Use decomposed manure.2, physical measure：It in summer plowed soils, is covered with film, ground temperature is made to rise to 50 DEG C, disease is killed with high temperature
Bacterium.3, chemical measure：1. 3 grams of 1.5% gram of every square metre of 50% carbendazim of use or 75% pentachloronitrobenzene, are ploughed under in soil.②40%
Root-rot peaceful 1000 times or 80%402 missible oil, 1500 times of pouring roots.3. using chloropicrin fumigant, all bacteriums, true can be almost killed
Bacterium, virus, pest and mouse prevent a variety of soil-borne diseases, and prevention returns to stubble, improve native border environment.4, biological control measure：1. using
500 times of+3% nimbin of " magic square " board chitosan oligosaccharide citrate seed soaking, leaching bacterium.Anti- pathogen, virus, harmful fungoid, pest increase
Add the beneficial bacteriums flora such as lactic acid bacteria in soil, actinomyces, improve soil aggregate structure, enhance soil permeability, it is flat to restore ecology
Weighing apparatus.2. largely killing harmful fungoid, bacterium, virus using 1000 times of algae essence pouring roots, enhance soil fertility, promotes root system life
It is long.Although the above method has certain effect to treatment apple tree root rot, the effect is unsatisfactory.
Invention content
The present invention provides a kind of simple and practicable, excellent effect, free of contamination there is prevention and control apple tree root rot microorganism
Microbial inoculum.
The present invention also provides the preparation methods with prevention and control apple tree root rot microbial bacterial agent.
Technical scheme is as follows：
One kind having prevention and control apple tree root rot microbial bacterial agent, is made from the raw material by following parts by weight：Trichoderma harzianum
20-30 parts, 10-20 parts of bafillus natto, 15-25 parts of bacillus subtilis, 20-30 parts of Hansenula anomala, the brown fixed nitrogen of circle
5-15 parts of bacterium, 5-10 parts of porous magnetic chitosan microball.
Trichoderma harzianum, bafillus natto and the bacillus subtilis of the present invention is commercially available, living bacteria count 10-
10000000000/gram.
Hansenula anomala and azotobacter chroococcum used in the present invention is purchased from China General Microbiological culture presevation management
The heart（Address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica）, deposit number is respectively：
CGMCC No.3435 and CGMCC No.12402.
Described, porous magnetic chitosan microball is made from following steps：
1）Ammonium hydroxide, nano ferriferrous oxide, carboxymethyl chitosan and Tween-20 are uniformly mixed, sealed soak 2-4h, 50-55
DEG C heating water bath and stirring to carboxymethyl chitosan is completely dissolved, and it is static escaped completely to bubble, obtain carboxymethyl chitosan phase；
2）Span-80 and atoleine are added in flask, 50-60 DEG C of heating water bath, stirring is to being uniformly mixed, as paraffin
Phase；
3）Under 50-60 DEG C of condition of water bath heating, carboxymethyl chitosan is mutually added drop-wise in paraffin phase while stirring, be added dropwise after
Continuous stirring 5min；At the uniform velocity change 50-60 DEG C of water-bath into ice-water bath, ice-water bath is removed in 5-10min recession, and addition is mixed under room temperature
Solution is closed, 5-9h is reacted, stops stirring, is washed for several times with petroleum ether, absolute ethyl alcohol successively, vacuum drying, it is 5-10 μ to obtain grain size
The porous magnetic chitosan microball of m.
It is described, ammonium hydroxide, nano ferriferrous oxide, carboxymethyl chitosan and Tween-20 weight ratio be 100：5-15：
10-15：10-20；A concentration of the 20% of the ammonium hydroxide.
Described, the weight ratio of Span-80 and atoleine is 1：25-30.
Described, the weight ratio of carboxymethyl chitosan phase, paraffin phase and mixed solution is 1：6-8：0.4-0.6.
Described, mixed solution is by ethyl orthosilicate and absolute ethyl alcohol according to 1：What the weight ratio of 10-20 was mixed to get.
A kind of preparation method with prevention and control apple tree root rot microbial bacterial agent, is made from following steps：
1）The Hansenula anomala and each two ring of azotobacter chroococcum for taking inclined-plane conservation respectively are inoculated in beef extract-peptone training respectively
Base is supported, 37 DEG C, incubation time 20h of cultivation temperature obtains the seed liquor of Hansenula anomala and azotobacter chroococcum respectively；
2）The seed liquor of Hansenula anomala and azotobacter chroococcum is accessed by 5% inoculum concentration in 500L seed tank culture bases respectively,
35 DEG C, cultivate for 24 hours under the conditions of 180r/min, respectively Hansenula anomala and azotobacter chroococcum bacterium solution；
3）Respectively by the bacterium solution kieselguhr adsorption of Hansenula anomala and azotobacter chroococcum, plate and frame filter press pressure after the completion of absorption
Filter, spray drying, 140-180 DEG C of inlet air temperature, feed rate 13-17ml/min obtain Hansenula anomala agent and circle respectively
Brown fixed nitrogen bacteria agent；
4）By trichoderma harzianum, bafillus natto, bacillus subtilis, Hansenula anomala agent, azotobacter chroococcum microbial inoculum
It is uniformly mixed with porous magnetic chitosan microball, airslide disintegrating mill is added, fineness controls 300-400 mesh, you can is made with anti-
Control apple tree root rot microbial bacterial agent.
Described, the concrete composition of beef-protein medium：Beef extract 5g/L, peptone 10g/L, NaCl5g/L,
Agar 15g/L, remaining is water.
Described, the concrete composition of seed tank culture base：Beef extract 0.3%, peptone 1%, sodium chloride 0.5%, glucose
0.5%。
Beneficial effects of the present invention：
1. the present invention has prevention and control apple tree root rot microbial bacterial agent, do not change caused by the use because of chemical agent completely
The problems such as learning drug resistance, thus be conducive to the No-harmful apple orchard of apple.One is formed by mutual synergistic effect, Relationship With Proliferation
Complexity, stable structure, function widely have the biological flora of varied bacterial micro-organism group, have to apple tree root rot
There are good prevention and control to act on.Porous magnetic chitosan microball can provide habitat for microbial bacterial agent, enable microbial bacterial agent
The extraneous poor environment of tolerance.
2. the present invention has, prevention and control apple tree root rot microbial bacterial agent is simple and practicable, excellent effect, pollution-free, can be used for
Any growth period of apple tree, especially rest period, any resistance is not generated.
3. the manufacturing method of the present invention is simple and practicable, operating cost is low, has a vast market application prospect.
Specific implementation mode
The present invention is further elaborated in conjunction with the embodiments, does not constitute and the scope of the present invention is limited.
Trichoderma harzianum, bafillus natto and the bacillus subtilis of the present invention is commercially available, living bacteria count 10-
10000000000/gram.
Hansenula anomala and azotobacter chroococcum used in the present invention is purchased from China General Microbiological culture presevation management
The heart（Address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica）, deposit number is respectively：
CGMCC No.3435 and CGMCC No.12402.
Embodiment 1
One kind having prevention and control apple tree root rot microbial bacterial agent, is made from the raw material by following parts by weight：Trichoderma harzianum
20 parts, 10 parts of bafillus natto, 25 parts of bacillus subtilis, 30 parts of Hansenula anomala, 5 parts of azotobacter chroococcum, porous magnetic
10 parts of chitosan microball of property.
Described, porous magnetic chitosan microball is made from following steps：
1）Ammonium hydroxide, nano ferriferrous oxide, carboxymethyl chitosan and Tween-20 are uniformly mixed, sealed soak 2h, 55 DEG C of water-baths
Heat and stir to carboxymethyl chitosan and be completely dissolved, and it is static escaped completely to bubble, obtain carboxymethyl chitosan phase；
2）Span-80 and atoleine are added in flask, 50 DEG C of heating water baths, stirring is to being uniformly mixed, as paraffin phase；
3）Under 50 DEG C of condition of water bath heating, carboxymethyl chitosan is mutually added drop-wise in paraffin phase while stirring, continuation is added dropwise
Stir 5min；At the uniform velocity changing 50 DEG C of water-bath into ice-water bath, 10min recession removes ice-water bath, adds mixed solution under room temperature,
5h is reacted, stirring is stopped, being washed for several times with petroleum ether, absolute ethyl alcohol successively, vacuum drying obtains the porous magnetic that grain size is 5-10 μm
Property chitosan microball.
It is described, ammonium hydroxide, nano ferriferrous oxide, carboxymethyl chitosan and Tween-20 weight ratio be 100：5：10：
20；A concentration of the 20% of the ammonium hydroxide.
Described, the weight ratio of Span-80 and atoleine is 1：25.
Described, the weight ratio of carboxymethyl chitosan phase, paraffin phase and mixed solution is 1：6：0.6.
Described, mixed solution is by ethyl orthosilicate and absolute ethyl alcohol according to 1：What 10 weight ratio was mixed to get.
A kind of preparation method with prevention and control apple tree root rot microbial bacterial agent, is made from following steps：
1）The Hansenula anomala and each two ring of azotobacter chroococcum for taking inclined-plane conservation respectively are inoculated in beef extract-peptone training respectively
Base is supported, 37 DEG C, incubation time 20h of cultivation temperature obtains the seed liquor of Hansenula anomala and azotobacter chroococcum respectively；
2）The seed liquor of Hansenula anomala and azotobacter chroococcum is accessed by 5% inoculum concentration in 500L seed tank culture bases respectively,
35 DEG C, cultivate for 24 hours under the conditions of 180r/min, respectively Hansenula anomala and azotobacter chroococcum bacterium solution；
3）Respectively by the bacterium solution kieselguhr adsorption of Hansenula anomala and azotobacter chroococcum, plate and frame filter press pressure after the completion of absorption
Filter, spray drying, 140 DEG C of inlet air temperature, feed rate 17ml/min obtain Hansenula anomala agent and azotobacter chroococcum respectively
Microbial inoculum；
4）By trichoderma harzianum, bafillus natto, bacillus subtilis, Hansenula anomala agent, azotobacter chroococcum microbial inoculum
It is uniformly mixed with porous magnetic chitosan microball, airslide disintegrating mill is added, fineness controls 300-400 mesh, you can is made with anti-
Control apple tree root rot microbial bacterial agent.
Described, the concrete composition of beef-protein medium：Beef extract 5g/L, peptone 10g/L, NaCl5g/L,
Agar 15g/L, remaining is water.
Described, the concrete composition of seed tank culture base：Beef extract 0.3%, peptone 1%, sodium chloride 0.5%, glucose
0.5%。
There is prevention and control apple tree root rot microbial bacterial agent to be diluted with water 10 times the present embodiment, be uniformly mixed, is made micro-
Biological bacterium solution；By microbial bacteria liquid irrigating root, each tree pours 100 jin.The pouring root time is selected in apple tree root young fruit expanding stage（April-
May）It carries out, observation detection is carried out within 75 days after pouring root and is found, average every plant of apple tree increases by 18 new roots, every gram of chlorophyll content
It it is 7.69 milligrams, volume increase 9.4%, P in soil H values become 5.5 from 4.5 before non-pouring root, have prevention and control apple tree using the present embodiment
Root rot microbial bacterial agent handles apple tree root rot index by untreated 32%, drops to 16%, prevention and control apple tree root rot
Significant effect.
Embodiment 2
One kind having prevention and control apple tree root rot microbial bacterial agent, is made from the raw material by following parts by weight：Trichoderma harzianum
25 parts, 15 parts of bafillus natto, 20 parts of bacillus subtilis, 25 parts of Hansenula anomala, 10 parts of azotobacter chroococcum, porous magnetic
8 parts of chitosan microball of property.
Described, porous magnetic chitosan microball is made from following steps：
1）Ammonium hydroxide, nano ferriferrous oxide, carboxymethyl chitosan and Tween-20 are uniformly mixed, sealed soak 3h, 53 DEG C of water-baths
Heat and stir to carboxymethyl chitosan and be completely dissolved, and it is static escaped completely to bubble, obtain carboxymethyl chitosan phase；
2）Span-80 and atoleine are added in flask, 55 DEG C of heating water baths, stirring is to being uniformly mixed, as paraffin phase；
3）Under 55 DEG C of condition of water bath heating, carboxymethyl chitosan is mutually added drop-wise in paraffin phase while stirring, continuation is added dropwise
Stir 5min；At the uniform velocity changing 55 DEG C of water-bath into ice-water bath, 8min recession removes ice-water bath, adds mixed solution under room temperature,
7h is reacted, stirring is stopped, being washed for several times with petroleum ether, absolute ethyl alcohol successively, vacuum drying obtains the porous magnetic that grain size is 5-10 μm
Property chitosan microball.
It is described, ammonium hydroxide, nano ferriferrous oxide, carboxymethyl chitosan and Tween-20 weight ratio be 100：10：13：
15；A concentration of the 20% of the ammonium hydroxide.
Described, the weight ratio of Span-80 and atoleine is 1：28.
Described, the weight ratio of carboxymethyl chitosan phase, paraffin phase and mixed solution is 1：7：0.5.
Described, mixed solution is by ethyl orthosilicate and absolute ethyl alcohol according to 1：What 15 weight ratio was mixed to get.
A kind of preparation method with prevention and control apple tree root rot microbial bacterial agent, is made from following steps：
1）The Hansenula anomala and each two ring of azotobacter chroococcum for taking inclined-plane conservation respectively are inoculated in beef extract-peptone training respectively
Base is supported, 37 DEG C, incubation time 20h of cultivation temperature obtains the seed liquor of Hansenula anomala and azotobacter chroococcum respectively；
2）The seed liquor of Hansenula anomala and azotobacter chroococcum is accessed by 5% inoculum concentration in 500L seed tank culture bases respectively,
35 DEG C, cultivate for 24 hours under the conditions of 180r/min, respectively Hansenula anomala and azotobacter chroococcum bacterium solution；
3）Respectively by the bacterium solution kieselguhr adsorption of Hansenula anomala and azotobacter chroococcum, plate and frame filter press pressure after the completion of absorption
Filter, spray drying, 160 DEG C of inlet air temperature, feed rate 15ml/min obtain Hansenula anomala agent and azotobacter chroococcum respectively
Microbial inoculum；
4）By trichoderma harzianum, bafillus natto, bacillus subtilis, Hansenula anomala agent, azotobacter chroococcum microbial inoculum
It is uniformly mixed with porous magnetic chitosan microball, airslide disintegrating mill is added, fineness controls 300-400 mesh, you can is made with anti-
Control apple tree root rot microbial bacterial agent.
Described, the concrete composition of beef-protein medium：Beef extract 5g/L, peptone 10g/L, NaCl5g/L,
Agar 15g/L, remaining is water.
Described, the concrete composition of seed tank culture base：Beef extract 0.3%, peptone 1%, sodium chloride 0.5%, glucose
0.5%。
There is prevention and control apple tree root rot microbial bacterial agent to be diluted with water 10 times the present embodiment, be uniformly mixed, is made micro-
Biological bacterium solution；By microbial bacteria liquid irrigating root, each tree pours 100 jin.The pouring root time is selected in apple tree root young fruit expanding stage（April-
May）It carries out, observation detection is carried out within 75 days after pouring root and is found, average every plant of apple tree increases by 16 new roots, every gram of chlorophyll content
It it is 7.71 milligrams, volume increase 9.1%, P in soil H values become 5.4 from 4.5 before non-pouring root, have prevention and control apple tree using the present embodiment
Root rot microbial bacterial agent handles apple tree root rot index by untreated 30%, drops to 16%, prevention and control apple tree root rot
Significant effect.
Embodiment 3
One kind having prevention and control apple tree root rot microbial bacterial agent, is made from the raw material by following parts by weight：Trichoderma harzianum
30 parts, 20 parts of bafillus natto, 15 parts of bacillus subtilis, 20 parts of Hansenula anomala, 15 parts of azotobacter chroococcum, porous magnetic
5 parts of chitosan microball of property.
Described, porous magnetic chitosan microball is made from following steps：
1）Ammonium hydroxide, nano ferriferrous oxide, carboxymethyl chitosan and Tween-20 are uniformly mixed, sealed soak 4h, 50 DEG C of water-baths
Heat and stir to carboxymethyl chitosan and be completely dissolved, and it is static escaped completely to bubble, obtain carboxymethyl chitosan phase；
2）Span-80 and atoleine are added in flask, 60 DEG C of heating water baths, stirring is to being uniformly mixed, as paraffin phase；
3）Under 60 DEG C of condition of water bath heating, carboxymethyl chitosan is mutually added drop-wise in paraffin phase while stirring, continuation is added dropwise
Stir 5min；At the uniform velocity changing 60 DEG C of water-bath into ice-water bath, 5min recession removes ice-water bath, adds mixed solution under room temperature,
9h is reacted, stirring is stopped, being washed for several times with petroleum ether, absolute ethyl alcohol successively, vacuum drying obtains the porous magnetic that grain size is 5-10 μm
Property chitosan microball.
It is described, ammonium hydroxide, nano ferriferrous oxide, carboxymethyl chitosan and Tween-20 weight ratio be 100：15：15：
10；A concentration of the 20% of the ammonium hydroxide.
Described, the weight ratio of Span-80 and atoleine is 1：30.
Described, the weight ratio of carboxymethyl chitosan phase, paraffin phase and mixed solution is 1：8：0.4.
Described, mixed solution is by ethyl orthosilicate and absolute ethyl alcohol according to 1：What 20 weight ratio was mixed to get.
A kind of preparation method with prevention and control apple tree root rot microbial bacterial agent, is made from following steps：
1）The Hansenula anomala and each two ring of azotobacter chroococcum for taking inclined-plane conservation respectively are inoculated in beef extract-peptone training respectively
Base is supported, 37 DEG C, incubation time 20h of cultivation temperature obtains the seed liquor of Hansenula anomala and azotobacter chroococcum respectively；
2）The seed liquor of Hansenula anomala and azotobacter chroococcum is accessed by 5% inoculum concentration in 500L seed tank culture bases respectively,
35 DEG C, cultivate for 24 hours under the conditions of 180r/min, respectively Hansenula anomala and azotobacter chroococcum bacterium solution；
3）Respectively by the bacterium solution kieselguhr adsorption of Hansenula anomala and azotobacter chroococcum, plate and frame filter press pressure after the completion of absorption
Filter, spray drying, 180 DEG C of inlet air temperature, feed rate 13ml/min obtain Hansenula anomala agent and azotobacter chroococcum respectively
Microbial inoculum；
4）By trichoderma harzianum, bafillus natto, bacillus subtilis, Hansenula anomala agent, azotobacter chroococcum microbial inoculum
It is uniformly mixed with porous magnetic chitosan microball, airslide disintegrating mill is added, fineness controls 300-400 mesh, you can is made with anti-
Control apple tree root rot microbial bacterial agent.
Described, the concrete composition of beef-protein medium：Beef extract 5g/L, peptone 10g/L, NaCl5g/L,
Agar 15g/L, remaining is water.
Described, the concrete composition of seed tank culture base：Beef extract 0.3%, peptone 1%, sodium chloride 0.5%, glucose
0.5%。
There is prevention and control apple tree root rot microbial bacterial agent to be diluted with water 10 times the present embodiment, be uniformly mixed, is made micro-
Biological bacterium solution；By microbial bacteria liquid irrigating root, each tree pours 100 jin.The pouring root time is selected in apple tree root young fruit expanding stage（April-
May）It carries out, observation detection is carried out within 75 days after pouring root and is found, average every plant of apple tree increases by 15 new roots, every gram of chlorophyll content
It it is 7.62 milligrams, volume increase 9.3%, P in soil H values become 5.3 from 4.5 before non-pouring root, have prevention and control apple tree using the present embodiment
Root rot microbial bacterial agent handles apple tree root rot index by untreated 32%, drops to 18%, prevention and control apple tree root rot
Significant effect.
Comparative example 1
One kind having prevention and control apple tree root rot microbial bacterial agent, is made from the raw material by following parts by weight：Trichoderma harzianum
20 parts, 10 parts of bafillus natto, 25 parts of bacillus subtilis, 30 parts of Hansenula anomala, 5 parts of azotobacter chroococcum.
Remaining is the same as embodiment 1.
There is prevention and control apple tree root rot microbial bacterial agent to be diluted with water 10 times the present embodiment, be uniformly mixed, is made micro-
Biological bacterium solution；By microbial bacteria liquid irrigating root, each tree pours 100 jin.The pouring root time is selected in apple tree root young fruit expanding stage（April-
May）It carries out, observation detection is carried out within 75 days after pouring root and is found, average every plant of apple tree increases by 10 new roots, every gram of chlorophyll content
It it is 7.21 milligrams, volume increase 5.5%, P in soil H values become 4.9 from 4.5 before non-pouring root, have prevention and control apple tree using the present embodiment
Root rot microbial bacterial agent handles apple tree root rot index by untreated 31%, drops to 26%, prevention and control apple tree root rot
Effect is general.
Comparative example 2
One kind having prevention and control apple tree root rot microbial bacterial agent, is made from the raw material by following parts by weight：Trichoderma harzianum
20 parts, 10 parts of bafillus natto, 25 parts of bacillus subtilis, 10 parts of porous magnetic chitosan microball.
Remaining is the same as embodiment 1.
There is prevention and control apple tree root rot microbial bacterial agent to be diluted with water 10 times the present embodiment, be uniformly mixed, is made micro-
Biological bacterium solution；By microbial bacteria liquid irrigating root, each tree pours 100 jin.The pouring root time is selected in apple tree root young fruit expanding stage（April-
May）It carries out, observation detection is carried out within 75 days after pouring root and is found, average every plant of apple tree increases by 7 new roots, every gram of chlorophyll content
It it is 6.56 milligrams, volume increase 4.7%, P in soil H values become 4.8 from 4.5 before non-pouring root, have prevention and control apple tree using the present embodiment
Root rot microbial bacterial agent handles apple tree root rot index by untreated 32%, drops to 27%, prevention and control apple tree root rot
Effect is general.
The above is only a preferred embodiment of the present invention, it should be pointed out that：For the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (9)
1. one kind having prevention and control apple tree root rot microbial bacterial agent, which is characterized in that be made by the raw material of following parts by weight
's：20-30 parts of trichoderma harzianum, 10-20 parts of bafillus natto, 15-25 parts of bacillus subtilis, Hansenula anomala 20-
30 parts, 5-15 parts of azotobacter chroococcum, 5-10 parts of porous magnetic chitosan microball.
2. having prevention and control apple tree root rot microbial bacterial agent according to claim 1, which is characterized in that the porous magnetic
Chitosan microball is made from following steps：
1）Ammonium hydroxide, nano ferriferrous oxide, carboxymethyl chitosan and Tween-20 are uniformly mixed, sealed soak 2-4h, 50-55
DEG C heating water bath and stirring to carboxymethyl chitosan is completely dissolved, and it is static escaped completely to bubble, obtain carboxymethyl chitosan phase；
2）Span-80 and atoleine are added in flask, 50-60 DEG C of heating water bath, stirring is to being uniformly mixed, as paraffin
Phase；
3）Under 50-60 DEG C of condition of water bath heating, carboxymethyl chitosan is mutually added drop-wise in paraffin phase while stirring, be added dropwise after
Continuous stirring 5min；At the uniform velocity change 50-60 DEG C of water-bath into ice-water bath, ice-water bath is removed in 5-10min recession, and addition is mixed under room temperature
Solution is closed, 5-9h is reacted, stops stirring, is washed for several times with petroleum ether, absolute ethyl alcohol successively, vacuum drying, it is 5-10 μ to obtain grain size
The porous magnetic chitosan microball of m.
3. having prevention and control apple tree root rot microbial bacterial agent according to claim 2, which is characterized in that the ammonium hydroxide is received
The weight ratio of rice ferroso-ferric oxide, carboxymethyl chitosan and Tween-20 is 100：5-15：10-15：10-20；The ammonium hydroxide it is dense
Degree is 20%.
4. having prevention and control apple tree root rot microbial bacterial agent according to claim 2, which is characterized in that the Span-80
Weight ratio with atoleine is 1：25-30.
5. having prevention and control apple tree root rot microbial bacterial agent according to claim 2, which is characterized in that the carboxymethyl shell
The weight ratio of glycan phase, paraffin phase and mixed solution is 1：6-8：0.4-0.6.
6. having prevention and control apple tree root rot microbial bacterial agent according to claim 2, which is characterized in that the mixed solution
It is by ethyl orthosilicate and absolute ethyl alcohol according to 1：What the weight ratio of 10-20 was mixed to get.
7. the preparation method with prevention and control apple tree root rot microbial bacterial agent described in a kind of any one of claim 1-6,
It is characterized in that, is made from following steps：
1）The Hansenula anomala and each two ring of azotobacter chroococcum for taking inclined-plane conservation respectively are inoculated in beef extract-peptone training respectively
Base is supported, 37 DEG C, incubation time 20h of cultivation temperature obtains the seed liquor of Hansenula anomala and azotobacter chroococcum respectively；
2）The seed liquor of Hansenula anomala and azotobacter chroococcum is accessed by 5% inoculum concentration in 500L seed tank culture bases respectively,
35 DEG C, cultivate for 24 hours under the conditions of 180r/min, respectively Hansenula anomala and azotobacter chroococcum bacterium solution；
3）Respectively by the bacterium solution kieselguhr adsorption of Hansenula anomala and azotobacter chroococcum, plate and frame filter press pressure after the completion of absorption
Filter, spray drying, 140-180 DEG C of inlet air temperature, feed rate 13-17ml/min obtain Hansenula anomala agent and circle respectively
Brown fixed nitrogen bacteria agent；
4）By trichoderma harzianum, bafillus natto, bacillus subtilis, Hansenula anomala agent, azotobacter chroococcum microbial inoculum
It is uniformly mixed with porous magnetic chitosan microball, airslide disintegrating mill is added, fineness controls 300-400 mesh, you can is made with anti-
Control apple tree root rot microbial bacterial agent.
8. the preparation method with prevention and control apple tree root rot microbial bacterial agent according to claim 7, which is characterized in that institute
State the concrete composition of beef-protein medium：Beef extract 5g/L, peptone 10g/L, NaCl5g/L, agar 15g/L, remaining
For water.
9. the preparation method with prevention and control apple tree root rot microbial bacterial agent according to claim 7, which is characterized in that institute
State the concrete composition of seed tank culture base：Beef extract 0.3%, peptone 1%, sodium chloride 0.5%, glucose 0.5%.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109136155A (en) * | 2018-09-30 | 2019-01-04 | 山西永丰生物工程技术有限公司 | A kind of preparation method of microbial bacterial agent that preventing and treating fruit tree root rot |
| CN109287679A (en) * | 2018-11-23 | 2019-02-01 | 王变芝 | Biological agent and preparation method thereof for preventing and treating peanut root rot |
| CN109997875A (en) * | 2019-04-23 | 2019-07-12 | 山东中创亿丰肥料集团有限公司 | A kind of microbial bacterial agent and preparation method thereof with pest-resistant sterilizing function |
| CN111302864A (en) * | 2020-04-23 | 2020-06-19 | 史丹利农业集团股份有限公司 | Special fertilizer for apples and fertilizing method |
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| CN108485990B (en) | 2021-06-29 |
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