CN110477021A - The solid microbial and preparation method of a kind of pair of potato disease-preventing and yield-increasing - Google Patents
The solid microbial and preparation method of a kind of pair of potato disease-preventing and yield-increasing Download PDFInfo
- Publication number
- CN110477021A CN110477021A CN201910832008.7A CN201910832008A CN110477021A CN 110477021 A CN110477021 A CN 110477021A CN 201910832008 A CN201910832008 A CN 201910832008A CN 110477021 A CN110477021 A CN 110477021A
- Authority
- CN
- China
- Prior art keywords
- yield
- potato
- increasing
- preventing
- solid microbial
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- 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
-
- 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
- C12N1/04—Preserving or maintaining viable microorganisms
-
- 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
- C12N1/20—Bacteria; Culture media therefor
Abstract
The present invention relates to the solid microbial and preparation method of a kind of pair of potato disease-preventing and yield-increasing, the active constituent of the solid microbial to potato disease-preventing and yield-increasing is bacillus amyloliquefaciens PHODG36, additionally contains dispersing agent, wetting agent and filler.The invention also discloses the methods for preparing the microbial bacterial agent: fermentation prepares the bacillus amyloliquefaciens PHODG36 fermentation liquid, will prepare resulting fermentation liquid and other raw materials are sufficiently mixed uniformly to get the solid microbial in a manner of mechanical mixture.The invention also discloses the solid microbials to the specific effect in terms of prevention and treatment potato verticillium wilt and yield.The generation of potato verticillium wilt can be greatly lowered using solid microbial of the present invention, and the yield of potato can be significantly improved.
Description
Technical field
The invention belongs to technical field of microbe application, and in particular to the micro- life of the solid of a kind of pair of potato disease-preventing and yield-increasing
Object microbial inoculum and preparation method.
Background technique
Potato is the important cereal crops in China, is had great importance to Ensuring Food Safety.China is by Ma Ling
Potato is classified as the fourth-largest staple food grain after rice, wheat, corn, and potato is ensureing China's grain security, accurate poverty alleviation, kind
It plants in adjustment of agricultural stracture and agricultural industry transition and upgrade and plays vital effect.The cultivated area of China potato
It is ranked the first in the world with total output, but unit yield is because the restriction of the bottleneck factors such as pest and disease damage is lower than world average level.Closely
Nian Lai directly affects potato yield and quality since potato continuous cropping causes soil-borne disease to have the tendency that aggravating year by year,
Biggish economic loss is brought to Potato Industry.
Potato verticillium wilt (Verticillium wilt of potato) is also known as early dead disease or precocious disease, gradually at
For one of the Major Diseases in potato production, it is that simultaneous kind of typical soil biography passes vascular bundle diseases, causes potato early dead, gently
Person underproduction 20%-30%, severe one are lost up to 50% or more, and the yield and quality of potato is seriously affected.It is common in the art
Using the method for chemical bactericide prevention and treatment soil-borne disease, however existing chemical bactericide effect is single, and long-time service leads to cause of disease
The enhancing of bacterium resistance, while also will cause ecological environment and seriously polluting, directly increase the residual in agricultural product, to human health band
To seriously endanger.Microbial bactericide is with its environment friendly and adaptable is more and more closed in agricultural production
Note.Therefore, the microbial bactericide that targetedly development prevents and treats potato verticillium wilt has become urgent in agricultural production
Demand.
At this stage, correlative study and product of the microbial bactericide applied to potato verticillium wilt are less, and lack correct
The operation standard effectively prevented is directed to potato verticillium wilt safely and effectively microorganism so needing us and developing one kind
Microbial inoculum, to cope with the present market demand.
Summary of the invention
In order to solve the above problems existing in the present technology, potato disease-preventing and yield-increasing is consolidated the present invention provides a kind of
Body microbial bacterial agent, the active constituent of the solid microbial to potato disease-preventing and yield-increasing are bacillus amyloliquefaciens
PHODG36 additionally contains dispersing agent, wetting agent and filler, each material component is carried out appropriate weight proportion, finally
The solid microbial that there is prevention and treatment verticillium wilt and production-increasing function to potato is prepared.
Bacillus amyloliquefaciens PHODG36 of the present invention was preserved in China Microbiological bacterium on September 26th, 2016
Kind preservation administration committee common micro-organisms center, depositary institution address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 China
Institute of microbiology, the academy of sciences, deposit number are CGMCC No.13041；Classification system are as follows: Bacillus
amyloliquefaciens PHODG36.Patent (the application number that the bacterial strain has been announced on 2 15th, 2017
201610876803.2) it is disclosed, is not related to the preservation of the bacterial strain in the application.
The technical scheme adopted by the invention is as follows:
The solid microbial of a kind of pair of potato disease-preventing and yield-increasing, raw material components include:
Bacillus amyloliquefaciens PHODG36 10-80 parts by weight, auxiliary agent 2-16 parts by weight, filler 10-80 parts by weight.
Further, the auxiliary agent includes dispersing agent and wetting agent.
Further, the filler is one of calcium carbonate, soluble starch or talcum powder or a variety of mixtures.
Further, the mass ratio of the dispersing agent and wetting agent is (1-8): (1-8).
Further, the dispersing agent is lignin sulfonic acid sodium salt, sanlose, desugar lignin sulfonic acid
One of sodium, ethylene glycol monobutyl ether, polyoxyethanyl alkyl ether, neopelex or a variety of mixtures.
Further, the wetting agent is nekal, fatty alcohol polyoxyethylene ether, detergent alkylate sulphur
One of acid sodium-salt or a variety of mixtures.
Further, the dosage form of the solid microbial is wettable powder.
The solid microbial to potato disease-preventing and yield-increasing is made with the following method:
(1) the bacillus amyloliquefaciens PHODG36 bacterial strain frozen is gone into activation culture on LB solid medium, obtained
Activated strains；
(2) activated strains are seeded in LB liquid medium, carry out shaking table culture, obtains primary seed solution；
(3) primary seed solution is transferred in seed culture medium, is expanded culture under agitation, obtained
Secondary seed solution；
(4) secondary seed solution is forwarded in fermentation medium, carries out fermented and cultured under agitation, obtains
The bacillus amyloliquefaciens PHODG36 fermentation liquid；
(5) filler is added in the bacillus amyloliquefaciens PHODG36 fermentation liquid, is spray-dried, obtain dry bacterium
Powder；
(6) the dry bacterium powder mixed with the auxiliary agent, stirred, after air-flow crushing and is sieved, it is described right to obtain
The solid microbial of potato disease-preventing and yield-increasing.
Further, in step (1), the cultivation temperature is 36-38 DEG C, and the incubation time is 20-28h.
Further, in step (2), the cultivation temperature is 35-38 DEG C, and the incubation time is 20-24h, described to shake
Bed revolving speed is 150-200rpm.
Further, in step (3), the volume ratio of the primary seed solution and seed culture medium is (1-5): 100, institute
Stating cultivation temperature is 36-38 DEG C, and the incubation time is 18-22h, and the dissolved oxygen amount is 10%-20%, the mixing speed
For 170-190rpm, the obtained thallus in the secondary seed solution is logarithmic growth phase.
Further, in step (4), the volume ratio of the secondary seed solution and fermentation medium is (4-6): 100, institute
Stating cultivation temperature is 36-38 DEG C, and the incubation time is 24-32h, and the dissolved oxygen amount is 15%-18%, the mixing speed
For 170-190rpm.
Further, in step (4), thalline quantity >=6,000,000,000 in the bacillus amyloliquefaciens PHODG36 fermentation liquid/
ML, spore forming rate >=95%.
Further, in step (5), the inlet temperature of the spray drying is 150-200 DEG C, the spray drying
Outlet temperature is 50-90 DEG C.
Further, in step (6), the partial size of the solid microbial after the sieving is 300-350 mesh.
Further, the usage mode of the solid microbial to potato disease-preventing and yield-increasing be trickle irrigation, punching apply or
Seed dressing.
The invention has the benefit that
(1) solid microbial of the present invention to potato disease-preventing and yield-increasing, by using solution starch gemma bar
Bacterium PHODG36 enables the solid microbial to colonize plant rhizosphere rapidly as active constituent, by with Huang
The effects of competition of disease of withering pathogen infects site, secretion Substance and plant nutrient is promoted to absorb, farthest sends out
Microbial bacterial agent induction disease resistance of plant and the persistently ability of control disease are waved, is had to potato verticillium wilt pathogen relatively strong
Inhibiting effect, while can simultaneously significantly improve the yield of potato.
(2) solid microbial of the present invention can reduce the access times and dosage of chemical bactericide, reduce
Cost；Delay the generation of drug resistance, nontoxic, non agricultural chemical residuum is environmental-friendly；Ecological Distribution of Soil Microorganisms fauna is adjusted, promotes to make
Object growth, and there is production-increasing function；In the selection of dispersing agent and wetting agent, fully takes into account the antagonism between microorganism, assists
The requirement handled with seed potato in effect and production practices；Can be used for dressing seed, trickle irrigation and punching apply use, can be according to not
With situation reasonable employment, manpower and use cost are saved.
(3) preparation method of solid microbial of the present invention, by sending out bacillus amyloliquefaciens PHODG36
Zymotic fluid is prepared as solid microbial, reach bacterial content is high, can save at normal temperature and not easy in inactivation, reduce finished product packet
Dress up this, be easy to transport and other effects, solid microbial of the present invention, the total viable count of effective component between 1.5 ×
109-2.0×1010CFU/g, concentration with higher.The solid microbial preparation method is simple, at low cost, raw simultaneously
Object is highly-safe, is suitable for industrialized production.
Specific embodiment
To make the object, technical solutions and advantages of the present invention clearer, technical solution of the present invention will be carried out below
Detailed description.Obviously, described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.Base
Embodiment in the present invention, those of ordinary skill in the art obtained institute without making creative work
There is other embodiment, belongs to the range that the present invention is protected.
Embodiment 1
The present embodiment provides the solid microbial of a kind of pair of potato disease-preventing and yield-increasing, raw material composition includes:
Bacillus amyloliquefaciens PHODG36 10g, dispersing agent 1g, wetting agent 1g, talcum powder 10g；
Dispersing agent 1g, specially lignin sulfonic acid sodium salt 0.3g, sanlose 0.3g, desugar sulfomethylated lignin
Sour sodium 0.4g；
Wetting agent 1g, specially nekal 0.5g, fatty alcohol polyoxyethylene ether 0.5g.
The preparation method of the solid microbial to potato disease-preventing and yield-increasing, comprising the following steps:
(1) constituent of every liter of seed culture medium are as follows: corn flour 30g, yeast extract 3.5g, soybean powder 15g, phosphorus
Sour disodium hydrogen 2.0g, potassium dihydrogen phosphate 0.2g, calcium carbonate 2.0g, potassium chloride 0.5g, magnesium sulfate 0.5g, ammonium nitrate 1.50g, sulphur
Sour manganese 0.2g, organosilicon 0.3g, pH 7.0-7.5,121 DEG C of sterilizing 30min；The constituent of every liter of fermentation medium are as follows: beautiful
Rice flour 30g, yeast extract 3.5g, soybean powder 15g, disodium hydrogen phosphate 2.0g, potassium dihydrogen phosphate 0.2g, calcium carbonate 2.0g, chlorination
Potassium 0.5g, magnesium sulfate 0.5g, ammonium nitrate 1.50g, manganese sulfate 0.2g, organosilicon 0.3g, pH 7.0-7.5,121 DEG C of sterilizings
30min；
(2) the bacillus amyloliquefaciens PHODG36 bacterial strain frozen is gone on LB solid medium, is being cultivated at 36 DEG C
20h obtains activated strains；
(3) activated strains are seeded in LB liquid medium, 20h is cultivated at 35 DEG C, shaking speed is
150rpm obtains primary seed solution；
(4) primary seed solution is transferred in seed culture medium according to percent by volume for 1%, dissolved oxygen amount is
10%, mixing speed 170rpm, 36 DEG C of culture 18h to thallus are logarithmic growth phase, obtain secondary seed solution；
(5) secondary seed solution is forwarded in fermentation medium according to percent by volume for 4%, dissolved oxygen amount is
15%, mixing speed 170rpm, 36 DEG C are cultivated for 24 hours, until cell concentration >=6,000,000,000/mL, spore forming rate >=95% obtain
The bacillus amyloliquefaciens PHODG36 fermentation liquid；
(6) talcum powder is added in the bacillus amyloliquefaciens PHODG36 fermentation liquid and is spray-dried, import
Temperature is 150 DEG C, and outlet temperature is 50 DEG C, obtains dry bacterium powder；
(7) the dry bacterium powder mixed with the dispersing agent and wetting agent, stirred, after air-flow crushing and cross 300
Mesh obtains the solid microbial to potato disease-preventing and yield-increasing.
Embodiment 2
The present embodiment provides the solid microbial of a kind of pair of potato disease-preventing and yield-increasing, raw material composition includes:
Bacillus amyloliquefaciens PHODG36 80g, dispersing agent 8g, wetting agent 8g, soluble starch 80g；
Dispersing agent 8g, specially ethylene glycol monobutyl ether 3g, polyoxyethanyl alkyl ether 3g, neopelex 2g；
Wetting agent 8g, specially fatty alcohol polyoxyethylene ether 4g, pelopon A 4g.
The preparation method of the solid microbial to potato disease-preventing and yield-increasing, comprising the following steps:
(1) constituent of every liter of seed culture medium are as follows: corn flour 30g, yeast extract 3.5g, soybean powder 15g, phosphorus
Sour disodium hydrogen 2.0g, potassium dihydrogen phosphate 0.2g, calcium carbonate 2.0g, potassium chloride 0.5g, magnesium sulfate 0.5g, ammonium nitrate 1.50g, sulphur
Sour manganese 0.2g, organosilicon 0.3g, pH 7.0-7.5,121 DEG C of sterilizing 30min；The constituent of every liter of fermentation medium are as follows: beautiful
Rice flour 30g, yeast extract 3.5g, soybean powder 15g, disodium hydrogen phosphate 2.0g, potassium dihydrogen phosphate 0.2g, calcium carbonate 2.0g, chlorination
Potassium 0.5g, magnesium sulfate 0.5g, ammonium nitrate 1.50g, manganese sulfate 0.2g, organosilicon 0.3g, pH 7.0-7.5,121 DEG C of sterilizings
30min；
(2) the bacillus amyloliquefaciens PHODG36 bacterial strain frozen is gone on LB solid medium, is cultivated at 38 DEG C
28h obtains activated strains；
(3) activated strains are seeded in LB liquid medium, are cultivated at 38 DEG C for 24 hours, shaking speed is
200rpm obtains primary seed solution；
(4) primary seed solution is transferred in seed culture medium according to percent by volume for 5%, dissolved oxygen amount is
20%, mixing speed 190rpm, 38 DEG C of culture 22h to thallus are logarithmic growth phase, obtain secondary seed solution；
(5) secondary seed solution is forwarded in fermentation medium according to percent by volume for 6%, dissolved oxygen amount is
18%, mixing speed 190rpm, 38 DEG C of culture 32h, until cell concentration >=6,000,000,000/mL, spore forming rate >=95% obtain
The bacillus amyloliquefaciens PHODG36 fermentation liquid；
(6) soluble starch is added in the bacillus amyloliquefaciens PHODG36 fermentation liquid and is spray-dried, into
Mouth temperature is 200 DEG C, and outlet temperature is 90 DEG C, obtains dry bacterium powder；
(7) the dry bacterium powder mixed with the dispersing agent and wetting agent, stirred, after air-flow crushing and cross 350
Mesh obtains the solid microbial to potato disease-preventing and yield-increasing.
Embodiment 3
The present embodiment provides the solid microbial of a kind of pair of potato disease-preventing and yield-increasing, raw material composition includes:
Bacillus amyloliquefaciens PHODG36 45g, dispersing agent 4g, wetting agent 4g, calcium carbonate 45g；
Dispersing agent 4g, lignin sulfonic acid sodium salt 1g, sanlose 1g, desugar sodium lignin sulfonate 1g, second two
Alcohol single-butyl ether 0.2g, polyoxyethanyl alkyl ether 0.2g, neopelex 0.6g；
Wetting agent 4g, nekal 1.5g, fatty alcohol polyoxyethylene ether 1.5g, pelopon A
1g。
The preparation method of the solid microbial to potato disease-preventing and yield-increasing, comprising the following steps:
(1) constituent of every liter of seed culture medium are as follows: corn flour 30g, yeast extract 3.5g, soybean powder 15g, phosphorus
Sour disodium hydrogen 2.0g, potassium dihydrogen phosphate 0.2g, calcium carbonate 2.0g, potassium chloride 0.5g, magnesium sulfate 0.5g, ammonium nitrate 1.50g, sulphur
Sour manganese 0.2g, organosilicon 0.3g, pH 7.0-7.5,121 DEG C of sterilizing 30min；The constituent of every liter of fermentation medium are as follows: beautiful
Rice flour 30g, yeast extract 3.5g, soybean powder 15g, disodium hydrogen phosphate 2.0g, potassium dihydrogen phosphate 0.2g, calcium carbonate 2.0g, chlorination
Potassium 0.5g, magnesium sulfate 0.5g, ammonium nitrate 1.50g, manganese sulfate 0.2g, organosilicon 0.3g, pH 7.0-7.5,121 DEG C of sterilizings
30min；
(2) the bacillus amyloliquefaciens PHODG36 bacterial strain frozen is gone on LB solid medium, is cultivated at 37 DEG C
For 24 hours, activated strains are obtained；
(3) activated strains are seeded in LB liquid medium, 22h is cultivated at 36.5 DEG C, shaking speed is
175rpm obtains primary seed solution；
(4) primary seed solution is transferred in seed culture medium according to percent by volume for 3%, dissolved oxygen amount is
15%, mixing speed 180rpm, 37 DEG C of culture 20h obtain secondary seed solution until thallus is logarithmic growth phase；
(5) secondary seed solution is forwarded in fermentation medium according to percent by volume for 5%, dissolved oxygen amount is
16.5%, mixing speed 180rpm, 37 DEG C of culture 28h, until cell concentration >=6,000,000,000/mL, spore forming rate >=95% obtain
To the bacillus amyloliquefaciens PHODG36 fermentation liquid；
(6) calcium carbonate is added in the bacillus amyloliquefaciens PHODG36 fermentation liquid and is spray-dried, import temperature
Degree is 175 DEG C, and outlet temperature is 70 DEG C, obtains dry bacterium powder；
(7) the dry bacterium powder mixed with the dispersing agent and wetting agent, stirred, after air-flow crushing and cross 325
Mesh obtains the solid microbial to potato disease-preventing and yield-increasing.
Experimental example
One, influence-potted plant experiment that 3 obtained solid microbial bacterial agent of the embodiment of the present invention grows potato plant
It is micro- in 3 obtained solid of the embodiment of the present invention of same amount difference bacteria suspension concentration by greenhouse pot culture test development
Safety research of the bacteria agent to potato.It is different according to solid microbial and potato seed amount (4:150, w/w) ratio
The solid microbial (1,500,000,000,5,000,000,000,10,000,000,000,20,000,000,000) of concentration and potato seed carry out Dressing, are with not Dressing
Blank control (CK).Each processing dresses seed postposition in a cool and dry place, plant in flowerpot within second day (upper diameter 135mm × under
Diameter 90mm × high 115mm), it is cultivated in the greenhouse, each concentration processing 10 basins of plantation, 2 pieces of every basin is being cultivated later
Potato emergence rate and plant height are recorded at 30 days and 60 days, are calculated correction emergence rate according to the following formula, are emerged by correction
The influence that rate and plant height two indices evaluation microbial bacterial agent grow potato plant, the results are shown in Table 1 and table 2.
Emergence rate (%)=(emergence quantity/20) × 100
Correct emergence rate (%)=(emergence rate/blank control emergence rate of processing) × 100.
The influence that table 1- solid microbial application dosage emerges to potato
As can be seen from Table 1,3 gained microbial bacterial agent dosage of the embodiment of the present invention is right in hundred million range of 15-200
Potato emergence has differences, and wherein dosage can be improved emergence rate in hundred million range of 15-50, with the increase of concentration,
Emergence rate is on a declining curve, and when the microbial bacterial agent dosage is greater than 20,000,000,000, correction emergence rate is only 81.73%,
And it is significantly lower than comparison medicament group, it can be deduced that, 3 obtained solid microbial bacterial agent dosage of embodiment is in hundred million range of 15-50
It inside can be improved emergence rate, and unsuitable too high or too low using concentration.
The influence that table 2- solid microbial application dosage grows potato plant
As can be seen from Table 2,3 obtained solid microbial bacterial agent dosage of the embodiment of the present invention is in hundred million range of 15-200
Interior to have facilitation to potato plant growth, solid microbial dosage is in hundred million range of 15-100, with it
The increase of concentration, facilitation also gradually increase.
Two, the field trial of 3 obtained solid microbial bacterial agent of embodiment
1, experimental field basic condition
Test is respectively at 2018 and 2019 in the village Xi Zhangtou, the Handan in Hebei province Quzhou County township Huai Qiao, height above sea level 40m.
Selection topography is flat, the relatively uniform plot of fertility is tested field.
2, for studying object and controlling object
It is potato for studying object, potato seed dosage is 150kg/667m2, kind is Holland 15；Controlling object is potato
Verticillium wilt.
3, reagent agent
Solid microbial prepared by the embodiment of the present invention 3；Comparison medicament is 70% thiophanate methyl wettable powder
(thiophanate methyl, Qingdao KYX Chemical Co., Ltd.), 3% Zhongshengmycin wettable powder
(zhongshengmycin, Shenzhen Nuopuxin Agricultural Chemical Co., Ltd).
4, field plot trial designs
(1) 2018 year field trial
It is 1,500,000,000 gemma/gram test that solid microbial, which is prepared into concentration,.Three processing of test setting, tool
Body is as follows, T1:4kg/ mus of seed dressings, and operating procedure is as follows: by seed potato stripping and slicing, guaranteeing 1-2 bud eye of each potato wedge, presses
According to potato seed amount and microbial inoculum (150:4) ratio, it is uniformly spread to potato seed, stirs and stirs and evenly mixs, seed dressing uniform adhesion is made to exist
In potato seed, it is placed in ventilating and cooling place；T2:150g thiophanate methyl+100g Zhongshengmycin+4kg talcum powder handles potato seed；T3: kind
Potato is blank control without any processing.
Potato verticillium wilt grade scale is as follows:
0 grade: healthy plant；
The plant of 1 grade: 25% blade morbidity；
The plant of 2 grades: 25%-50% blade morbidity；
The plant of 3 grades: 50%-75% blade morbidity；
4 grades: 75% withered with the plant of blade morbidity or plant.
Different disposal plant incidence is investigated, calculates disease index and control efficiency, the results are shown in Table 3.
3 obtained solid microorganism formulation of table 3- embodiment evaluates the control efficiency and effect of increasing production of potato verticillium wilt
(Handan Quzhou, 2018)
From table 3 it is observed that the microbial bacterial agent is 39.02% to the preventive effect of potato verticillium wilt, lower than control
Medicament, but microbial bacterial agent mu rate of growth is higher than comparison medicament.Illustrate 3 obtained solid microorganism formulation pair of the embodiment of the present invention
Potato has good effect of increasing production, and effect of increasing production is better than its protection effect.
(2) 2019 years field trials
It is 1,500,000,000,5,000,000,000,10,000,000,000,20,000,000,000 gemma/gram test that solid microbial, which is prepared into concentration,.Examination
Test setting six processing, it is specific as follows, hundred million gemma of T1:15/gram microbial bacterial agent seed dressing；Operating procedure is as follows: by potato
Potato seed stripping and slicing guarantees 1-2 bud eye of each potato wedge, and according to potato seed amount and seed dressing (150:4) ratio, it is uniformly spread to kind
It on potato, stirs and stirs and evenly mixs, make seed dressing uniform adhesion in potato seed, be placed in ventilating and cooling place；Hundred million gemma of T2:50/gram it is micro-
Bacteria agent seed dressing；Hundred million gemma of T3:100/gram microbial bacterial agent seed dressing；Hundred million gemma of T4:200/gram microbial bacterial agent mix
Kind；T5:150g thiophanate methyl+100g Zhongshengmycin+4kg talcum powder handles potato seed；T6: potato seed is sky without any processing
White control.Test result is as shown in table 4 and table 5.
3 obtained solid microorganism formulation of table 4- embodiment to the evaluation of the effect of increasing production of potato (Handan Quzhou plot A,
2019)
| Processing | Per mu yield (kg) | Rate of growth (%) |
| 3 obtained solid microorganism formulation (1,500,000,000) of embodiment | 3220.55 | 8.87 |
| 3 obtained solid microorganism formulation (5,000,000,000) of embodiment | 3495.69 | 18.18 |
| 3 obtained solid microorganism formulation (10,000,000,000) of embodiment | 3856.47 | 30.37 |
| 3 obtained solid microorganism formulation (20,000,000,000) of embodiment | 3251.63 | 9.92 |
| First support+Zhongshengmycin | 2871.89 | -2.91 |
| Blank control | 2958.04 | —— |
As can be seen from Table 4, the 3 obtained solid microbial bacterial agent of embodiment has volume increase well to potato
Effect；For solid microbial when concentration is between 1,500,000,000-100 hundred million, rate of growth is between 8.87%-30.37%；Wherein when
Using concentration be 10,000,000,000 3 obtained solid microbial bacterial agent of embodiment effect of increasing production highest, rate of growth 30.37%, into
One step illustrate 3 obtained solid microbial bacterial agent of embodiment to potato have good effect of increasing production, and effect of increasing production better than pair
According to medicament.
3 obtained solid microorganism formulation of table 5- embodiment to the evaluation of the effect of increasing production of potato (Handan Quzhou plot B,
2019)
| Processing | Per mu yield (kg) | Rate of growth (%) |
| 3 obtained solid microorganism formulation (1,500,000,000) of embodiment | 3220.55 | 14.75 |
| 3 obtained solid microorganism formulation (5,000,000,000) of embodiment | 3495.69 | 21.70 |
| 3 obtained solid microorganism formulation (10,000,000,000) of embodiment | 3856.47 | 28.04 |
| 3 obtained solid microorganism formulation (20,000,000,000) of embodiment | 3251.63 | 22.81 |
| First support+Zhongshengmycin | 2871.89 | -3.66 |
| Blank control | 2958.04 | —— |
As can be seen from Table 5, the resulting solid microbial of embodiment 3 has volume increase effect well to potato
Fruit；For solid microbial when concentration is between 1,500,000,000-100 hundred million, rate of growth is between 14.75%-28.04%；Wherein when making
With the effect of increasing production highest for the 3 obtained solid microbial bacterial agent of embodiment that concentration is 10,000,000,000, rate of growth 28.04%.
Three, the field trial of 3 obtained solid microbial bacterial agent of embodiment
1, experimental field basic condition
It tests and was carried out in 2019 in Hong-shi-la-zi type village, decorated archway township, Chengde City, Hebei Province Weichang Manchu and Mongolian Autonomous County.
Experimental plot soil is sandy loam, and topography is smooth, and water and fertilizer condition is preferable.
2, for studying object and controlling object
It is potato for studying object, potato seed dosage is 150kg/667m2, kind is Holland 15；Controlling object is potato
Verticillium wilt.
3, reagent agent
Solid microbial prepared by the embodiment of the present invention 3；Comparison medicament is 70% thiophanate methyl wettable powder
(thiophanate methyl, Qingdao KYX Chemical Co., Ltd.), 3% Zhongshengmycin wettable powder
(zhongshengmycin, Shenzhen Nuopuxin Agricultural Chemical Co., Ltd).
4, field plot trial designs
It is 1,500,000,000,5,000,000,000,10,000,000,000,20,000,000,000 gemma/gram test that solid microbial, which is prepared into concentration,.Examination
Test setting six processing, it is specific as follows, hundred million gemma of T1:15/gram microbial bacterial agent seed dressing；Operating procedure is as follows: by potato
Potato seed stripping and slicing guarantees 1-2 bud eye of each potato wedge, and according to potato seed amount and seed dressing (150:4) ratio, it is uniformly spread to kind
It on potato, stirs and stirs and evenly mixs, make seed dressing uniform adhesion in potato seed, be placed in ventilating and cooling place；Hundred million gemma of T2:50/gram it is micro-
Bacteria agent seed dressing；Hundred million gemma of T3:100/gram microbial bacterial agent seed dressing；Hundred million gemma of T4:200/gram microbial bacterial agent mix
Kind；T5:150g thiophanate methyl+100g Zhongshengmycin+4kg talcum powder handles potato seed；T6: potato seed is sky without any processing
White control.Test result is as shown in table 6.
3 obtained solid microorganism formulation of table 6- embodiment evaluates (Chengde Weichang) to the effect of increasing production of potato
As can be seen from Table 6, the resulting solid microbial of embodiment 3 increases potato with good diseases prevention
Produce effect；For solid microbial when concentration is between 1,500,000,000-100 hundred million, rate of growth is between 10.55%-26.80%；Wherein
When the preventive effect highest for the 3 obtained solid microbial bacterial agent of embodiment for the use of concentration being 5,000,000,000, control efficiency 52.31%；When making
With the effect of increasing production highest for the 3 obtained solid microbial bacterial agent of embodiment that concentration is 10,000,000,000, rate of growth 26.80%.
It is good that the comprehensive test result of ground more than 2 years illustrates that 3 obtained solid microbial bacterial agent of embodiment has potato
Preventing disease production-increasing effect, and effect of increasing production is better than comparison medicament.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, appoints
What those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, answer
It is included within the scope of the present invention.Therefore, protection scope of the present invention should be with the scope of protection of the claims
It is quasi-.
Claims (10)
1. the solid microbial of a kind of pair of potato disease-preventing and yield-increasing, which is characterized in that raw material components include:
Bacillus amyloliquefaciens PHODG36 10-80 parts by weight, auxiliary agent 2-16 parts by weight, filler 10-80 parts by weight.
2. the solid microbial according to claim 1 to potato disease-preventing and yield-increasing, which is characterized in that the auxiliary agent
Including dispersing agent and wetting agent；The filler is one of calcium carbonate, soluble starch or talcum powder or a variety of mixtures.
3. the solid microbial according to claim 2 to potato disease-preventing and yield-increasing, which is characterized in that the dispersion
The mass ratio of agent and wetting agent is (1-8): (1-8).
4. the solid microbial according to claim 2 to potato disease-preventing and yield-increasing, which is characterized in that the dispersion
Agent is lignin sulfonic acid sodium salt, sanlose, desugar sodium lignin sulfonate, ethylene glycol monobutyl ether, polyoxyethanyl
One of alkyl ether, neopelex or a variety of mixtures；
The wetting agent is one of nekal, fatty alcohol polyoxyethylene ether, pelopon A
Or a variety of mixture.
5. the solid microbial according to claim 1 to potato disease-preventing and yield-increasing, which is characterized in that the solid
The dosage form of microbial bacterial agent is wettable powder.
6. a kind of method for preparing the solid microbial described in claim 1 to potato disease-preventing and yield-increasing, feature exist
In, comprising the following steps:
(1) the bacillus amyloliquefaciens PHODG36 bacterial strain frozen is gone into activation culture on LB solid medium, obtains activation bacterium
Strain；
(2) activated strains are seeded in LB liquid medium, carry out shaking table culture, obtains primary seed solution；
(3) primary seed solution is transferred in seed culture medium, is expanded culture under agitation, obtain second level kind
Sub- liquid；
(4) secondary seed solution is forwarded in fermentation medium, carries out fermented and cultured under agitation, obtain the solution
Bacillus amyloliquefaciens PHODG36 fermentation liquid；
(5) filler is added in the bacillus amyloliquefaciens PHODG36 fermentation liquid, is spray-dried, obtain dry bacterium powder；
(6) the dry bacterium powder mixed with the auxiliary agent, stirred, after air-flow crushing and is sieved, being obtained described to Ma Ling
The solid microbial of potato disease-preventing and yield-increasing.
7. the preparation method of the solid microbial according to claim 6 to potato disease-preventing and yield-increasing, feature exist
In in step (1), the cultivation temperature is 36-38 DEG C, and the incubation time is 20-28h.
8. the preparation method of the solid microbial according to claim 6 to potato disease-preventing and yield-increasing, feature exist
In in step (2), the cultivation temperature is 35-38 DEG C, and the incubation time is 20-24h, and the shaking speed is 150-
200rpm；
In step (3), the volume ratio of the primary seed solution and seed culture medium is (1-5): 100, the cultivation temperature is 36-
38 DEG C, the incubation time is 18-22h, and the dissolved oxygen amount is 10%-20%, and the mixing speed is 170-190rpm, is obtained
The secondary seed solution in thallus be logarithmic growth phase.
9. the preparation method of the solid microbial according to claim 6 to potato disease-preventing and yield-increasing, feature exist
In in step (4), the volume ratio of the secondary seed solution and fermentation medium is (4-6): 100, the cultivation temperature is 36-
38 DEG C, the incubation time is 24-32h, and the dissolved oxygen amount is 15%-18%, and the mixing speed is 170-190rpm；
Thalline quantity >=6,000,000,000/mL in the bacillus amyloliquefaciens PHODG36 fermentation liquid, spore forming rate >=95%.
10. the preparation method of the solid microbial according to claim 6 to potato disease-preventing and yield-increasing, feature exist
In in step (5), the inlet temperature of the spray drying is 150-200 DEG C, and the outlet temperature of the spray drying is 50-90
℃；
In step (6), the partial size of the solid microbial after the sieving is 300-350 mesh.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910832008.7A CN110477021A (en) | 2019-09-04 | 2019-09-04 | The solid microbial and preparation method of a kind of pair of potato disease-preventing and yield-increasing |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910832008.7A CN110477021A (en) | 2019-09-04 | 2019-09-04 | The solid microbial and preparation method of a kind of pair of potato disease-preventing and yield-increasing |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110477021A true CN110477021A (en) | 2019-11-22 |
Family
ID=68556423
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910832008.7A Pending CN110477021A (en) | 2019-09-04 | 2019-09-04 | The solid microbial and preparation method of a kind of pair of potato disease-preventing and yield-increasing |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110477021A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114958675A (en) * | 2022-06-14 | 2022-08-30 | 河北省农林科学院植物保护研究所 | Bacillus composition, microbial agent and application |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102925386A (en) * | 2012-10-24 | 2013-02-13 | 山东省林业科学研究院 | Bacillus amyloliquefaciens and application thereof in prevention and treatment of walnut anthracnose |
| CN106399177A (en) * | 2016-10-08 | 2017-02-15 | 河北省农林科学院植物保护研究所 | Bacillus amyloliquefaciens with inorganic phosphorus degrading and bacteria restraining effects and bacterial agent thereof |
-
2019
- 2019-09-04 CN CN201910832008.7A patent/CN110477021A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102925386A (en) * | 2012-10-24 | 2013-02-13 | 山东省林业科学研究院 | Bacillus amyloliquefaciens and application thereof in prevention and treatment of walnut anthracnose |
| CN106399177A (en) * | 2016-10-08 | 2017-02-15 | 河北省农林科学院植物保护研究所 | Bacillus amyloliquefaciens with inorganic phosphorus degrading and bacteria restraining effects and bacterial agent thereof |
Non-Patent Citations (3)
| Title |
|---|
| 方中达: "《中国农业植物病害》", 30 November 1996, 中国农业出版社，第1版 * |
| 江洪: "《智慧农业导论：理论、技术和应用》", 30 September 2015, 上海交通大学出版社，第1版 * |
| 赵卫松 等: "解淀粉芽胞杆菌PHODG36菌剂的研制及其对马铃薯黄萎病的防病增产效果", 《中国生物防治学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114958675A (en) * | 2022-06-14 | 2022-08-30 | 河北省农林科学院植物保护研究所 | Bacillus composition, microbial agent and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103964946B (en) | A kind of paddy rice Special compound microbial fertilizer and preparation method thereof | |
| US8518428B2 (en) | Antagonistic bacteria for controlling the Fusarium wilt of continuous cropping banana and their microbial organic fertilizer | |
| CN103553828B (en) | Liquid compound biological fertilizer with long shelf life as well as preparation method and application thereof | |
| CN106591193B (en) | One plant of bacillus amyloliquefaciens with the degeneration-resistant effect of wide spectrum growth-promoting | |
| CN106883063A (en) | A kind of biological organic fertilizer and its production technology | |
| CN108753664A (en) | A kind of biocontrol agent, biological organic fertilizer and its application | |
| CN101555459B (en) | Strain SB177 for controlling strawberry replant diseases and preparation thereof | |
| CN109650986B (en) | Carbon-based bio-organic fertilizer for improving degraded vegetable field soil and application thereof | |
| CN106495933B (en) | A kind of cucumber seedling-raising substrate and preparation method thereof | |
| CN107619308A (en) | Prevent and treat root-knot nematode biological organic fertilizer and preparation method and application | |
| CN107188674B (en) | Solanaceous vegetable seedling culture substrate and preparation method thereof | |
| CN106495951A (en) | A kind of high concentration microorganism microbial inoculum containing polyglutamic acid and preparation method and application | |
| CN106190929B (en) | A kind of new strain of Bacillus licheniformis and its application | |
| CN106336295B (en) | Disease-resistant type fertilizer synergist of a kind of combination containing waxy Bacillus, manganese sulfate and polyglutamic acid and preparation method thereof | |
| CN106305792A (en) | Clonostachys rosea-bacillus subtilis composite wettable powder | |
| CN108450265A (en) | A kind of paddy rice nursery method | |
| CN101812410B (en) | Streptomycesvinaceus-drappus microbial inoculum and method for preparing same | |
| CN106977293A (en) | It is a kind of to improve complex microorganism organic fungi-manure of seedling salt tolerance and preparation method thereof | |
| CN104830706B (en) | One plant of bacillusmusilaginosiengineering and the organic sustained-controll-release fertiliser of potato special bio | |
| CN105175159A (en) | Production method of rice seedling dry raising and strengthening biological medium and application thereof | |
| CN106035378A (en) | Compound bacterium agent for preventing and treating bacterial diseases of tobacco | |
| CN110477021A (en) | The solid microbial and preparation method of a kind of pair of potato disease-preventing and yield-increasing | |
| CN106520595B (en) | A kind of arthrobacterium and its application in terms of biological control bacterial wilt of tomato | |
| CN108409411A (en) | A kind of biology of enhancing paddy rice anti contravariance strengthens seedling agent and preparation method thereof | |
| CN109734484A (en) | A kind of microorganism formulation and preparation method thereof conducive to salt-soda soil plant growth |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20191122 |
|
| WD01 | Invention patent application deemed withdrawn after publication |