CN107509744A - Prevent and treat complex biological agricultural chemicals of wheat base rot disease and preparation method and application - Google Patents
Prevent and treat complex biological agricultural chemicals of wheat base rot disease and preparation method and application Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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Abstract
The invention discloses a kind of complex biological agricultural chemicals for preventing and treating wheat base rot disease and preparation method thereof and its application in wheat increase yield.The biological pesticide is the complex biological agricultural chemicals being uniformly mixed to get in proportion by bacillus subtilis microbial agent and shenqinmycin.The biological pesticide is efficient, green, safety, reduces the use of agrochemical, is advantageous to the No-harmful apple orchard of wheat crop；The prevention effect to wheat base rot disease caused by false Fusarium graminearum is significantly improved, and with volume increase function, can be that peasant increases income, there is very big developing and utilizingpotentiality.
Description
Technical field
The present invention relates to technical field of pesticide combination, and in particular to a kind of complex biological agricultural chemicals for preventing and treating wheat base rot disease
And preparation method thereof with its application in wheat increase yield.
Background technology
Wheat base rot disease is a kind of soil-borne disease as caused by a variety of disease funguses, is commonly called as " non-irrigated pedopathy ", existed in recent years
Getting worse occurs for China Yellow River-Huai River region, and the wheat safety in production to China brings serious threat.Wherein, false Fusarium graminearum
(Fusariumpseudograminearum) it is newfound a kind of pathogen on China's wheat base rot disease in recent years, its
Pathogenicity is strong, propagates the Main Pathogenic Bacteria for soon, having become wheat base rot disease at present, has a strong impact on the yield and product of wheat
Matter, serious economic loss is brought to China's agricultural production.In addition, false Fusarium graminearum can also produce some toxin, to people and animals
Health causes to seriously endanger.Although using Agro-chemicals control wheat base rot disease with certain effect, chemical pesticide
High poison high residue, environment is polluted, causes pathogen to develop immunity to drugs, serious threat the sustainable development of China's agricultural, research and development
Environmentally safe friendly biological pesticide is particularly important.
Bacillus subtilis is a kind of biocontrol microorganisms for having developing and utilizingpotentiality, has very wide antimicrobial spectrum.At present
It was found that it is to gaeumannomyces graminis, rhizoctonia cerealis, fusarium graminearum, botrytis cinerea, botrytis cinerea pers, apple
The pathogens such as brown patch germ, Chinese yam anthrax bacteria, withered germ of water-melon, Causal Organism of Cucumber Blight, cotton boll rot pathogens have very strong
Inhibition, and biological agent is had been developed that into applied in the biological control of take-all.Shenqinmycin is fluorescence vacation
A kind of antibiotic of bacillus secretion is assured, there are stronger disease-resistant growth-promoting functions, shenqinmycin has been used for wheat total eclipse at present
In the preventing and treating of disease and wheat scab, but not yet find that it can be used for anti-wheat brown foot rot caused by Fusarium graminearum of cracking down on counterfeiting
Disease.
The content of the invention
Obtained the technical problem to be solved in the present invention is to provide a kind of bacillus subtilis and shenqinmycin decrement compounding
Biological pesticide and its in the anti-application cracked down on counterfeiting caused by Fusarium graminearum on wheat base rot disease and wheat increase yield.
In order to solve the above technical problems, the present invention adopts the following technical scheme that：
A kind of complex biological agricultural chemicals for preventing and treating wheat base rot disease is designed, it is mould containing bacillus subtilis microbial agent and Shen piperazine
Element.
Bacillus subtilis are the bacillus subtilis YB-05 bacterial strains that deposit number is CGMCC NO.6498.
Bacillus subtilis microbial agent is made via following methods：
It will be rule in bacillus subtilis YB-05 inoculations to LA culture mediums, 30-36h cultivated extremely under the conditions of 28-32 DEG C
Grow single bacterium colony；Picking single bacterium colony is inoculated into LB nutrient solutions, 30-32 DEG C, 160rpm, concussion and cultivate 12-16h, seed liquor is made；
Seed liquor is taken to be inoculated in 2% inoculum concentration in fermentation medium, the concussion and cultivate 48-54h under the conditions of 30-32 DEG C, 160rpm,
Zymotic fluid is diluted 10 times with sterilized water.
Preferably, it is with volume basis, the ratio for the shenqinmycin that bacillus subtilis microbial agent and percentage by weight are 1%
20~17:5~1.
Preferably, the concentration of shenqinmycin is 0.5mg/ml in complex biological agricultural chemicals.
Preferably, the bacterium colony concentration of bacillus is 10 in complex biological agricultural chemicals7~108CFU/ml。
A kind of preparation method for the complex biological agricultural chemicals for preventing and treating wheat base rot disease is designed, is comprised the following steps：
(1) bacillus subtilis is inoculated on LA culture mediums and rule, 30-36h is cultivated under the conditions of 28-32 DEG C to growing list
Bacterium colony；Single bacterium colony described in picking is inoculated into LB nutrient solutions, 30-32 DEG C, 160rpm, concussion and cultivate 12-16h, seed liquor is made；Take
The seed liquor is inoculated in fermentation medium with 2% inoculum concentration, the concussion and cultivate 48- under the conditions of 30-32 DEG C, 160rpm
54h, zymotic fluid is diluted 10 times with sterilized water, produces bacillus subtilis microbial agent；
(2) bacillus subtilis microbial agent is taken uniformly to be mixed with the shenqinmycin.
Preferably, fermentation medium contains 1% sucrose, 1.5% corn flour, 3.5% analysis for soybean powder, 0.2% dipotassium hydrogen phosphate, pH
For 7.2.
Prevent and treat application of the complex biological agricultural chemicals of wheat base rot disease in wheat increase yield.
The advantageous effects of the present invention are：
1. complex biological agricultural chemicals of the present invention has the characteristics of efficient, green, safe and environmentally friendly, agricultural chemicals is reduced
The use of fertilizer, is advantageous to the No-harmful apple orchard of wheat crop, is advantageous to lift grain quality；Biological pesticide cost is cheap, is agriculture
The people reduce expenses.
2. complex biological agricultural chemicals of the present invention significantly improves the preventing and treating to wheat base rot disease caused by false Fusarium graminearum
Effect.
3. complex biological agricultural chemicals of the present invention has production-increasing function, it can be that peasant increases income, there are wide market prospects.
Brief description of the drawings
Fig. 1 is the influence figure that bacillus subtilis YB-05 grows to false Fusarium graminearum Fp001；
Fig. 2 is bacillus subtilis YB-05 ferment filtrates to false Fusarium graminearum Fp001 growth inhibition effect figures；
Fig. 3 is influence figure of the bacillus subtilis YB-05 ferment filtrates to false Fusarium graminearum Fp001 hypha forms；
Fig. 4 is inhibitory action figure of the shenqinmycin to false Fusarium graminearum Fp001 of various concentrations；
Fig. 5 is inhibitory action figure of the shenqinmycin to bacillus subtilis YB-05 of various concentrations；
Fig. 6 is indoor control design sketch of the different biological prevention and control agents to wheat base rot disease；
Fig. 7 is field control effect figure of the complex biological agricultural chemicals to wheat base rot disease.
Embodiment
Illustrate the embodiment of the present invention with reference to the accompanying drawings and examples, but following examples are used only in detail
Describe the bright present invention in detail, the scope not limiting the invention in any way.Involved instrument and equipment is such as in the examples below
It is routine instrument device without special instruction；Involved raw material and reagent are commercially available conventional products unless otherwise instructed；
Involved experimental method, it is normal experiment method unless otherwise instructed.
Embodiment 1：
Growth effect of the bacillus subtilis to false Fusarium graminearum Fp00：
(1) bacillus subtilis is the YB-05 bacterial strains that Patent publication No is protected by the A of CN 103074271；
(2) beaten with card punch and take false Fusarium graminearum bacteria cake to move to PDA centers, from the bacteria cake about at 2.5cm, by YB-05
Bacterial strain is symmetrically inoculated in false Fusarium graminearum Fp001 bacteria cakes both sides, and the flat board only to connect pathogen is used as control, 3 repetitions.25
Cultivated in DEG C incubator, after control group mycelia covers with flat board, measure antibacterial bandwidth, while measure wheat pathogen causing the basilar stem rotting
The colony growth diameter of bacterium, and colony growth inhibiting rate is calculated, calculation formula is as follows：
Colony growth bacteriostasis rate (%)=(control colony growth diameter-net growth diameter of processing bacterium colony)/control bacterium colony is net
Growth diameter × 100%
Experimental result is as shown in figure 1, bacillus subtilis YB-05 reaches to false Fusarium graminearum Fp001 antibacterial band
14mm, colony growth inhibiting rate are 75%.Therefore, there is YB-05 bacterial strains strong suppression to make to false Fusarium graminearum Fp001
With.
Embodiment 2：
The influence that bacillus subtilis YB-05 ferment filtrate grows to false Fusarium graminearum Fp001, including following step
Suddenly：
Bacillus subtilis YB-05 is inoculated in LB culture mediums, and 30 DEG C, after 180r/min cultivates 36h, 12000r/min is centrifuged
5min, supernatant is taken to filter out thalline with 0.22 μm of biofilter, then by obtained ferment filtrate and PDA culture medium by 1:10
Plate is down flat after ratio is well mixed, to add the PDA plate of clear water as control group, 3 repetitions, by a diameter of 0.9cm false cereal
Reaping hook bacteria cake is inoculated in flat board center, cultivates and determines colony diameter with crossing method after 7d, calculates colony growth inhibiting rate.
As shown in Fig. 2 compared with clear water compares, the ferment filtrate of bacillus subtilis YB-05 bacterial strains substantially inhibits vacation
Fusarium graminearum Fp001 growth, inhibiting rate reach 83%.Pass through micro- sem observation bacillus subtilis YB-05 strain fermentations
Influence of the filtrate to false Fusarium graminearum Fp001 hypha forms, as shown in figure 3, after the processing of fermented filtrate, mycelia occurs obvious
Mycelia is expanded, mycelia is thicker, mycelia node shorten etc. abnormal phenomenon, and undressed hyphal surface is smooth, even thickness, life
Long point is normal.Illustrate that the secondary metabolite in strain fermentation filtrate has obvious shadow to false Fusarium graminearum Fp001 mycelial growths
Ring, it is suppressed that false Fusarium graminearum Fp001 growth.
Embodiment 3：
Bacillus subtilis YB-05 microbial bacterial agent preparation methods, comprise the following steps：
(1) strain culturing is with preparing：It will rule in YB-05 inoculations to LA culture mediums, cultivated under the conditions of 28-32 DEG C
30-36h is to growing single bacterium colony；
(2) prepared by seed liquor：The single bacterium colony that step (1) obtains is inoculated into LB nutrient solutions, 30-32 DEG C, 160rpm, shaken
12-16h is cultivated, seed liquor is made；
(3) preparation of fermentation medium：Sucrose, corn flour, analysis for soybean powder, dipotassium hydrogen phosphate is taken to add water to be prepared into containing sugarcane respectively
Sugar 1%, corn flour 1.5%, analysis for soybean powder 3.5%, the fermentation medium of dipotassium hydrogen phosphate 0.2%, pH value 7.2, to the base that ferments
121 DEG C are carried out, 30min autoclavings；
(4) liquid fermentation produces：Seed liquor prepared by aspiration step (2) is inoculated in described in step (3) by 2% inoculum concentration
Fermentation medium, after concussion and cultivate 48-54h under the conditions of 30-32 DEG C, 160rpm, collect nutrient solution, its cellular biomass is
109-1010CFU/mL, 10 times are diluted with sterilized water, produces YB-05 microbial bacterial agents.
Embodiment 4：
Shenqinmycin determines to the inhibition of false Fusarium graminearum, the shenqinmycin that wherein percentage by weight is 1% by
Hebei Sannong Agricultural Chemical Co., Ltd. provides, and specifically includes following steps：
1% shenqinmycin constant gradient is diluted using sterilized water, mixed with PDA culture medium, make final concentration be respectively 1,
0.5th, 0.1,0.05,0.01,0.005,0.001,0.0005,0.0001 and 0.00005mg/ml is then straight in the inoculation of ware center
Footpath is 0.9cm false Fusarium graminearum cake, using the processing of PDA plus equivalent clear water as control.After control group mycelia covers with flat board,
The colony diameter in each processing is measured with crossing method, calculates suppression of the shenqinmycin of each concentration to false Fusarium graminearum
Effect processed.
Drawn with reference to Fig. 4 and table 1,1,0.5,0.1,0.05 and 0.01mg/ml shenqinmycin completely inhibit false cereal
The growth of sickle-like bacteria mycelia, inhibiting rate reach 100%, 0.005,0.001,0.0005 and 0.0001mg/ml shenqinmycin pair
The inhibiting rate of false Fusarium graminearum mycelia also all reaches more than 80%, respectively 89.44%, 87.78%, 83.33% and 80%,
0.00005mg/ml shenqinmycin has also reached 77.78% to the inhibiting rate of false Fusarium graminearum.Therefore, shenqinmycin is to vacation
Fusarium graminearum has stronger inhibitory action.
Inhibitory action of the shenqinmycin of the various concentrations of table 1 to false Fusarium graminearum
| Concentration (mg/ml) | Inhibiting rate (%) | Concentration (mg/ml) | Inhibiting rate (%) |
| 1 | 100 | 0.005 | 89.4 |
| 0.5 | 100 | 0.001 | 87.8 |
| 0.1 | 100 | 0.0005 | 83.3 |
| 0.05 | 100 | 0.0001 | 80.0 |
| 0.01 | 100 | 0.00005 | 77.8 |
Embodiment 5：
Shenqinmycin determines to the safe concentration of bacillus subtilis YB-05 bacterial strains, comprises the following steps：
1% shenqinmycin constant gradient is diluted using sterilized water, mixed with PDA culture medium, make its final concentration be respectively 1,
0.5th, 0.1 and 0.05mg/ml, dry after a kind of rhyme scheme in Chinese operas serving as the prelude to a complete score for voices, picking bacillus subtilis YB-05 bacterium colonies are drawn on the flat board of each concentration
Line, not add the culture medium of shenqinmycin to observe the growing state of bacterial strain as control.
As shown in figure 5, compared with the control, 1mg/ml shenqinmycin has certain to bacillus subtilis YB-05 bacterial strains
Inhibitory action, the growth of 0.5,0.1 and 0.05mg/ml shenqinmycin to bacillus subtilis YB-05 bacterial strains be not obvious
Inhibitory action.
Embodiment 6：
The preparation of bacillus subtilis YB-05 bacterial strains and the complex biological agricultural chemicals of shenqinmycin, comprises the following steps：
The bacillus subtilis YB-05 microbial bacterial agents of Example 3 are pressed with 1% shenqinmycin that concentration is 0.5mg/ml
20:3 (vt/vt) ratio is fully uniformly mixed so as to obtain complex biological agricultural chemicals；The biological pesticide of compounding is taken to determine wherein bacillus subtilis
Bacterium YB-05 bacterium colony concentration, it is 10 through diluting coated plate measure concentration7-108CFU/mL。
Embodiment 7：
Difference from Example 6 is, the ratio of bacillus subtilis YB-05 microbial inoculums and shenqinmycin based on vt/vt
For 18:5.
Embodiment 8：
Difference from Example 7 is, the ratio of bacillus subtilis YB-05 microbial inoculums and shenqinmycin based on vt/vt
For 19:1.
Embodiment 9：
Difference from Example 8 is, the ratio of bacillus subtilis YB-05 microbial inoculums and shenqinmycin based on vt/vt
For 17:2.
Embodiment 10：
Contrast the interior of bacillus subtilis YB-05 bacterial strains, shenqinmycin and complex biological agricultural chemicals to wheat base rot disease
Prevention effect, including herein below：
This experiment is carried out in the greenhouse of Henan Academy of Agricultural Sciences, and host material used in indoor pot experiment is will
Soil：Turf：Vermiculite=7：2：1 ratio mixes.False Fusarium graminearum Fp001 bacterial strains fresh in culture dish are broken into bacterium
Cake, millet cool down bottling after boiling to maturation, every bottle is inoculated with false 2 pieces of Fusarium graminearum bacteria cake, is cultivated 7-10 days in 25 DEG C of incubators
Afterwards, native use is mixed according to 0.5% inoculum concentration, is Zheng wheat 366 for examination wheat breed, comparison medicament is 2% Tebuconazole 1ml/kg.Place
Reason method is as shown in table 2：
Indoor control test process method of the 2 each experimental group of table to wheat base rot disease
| Processing numbering | Handle title |
| 1 | YB-05 microbial inoculums 30ml/kg dresses seed |
| 2 | 1% shenqinmycin 2ml/kg dresses seed |
| 3 | YB-05 and 1% shenqinmycin 19:1 (vt/vt) biological pesticide 20ml/kg dresses seed |
| 4 | 2% Tebuconazole 1ml/kg Dressings |
| 5 | CK |
After inoculation, routine observation is potted plant to be watered, and wheat root is rinsed with water after 30 days, investigates disease incidence,
According to disease, a situation arises calculates disease index and protection effect.
Grade scale：
0：It is asymptomatic；1：Blastodisc section browning；2：Blastodisc section, in stem Huo Ye Pins brownings；3；Blastodisc section, in stem or leaf pin
Equal browning；4：Stem, leaf pin and bottom cane browning in ground；5；There is withered rotten symptom in cane；6；Cause to plant by Stalk rot
Strain is dead；7：Seedling is withered or seed decay.
Disease index and protection effect are calculated according to following equation:
As a result as shown in Fig. 6 and table 3, bacillus subtilis YB-0530ml/kg Dressings and 1% shenqinmycin 2ml/
Kg Dressings all have preferable prevention effect to wheat base rot disease, and preventive effect is respectively up to 48.1% and 54.5%；In addition,
Bacillus subtilis YB-05 and 1% shenqinmycin 19:1 decrement compounding 20ml/kg Dressing morbidities are most light, and prevention effect is most
Good, preventive effect reaches 58.9%, has exceeded single dose processing and the preventing and treating of the Tebuconazole 1ml/kg Dressings of comparison medicament 2% effect
Fruit.As can be seen here, bacillus subtilis YB-05 microbial inoculums and 1% shenqinmycin compounding are significantly improved to wheat base rot disease
Prevention effect, there is very big value of exploiting and utilizing.
The indoor control effect of the YB-05 of table 3, shenqinmycin and its complex biological agricultural chemicals to wheat base rot disease
| Numbering | Handle title | Disease index | Prevention effect (%) |
| 1 | YB-05 microbial inoculums 30ml/kg dresses seed | 40.1b | 48.1 |
| 2 | 1% shenqinmycin 2ml/kg dresses seed | 35.2bc | 54.5 |
| 3 | YB-05 and 1% shenqinmycin 19:1 compounding 20ml/kg seed dressings | 31.8c | 58.9 |
| 4 | 2% Tebuconazole 1ml/kg Dressings | 33.5c | 56.7 |
| 5 | CK | 77.3a | / |
Embodiment 11：
The field control effect of bacillus subtilis YB-05 and 1% shenqinmycin complex biological agricultural chemicals to wheat base rot disease
Fruit, including herein below：
Test site：Henan Province Jiaozuo City Wuzhi County Wang Zhao townshiies, wheat is planted in experiment plot throughout the year and base rot disease occurs
It is heavier, applied fertilizer to the subsoil and top dressing according to conventional, whole growth period is in addition to experiment without using other bactericide.
Wheat breed：It is short by anti-58,15 kgs/acre of the amount of broadcasting.
Test period：In June, -2017 in October, 2016
Test process：Handle and mixed for bacillus subtilis YB-05 microbial inoculums 30ml/kg seed dressings, 1% shenqinmycin 2ml/kg
Kind, bacillus subtilis YB-05 microbial inoculums and 1% shenqinmycin 19:1 (vt/vt) complex biological agricultural chemicals 20ml/kg dresses seed, control
Medicament is 4.8% suitable wheat pellet aqueous suspension agent 2ml/kg seed dressings, and blank control is that seed is without any processing, 3 repetitions.
Efficacy survey is divided into 3 times, seedling stage, jointing stage and pustulation period respectively before the winter.Each cell uses five point sampling,
Each point takes 25 young plants, and plant is connected into root digs out, and keeps root system complete, classification tune is carried out according to the grade scale of different times
Look into, prevention effect is calculated according to disease index.
Field seedling stage and the jointing stage of turning green grade scale：
0 grade：Whole strain is without browning symptom；1 grade：Stem browning in plant plumule or ground；3 grades：First leaf sheath browning；5 grades：The
Two leaf sheath brownings；7 grades：The leaf sheath browning of plant the 3rd or whole strain stalk browning feel like jelly；9 grades：Plant is dead.
Pustulation period grade scale：
0 grade：Whole strain is without browning symptom；1 grade：The first stipes of aerial part part browning；3 grades：The stipes of aerial part first
Complete browning or the second stipes part browning；5 grades：The complete browning of the stipes of aerial part second or the 3rd stipes part browning；7
Level：The complete browning of the stipes of aerial part the 3rd or the 3rd stipes above browning, but plant is not dead；9 grades：Plant is withered.
In addition, carrying out survey production in harvest time, per 5 points of samplings of cell, every takes 1m2Wheat plant, it is paid to survey production, and convert into
Output per hectare, and calculate rate of growth.
As a result as shown in Fig. 7 and table 4, bacillus subtilis YB-05 and 1% shenqinmycin 19:1 (vt/vt) complex biological
Agricultural chemicals is best in the prevention effect of Wheat Seedling, reaches 62.1%, at 56.2% and single dose more than suitable wheat pellet Dressing
The prevention effect of reason, in wheat during jointing stage and Adult plant, prevention effect also respectively reaches 57.6% and 43.1%, is above single dose
With the prevention effect of suitable wheat pellet Dressing.
Field control effect of each experimental group agricultural chemicals of table 4 to wheat base rot disease caused by false Fusarium graminearum
Bacillus subtilis YB-05 and 1% shenqinmycin 19 are determined in wheat harvest:1 (vt/vt) complex biological agriculture
Influence of the medicine to wheat yield, as shown in table 5, the average yield per mu of complex biological pesticide-treated is 631.1kg, is increased than control
11.3%, higher than YB-05 microbial inoculums Dressing and the 608.8kg and 599.2kg of shenqinmycin Dressing.As can be seen here, it is withered
Careless bacillus YB-05 and 1% shenqinmycin 19:1 (vt/vt) complex biological agricultural chemicals is under field condition not only to wheat stem foot
Maize ear rot has more stable prevention effect, also with certain production-increasing function, has very big value of exploiting and utilizing.
Influence of each experimental group agricultural chemicals of table 5 to wheat yield
| Handle title | Average yield per mu (kg) | Increase and decrease (%) than control |
| YB-05 dresses seed | 608.8±67.1 | 7.4 |
| Shenqinmycin is dressed seed | 599.2±33.6 | 5.7 |
| This example complex biological agricultural chemicals | 631.1±20.3 | 11.3 |
| Suitable Mai Dan | 595.1±52.7 | 5.0 |
| Control | 566.9±58.0 | / |
The present invention is described in detail above in conjunction with drawings and examples, still, those of skill in the art
Member can also be carried out it is understood that on the premise of present inventive concept is not departed to each design parameter in above-described embodiment
Change, forms multiple specific embodiments, is the common excursion of the present invention, is no longer described in detail one by one herein.
Claims (10)
1. a kind of complex biological agricultural chemicals for preventing and treating wheat base rot disease, it is characterised in that contain bacillus subtilis microbial agent and Shen
Piperazine mycin.
2. the complex biological agricultural chemicals of preventing and treating wheat base rot disease according to claim 1, it is characterised in that the withered grass
Bacillus is the bacillus subtilis YB-05 bacterial strains that deposit number is CGMCC NO.6498.
3. the complex biological agricultural chemicals of preventing and treating wheat base rot disease according to claim 2, it is characterised in that the withered grass bud
Spore bacillus microbial inoculum is made via following methods：
It will be rule in the bacillus subtilis YB-05 inoculations to LA culture mediums, 30-36 h cultivated under the conditions of 28-32 DEG C
To growing single bacterium colony；Single bacterium colony described in picking is inoculated into LB nutrient solutions, 30-32 DEG C, 160 rpm, concussion and cultivate 12-16 h, makes
Into seed liquor；Take the seed liquor to be inoculated in 2% inoculum concentration in fermentation medium, shaken under the conditions of 30-32 DEG C, 160 rpm
Culture 48-54 h are swung, zymotic fluid is diluted 10 times with sterilized water.
4. the complex biological agricultural chemicals of preventing and treating wheat base rot disease according to claim 3, it is characterised in that with volume ratio
Meter, the ratio for the shenqinmycin that the bacillus subtilis microbial agent and percentage by weight are 1% is 20~17:5~1.
5. the complex biological agricultural chemicals of preventing and treating wheat base rot disease according to claim 2, it is characterised in that described multiple
Concentration with shenqinmycin in biological pesticide is 0.5 mg/ml.
6. the complex biological agricultural chemicals of preventing and treating wheat base rot disease according to claim 2, it is characterised in that the complex biological
The bacterium colony concentration of bacillus is 10 in agricultural chemicals7~108CFU/ml。
7. described in claim 1 prevent and treat wheat base rot disease complex biological agricultural chemicals preparation method, it is characterised in that including with
Lower step：
（1）Bacillus subtilis is inoculated on LA culture mediums and rule, 30-36 h are cultivated under the conditions of 28-32 DEG C to growing single bacterium
Fall；Single bacterium colony described in picking is inoculated into LB nutrient solutions, 30-32 DEG C, 160 rpm, concussion and cultivate 12-16 h, seed liquor is made；Take
The seed liquor is inoculated in fermentation medium with 2% inoculum concentration, the concussion and cultivate 48-54 under the conditions of 30-32 DEG C, 160 rpm
H, zymotic fluid is diluted 10 times with sterilized water, produces bacillus subtilis microbial agent；
（2）The bacillus subtilis microbial agent is taken uniformly to be mixed with the shenqinmycin.
8. the preparation method of the complex biological agricultural chemicals of preventing and treating wheat base rot disease according to claim 7, it is characterised in that
The fermentation medium contains 1% sucrose, 1.5% corn flour, 3.5% analysis for soybean powder, 0.2% dipotassium hydrogen phosphate, pH 7.2.
9. the preparation method of the complex biological agricultural chemicals of preventing and treating wheat base rot disease according to claim 7, its feature exist
In the bacillus subtilis are the bacillus subtilis YB-05 bacterial strains that deposit number is CGMCC NO.6498.
10. application of the complex biological agricultural chemicals of the preventing and treating wheat base rot disease described in claim 1 in wheat increase yield.
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| CN108402093A (en) * | 2018-04-28 | 2018-08-17 | 信阳农林学院 | A kind of prevention rice sheath blight disease microorganism and plant source compound pesticide |
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| CN110200014A (en) * | 2019-05-17 | 2019-09-06 | 河南省农业科学院植物保护研究所 | The microorganism formulation and its preparation method and application for preventing and treating fusarium disease |
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| CN108676735B (en) * | 2018-05-31 | 2021-05-28 | 河南农业大学 | Fusarium pseudograminearum strain FC136-2A containing giant virus FpgMBV1 |
| CN110200014A (en) * | 2019-05-17 | 2019-09-06 | 河南省农业科学院植物保护研究所 | The microorganism formulation and its preparation method and application for preventing and treating fusarium disease |
| CN114946484A (en) * | 2022-04-21 | 2022-08-30 | 辽宁省农业科学院 | Application of bacillus methylotrophicus CB-12 in prevention and treatment of rice false smut |
| CN115537350A (en) * | 2022-08-25 | 2022-12-30 | 河南省农业科学院植物保护研究所 | Stenotrophomonas maltophilia YB-1573, biocontrol microbial inoculum and preparation and application thereof |
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| CN116172015A (en) * | 2022-11-01 | 2023-05-30 | 邹城市农业农村局 | Seed dressing agent for preventing and treating wheat stem-based rot, seed coating agent and comprehensive prevention and treatment method |
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