CN1245503C - Microecological preparation, making method and specific strain thereof - Google Patents
Microecological preparation, making method and specific strain thereof Download PDFInfo
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- CN1245503C CN1245503C CN 200310113737 CN200310113737A CN1245503C CN 1245503 C CN1245503 C CN 1245503C CN 200310113737 CN200310113737 CN 200310113737 CN 200310113737 A CN200310113737 A CN 200310113737A CN 1245503 C CN1245503 C CN 1245503C
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Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention discloses a micro-ecological preparation and a making method thereof, and specific strains of the micro-ecological preparation. The purpose of the present invention is to provide a strain which can effectively prevent and cure wheat sharp eyespot, a micro-ecological preparation which is prepared from the strains and is used for preventing and curing the wheat sharp eyespot, and a preparation method thereof. The strain which is provided by the present invention and is used for preventing and curing the wheat sharp eyespot is bacillus subtilis B946 CGMCC NO. 1032 and bacillus cereus B932 CGMCC NO. 1031. The micro-ecological preparation which is provided by the present invention and is used for preventing and curing the wheat sharp eyespot takes at least one strain body of bacillus cereus B932 CGMCC NO. 1031 and bacillus subtilis B946 CGMCC NO. 1032 as active ingredients, and the strain body is a trophosome and/or a spore. The prevention and cure efficiency of the micro-ecological preparation of the present invention is more than 70 %, the micro-ecological preparation plays an obvious role of promoting the growth of wheat, and the plant height, the wet weight, etc. of crops which are treated by the micro-ecological preparation are improved.
Description
Technical field
The present invention relates to a kind of probiotics of preventing and treating wheat hypochnus and preparation method thereof and special strain therefore in the plant biological prevention and control field.
Background technology
Wheat hypochnus is the disease due to mainly being infected by cereal rhizoctonia (Rhizoctonia cerealis), and distribution range is wider.Since the nineties in 20th century, each winter wheat district generally takes place, and increases the weight of year by year, becomes one of main factor of restriction China Wheat Production.For the control of this disease,, there are problems such as effect instability, contaminate environment, input cost height though the part chemical pesticide has certain preventive effect.Biological control is to be proved to be very fruitful new way in recent years, and it mainly is the antagonistic action of utilizing between the microorganism, and the microorganism of selecting agricultural-food are not worked the mischief suppresses growth of pathogenic bacteria.Use antagonism bacterium control corps diseases, can reduce the consumption of chemical pesticide and the residual hazard on agricultural-food, to protecting HUMAN HEALTH and preventing the pollution of the environment all very important.
The innovation and creation content
The purpose of this invention is to provide the bacterial strain that effectively to prevent and treat wheat hypochnus and have facilitating effects.
The bacterial strain that can prevent and treat wheat hypochnus is bacillus cereus (Bacillus cereus) B932 and subtilis (Bacillus subtilis) B946, above-mentioned two strain bacterium all are preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center (being called for short CGMCC) on November 17th, 2003, and preserving number is respectively CGMCC № 1031 and CGMCC № 1032.
Bacillus cereus (Bacillus cereus) B932 CGMCC № 1031 and subtilis (Bacillus subtilis) B946 CGMCC № 1032, all the healthy plant separation from wheat hypochnus grave illness field obtains.Bacillus cereus (Bacillus cereus) B932 CGMCC № 1031, it is shaft-like that thalline is, long 4-7 micron, wide 1.0-1.2 micron, the Gram-reaction positive is given birth in the gemma or nearly middle giving birth to, ellipse or column, and sporangium is not expanded, the energy anaerobic growth, V.P reacting positive, V.P liquid pH4.3-5.5; Growth temperature 10-45 ℃, grow among the 7%NaCl; PH5.7 grows, and produces acid from glucose, maltose, does not produce acid from wood sugar, lactose, N.F,USP MANNITOL, and the energy hydrolyzed starch utilizes Citrate trianion; Can reductive NO
3 -And NO
2 -To compare thalline longer with other bacterial strain of the same race.Subtilis (Bacillus subtilis) B946 CGMCC № 1032, it is shaft-like that thalline is, long 2-3 micron, wide 0.8-0.9 micron, the Gram-reaction positive, gemma ellipse or column, middle giving birth to or nearly middle giving birth to, sporangiocyst does not expand, and does not contain the beta-hydroxy-butanoic acid salt particle in the protoplasma; Grow among the 7%NaCl, stone core milk does not produce acid, pH5.7 growth, the v.p positive, hydrolyzed starch, reductive NO
3 -And NO
2 -, can not anaerobic growth, the catalase positive.Compare the thalline broad with other bacterial strain of the same race.
Another object of the present invention provides a kind of probiotics of preventing and treating wheat hypochnus and preparation method thereof.
The probiotics of control wheat hypochnus provided by the present invention, be that thalline with at least one strain among bacillus cereus (Bacilluscereus) B932 CGMCC № 1031 and subtilis (Bacillus subtilis) the B946 CGMCC № 1032 is an activeconstituents, described thalline is nourishing body and/or gemma.
Described probiotics can be solid-state or liquid, when being solid-state, it also comprises the absorption additive, as lime carbonate, the peat composed of rotten mosses, high calcium soil etc., wherein the diameter of calcium carbonate granule is preferably 0.09mm, its bacteria containing amount is hundred million thalline of 300-700/gram probiotics, is preferably 50,000,000,000 thalline/gram probiotics.The consumption that solid-state probiotics is every mu is the 30-70 gram, and can dress seed also can spray in period of seedling establishment, jointing stage, filling stage.
When probiotics was liquid state, its bacteria containing amount was 1.0 * 10
6-1.0 * 10
10Cfu/ml water is preferably 1.0 * 10
8Cfu/ml water.The consumption that liquid probiotics is every mu is the 0.15-5 liter, and can dress seed also can spray in period of seedling establishment, jointing stage, filling stage.
A kind of method for preparing the probiotics of above-mentioned control wheat hypochnus, be to cultivate bacillus cereus (Bacillus cereus) B932 CGMCC № 1031 and/or subtilis (Bacillus subtilis) B946 CGMCC № 1032, and fermented liquid carried out aftertreatment obtains.
That described aftertreatment comprises is concentrated, add steps such as stopping composition, press filtration, drying or pulverizing.
The present invention is directed to the problem that exists on seriousness that wheat hypochnus takes place and the Wheat Production, learn principle according to plant microecology, isolate 2 strains in the healthy plant body by wheat hypochnus grave illness field to the tangible genus bacillus of wheat hypochnus prevention effect, and be made into probiotics, the preventive effect of probiotics of the present invention reaches more than 70%, and wheat growth is had obvious facilitation, and indexs such as plant height, fresh weight are improved.
Embodiment
Separation of embodiment 1, endogenous spore bacillus and evaluation
1, the separation of endogenous spore bacillus
(1) separates
Select healthy plant in wheat hypochnus grave illness field, clean with distilled water flushing, 1%NaClO surface sterilization 10 minutes are used in whole then strain, and aseptic water washing 3 times shreds, and adds an amount of sterilized water and grinds, and is diluted to 10
-1, vibration is evenly put into 80 ℃ water and was handled gradient dilution to 10 10 minutes
-, to get 0.1ml and be inoculated on the beef extract-peptone nutrient agar flat board, coating is evenly.Place the dark 3d of cultivation of 30 ℃ of thermostat containers, treat that single bacterium colony grows after, purifying, tube is preserved.
(2) screening
Inoculation pathogenic bacteria cereal rhizoctonia (Rhizoctonia cerealis) is in the dull and stereotyped central authorities of PDA, and genus bacillus to be measured is inoculated in around the flat board, and every ware is inoculated 1 strain in 4 orientation, repeats 3 times, carries out primary dcreening operation; Get the tangible bacterial strain of antagonism, carry out the face-off of individual plant and pathogenic bacteria again and cultivate, Yi Bian the spore bacillus of taking to budder, an edge joint pathogenic bacteria cereal rhizoctonia (Rhizoctonia cerealis), or with the centre line spore bacillus of buddering, two kinds of face-off culture methods of two edge joint pathogenic bacterias carry out multiple sieve.The result obtains the bacterial strain that 32 strains have antagonistic action to cereal rhizoctonia (Rhizoctoniacerealis).
(3) potted plant diseases prevention test
The bacterial strain (32 strain) that will have antagonism carries out pot experiment, is inoculated in the beef extract-peptone nutrient solution after the bacterial strain activation, and 30 ℃ of shaking tables were cultivated 30 hours, detects bacterium liquid bacteria containing amount, and sterilized water is diluted to 1.0 * 10
8Cfu/ml.
Wheat seed was through 1%NaClO surface sterilization 10 minutes, and sterilized water washes 5 times repeatedly, 25 ℃ of following vernalization.After the seed that will sprout was soaked seed 2 hours with the above-mentioned bacterium liquid for preparing, 250 gram composite soils are (native naturally: as to sow in flowerpot turfy soil=2: 1) being equipped with, 15 seeds of every basin, treat that wheat was emerged 4-5 days after, during the long 1-2 sheet of wheat leaf, with punch tool length there is the dull and stereotyped punching of sheath blight fungus, get pathogenic bacteria and be affixed on the stem and leaf of Wheat base portion, and cover mixed with little amount soil, keep about 25 ℃ temperature and higher levels of humidity, the statistics disease index calculates preventive effect after waiting to fall ill.Be treated to contrast with clear water.
State of an illness grade scale:
0 grade: complete stool is anosis:
1 grade: the leaf sheath morbidity, cane does not have scab:
2 grades: the scab width is less than 1/2 of cane on the cane;
3 grades: the scab width is greater than 1/2 of cane on the cane; Less than girth 3/4, cane is not softening;
4 grades: the scab width is greater than 3/4 of the cane girth on the cane, and cane is softening, forms dead ears.
With the bacterial strain that 32 strains have antagonistic action to cereal rhizoctonia (Rhizoctonia cerealis), through three batches of potted plant mensuration, the result is as shown in table 1, and showing has 2 bacillus B932 and the higher prevention effect of B946 performance, and all more than 70%.
Table 1 genus bacillus control wheat hypochnus results from pot experiment test
| Bacterial strain | Disease index | Disease index mean value | Preventive effect % | ||
| I | II | III | |||
| B932 B946 contrast | 7.3 6.5 23.5 | 7.6 6.8 26.7 | 6.7 7.4 24.2 | 7.2 6.9 24.8 | 71.0 72.2 - |
2, the evaluation of endogenous spore bacillus B932 and B946
Through identifying that B932 is bacillus cereus (Bacillus cereus), B946 is subtilis (Bacillus subtilis).
The preparation of embodiment 2, probiotics
1. production technique is for to make pulvis through the liquid submerged fermentation method, and the technology key link is that former actication of culture → strain expanded culture → seed tank culture → fermentor cultivation → interpolation stopping composition → press filtration → drying → pulverizing → pulvis quality examination → packing → product detects.
2. technical indicator
Various technical indicators are as shown in table 2.
Table 2 technical indicator
| 1 | The sterilization requirement | Temperature | 121-125℃ |
| Pressure | The pressure 0.11MPa sky that disappears the in fact pressure 0.11Mpa that disappears | ||
| Time | 30min | ||
| 2 | Culture condition | Temperature | 28-30℃ |
| Pressure | 0.05Mpa | ||
| Air flow | 1: 1.3 (per minute turnover volume ratio) | ||
| pH | 7.0-7.5 | ||
| 3 | Put a jar index | Bacteria containing amount | ＞100 hundred million thalline/g |
| Gemma formation amount | ＞70％ | ||
| Assorted bacterium amount | ≤0.3％ | ||
| 4 | The finished product standard | Bacteria containing amount | 50,000,000,000 thalline/g probioticses |
| Water content | ≤5％ | ||
| Fineness | 0.09mm aperture sieve | ||
| Assorted bacterium amount | ≤1％ |
3. bacterial classification production operation rules
Medium preparation: press following formulation substratum: extractum carnis 0.3%, peptone 0.5%, NaCl 0.5%, agar 2%.With agar heating dissolve the back with NaOH or Hcl adjust pH to 7.0-7.5, packing Kolle flask or test tube place in the Autoclave then, the 30min that under 0.11MPa pressure, sterilizes, cooling back bevel is standby.
Actication of culture and enlarged culturing: with substratum under aseptic condition from the former strain inclined plane a little lawn of picking, put in the test tube slant and rule, put then and cultivate 24h in the 28-30 ℃ of incubator, do not have assorted bacterium on inspection, thalli growth is neat, put again and cultivate 24-28h, visual inspection in the 28-30 ℃ of incubator, lawn is plentiful, smear, chromoscopy do not have assorted bacterium, and gemma is of the same size, can put to deposit standby (being advisable about 4 ℃) in the refrigerator.
4. seed tank culture
(1) substratum: seed tank culture based formulas: soybean cake powder 0.5%, fish meal 0.3%, Semen Maydis powder 0.5%, glucose 0.3%, soya-bean oil 0.2%-0.3% (or bubble enemy 0.3%), pH value 7.0-7.5.
(2) empty fermentor tank sterilization: fermentor tank cleans up before application, gets rid of dirt, and under 0.14MPa pressure, sterilization 30min.
(3) seeding tank charging: with reference to the formula rate charging of (1), charging capacity must not surpass tank volume 70%.
(4) seeding tank sterilization: after installing material, the 30min that sterilizes under 0.11MPa pressure can inoculate after being cooled to 30 ℃.
(5) suspension preparation: under aseptic condition each Kolle flask slant strains is added the 100ml sterilized water, lawn is scraped make spore suspension, bacteria containing amount is 1,000,000,000 thalline/ml, and handles 20min in 80 ℃ of constant temperature kettles.
(6) inoculation: with bacillus cereus (Bacillus cereus) B932 that makes or the bacteria suspension of subtilis (Bacillus subtilis) B946, the ratio in 1% is inoculated in the seeding tank that is cooled to 30 ℃ and cultivates.
(7) seed tank culture condition control: the inoculation back is 28-30 ℃ in temperature, under pressure 0.05MPa air flow 1: 1.3 (every min volume ratio) and the 220 commentaries on classics/min agitation conditions, and cultivation 8-10h.
5. ferment tank
(1) fermentation tank culture medium:
Soybean cake powder 2%, fish meal 0.5%, corn steep liquor 1.8%, starch 0.4%, sal epsom 0.06%, lime carbonate 0.5%, potassium primary phosphate 0.03%, soya-bean oil 0.1% or bubble enemy 0.05% transfer about pH value 7.0-7.5 with NaOH or HCl, install to be 70% of tank volume.
(2) fermentation culture conditions
Stirring velocity 180-200 commentaries on classics/min, tank pressure 0.05MPa; Air flow: the interior air flow of 4h has 1 behind the culture transferring: 0.7-0.8 (ratio of per minute turnover volume) increases to 1: 1.3 by wash rice later on.The general fermentation culture cycle is 12-18 hour, forms at gemma 70%, when indivedual gemma begin to come off, can put jar, at this moment living contaminants＜0.3% bacteria containing amount＞10,000,000,000 thalline/ml.
6. aftertreatment
(1) equipment: stirrer, sheet frame, drying machine (device), ultrafine pulverizer etc.
(2) sorbent material: the light calcium carbonate that the general granularity that adopts is 0.09mm is the absorption additive, and add-on is calculated by following formula:
(3) press filtration: by (2) regulation adds sorbent material in the fermented liquid that step 5 obtains after, mechanical stirring 3-6h all is adsorbed gemma, and then uses filter press.
(4) drying: after the filter cake behind the filter press smashed to pieces.Place in dryer or the drying unit and dry.Temperature during oven dry is no more than 65 ℃ (in order to prevent thalline death), constantly stir, and makes to be heated evenly.When water ratio≤5%, can pulverize, pack.
(5) pulverize: utilize high fineness pulverizer to pulverize, 100% by the 0.09mm aperture sieve.
7. inspection after construction
Water ratio is measured with dry weight-loss method (weighting method), and water ratio≤5% be qualified, and fineness adopts wet screening to measure, and 100% is qualified by the 0.09mm sieve, and concrete grammar is pressed HG
2The wet screening of-896 standards is measured.
8. pack
25Kg packs with bag weaved (in plastics bag is arranged), stores preservation period 3 years in dry ventilation.
The field disease-preventing effect of embodiment 3, probiotics
1, field diseases prevention test
(1) preparation of probiotics
According to the technology of embodiment 2, be to be mixed with the control wheat hypochnus probiotics that bacteria containing amount is 50,000,000,000 thalline of every gram at 1: 1 by the bacteria containing amount ratio by bacillus cereus (Bacillus cereus) B932 and subtilis (Bacillus subtilis) B946.
(2) sub-district is provided with
Select the wheat hypochnus even plot of falling ill in former years, whenever be treated to a sub-district, repeat four times, the sub-district area is 67m
2, random alignment is contrast with the clear water.
(3) treatment process
During sowing, the consumption that probiotics is every mu is 50 grams, according to the required grain weight in sub-district probiotics is added and dresses seed after suitable quantity of water is diluted, and can sow behind the airing slightly.Respectively carry out 1 spraying in period of seedling establishment, jointing stage, filling stage then, every mu of consumption 50 grams spray probiotics dissolving back according to sub-district institute water requirement.
(4) investigation method and result
Adopt five point samplings, each treatment zone and check plot are carried out sampling survey.Jointing stage by the strain sampling, was got 50 strains at every in the past; Press the stem sampling after jointing stage, get 50 canes at every.The classification investigation, state of an illness grade scale is the same, calculates preventive effect.
Wheat hypochnus just begins morbidity in seedling stage, this probiotics is as shown in table 3 at the protection effect in seedling stage, show that this probiotics has obvious control effect by seed dressing to wheat hypochnus, protection effect reaches more than 80%, and remarkable at the a=0.05 level difference.
Table 3 control wheat hypochnus probiotics protection effect in seedling stage
| Handle | Sickness rate (%) | Disease refers to | Preventive effect (%) |
| Diseases prevention probiotics CK | 2.40 9.00 | 1.25 6.40 | 80.4 - |
This probiotics is as shown in table 4 at the protection effect of jointing stage, shows by seed dressing+spraying to handle for 1 time, and in the jointing stage investigation, preventive effect surpasses 70%, reaches 75.1%, and remarkable at the a=0.05 level difference.
Table 4 control wheat hypochnus probiotics jointing stage protection effect
| Handle | Sickness rate (%) | Disease refers to | Preventive effect (%) |
| Diseases prevention probiotics CK | 7.15 27.36 | 2.10 8.43 | 75.1 - |
This probiotics protection effect in the watery stage is as shown in table 5, shows by seed dressing+spraying to handle for 2 times, and the filling stage investigation, this probiotics all has higher control effect to wheat hypochnus, and preventive effect surpasses 70%, and remarkable at the a=0.05 level difference.
Table 5 control wheat hypochnus probiotics filling stage protection effect
| Handle | Sickness rate (%) | Disease refers to | Preventive effect (%) |
| Diseases prevention probiotics CK | 30.18 60.70 | 10.38 38.40 | 73.0 ----- |
Embodiment 4, probiotics are preventing that wheat hypochnus from the long volume increase test of simultaneously short taking place
(1) short long mensuration the in the greenhouse of bacillus cereus (Bacillus cereus) B932 probiotics and subtilis (Bacillus subtilis) B946 probiotics
Be inoculated into respectively in the beef extract-peptone nutrient solution after bacillus cereus (Bacillus cereus) B932 and subtilis (Bacillus subtilis) B946 activated respectively, 30 ℃ of shaking tables were cultivated 30 hours, detect bacterium liquid bacteria containing amount, sterilized water is diluted to 1.0 * 10
8Cfu/ml obtains bacillus cereus (Bacilluscereus) B932 probiotics and subtilis (Bacillus subtilis) B946 probiotics respectively.
Wheat seed was through 1%NaClO surface sterilization 10 minutes, and sterilized water washes 5 times repeatedly, 25 ℃ of following vernalization.The seed that will sprout is divided into three groups: control group, B932 group and B946 group, the control group water, after B932 group and B946 group were soaked seed 2 hours with the above-mentioned probiotics for preparing respectively, 250 gram composite soils are (native naturally: as to sow in flowerpot turfy soil=2: 1) being equipped with, 15 seeds of every basin, after treating that wheat was emerged 4-5 days, during the long 1-2 sheet of wheat leaf, with punch tool length there is the dull and stereotyped punching of sheath blight fungus, get pathogenic bacteria and be affixed on the stem and leaf of Wheat base portion, and cover mixed with little amount soil, keep about 25 ℃ temperature and higher levels of humidity, promote growth result to measure to bacillus cereus (Bacillus cereus) B932 probiotics and subtilis (Bacillus subtilis) B946 probiotics, the investigation plant height, indexs such as fresh weight.
Plant height, fresh weight result are shown in table 6, table 7, show by seed soaking and handle, bacillus cereus (Bacilluscereus) B932 probiotics and subtilis (Bacillus subtilis) B946 probiotics has the growth of promotion effect, plant height increases 17.9-25.6%, fresh weight increases 27.5-34.1%, and on the a=0.01 level significant difference.
Table 6 genus bacillus is to the influence of wheat plant height
| Bacterial strain | Plant height (cm) | Plant height mean value (cm) | Increase rate (%) | ||
| I | II | III | |||
| B932 B946 contrast | 30.4 32.9 25.4 | 30.0 29.5 23.7 | 26.7 28.8 24.7 | 29.0b 30.4a 24.6c | 17.9 23.6 - |
Table 7 genus bacillus is to the influence of wheat fresh weight
| Bacterial strain | Fresh weight (g/100 strain) | Fresh weight mean value (g/100 strain) | Rate of body weight gain (%) | ||
| I | II | III | |||
| B932 B946 contrast | 8.2 11.5 7.0 | 11.8 11.6 9.0 | 16.7 11.5 11.2 | 12.2b 11.6a 9.1c | 34.1 27.5 - |
(2) effect of increasing production of probiotics is measured
Probiotics in the present embodiment is the probiotics among the embodiment 3.
Select the wheat hypochnus even plot of falling ill in former years, whenever be treated to a sub-district, repeat four times, the sub-district area is 67m
2, random alignment is contrast with the clear water.
During sowing, the consumption that probiotics is every mu is 50 grams, according to the required grain weight in sub-district probiotics is added and dresses seed after suitable quantity of water is diluted, and can sow behind the airing slightly.Respectively carry out 1 spraying in period of seedling establishment, jointing stage, filling stage then, every mu of consumption 50 grams spray probiotics dissolving back according to sub-district institute water requirement.
At wheat harvest, every sub-district 5 points are got 1 square metre at every and are measured mu spike number, grain number per spike and thousand seed weight, calculate output.In addition, each sub-district is carried out real receipts and is measured actual output, calculates effect of increasing production.This probiotics effect of increasing production is as shown in table 8, shows that this probiotics has effect of increasing production, can make seedling mu volume increase 10.86%.
Table 8 control wheat hypochnus probiotics effect of increasing production is measured
| Handle | Measure output (Kg/ mu) | Stimulation ratio (%) | Actual output (Kg/ mu) | Stimulation ratio (%) |
| Diseases prevention probiotics CK | 579.85 522.92 | 10.89 ----- | 539.75 486.87 | 10.86 ----- |
Claims (10)
1, subtilis (Bacillus subtilis) B946 CGMCC № 1032.
2, bacillus cereus (Bacillus cereus) B932 CGMCC № 1031.
3, a kind of probiotics of preventing and treating wheat hypochnus, be that thalline with at least one strain among bacillus cereus (Bacillus cereus) B932 CGMCC № 1031 and subtilis (Bacillus subtilis) the B946 CGMCC № 1032 is an activeconstituents, described thalline is nourishing body and/or gemma.
4, probiotics according to claim 3 is characterized in that: described probiotics is solid-state.
5, probiotics according to claim 4 is characterized in that: described probiotics also comprises the absorption additive.
6, probiotics according to claim 5 is characterized in that: described absorption additive is a lime carbonate.
7, probiotics according to claim 6 is characterized in that: the bacteria containing amount of described probiotics is hundred million thalline/grams of 300-700.
8, probiotics according to claim 3 is characterized in that: described probiotics is for liquid.
9, probiotics according to claim 8 is characterized in that: the bacteria containing amount of described probiotics is 1.0 * 10
6-1.0 * 10
10Cfu/ml water.
10, a kind of method for preparing the probiotics of the described control wheat hypochnus of claim 3, be to cultivate bacillus cereus (Bacillus cereus) B932 CGMCC № 1031 and/or subtilis (Bacillussubtilis) B946 CGMCC № 1032, and fermented liquid carried out aftertreatment obtains.
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| CN105274078B (en) * | 2015-11-02 | 2019-01-29 | 古吉生物科技（大连）有限公司 | A method of Nattokinase is extracted using deep fermentation |
| CN105543138B (en) * | 2016-01-08 | 2020-02-07 | 河南工业大学 | Bacillus subtilis and application thereof in prevention and control of wheat sharp eyespot |
| CN105794845A (en) * | 2016-04-15 | 2016-07-27 | 佛山市聚成生化技术研发有限公司 | Microbial pesticide and preparing method thereof |
| CN107646873A (en) * | 2017-10-12 | 2018-02-02 | 泰克美生物科技（喀左）有限公司 | A kind of microbial bacterial agent for preventing and treating bacterial plant disease and application |
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