CN103773711B - Bacillus amyloliquefaciens and microbial bacterial agent and their application - Google Patents
Bacillus amyloliquefaciens and microbial bacterial agent and their application Download PDFInfo
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Abstract
The invention provides a kind of bacillus amyloliquefaciens (Bacillus amyloliquefaciens), its deposit number is CCTCC NO.M2013501.Present invention also offers a kind of microbial bacterial agent, this microbial bacterial agent comprises culture medium and thalline, described thalline includes the bacillus amyloliquefaciens that deposit number is CCTCC NO.M2013501, and described thalline also includes Bacillus subtillis (Bacillus subtilis) and bacillus licheniformis (Bacillus lincheniformis).Present invention also offers the application in alkaline land improving, in increase drought resistance in plants, in preventing and treating wheat sharp eyespot and in preventing and treating cucumber fusarium axysporum of bacillus amyloliquefaciens as above and microbial bacterial agent as above.
Description
Technical field
The present invention relates to agricultural biological technical field, in particular it relates to a kind of bacillus amyloliquefaciens, one
Plant microbial bacterial agent and their application.
Background technology
Saline Land is the serious problems affecting agricultural production and ecological environment.The whole world there are about 20%
Arable land and close to 50% irrigated farmland yield seriously affected by soil high concentration is saline and alkaline.In China,
There are about 100,000,000 mu of secondary salinization farmlands, account for the 10% of total cultivated area.The most really improve and effectively
Utilize salt-soda soil, improve economy, society and ecological benefits that agriculture and forestry produce, be that Modern Agriculture forestry is closed
The significant problem of note.
Alkaline land improving research field at home and abroad, successively proposes " plant rice and change alkali " agricultural improvement measure,
" washing salinity by irrigation " engineering ameliorative measure, and utilize gypsum, calcium chloride, industrial waste acid, coal-fired flue-gas
The chemical modifying measures such as desulfurization thing.These measure instant effects, achieve notable achievement.But improvement cost
Height, resource cost is big, and produces nutrient loss, pollutes downstream, the improved effect accumulation of salt in the surface soil unstable, easy
Etc. problem.
In recent years, on the basis of the achievement that studies for a long period of time, people gradually recognize from recovering ecological angle
To the improvement salt-soda soil with in conjunction with getting up, i.e. " during residence is improved to utilize, improves and utilize parallel ",
Ecological recovery technology with biological utilisation as core is the breach that following alkaline land improving is repaired.At present,
Biological modification includes planting saline alkali tolerant plant, using microbial-bacterial fertilizer etc..And develop efficient microbial bacteria
Fertile preparation, first will obtain tolerance saline and alkaline and to the saline and alkaline microbial strains with removal effect, but,
It is the most poor that existing microbial strains removes saline and alkaline effect.
Summary of the invention
In order to overcome existing microbial strains to remove the defect that saline and alkaline effect is poor, the invention provides
A kind of bacillus amyloliquefaciens, the guarantor of this bacillus amyloliquefaciens (Bacillus amyloliquefaciens)
Hide numbered CCTCC NO.M2013501.
Present invention also offers a kind of microbial bacterial agent, this microbial bacterial agent comprises culture medium and thalline, institute
State the bacillus amyloliquefaciens that thalline includes that deposit number is CCTCC NO.M2013501, and described
Thalline also includes Bacillus subtillis (Bacillus subtilis) and bacillus licheniformis (Bacillus
Lincheniformis).
Present invention also offers bacillus amyloliquefaciens as above and microbial bacterial agent as above
Application in alkaline land improving.
Present invention also offers bacillus amyloliquefaciens as above and microbial bacterial agent as above
Application in increasing drought resistance in plants.
Present invention also offers bacillus amyloliquefaciens as above and microbial bacterial agent as above
Application in preventing and treating wheat sharp eyespot.
Present invention also offers bacillus amyloliquefaciens as above and microbial bacterial agent as above
Application in preventing and treating cucumber fusarium axysporum.
By technique scheme, the present invention can more efficiently remove saline and alkaline, such as can be by soil
Salt rejection rate improves to more than 55%.
Other features and advantages of the present invention will be described in detail in detailed description of the invention part subsequently.
Biomaterial preservation
The bacillus amyloliquefaciens (Bacillus amyloliquefaciens) of the present invention is the invention of the present invention
The pure culture that people separates from the soil of Ling County, Shandong, its deposit number is CCTCC NO.M
2013501, preservation date is on October 24th, 2013, and depositary institution is China typical culture collection
Center, address is Wuhan, China Wuhan University, and Classification And Nomenclature is bacillus amyloliquefaciens (Bacillus
Amyloliquefaciens).
Detailed description of the invention
Hereinafter the detailed description of the invention of the present invention is described in detail.It should be appreciated that this place is retouched
The detailed description of the invention stated is merely to illustrate and explains the present invention, is not limited to the present invention.
The invention provides a kind of bacillus amyloliquefaciens, this bacillus amyloliquefaciens (Bacillus
Amyloliquefaciens) deposit number is CCTCC NO.M2013501.
Present invention also offers a kind of microbial bacterial agent, this microbial bacterial agent comprises culture medium and thalline, institute
State the bacillus amyloliquefaciens that thalline includes that deposit number is CCTCC NO.M2013501, and described
Thalline also includes Bacillus subtillis (Bacillus subtilis) and bacillus licheniformis (Bacillus
Lincheniformis).
Wherein, the amount of the thalline contained in described microbial bacterial agent can in very large range change, preferably
In the case of, the total viable count contained by every gram of described microbial bacterial agent can be 2-20 × 107CFU。
Preferably, in terms of the number of viable bacteria body, relative to bacillus amyloliquefaciens every part described, described withered
The content of grass bacillus is 0.05-0.2 part, and the content of described bacillus licheniformis is 0.05-0.2 part.
Under this preferable case, described microbial bacterial agent has the ability that more excellent removal is saline and alkaline.
According to the present invention, the kind of described culture medium can in very large range change, and can be various energy
It is enough in and cultivates bacillus amyloliquefaciens, Bacillus subtillis, the culture medium of bacillus licheniformis, such as,
Can be that the conventional culture mediums such as beef-protein medium, broth bouillon, LB culture medium are as kind
Sub-culture medium, for the preservation of bacterial classification；And the culture medium fermented is generally used for producing, these culture mediums
Kind is the most known to those skilled in the art.Above-mentioned culture medium can be commercially available or according to " micro-life
Thing culture medium handbook " record of (Microbiology Culture Media Manual) prepares.Example
As, beef-protein medium: beef extract 3g, peptone 10g, sodium chloride 5g, agar 15-20g,
Distilled water 1000ml, pH value 7.0-7.2,121 DEG C of sterilizing 20min；Fermentation medium: glucose 15g,
Starch 1g, beancake powder 25g, manganese sulfate 1g, potassium dihydrogen phosphate 1.5g, magnesium sulfate 0.5g, yeast extract
0.2g, iron chloride 0.1g, calcium carbonate 0.1g, pH value 7.0-7.2,115 DEG C of sterilizing 20min.
Wherein, the preparation method of described microbial bacterial agent may include that bacillus amyloliquefaciens, withered grass
Bacillus and bacillus licheniformis are inoculated in culture medium respectively and cultivate.Wherein it is possible to each
Independent cultivating system is cultivated bacillus amyloliquefaciens, Bacillus subtillis and lichens brood cell's bar respectively
Bacterium, and proportionally mix cultivating the microorganism obtained respectively.Under preferable case, by the most solely
The vertical cultivation in cultivating system and the mixing after cultivating so as to get total viable bacteria of every gram of microbial bacterial agent
Number is 2-20 × 107CFU。
According to the present invention, the cultural method of described bacillus subtilis (Bacillus subtilis) does not has spy
Other be limited to known to one of skill in the art, for example, it is possible to first by bacillus subtilis seed train
Support in base (LB culture medium) and cultivate to strain density OD600Value is 0.6-0.8, obtains bacterium solution, then exists
The production medium of 100 weight portions (glucose 5g, dregs of beans 30g, peptone 2g, distilled water 1000ml,
PH7.0-7.2) bacterium solution of the bacillus subtilis (Bacillus subtilis) of inoculation 2-5 weight portion in,
37 DEG C of cultivations, until the viable count of bacillus subtilis (Bacillus subtilis) is 2-20 × 107Individual/
Gram culture medium.
The cultural method of described bacillus licheniformis (Bacillcus lincheniformis) the most particularly limits
It is made as known to one of skill in the art, for example, it is possible to first by bacillus licheniformis at seed culture medium (LB
Culture medium) in cultivate to strain density OD600Value is 0.6-0.8, obtains bacterium solution, then at 100 weight portions
Culture medium (corn steep liquor 0.45g, peptone 1.50g, beancake powder leaching juice be 1:15, glucose 1.50g,
PH value 7.5) the middle bacillus licheniformis (Bacillcus lincheniformis) inoculating 2-5 weight portion
Bacterium solution, 37 DEG C of cultivations, until the viable count of bacillus licheniformis (Bacillcus lincheniformis) is
2-20×107Individual/gram culture medium.
The cultural method of described bacillus amyloliquefaciens (Bacillus amyloliquefaciens) is the most especially
Restriction, for example, it is possible to first by bacillus amyloliquefaciens training in the seed culture medium (LB culture medium)
Support to strain density OD600Value is 0.6-0.8, obtains bacterium solution, then at 100 weight portion culture medium (soya-bean cakes
Powder 50g, sucrose 20g, ammonium sulfate 5g, trisodium citrate 2.5g, potassium dihydrogen phosphate 0.3g, magnesium sulfate
0.5g, ferrous sulfate 0.05g, pH7.0-7.2) the middle bacillus amyloliquefaciens inoculating 2-5 weight portion
The bacterium solution of (Bacillus amyloliquefaciens), 37 DEG C of cultivations, until bacillus amyloliquefaciens
The viable count of (Bacillus amyloliquefaciens) is 2-20 × 107Individual/gram culture medium.
By the cultivation in the most independent cultivating system and mixing proportionally after cultivating, described mixed
Close condition have no particular limits, preferably such that to microbial bacterial agent meet following condition: with
The number meter of viable bacteria body, relative to bacillus amyloliquefaciens every part described, containing of described Bacillus subtillis
Amount is 0.05-0.2 part, and the content of described bacillus licheniformis is 0.05-0.2 part.
According to the present invention, described Bacillus subtillis (Bacillus subtilis) and bacillus licheniformis
The bacterial classification of (Bacillus lincheniformis) can be commercially available.Such as, described Ko subtilis
Bacillus is purchased from Chinese agriculture Culture Collection and numbered ACCC03221, described lichens bud
Born of the same parents bacillus is purchased from Chinese agriculture Culture Collection and numbered ACCC02698.
Present invention also offers bacillus amyloliquefaciens as above and microbial bacterial agent as above
Application in alkaline land improving.
Wherein, Cl in described alkaline land soil-Concentration can be at least 0.15mol/kg, pH value is permissible
Being more than 8, the bacillus amyloliquefaciens of the present invention can have more than 0.15mol/kg Cl-Concentration and 8
The alkaline land soil of above pH value well grows and plays reduction Cl-Effect.
Present invention also offers bacillus amyloliquefaciens as above and microbial bacterial agent as above
Application in increasing drought resistance in plants.
Wherein, described plant can be watermelon, muskmelon, cucumber, tomato, capsicum, eggplant, clover,
Paddy rice, apple, pears, grape, peach, apricot or strawberry etc..
Present invention also offers bacillus amyloliquefaciens as above and microbial bacterial agent as above
Application in preventing and treating wheat sharp eyespot.
Present invention also offers bacillus amyloliquefaciens as above and microbial bacterial agent as above
Application in preventing and treating cucumber fusarium axysporum.
Further describe the present invention by the following examples:
Embodiment 1
The present embodiment is for illustrating bacillus amyloliquefaciens and the performance thereof of the present invention.
By the bacillus amyloliquefaciens (Bacillus that deposit number is CCTCC NO.M2013501
Amyloliquefaciens) be inoculated into LB culture medium (containing tryptone 10g/L, yeast extract 5g/L,
NaCl10g/L), in, cultivate to strain density OD600Value is 0.6-0.8, obtains bacterium solution.
The concentration adding NaCl adjustment NaCl in LB culture medium is 220g/L, and adds NaOH
Adjusting pH value is 9.5, obtains Saline Alkali Stress culture medium.
Above-mentioned for 3mL bacterium solution is added in Saline Alkali Stress culture medium, makes initial OD600It is 0.1, in 30 DEG C,
180 turns of every point of lower shaken cultivation, strain density (OD in 4h sampling and measuring zymotic fluid600).Result shows
Show, the zymotic fluid OD of the 0th, 4,8,12,16,20,24 hours600Be respectively 0.1,0.12,0.17,
0.22、0.37、0.54.Thus illustrating, deposit number is the solution starch of CCTCC NO.M2013501
Bacillus can be in the Saline Alkali Stress culture medium that the concentration of NaCl is 220g/L and pH value is 9.5
Good growth.
The concentration adding anhydrous calcium chloride adjustment calcium chloride in LB culture medium is 0.5g/L, adds
MgSO4Adjust MgSO4Concentration be 30g/L, and pH value is adjusted to 9.5, obtains saline and alkaline removing
Test media.
Above-mentioned for 3mL bacterium solution is added in the saline and alkaline removing test media of 150mL, in 30 DEG C, 180
Turn every point of lower shaken cultivation, Cl in sampling and measuring zymotic fluid after 24h-Concentration and pH value, result show
Show, Cl-Concentration have dropped 21.2%, pH value drops to 8.2, it was demonstrated that deposit number is CCTCC NO.
The bacillus amyloliquefaciens of M2013501 has the saline and alkaline Scavenging activity of excellence.
Comparative example 1
By the solution starch bud purchased from Chinese agriculture Culture Collection and numbered ACCC01855
Born of the same parents bacillus (Bacillus amyloliquefaciens) be inoculated into LB culture medium (containing tryptone 10g/L,
Yeast extract 5g/L, NaCl10g/L) in, cultivate to strain density OD600Value is 0.6-0.8, obtains
Bacterium solution.
The concentration adding NaCl adjustment NaCl in LB culture medium is 220g/L, and adds NaOH
Adjusting pH value is 9.5, obtains Saline Alkali Stress culture medium.
Above-mentioned for 3mL bacterium solution is added in Saline Alkali Stress culture medium, makes initial OD600It is 0.1, in 30 DEG C,
180 turns of every point of lower shaken cultivation, strain density (OD in 4h sampling and measuring zymotic fluid600).Result shows
Show, the zymotic fluid OD of the 0th, 4,8,12,16,20,24 hours600Be respectively 0.1,0.1,0.1,
0.1、0.1、0.1.Thus illustrate, purchased from Chinese agriculture Culture Collection and numbered ACCC
The bacillus amyloliquefaciens of 01855 can not the concentration of NaCl be 220g/L and pH value be 9.5 saline and alkaline
Coerce in culture medium and grow.
The concentration adding anhydrous calcium chloride adjustment calcium chloride in LB culture medium is 0.5g/L, adds
MgSO4Adjust MgSO4Concentration be 30g/L, and pH value is adjusted to 9.5, obtains saline and alkaline removing
Test media.
Above-mentioned for 3mL bacterium solution is added in the saline and alkaline removing test media of 150mL, in 30 DEG C, 180
Turn every point of lower shaken cultivation, Cl in sampling and measuring zymotic fluid after 24h-Concentration and pH value, result show
Show, Cl-Concentration have dropped 1.5%, pH value drops to 9.3, it was demonstrated that purchased from Chinese agriculture microbial bacteria
The bacillus amyloliquefaciens planting preservation center and numbered ACCC01855 does not the most have saline and alkaline removing energy
Power.
Embodiment 2
The present embodiment is for illustrating microbial bacterial agent and the performance thereof of the present invention.
(1) in culture medium (glucose 5g, dregs of beans 30g, peptone 2g, the distillation of 100 weight portions
Water 1000ml, pH7.0-7.2) in inoculation 5 weight portions Bacillus subtillis bacterium solution (purchased from China agriculture
Industry Culture Collection and numbered ACCC03221, train in the most identical LB culture medium
Support to strain density OD600Value is 0.6-0.8, obtains bacterium solution), 37 DEG C of cultivations, carry out in incubation
Sample and observed by ascites method, until the viable count of Bacillus subtillis is
2×107The culture medium of CFU/ gram.
(2) in culture medium (corn steep liquor 0.45g, peptone 1.50g, the beancake powder leaching of 100 weight portions
Juice is 1:15, glucose 1.50g, pH value 7.5) the middle bacillus licheniformis bacterium solution inoculating 5 weight portions
(purchased from Chinese agriculture Culture Collection and numbered ACCC02698, at the most identical LB
Culture medium is cultivated to strain density OD600Value is 0.6-0.8, obtains bacterium solution), 37 DEG C of cultivations, cultivating
During be sampled and observed by ascites method, until the work of bacillus licheniformis
Bacterium number is 2 × 107The culture medium of CFU/ gram.
(3) at culture medium (glucose 15g, starch 1g, beancake powder 25g, the sulfuric acid of 100 weight portions
Manganese 1g, potassium dihydrogen phosphate 1.5g, magnesium sulfate 0.5g, yeast extract 0.2g, iron chloride 0.1g, calcium carbonate
0.1g, pH7.0-7.2) in inoculation 5 weight portions bacillus amyloliquefaciens bacterium solution (deposit number is CCTCC
NO.M2013501, cultivates to strain density OD600 value as 0.6-0.8 in the most identical LB culture medium,
Obtain bacterium solution), 37 DEG C of cultivations, it is sampled in incubation and is entered by ascites method
Row is observed, until the viable count of bacillus amyloliquefaciens is 2 × 107The culture medium of CFU/ gram.
(4) by the Bacillus subtillis respectively obtained in step (1) to (3), bacillus licheniformis,
Bacillus amyloliquefaciens proportionally mixes, and mixed proportion makes in the microbial bacterial agent obtained, with work
The number meter of thalline, relative to bacillus amyloliquefaciens every part described, the content of described Bacillus subtillis
Being 0.1 part, the content of described bacillus licheniformis is 0.1 part, obtains the microbial bacterial agent of the present embodiment.
Embodiment 3
The present embodiment is for illustrating microbial bacterial agent and the performance thereof of the present invention.
According to the method for embodiment 2, by the Bacillus subtillis respectively obtained in step (1) to (3),
Bacillus licheniformis, bacillus amyloliquefaciens proportionally mix, except that, mixed proportion
Make in the microbial bacterial agent obtained, in terms of the number of viable bacteria body, relative to solution starch brood cell's bar every part described
Bacterium, the content of described Bacillus subtillis is 0.2 part, and the content of described bacillus licheniformis is 0.5 part,
Obtain the microbial bacterial agent of the present embodiment.
Embodiment 4
The present embodiment is for illustrating microbial bacterial agent and the performance thereof of the present invention.
According to the method for embodiment 2, by the Bacillus subtillis respectively obtained in step (1) to (3),
Bacillus licheniformis, bacillus amyloliquefaciens, bacillus amyloliquefaciens proportionally mix, and institute is not
With, mixed proportion makes in the microbial bacterial agent obtained, in terms of the number of viable bacteria body, relative to every part
Described bacillus amyloliquefaciens, the content of described Bacillus subtillis is 0.2 part, described lichens brood cell's bar
The content of bacterium is 0.05 part, obtains the microbial bacterial agent of the present embodiment.
Embodiment 5
The present embodiment is for illustrating microbial bacterial agent and the performance thereof of the present invention.
According to the method for embodiment 2, by the Bacillus subtillis respectively obtained in step (1) to (3),
Bacillus licheniformis, bacillus amyloliquefaciens, bacillus amyloliquefaciens proportionally mix, and institute is not
With, mixed proportion makes in the microbial bacterial agent obtained, in terms of the number of viable bacteria body, relative to every part
Described bacillus amyloliquefaciens, the content of described Bacillus subtillis is 2 parts, described bacillus licheniformis
Content be 5 parts, obtain the microbial bacterial agent of the present embodiment.
Embodiment 6
The present embodiment is for illustrating microbial bacterial agent and the performance thereof of the present invention.
Microbial bacterial agent is prepared, except for the difference that, by step (2) to (3) according to the method for embodiment 2
In the bacillus licheniformis that respectively obtains, bacillus amyloliquefaciens proportionally mix, but be added without
Bacillus subtillis, obtains the microbial bacterial agent of the present embodiment.
Comparative example 2
Microbial bacterial agent is prepared according to the method for embodiment 2, except for the difference that, solution starch brood cell's bar used
Bacterium is the solution starch brood cell purchased from Chinese agriculture Culture Collection and numbered ACCC01855
Bacillus.
Testing example 1
This testing example measures embodiment 2-6 and the microbial bacteria of comparative example 2 by outdoor potted plant experiment
Agent effect in alkaline land improving.
The matrix soil of outdoor potted plant use is purchased from the flower cultivating soil of flowers market, Beijing, soil organic matter content
10.24g/kg, full nitrogen 0.561g/kg, full phosphorus 1.14g/kg, full potassium 14.98g/kg, available nitrogen 39.19mg/kg,
Rapid available phosphorus 16.26mg/kg, available potassium 160.07mg/kg, pH6.5.Further, add in matrix soil
NaCl solution so that Cl-Concentration be 0.19mol/kg, add NaOH so that pH value is 9.0,
Obtain testing soil.Pot experiment basin height 21cm, upper diameter 20cm, lower diameter 13cm, every basin adds
Soil 3.8kg, is separately added into the microbial bacterial agent 0.05kg of embodiment 2-6 and comparative example 2.
Basin is planted into alfalfa, when the average plant height of alfalfa reaches 20cm, according to " NY/T
1121.17-2006 Soil K+adsorption the 17th part: the mensuration of Soil Chlorine ion concentration " in method measure
Cl in soil-Concentration, and calculate Cl-Clearance, result is as shown in table 1.
Table 1
| Microbial bacterial agent | Average Cl-Clearance |
| Embodiment 2 | 77% |
| Embodiment 3 | 74% |
| Embodiment 4 | 75% |
| Embodiment 5 | 67% |
| Embodiment 6 | 64% |
| Comparative example 2 | 48% |
From the data of table 1 it can be seen that the microbial bacterial agent of the present invention has excellent alkaline land improving effect
Really, and, preferably in terms of the number of viable bacteria body, relative to bacillus amyloliquefaciens every part described, institute
The content stating Bacillus subtillis is 0.05-0.2 part, and the content of described bacillus licheniformis is 0.05-0.2
In the case of Fen, described microbial bacterial agent has the ability that more excellent removal is saline and alkaline.
Testing example 2
This testing example measures embodiment 2-6 and the microbial bacteria of comparative example 2 by outdoor potted plant experiment
Agent effect in increasing drought resistance in plants.
The matrix soil using testing example 1 makes Nutrition Soil basin alms bowl, by tomato seeds vernalization 48h
After (25 DEG C), will move in Nutrition Soil basin alms bowl, transplant seedlings to new Nutrition Soil as after greenhouse grows 15 days
In basin alms bowl (each basin alms bowl 3.8kg soil, transplant 4 strain tomatoes), water 200mL water every other day.Transplant seedlings
After 15 days, each basin alms bowl is separately added into the microbial bacterial agent 0.05kg of embodiment 2-6 and comparative example 2, just
After often watering 6 days, after cutting off the water supply 20 days, measure tomato leaf relative water content, result such as table 2 institute
Show.
Table 2
| Microbial bacterial agent | The relative water content of tomato leaf after cutting off the water supply 20 days |
| Embodiment 2 | 87% |
| Embodiment 3 | 84% |
| Embodiment 4 | 85% |
| Embodiment 5 | 77% |
| Embodiment 6 | 74% |
| Comparative example 2 | 54% |
From the data of table 2 it can be seen that the microbial bacterial agent of the present invention has excellent increase drought tolerance in plants
Property effect, and, preferably in terms of the number of viable bacteria body, relative to bacillus amyloliquefaciens every part described,
The content of described Bacillus subtillis is 0.05-0.2 part, and the content of described bacillus licheniformis is 0.05-0.2
In the case of Fen, described microbial bacterial agent has the more excellent ability increasing drought resistance in plants.
Testing example 3
This testing example measures embodiment 2-6 and the microbial bacteria of comparative example 2 by outdoor potted plant experiment
Agent effect in preventing and treating wheat sharp eyespot.From the wheat planting having infected banded sclerotial blight, big Tanaka takes out soil
Earth, as the susceptible soil of banded sclerotial blight.
The susceptible soil of above-mentioned banded sclerotial blight is used to make Nutrition Soil basin alms bowl, by wheat seed vernalization 48h (25 DEG C)
After, its kind enters Nutrition Soil basin alms bowl, and (each basin alms bowl 3.8kg susceptible soil of above-mentioned banded sclerotial blight plants 20
Strain wheat, and it is separately added into embodiment 2-6 and the microbial bacterial agent of comparative example 2 at each basin alms bowl
0.05kg), normal plantation wheat, after 90 days, measures the preventive effect of banded sclerotial blight sense, and result is as shown in table 3.
Table 3
| Microbial bacterial agent | Plantation wheat preventive effect of banded sclerotial blight after 90 days |
| Embodiment 2 | 80% |
| Embodiment 3 | 78% |
| Embodiment 4 | 79% |
| Embodiment 5 | 69% |
| Embodiment 6 | 63% |
| Comparative example 2 | 44% |
From the data of table 3 it can be seen that the microbial bacterial agent of the present invention to have excellent preventing and treating wheat line withered
Sick effect, and, preferably in terms of the number of viable bacteria body, relative to solution starch brood cell's bar every part described
Bacterium, the content of described Bacillus subtillis is 0.05-0.2 part, and the content of described bacillus licheniformis is
In the case of 0.05-0.2 part, described microbial bacterial agent has more excellent preventing and treating wheat sharp eyespot
Ability.
Testing example 4
This testing example measures embodiment 2-6 and the microbial bacteria of comparative example 2 by outdoor potted plant experiment
Agent effect in preventing and treating cucumber fusarium axysporum.From the green cucumber having infected droop, big Tanaka takes out soil
Earth, as the susceptible soil of droop.
The matrix soil using testing example 1 makes Nutrition Soil basin alms bowl, by cucumber seeds vernalization 48h
After (25 DEG C), move it in Nutrition Soil basin alms bowl, transplant seedlings to new nutrition as after greenhouse grows 15 days
Soil basin alms bowl (each basin alms bowl 3.8kg susceptible soil of above-mentioned droop, transplants 6 strain cucumber, and each
Basin alms bowl is separately added into the microbial bacterial agent 0.05kg of embodiment 2-6 and comparative example 2), normally plant cucumber
After 60 days, measuring the preventive effect of droop, result is as shown in table 4.
Table 4
| Microbial bacterial agent | Plantation cucumber preventive effect of droop after 60 days |
| Embodiment 2 | 74% |
| Embodiment 3 | 72% |
| Embodiment 4 | 73% |
| Embodiment 5 | 67% |
| Embodiment 6 | 62% |
| Comparative example 2 | 33% |
From the data of table 4 it can be seen that the microbial bacterial agent of the present invention have excellent preventing and treating cucumber wither
Sick effect, and, preferably in terms of the number of viable bacteria body, relative to solution starch brood cell's bar every part described
Bacterium, the content of described Bacillus subtillis is 0.05-0.2 part, and the content of described bacillus licheniformis is
In the case of 0.05-0.2 part, described microbial bacterial agent has the energy of more excellent preventing and treating cucumber fusarium axysporum
Power.
The preferred embodiment of the present invention described in detail above, but, the present invention is not limited to above-mentioned reality
Execute the detail in mode, in the technology concept of the present invention, can be to the technical side of the present invention
Case carries out multiple simple variant, and these simple variant belong to protection scope of the present invention.
It is further to note that each the concrete technology described in above-mentioned detailed description of the invention is special
Levy, in the case of reconcilable, can be combined by any suitable means, in order to avoid need not
The repetition wanted, various possible combinations are illustrated by the present invention the most separately.
Additionally, can also be combined between the various different embodiment of the present invention, as long as its
Without prejudice to the thought of the present invention, it should be considered as content disclosed in this invention equally.
Claims (9)
1. a bacillus amyloliquefaciens, it is characterised in that: this bacillus amyloliquefaciens (Bacillus amyloliquefaciens) deposit number be CCTCC NO.M 2013501.
2. a microbial bacterial agent, this microbial bacterial agent comprises culture medium and thalline, it is characterised in that: described thalline includes the bacillus amyloliquefaciens that deposit number is CCTCC NO.M 2013501, and described thalline also include Bacillus subtillis (Bacillus subtilis) ACCC 03221 and bacillus licheniformis (Bacillus lincheniformis)ACCC 02698。
Microbial bacterial agent the most according to claim 2, it is characterised in that: the total viable count contained by every gram of described microbial bacterial agent is 2-20 × 107CFU。
4. according to the microbial bacterial agent described in Claims 2 or 3, it is characterized in that: in terms of the number of viable bacteria body, relative to bacillus amyloliquefaciens every part described, the content of described Bacillus subtillis is 0.05-0.2 part, and the content of described bacillus licheniformis is 0.05-0.2 part.
5. according to the microbial bacterial agent described in Claims 2 or 3, it is characterised in that: described culture medium is at least one in beef-protein medium, broth bouillon and LB culture medium.
6. microbial bacterial agent described in any one application in alkaline land improving in bacillus amyloliquefaciens described in claim 1 and claim 2-5.
7. microbial bacterial agent described in any one application in increasing drought resistance in plants in bacillus amyloliquefaciens described in claim 1 and claim 2-5.
8. microbial bacterial agent described in any one application in preventing and treating wheat sharp eyespot in bacillus amyloliquefaciens described in claim 1 and claim 2-5.
9. microbial bacterial agent described in any one application in preventing and treating cucumber fusarium axysporum in bacillus amyloliquefaciens described in claim 1 and claim 2-5.
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| CN104630103B (en) * | 2015-01-29 | 2019-07-09 | 深圳市润田生物科技有限公司 | A kind of microbial bacterial agent and its preparation method and application |
| CN105670976B (en) * | 2016-03-18 | 2021-07-06 | 中化农业生态科技(湖北)有限公司 | Fermentation medium of bacillus amyloliquefaciens and application thereof |
| CN107699245A (en) * | 2017-10-09 | 2018-02-16 | 广西宾德利生物科技有限公司 | Microbe soil repairs insecticide and preparation method and application |
| CN108977373B (en) * | 2018-07-04 | 2022-06-28 | 中国热带农业科学院热带生物技术研究所 | Bacillus terracotta cotta HMD9161 and microbial inoculum and application thereof |
| CN110684683A (en) * | 2019-09-10 | 2020-01-14 | 北方民族大学 | Bacillus amyloliquefaciens, microbial inoculum and application |
| CN113373092B (en) * | 2021-06-30 | 2023-09-15 | 海南金雨丰生物工程有限公司 | Composite microbial preparation and preparation method thereof |
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| CN1369552A (en) * | 2002-03-20 | 2002-09-18 | 刘铁军 | Composite microbial bacterim praparation and leaven and fertilizer prepared from it |
| CN102229900A (en) * | 2011-05-20 | 2011-11-02 | 周剑平 | Compound probiotics preparation for biological organic fertilizers and preparation method thereof |
| CN103131655A (en) * | 2013-03-06 | 2013-06-05 | 江苏苏滨生物农化有限公司 | Bacillus amyloliquefaciens K-8 and bactericide thereof |
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| CN1369552A (en) * | 2002-03-20 | 2002-09-18 | 刘铁军 | Composite microbial bacterim praparation and leaven and fertilizer prepared from it |
| CN102229900A (en) * | 2011-05-20 | 2011-11-02 | 周剑平 | Compound probiotics preparation for biological organic fertilizers and preparation method thereof |
| CN103131655A (en) * | 2013-03-06 | 2013-06-05 | 江苏苏滨生物农化有限公司 | Bacillus amyloliquefaciens K-8 and bactericide thereof |
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