CN103805534A - Bacillus methylotrophicus, microorganism bacterium agents and application of bacillus methylotrophicus and microorganism bacterium agents - Google Patents
Bacillus methylotrophicus, microorganism bacterium agents and application of bacillus methylotrophicus and microorganism bacterium agents Download PDFInfo
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Abstract
The invention provides bacillus methylotrophicus with preservation number-CCTCC NO. M2013500. The invention also provides a microorganism bacterium agent. The microorganism bacterium agent contains a culture medium and thallus, wherein the thallus comprise the bacillus methylotrophicus with preservation number-CCTCC NO. M2013500, bacillus subtilis, bacillus lincheniformis and bacillus amyloliquefaciens. The invention also provides an application of the bacillus methylotrophicus and the above microorganism bacterium agents to improvement of saline lands, improvement of plant drought tolerance, prevention of bacterial wilt of pepper and prevention of Citrus anthracnose.
Description
Technical field
The present invention relates to agricultural biological technical field, particularly, relate to a kind of methylotrophy type bacillus, a kind of microbiobacterial agent and their application.
Background technology
Saline Land is the serious problems that affect agriculture production and ecotope.Approximately there is 20% arable land in the whole world and approaches 50% irrigated farmland output and is seriously subject to the saline and alkaline impact of soil high density.In China, approximately there are 100,000,000 mu of secondary salinization farmlands, account for 10% of total cultivated area.How really improveing and effectively to utilize saltings, improve economy, society and ecological benefits that agroforestry are produced, is the significant problem that Modern Agriculture forestry is paid close attention to.
Alkaline land improving research field at home and abroad, has successively proposed " plant rice and change alkali " agricultural improvement measure, the ameliorative measure of " washing salinity by irrigation " engineering, and utilize the chemical modifying measures such as gypsum, calcium chloride, industrial waste acid, coal-fired flue gas desulfurization thing.These measure instant effects, have obtained remarkable achievement.But improvement cost is high, and resource cost is large, and produces nutrient loss, pollute the problems such as downstream, improved effect are unstable, the easy accumulation of salt in the surface soil.
In recent years, on the basis of achievement that studies for a long period of time, people recognize gradually the improvement in saltings and utilization are combined from the angle of ecology recovering, i.e. " residence is improved in utilization; improvement is parallel with utilization ", the ecological recovery technology take biological utilisation as core is the breach that following alkaline land improving is repaired.At present, biological modification comprises plantation saline alkali tolerant plant, uses microbial-bacterial fertilizer etc.And develop efficient microbial-bacterial fertilizer preparation, first will obtain tolerance saline and alkaline and to the saline and alkaline microorganism strains with removal effect, still, it is still poor that existing microorganism strains is removed saline and alkaline effect.
Summary of the invention
Remove the poor defect of saline and alkaline effect in order to overcome existing microorganism strains, the invention provides a kind of methylotrophy type bacillus, the deposit number of this methylotrophy type bacillus (Bacillus methylotrophicus) is CCTCC NO.M2013500.
The present invention also provides a kind of microbiobacterial agent, this microbiobacterial agent comprises substratum and thalline, described thalline comprises that deposit number is the methylotrophy type bacillus of CCTCC NO.M2013500, and described thalline also comprises Bacillus subtillis (Bacillus subtilis), bacillus licheniformis (Bacillus lincheniformis) and bacillus amyloliquefaciens (Bacillus amyloliquefaciens).
The present invention also provides methylotrophy type bacillus as above and the application of microbiobacterial agent as above in alkaline land improving.
The present invention also provides methylotrophy type bacillus as above and microbiobacterial agent as above in the application increasing in drought resistance in plants.
The present invention also provides methylotrophy type bacillus as above and the application of microbiobacterial agent as above in the bacterial wilt of control capsicum.
The present invention also provides methylotrophy type bacillus as above and the application of microbiobacterial agent as above in control citrus anthracnose.
By technique scheme, the present invention can remove saline and alkaline more effectively, for example, Soil Desalting rate can be increased to more than 55%.
Other features and advantages of the present invention are described in detail the embodiment part subsequently.
Biomaterial preservation
Methylotrophy type bacillus of the present invention (Bacillus methylotrophicus) is the pure growth that the present inventor separates from the soil of Ling County, Shandong, its deposit number is CCTCC NO.M2013500, preservation date is on October 24th, 2013, depositary institution is Chinese Typical Representative culture collection center, address is Wuhan, China Wuhan University, and Classification And Nomenclature is methylotrophy type bacillus (Bacillus methylotrophicus).
Embodiment
Below the specific embodiment of the present invention is elaborated.Should be understood that, embodiment described herein only, for description and interpretation the present invention, is not limited to the present invention.
The invention provides a kind of methylotrophy type bacillus, the deposit number of this methylotrophy type bacillus (Bacillus methylotrophicus) is CCTCC NO.M2013500.
The present invention also provides a kind of microbiobacterial agent, this microbiobacterial agent comprises substratum and thalline, described thalline comprises that deposit number is the methylotrophy type bacillus of CCTCC NO.M2013500, and described thalline also comprises Bacillus subtillis (Bacillus subtilis), bacillus licheniformis (Bacillus lincheniformis) and bacillus amyloliquefaciens (Bacillus amyloliquefaciens).
Wherein, the amount of the thalline containing in described microbiobacterial agent can in very large range change, and under preferable case, every gram of contained total viable count of described microbiobacterial agent can be 2-20 × 10
7cFU.
Preferably, in the number of viable bacteria body, with respect to methylotrophy type bacillus every part described, the content of described Bacillus subtillis is 0.05-0.2 part, the content of described bacillus licheniformis is 0.05-0.2 part, and the content of described bacillus amyloliquefaciens is 0.01-0.05 part.Under this preferable case, described microbiobacterial agent has the saline and alkaline ability of better removal.
According to the present invention, the kind of described substratum can in very large range change, can be the various substratum of cultivating methylotrophy type bacillus, Bacillus subtillis, bacillus licheniformis, bacillus amyloliquefaciens that can be used in, for example, can be that the conventional substratum such as beef-protein medium, broth culture, LB substratum are as seed culture medium, for the preservation of bacterial classification; And the substratum of fermentation is generally used for production, the kind of these substratum is also conventionally known to one of skill in the art.Above-mentioned substratum can be commercially available or prepare according to the record of " microbiological culture media handbook " (Microbiology Culture Media Manual).For example, beef-protein medium: extractum carnis 3g, peptone 10g, sodium-chlor 5g, agar 15-20g, distilled water 1000ml, pH value 7.0-7.2,121 ℃ of sterilizing 30min; Fermention medium: glucose 15g, starch 1g, soybean cake powder 25g, manganous sulfate 1g, potassium primary phosphate 1.5g, magnesium sulfate 0.5g, yeast extract paste 0.2g, iron(ic) chloride 0.1g, calcium carbonate 0.1g, pH value 7.0-7.2,115 ℃ of sterilizing 15min.
Wherein, the preparation method of described microbiobacterial agent can comprise: methylotrophy type bacillus, Bacillus subtillis, bacillus licheniformis and bacillus amyloliquefaciens are inoculated in substratum and are cultivated respectively.Wherein, can be independently cultivating respectively methylotrophy type bacillus, Bacillus subtillis, bacillus licheniformis and bacillus amyloliquefaciens in culture system separately, and proportionally mix cultivating respectively the microorganism obtaining.Under preferable case, by separately independently the cultivation in culture system and cultivate after mixing, the total viable count that makes the every gram of microbiobacterial agent obtaining is 2-20 × 10
7cFU.
According to the present invention, the cultural method of described subtilis (Bacillus subtilis) has no particular limits as conventionally known to one of skill in the art, for example, can first subtilis be cultured to nectar degree OD in seed culture medium (LB substratum)
600value is 0.6-0.8, obtain bacterium liquid, then at productive culture base (the glucose 5g of 100 weight parts, dregs of beans 30g, peptone 2g, distilled water 1000ml, pH7.0-7.2) in the bacterium liquid of subtilis (Bacillus subtilis) of inoculation 2-5 weight part, 37 ℃ of cultivations, until the viable count of subtilis (Bacillus subtilis) is 2-20 × 10
7individual/gram substratum.
The cultural method of described Bacillus licheniformis (Bacillcus lincheniformis) has no particular limits as conventionally known to one of skill in the art, for example, can first Bacillus licheniformis be cultured to nectar degree OD in seed culture medium (LB substratum)
600value is 0.6-0.8, obtain bacterium liquid, then at substratum (the corn steep liquor 0.45g of 100 weight parts, peptone 1.50g, it is 1:15 that soybean cake powder soaks juice, glucose 1.50g, pH value 7.5) in the bacterium liquid of Bacillus licheniformis (Bacillcus lincheniformis) of inoculation 2-5 weight part, 37 ℃ of cultivations, until the viable count of Bacillus licheniformis (Bacillcus lincheniformis) is 2-20 × 10
7individual/gram substratum.
The cultural method of described bacillus amyloliquefaciens (Bacillus amyloliquefaciens) has no particular limits, and for example, can first bacillus amyloliquefaciens be cultured to nectar degree OD in seed culture medium (LB substratum)
600value is 0.6-0.8, obtain bacterium liquid, then at 100 weight part substratum (soybean cake powder 50g, sucrose 20g, ammonium sulfate 5g, trisodium citrate 2.5g, potassium primary phosphate 0.3g, magnesium sulfate 0.5g, ferrous sulfate 0.05g, pH7.0-7.2) the bacterium liquid of the bacillus amyloliquefaciens (Bacillus amyloliquefaciens) of inoculation 2-5 weight part in, 37 ℃ of cultivations, until the viable count of bacillus amyloliquefaciens (Bacillus amyloliquefaciens) is 2-20 × 10
7individual/gram substratum.
The cultural method of described methylotrophy type genus bacillus has no particular limits, and for example, can first methylotrophy type genus bacillus be cultured to nectar degree OD in seed culture medium (LB substratum)
600value, for 0.6-0.8, obtains bacterium liquid, then at 100 weight part substratum (glucose 15g, starch 1g, soybean cake powder 25g, manganous sulfate 1g, potassium primary phosphate 1.5g, magnesium sulfate 0.5g, yeast extract paste 0.2g, iron(ic) chloride 0.1g, calcium carbonate 0.1g, pH7.0-7.2) the bacterium liquid of the methylotrophy type genus bacillus of inoculation 2-5 weight part in, 37 ℃ of cultivations, until the viable count of methylotrophy type genus bacillus is 2-20 × 10
7individual/gram substratum.
By separately independently the cultivation in culture system and cultivate after mixing proportionally, the condition of described mixing has no particular limits, preferably can make the microbiobacterial agent obtaining meet the following conditions: in the number of viable bacteria body, with respect to methylotrophy type bacillus every part described, the content of described Bacillus subtillis is 0.05-0.2 part, the content of described bacillus licheniformis is 0.05-0.2 part, and the content of described bacillus amyloliquefaciens is 0.01-0.05 part.
According to the present invention, the bacterial classification of described Bacillus subtillis (Bacillus subtilis), bacillus licheniformis (Bacillus lincheniformis), bacillus amyloliquefaciens (Bacillus amyloliquefaciens) can be commercially available.For example, described Bacillus subtillis is purchased from Chinese agriculture microbial strains preservation center and be numbered ACCC03221, described bacillus licheniformis is purchased from Chinese agriculture microbial strains preservation center and be numbered ACCC02698, and described bacillus amyloliquefaciens is purchased from Chinese agriculture microbial strains preservation center and be numbered ACCC10166.
The present invention also provides methylotrophy type bacillus as above and the application of microbiobacterial agent as above in alkaline land improving.
Wherein, in described alkaline land soil, the concentration of Cl-can be 0.15mol/kg at least, pH value can be more than 8, well grows and play the effect that reduces Cl-in the alkaline land soil of the pH value that methylotrophy type bacillus of the present invention can be more than having the above Cl-concentration of 0.15mol/kg and 8.
The present invention also provides methylotrophy type bacillus as above and microbiobacterial agent as above in the application increasing in 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.
The present invention also provides methylotrophy type bacillus as above and the application of microbiobacterial agent as above in the bacterial wilt of control capsicum.
The present invention also provides methylotrophy type bacillus as above and the application of microbiobacterial agent as above in control citrus anthracnose.
Further describe by the following examples the present invention:
Embodiment 1
The present embodiment is used for illustrating methylotrophy type bacillus of the present invention and performance thereof.
The methylotrophy type bacillus (Bacillus methylotrophicus) that is CCTCC NO.M2013500 by deposit number is inoculated in LB substratum (containing Tryptones 10g/L, yeast extract 5g/L, NaCl10g/L), is cultured to nectar degree OD
600value, for 0.6-0.8, obtains bacterium liquid.
In LB substratum, adding NaCl to adjust the concentration of NaCl is 220g/L, and adds NaOH to adjust pH value to be 9.5, to obtain Saline Alkali Stress substratum.
Above-mentioned 3mL bacterium liquid is added in Saline Alkali Stress substratum, make initial OD
600be 0.1, in 30 ℃, 180 turn shaking culture under per minute, nectar degree (OD in 4h sampling and measuring fermented liquid
600).Result shows, the fermented liquid OD of the 0th, 4,8,12,16,20,24 hours
600be respectively 0.1,0.13,0.16,0.23,0.39,0.55.Explanation thus, deposit number is that the methylotrophy type bacillus of CCTCC NO.M2013500 can be well to grow in 220g/L and the pH value Saline Alkali Stress substratum that is 9.5 in the concentration of NaCl.
The concentration that adds Calcium Chloride Powder Anhydrous to adjust calcium chloride in LB substratum is 0.5g/L, adds MgSO
4adjust MgSO
4concentration be 30g/L, and pH value is adjusted into 9.5, obtain saline and alkaline removing test media.
Above-mentioned 3mL bacterium liquid is added in the saline and alkaline removing test media of 150mL, in 30 ℃, 180 turn shaking culture under per minute, the concentration of Cl-and pH value in sampling and measuring fermented liquid after 24h, result shows, the density loss of Cl-21.2%, pH value drops to 8.2, proves that deposit number is that the methylotrophy type bacillus of CCTCC NO.M2013500 has excellent saline and alkaline removing ability.
Comparative example 1
By being inoculated in LB substratum (containing Tryptones 10g/L, yeast extract 5g/L, NaCl10g/L) purchased from Chinese agriculture microbial strains preservation center and the methylotrophy type bacillus (Bacillus methylotrophicus) that is numbered ACCC02647, be cultured to nectar degree OD
600value, for 0.6-0.8, obtains bacterium liquid.
In LB substratum, adding NaCl to adjust the concentration of NaCl is 220g/L, and adds NaOH to adjust pH value to be 9.5, to obtain Saline Alkali Stress substratum.
Above-mentioned 3mL bacterium liquid is added in Saline Alkali Stress substratum, make initial OD
600be 0.1, in 30 ℃, 180 turn shaking culture under per minute, nectar degree (OD in 4h sampling and measuring fermented liquid
600).Result shows, the fermented liquid OD of the 0th, 4,8,12,16,20,24 hours
600be respectively 0.1,0.1,0.1,0.1,0.1,0.1.Explanation thus, purchased from Chinese agriculture microbial strains preservation center and be numbered ACCC02076 methylotrophy type bacillus can not be to grow in 220g/L and the pH value Saline Alkali Stress substratum that is 9.5 in the concentration of NaCl.
The concentration that adds Calcium Chloride Powder Anhydrous to adjust calcium chloride in LB substratum is 0.5g/L, adds MgSO
4adjust MgSO
4concentration be 30g/L, and pH value is adjusted into 9.5, obtain saline and alkaline removing test media.
Above-mentioned 3mL bacterium liquid is added in the saline and alkaline removing test media of 150mL, in 30 ℃, 180 turn shaking culture under per minute, the concentration of Cl-and pH value in sampling and measuring fermented liquid after 24h, result shows, the density loss of Cl-1.5%, pH value drops to 9.4, proves substantially not have saline and alkaline removing ability purchased from Chinese agriculture microbial strains preservation center and the methylotrophy type bacillus that is numbered ACCC02647.
Embodiment 2
The present embodiment is used for illustrating microbiobacterial agent of the present invention and performance thereof.
(1) at substratum (the glucose 5g of 100 weight parts, dregs of beans 30g, peptone 2g, distilled water 1000ml, pH7.0-7.2) the Bacillus subtillis bacterium liquid of inoculation 5 weight parts in is (purchased from Chinese agriculture microbial strains preservation center and be numbered ACCC03221, being as above cultured to nectar degree OD in identical LB substratum
600value, for 0.6-0.8, obtains bacterium liquid), 37 ℃ of cultivations sample and observe by ascites method in culturing process, until the viable count of Bacillus subtillis is 2 × 10
7the substratum of CFU/ gram.
(2) at substratum (the corn steep liquor 0.45g of 100 weight parts, peptone 1.50g, it is 1:15 that soybean cake powder soaks juice, glucose 1.50g, pH value 7.5) in the bacillus licheniformis bacterium liquid of inoculation 5 weight parts (purchased from Chinese agriculture microbial strains preservation center and be numbered ACCC02698, being as above cultured to nectar degree OD in identical LB substratum
600value, for 0.6-0.8, obtains bacterium liquid), 37 ℃ of cultivations sample and observe by ascites method in culturing process, until the viable count of bacillus licheniformis is 2 × 10
7the substratum of CFU/ gram.
(3) at substratum (the soybean cake powder 50g of 100 weight parts, sucrose 20g, ammonium sulfate 5g, trisodium citrate 2.5g, potassium primary phosphate 0.3g, magnesium sulfate 0.5g, ferrous sulfate 0.05g, pH7.0-7.2) the bacillus amyloliquefaciens bacterium liquid of inoculation 4 weight parts is (purchased from Chinese agriculture microbial strains preservation center and be numbered ACCC10166, being as above cultured to nectar degree OD in identical LB substratum
600value, for 0.6-0.8, obtains bacterium liquid), 37 ℃ of cultivations sample and observe by ascites method in culturing process, until the viable count of bacillus amyloliquefaciens is 2 × 10
7the substratum of CFU/ gram.
(4) at substratum (the glucose 15g of 100 weight parts, starch 1g, soybean cake powder 25g, manganous sulfate 1g, potassium primary phosphate 1.5g, magnesium sulfate 0.5g, yeast extract paste 0.2g, iron(ic) chloride 0.1g, calcium carbonate 0.1g, pH7.0-7.2) in, (deposit number is CCTCC NO.M2013500 to the methylotrophy type bacillus bacteria liquid of inoculation 5 weight parts, being as above cultured to nectar degree OD in identical LB substratum
600value, for 0.6-0.8, obtains bacterium liquid), 37 ℃ of cultivations sample and observe by ascites method in culturing process, until the viable count of methylotrophy type bacillus is 2 × 10
7the substratum of CFU/ gram.
(5) Bacillus subtillis obtaining respectively in step (1) to (4), bacillus licheniformis, bacillus amyloliquefaciens, methylotrophy type bacillus are proportionally mixed, in the microbiobacterial agent that blending ratio makes to obtain, in the number of viable bacteria body, with respect to methylotrophy type bacillus every part described, the content of described Bacillus subtillis is 0.1 part, the content of described bacillus licheniformis is 0.1 part, the content of described bacillus amyloliquefaciens is 0.03 part, obtains the microbiobacterial agent of the present embodiment.
Embodiment 3
The present embodiment is used for illustrating microbiobacterial agent of the present invention and performance thereof.
According to the method for embodiment 2, the Bacillus subtillis obtaining respectively in step (1) to (4), bacillus licheniformis, bacillus amyloliquefaciens, methylotrophy type bacillus are proportionally mixed, difference is, in the microbiobacterial agent that blending ratio makes to obtain, in the number of viable bacteria body, with respect to methylotrophy type bacillus every part described, the content of described Bacillus subtillis is 0.2 part, the content of described bacillus licheniformis is 0.5 part, the content of described bacillus amyloliquefaciens is 0.05 part, obtains the microbiobacterial agent of the present embodiment.
Embodiment 4
The present embodiment is used for illustrating microbiobacterial agent of the present invention and performance thereof.
According to the method for embodiment 2, the Bacillus subtillis obtaining respectively in step (1) to (4), bacillus licheniformis, bacillus amyloliquefaciens, methylotrophy type bacillus are proportionally mixed, difference is, in the microbiobacterial agent that blending ratio makes to obtain, in the number of viable bacteria body, with respect to methylotrophy type bacillus every part described, the content of described Bacillus subtillis is 0.2 part, the content of described bacillus licheniformis is 0.05 part, the content of described bacillus amyloliquefaciens is 0.01 part, obtains the microbiobacterial agent of the present embodiment.
Embodiment 5
The present embodiment is used for illustrating microbiobacterial agent of the present invention and performance thereof.
According to the method for embodiment 2, the Bacillus subtillis obtaining respectively in step (1) to (4), bacillus licheniformis, bacillus amyloliquefaciens, methylotrophy type bacillus are proportionally mixed, difference is, in the microbiobacterial agent that blending ratio makes to obtain, in the number of viable bacteria body, with respect to methylotrophy type bacillus every part described, the content of described Bacillus subtillis is 2 parts, the content of described bacillus licheniformis is 5 parts, the content of described bacillus amyloliquefaciens is 1 part, obtains the microbiobacterial agent of the present embodiment.
Embodiment 6
The present embodiment is used for illustrating microbiobacterial agent of the present invention and performance thereof.
Method according to embodiment 2 is prepared microbiobacterial agent, different is, the bacillus licheniformis obtaining respectively in step (2) to (4), bacillus amyloliquefaciens, methylotrophy type bacillus are proportionally mixed, but do not add Bacillus subtillis, obtain the microbiobacterial agent of the present embodiment.
Comparative example 2
Method according to embodiment 2 is prepared microbiobacterial agent, different, and methylotrophy type bacillus used is purchased from Chinese agriculture microbial strains preservation center and is numbered the methylotrophy type bacillus of ACCC02647.
Test implementation example 1
The effect of the routine microbiobacterial agent by outdoor potted plant experiment mensuration embodiment 2-6 and comparative example 2 of this test implementation 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.And, in matrix soil, add NaCl solution, make Cl
-concentration be 0.19mol/kg, add NaOH, making pH value is 9.0, obtains testing soil.The high 21cm of pot experiment basin, upper diameter 20cm, lower diameter 13cm, every basin adds native 3.8kg, adds respectively the microbiobacterial agent 0.05kg of embodiment 2-6 and comparative example 2.
In basin, plant into alfalfa, in the time that the average plant height of alfalfa reaches 20cm, measure the concentration of the Cl-in soil according to the method in " NY/T1121.17-2006 soil detects the 17th part: the mensuration of Soil Chlorine ion content ", and calculate the clearance of Cl-, result is as shown in table 1.
Table 1
| Microbiobacterial agent | Average Cl -Clearance |
| Embodiment 2 | 77% |
| Embodiment 3 | 74% |
| Embodiment 4 | 75% |
| Embodiment 5 | 67% |
| Embodiment 6 | 64% |
| Comparative example 2 | 48% |
Can find out from the data of table 1, microbiobacterial agent of the present invention has good alkaline land improving effect, and, at the preferred number in viable bacteria body, with respect to methylotrophy type bacillus 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, the content of described bacillus amyloliquefaciens is in the situation of 0.01-0.05 part, and described microbiobacterial agent has the saline and alkaline ability of better removal.
Test implementation example 2
This test implementation example is measured the microbiobacterial agent of embodiment 2-6 and comparative example 2 in the effect increasing in drought resistance in plants by outdoor potted plant experiment.
The matrix soil of use test embodiment 1 is made Nutrition Soil basin alms bowl, by after tomato seeds vernalization 48h (25 ℃), to move in Nutrition Soil basin alms bowl, as for transplanting seedlings after growing in greenhouse 15 days to new Nutrition Soil basin alms bowl (each basin alms bowl 3.8kg soil, transplant 4 strain tomatoes) in, 200mL water watered every other day.Transplant seedlings after 15 days, each basin alms bowl adds respectively the microbiobacterial agent 0.05kg of embodiment 2-6 and comparative example 2, after normally watering 6 days, cuts off the water supply after 20 days, measures tomato leaf relative water content, and result is as shown in table 2.
Table 2
| Microbiobacterial 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% |
Can find out from the data of table 2, microbiobacterial agent of the present invention has good increase drought resistance in plants effect, and, at the preferred number in viable bacteria body, with respect to methylotrophy type bacillus 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, the content of described bacillus amyloliquefaciens is in the situation of 0.01-0.05 part, and described microbiobacterial agent has the ability of better increase drought resistance in plants.
Test implementation example 3
The effect of the routine microbiobacterial agent by outdoor potted plant experiment mensuration embodiment 2-6 and comparative example 2 of this test implementation in the bacterial wilt of control capsicum.Take out soil from the large Tanaka of pepper planting who has infected bacterial wilt, as the susceptible soil of bacterial wilt.
The matrix soil of use test embodiment 1 is made Nutrition Soil basin alms bowl, by after pepper seed vernalization 48h (25 ℃), moved in Nutrition Soil basin alms bowl, as for transplanting seedlings after growing in greenhouse 15 days to new Nutrition Soil basin alms bowl (the susceptible soil of the above-mentioned bacterial wilt of each basin alms bowl 3.8kg, transplant 4 strain capsicums, and add respectively the microbiobacterial agent 0.05kg of embodiment 2-6 and comparative example 2 at each basin alms bowl), normally plant capsicum after 30 days, the preventive effect of measuring bacterial wilt, result is as shown in table 3.
Table 3
| Microbiobacterial agent | The preventive effect of plantation capsicum bacterial wilt after 30 days |
| Embodiment 2 | 77% |
| Embodiment 3 | 74% |
| Embodiment 4 | 75% |
| Embodiment 5 | 67% |
| Embodiment 6 | 64% |
| Comparative example 2 | 34% |
Can find out from the data of table 3, microbiobacterial agent of the present invention has the effect of the bacterial wilt of good control capsicum, and, at the preferred number in viable bacteria body, with respect to methylotrophy type bacillus every part described, the content of described Bacillus subtillis is 0.05-0.2 part, the content of described bacillus licheniformis is 0.05-0.2 part, the content of described bacillus amyloliquefaciens is in the situation of 0.01-0.05 part, and described microbiobacterial agent has the ability of the bacterial wilt of better control capsicum.
Test implementation example 4
This test implementation example is measured the microbiobacterial agent of embodiment 2-6 and comparative example 2 in the effect of preventing and treating in citrus anthracnose by field experiment.
Before oranges and tangerines are by result, the microbiobacterial agent of embodiment 2-6 and comparative example 2 is administered in citrus orchard with the amount of application of every square metre of 5kg, after oranges and tangerines result, the anthrax incidence result of statistics citrusfruit is as shown in table 4.
Table 4
| Microbiobacterial agent | The anthrax incidence of citrusfruit |
| Embodiment 2 | 17% |
| Embodiment 3 | 20% |
| Embodiment 4 | 19% |
| Embodiment 5 | 25% |
| Embodiment 6 | 28% |
| Comparative example 2 | 58% |
Can find out from the data of table 4, microbiobacterial agent of the present invention has the effect of good control citrus anthracnose, and, at the preferred number in viable bacteria body, with respect to methylotrophy type bacillus 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, the content of described bacillus amyloliquefaciens is in the situation of 0.01-0.05 part, and described microbiobacterial agent has the ability of better control citrus anthracnose.
More than describe the preferred embodiment of the present invention in detail; but the present invention is not limited to the detail in above-mentioned embodiment, within the scope of technical conceive of the present invention; can carry out multiple simple variant to technical scheme of the present invention, these simple variant all belong to protection scope of the present invention.
It should be noted that in addition, each concrete technical characterictic described in above-mentioned embodiment, in reconcilable situation, can combine by any suitable mode, for fear of unnecessary repetition, the present invention is to the explanation no longer separately of various possible array modes.
In addition, also can carry out arbitrary combination between various embodiment of the present invention, as long as it is without prejudice to thought of the present invention, it should be considered as content disclosed in this invention equally.
Claims (9)
1. a methylotrophy type bacillus, is characterized in that: the deposit number of this methylotrophy type bacillus (Bacillus methylotrophicus) is CCTCC NO.M2013500.
2. a microbiobacterial agent, this microbiobacterial agent comprises substratum and thalline, it is characterized in that: described thalline comprises that deposit number is the methylotrophy type bacillus of CCTCC NO.M2013500, and described thalline also comprises Bacillus subtillis (Bacillus subtilis), bacillus licheniformis (Bacillus lincheniformis) and bacillus amyloliquefaciens (Bacillus amyloliquefaciens).
3. microbiobacterial agent according to claim 2, is characterized in that: every gram of contained total viable count of described microbiobacterial agent is 2-20 × 10
7cFU.
4. according to the microbiobacterial agent described in claim 2 or 3, it is characterized in that: in the number of viable bacteria body, with respect to methylotrophy type bacillus every part described, the content of described Bacillus subtillis is 0.05-0.2 part, the content of described bacillus licheniformis is 0.05-0.2 part, and the content of described bacillus amyloliquefaciens is 0.01-0.05 part.
5. according to the microbiobacterial agent described in claim 2 or 3, it is characterized in that: described substratum is at least one in beef-protein medium, broth culture and LB substratum.
6. the application of the microbiobacterial agent described in any one in alkaline land improving in methylotrophy type bacillus claimed in claim 1 and claim 2-6.
7. the application of the microbiobacterial agent described in any one in increase drought resistance in plants in methylotrophy type bacillus claimed in claim 1 and claim 2-6.
8. the application of the microbiobacterial agent described in any one in the bacterial wilt of control capsicum in methylotrophy type bacillus claimed in claim 1 and claim 2-6.
9. the application of the microbiobacterial agent described in any one in control citrus anthracnose in methylotrophy type bacillus claimed in claim 1 and claim 2-6.
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