CN104560768A - Microbial agent and preparation method and application thereof - Google Patents
Microbial agent and preparation method and application thereof Download PDFInfo
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- CN104560768A CN104560768A CN201310516559.5A CN201310516559A CN104560768A CN 104560768 A CN104560768 A CN 104560768A CN 201310516559 A CN201310516559 A CN 201310516559A CN 104560768 A CN104560768 A CN 104560768A
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- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
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Abstract
The invention provides a microbial agent. The microbial agent comprises a culture medium and thalli, wherein the thalli comprise Bacillus subtilis, Baclicus lincheniformis and Bacillus safensis. The microbial agent provided by the invention can obtain a good plant seedling index and further improve yield and quality of vegetables, thereby having broad application prospects in the field of vegetable seedling raising. In addition, by using the microbial agent provided by the invention, a plant rhizosphere microbial environment can be improved, and beneficial bacteria are increased, thereby inhibiting the growth of harmful bacteria and further improving the disease resistance of plants.
Description
Technical field
The present invention relates to a kind of microbiobacterial agent and its preparation method and application; Or rather, relate to a kind of microbiobacterial agent and preparation method thereof, also relate to the application of described bacteria agent in grow seedlings of vegetable.
Background technology
Along with the development of world's agricultural economy, agricultural planting structure also experiencings corresponding adjustment, changed to agricultural modernization gradually by traditional agriculture, the cultivated area of vegetables also increases year by year, nursery is then a very important sport technique segment in vegetable growing, grow seedlings of vegetable not only can make vegetables listing ahead of time, can also improve vegetable crop and improve quality.
Current grow seedlings of vegetable Problems existing is that grow seedlings of vegetable soil-borne disease is serious, and harmful bacteria amount reproduction, has a strong impact on grow seedlings of vegetable surviving rate; Modern agriculture facility cultivation causes vegetable continuous cropping to be hindered more and more seriously simultaneously, and soil easily hardens, poor air permeability, and soil moisture content is poor, thus affects vegetable crop, quality; Therefore, in the urgent need to developing the nursery correlation technique that can improve plant seedling vigorous index.
Summary of the invention
The object of the invention is to overcome the problem that the plant seedling vigorous index that exists in current grow seedlings of vegetable process is low, provide a kind of and can improve microbiobacterial agent of plant seedling vigorous index and its preparation method and application.
In order to realize first goal of the invention, the invention provides a kind of microbiobacterial agent, this microbiobacterial agent comprises substratum and thalline, wherein, described thalline comprises subtilis (Bacillus subtilis), Bacillus licheniformis (Baclicus lincheniformis) and husky good fortune genus bacillus (Bacillus safensis).
In order to realize second goal of the invention, present invention also offers a kind of preparation method of microbiobacterial agent, wherein, the method comprises: the method comprises: subtilis (Bacillus subtilis), Bacillus licheniformis (Baclicus lincheniformis) and husky good fortune genus bacillus (Bacillus safensis) are inoculated in substratum and are cultivated.
Present invention also offers described microbiobacterial agent in the application of described microbiobacterial agent in grow seedlings of vegetable.
Microbiobacterial agent provided by the invention can improve grow seedlings of vegetable surviving rate, and such as, the result detected in embodiment 5-8 can be found out, although seedling rate respectively processed and do not have notable difference with contrasting seedling stage; But seedling vigorous index A4 is up to 0.325, contrasting the poorest is 0.077, each process with contrast between reach difference pole conspicuous level, between respectively processing, A2, A3, A4 and A1 reach conspicuous level, do not have difference between A2, A3, A4; Sickness rate contrast is up to 5%, each process not morbidity; After watermelon seedlings field planting to land for growing field crops, flowering period is A2 the earliest, and being August 1, is secondly A1, A2, and being contrast the latest, is August 10, illustrates that using microbiobacterial agent nursery to improve yields positive results; Ultimate production A3 is up to 1441 kilograms, and minimum is contrast, 1092 kilograms, and each process reaches conspicuous level with contrasting, and does not have difference between process; Quality of watermelon, pol is up to A3 and A4, and minimum is contrast; Watermelon late onset rate is up to contrast, reaches 13.33%, and minimum is A3 and A4, and morbidity Major Diseases is blight; Comprehensively use microbiobacterial agent of the present invention can obtain good stand seedling vigorous index as can be seen from above, thus make the aspects such as final yield and quality all obviously be better than contrast, have broad application prospects in grow seedlings of vegetable field.In addition, owing to using microbiobacterial agent provided by the invention can improve plant rhizospheric microorganism environment, probiotics increases thus suppresses the growth of harmful bacteria, thus can also improve the disease resistance of plant.
Embodiment
The invention provides a kind of microbiobacterial agent, this microbiobacterial agent comprises substratum and thalline, wherein, described thalline comprises subtilis (Bacillus subtilis), Bacillus licheniformis (Baclicus lincheniformis) and husky good fortune genus bacillus (Bacillus safensis).
The amount of the thalline contained in described microbiobacterial agent can in very large range change, and under preferable case, the total viable count contained by every gram of described microbiobacterial agent can be 2-10 × 10
7individual.
Preferably, in described microbiobacterial agent, with the total viable count in described microbiobacterial agent for benchmark, the viable count of subtilis (Bacillus subtilis) can be the 10-50% of total viable count, the viable count of Bacillus licheniformis (Baclicus lincheniformis) can be able to be the 10-50% of total viable count for the viable count of the 10-50% of total viable count, husky good fortune genus bacillus (Bacillus safensis).
More preferably, with the total viable count in described microbiobacterial agent for benchmark, the viable count of subtilis (Bacillus subtilis) can be the 40-50% of total viable count, the viable count of Bacillus licheniformis (Baclicus lincheniformis) can be able to be the 15-35% of total viable count for the viable count of the 15-35% of total viable count, husky good fortune genus bacillus (Bacillus safensis).
According to the present invention, the kind of described substratum can in very large range change, the substratum cultivating subtilis, Bacillus licheniformis, bacillus amyloliquefaciens, husky good fortune genus bacillus can be can be used in for various, such as, can be the conventional substratum such as beef-protein medium, broth culture, LB substratum 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).Such as, beef-protein medium: extractum carnis 3g, peptone 10g, sodium-chlor 5g, agar 15-20g, distilled water 1000ml, pH value 7.0-7.2,121 DEG C of sterilizing 20min; 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 7.0-7.2,121 DEG C of sterilizing 20min.
Present invention also offers the preparation method of described microbiobacterial agent, wherein, the method comprises: subtilis (Bacillus subtilis), Bacillus licheniformis (Baclicus lincheniformis) and husky good fortune genus bacillus (Bacillus safensis) are inoculated in substratum and are cultivated.
According to the present invention, the method of described cultivation comprises: in respective independently culture system, cultivating subtilis (Bacillus subtilis), Bacillus licheniformis (Baclicus lincheniformis) and husky good fortune genus bacillus (Bacillus safensis) respectively, and proportionally mixing cultivating the microorganism obtained respectively.Under preferable case, by the cultivation in respective independently culture system and the mixing after cultivating, total viable count of the every gram of microbiobacterial agent obtained is made to be 2-10 × 10
7individual.
In a preferred embodiment, by in the cultivation separately independently in culture system and mixing proportionally after cultivating, the condition of described mixing has no particular limits, as long as the microbiobacterial agent obtained can be made to meet the following conditions: total to make viable count in described microbiobacterial agent for benchmark, the viable count of subtilis (Bacillus subtilis) is the 10-50% of total viable count, the viable count of Bacillus licheniformis (Baclicus lincheniformis) is the 10-50% of total viable count, the viable count of husky good fortune genus bacillus (Bacillus safensis) is the 10-50% of total viable count, preferably, with the total viable count in described microbiobacterial agent for benchmark, the viable count of subtilis (Bacillus subtilis) is the 40-50% of total viable count, the viable count of Bacillus licheniformis (Baclicus lincheniformis) is the 15-35% of total viable count, the viable count of husky good fortune genus bacillus (Bacillus safensis) is the 15-35% of total viable count.
According to the present invention, the bacterial classification of described subtilis (Bacillus subtilis), Bacillus licheniformis (Baclicus lincheniformis), husky good fortune genus bacillus (Bacillus safensis) can be commercially available, such as, the husky good fortune genus bacillus (CCTCCAB205598) of the subtilis (ACCC01266) of Chinese agriculture Culture Collection preservation, Bacillus licheniformis (ACCC01468), Beijing North Na Chuanlian Bioteknologisk Institut.
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, such as, can at the substratum of 100 weight parts (glucose 5g, dregs of beans 30g, peptone 2g, distilled water 1000ml, PH7.0-7.2) bacterial strain of the subtilis (Bacillus subtilis) of inoculation 2-5 weight part in, 37 DEG C of cultivations, until the viable count of subtilis (Bacillus subtilis) is 2-10 × 10
7individual/gram substratum.
The cultural method of described Bacillus licheniformis (Baclicus lincheniformis) has no particular limits as conventionally known to one of skill in the art, such as, can at the substratum of 100 weight parts (corn steep liquor 0.45g, peptone 1.50g, soybean cake powder leaching juice is 1:15, glucose 1.50g, pH value 7.5) the middle bacterial strain inoculating the Bacillus licheniformis (Baclicus lincheniformis) of 2-5 weight part, 37 DEG C of cultivations, until the viable count of Bacillus licheniformis (Baclicus lincheniformis) is 2-10 × 10
7individual/gram substratum.
The cultural method of described husky good fortune genus bacillus (Bacillus safensis) has no particular limits, such as, can at the substratum of 100 weight parts (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) bacterial strain of the husky good fortune genus bacillus (Bacillus safensis) of inoculation 2-5 weight part in, 37 DEG C of cultivations, until the viable count of husky good fortune genus bacillus (Bacillus safensis) is 2-10 × 10
7individual/gram substratum.
Present invention also offers the application of described microbiobacterial agent in grow seedlings of vegetable.
Described vegetables can be watermelon, muskmelon, cucumber, tomato, capsicum, eggplant etc., are preferably on the vegetables cash crop such as watermelon, muskmelon, cucumber.
Below by specific embodiment, further description is carried out to the present invention.
Embodiment 1
(1) at substratum (the glucose 5g of 100 weight parts, dregs of beans 30g, peptone 2g, distilled water 1000ml, PH7.0-7.2) subtilis (ACCC01266) of inoculation 5 weight parts in, 37 DEG C of cultivations, carry out sampling and being observed by ascites method in culturing process, until the viable count of subtilis is 2 × 10
7individual/gram substratum.
(2) at substratum (the corn steep liquor 0.45g of 100 weight parts, peptone 1.50g, soybean cake powder leaching juice is 1:15, glucose 1.50g, pH value 7.5) the middle Bacillus licheniformis (ACCC01468) inoculating 5 weight parts, 37 DEG C of cultivations, carry out sampling and being observed by ascites method in culturing process, until the viable count of Bacillus licheniformis is 2 × 10
7individual/gram substratum.
(3) at substratum (glucose 15g, starch 1g, the soybean cake powder 25g of 100 weight parts, 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 middle husky good fortune genus bacillus (CCTCCAB205598) inoculating 5 weight parts, 37 DEG C of cultivations, carry out sampling and being observed by ascites method in culturing process, until the viable count of husky good fortune genus bacillus is 2 × 10
7individual/gram substratum.
(4) subtilis (Bacillus subtilis) will obtained respectively in step (1) to (3), Bacillus licheniformis (Baclicus lincheniformis), husky good fortune genus bacillus (Bacillus safensis) proportionally mixes, make in the microbiobacterial agent obtained, the viable count of subtilis (Bacillus subtilis) is 35% of total viable count, the viable count of Bacillus licheniformis (Baclicus lincheniformis) is 35% of total viable count, the viable count of husky good fortune genus bacillus (Bacillus safensis) is 30% of total viable count, obtain microbiobacterial agent A1.
Embodiment 2
(1) at substratum (the glucose 5g of 100 weight parts, dregs of beans 30g, peptone 2g, distilled water 1000ml, PH7.0-7.2) subtilis (ACCC01266) of inoculation 4 weight parts in, 37 DEG C of cultivations, carry out sampling and being observed by ascites method in culturing process, until the viable count of subtilis is 8 × 10
7individual/gram substratum.
(2) at substratum (the corn steep liquor 0.45g of 100 weight parts, peptone 1.50g, soybean cake powder leaching juice is 1:15, glucose 1.50g, pH value 7.5) the middle Bacillus licheniformis (ACCC01468) inoculating 6 weight parts, 37 DEG C of cultivations, carry out sampling and being observed by ascites method in culturing process, until the viable count of Bacillus licheniformis is 8 × 10
7individual/gram substratum.
(3) at substratum (glucose 15g, starch 1g, the soybean cake powder 25g of 100 weight parts, 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 middle husky good fortune genus bacillus (CCTCCAB205598) inoculating 4 weight parts, 37 DEG C of cultivations, carry out sampling and being observed by ascites method in culturing process, until the viable count of husky good fortune genus bacillus is 8 × 10
7individual/gram substratum.
(4) subtilis (Bacillus subtilis) will obtained respectively in step (1) to (3), Bacillus licheniformis (Baclicus lincheniformis), husky good fortune genus bacillus (Bacillus safensis) proportionally mixes, make in the microbiobacterial agent obtained, the viable count of subtilis (Bacillus subtilis) is 20% of total viable count, the viable count of Bacillus licheniformis (Baclicus lincheniformis) is 40% of total viable count, the viable count of husky good fortune genus bacillus (Bacillus safensis) is 40% of total viable count, obtain microbiobacterial agent A2.
Embodiment 3
(1) at substratum (the glucose 5g of 100 weight parts, dregs of beans 30g, peptone 2g, distilled water 1000ml, PH7.0-7.2) subtilis (ACCC01266) of inoculation 3 weight parts in, 37 DEG C of cultivations, carry out sampling and being observed by ascites method in culturing process, until the viable count of subtilis is 5 × 10
7individual/gram substratum.
(2) at substratum (the corn steep liquor 0.45g of 100 weight parts, peptone 1.50g, soybean cake powder leaching juice is 1:15, glucose 1.50g, pH value 7.5) the middle Bacillus licheniformis (ACCC01468) inoculating 8 weight parts, 37 DEG C of cultivations, carry out sampling and being observed by ascites method in culturing process, until the viable count of Bacillus licheniformis is 5 × 10
7individual/gram substratum.
(3) at substratum (glucose 15g, starch 1g, the soybean cake powder 25g of 100 weight parts, 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 middle husky good fortune genus bacillus (CCTCCAB205598) inoculating 2 weight parts, 37 DEG C of cultivations, carry out sampling and being observed by ascites method in culturing process, until the viable count of husky good fortune genus bacillus is 5 × 10
7individual/gram substratum.
(4) subtilis (Bacillus subtilis) will obtained respectively in step (1) to (3), Bacillus licheniformis (Baclicus lincheniformis), husky good fortune genus bacillus (Bacillus safensis) proportionally mixes, make in the microbiobacterial agent obtained, the viable count of subtilis (Bacillus subtilis) is 40% of total viable count, the viable count of Bacillus licheniformis (Baclicus lincheniformis) is 30% of total viable count, the viable count of husky good fortune genus bacillus (Bacillus safensis) is 30% of total viable count, obtain microbiobacterial agent A3.
Embodiment 4
Microbiobacterial agent A4 is prepared according to the method identical with embodiment 3, difference is, by the subtilis (Bacillus subtilis) obtained respectively in step (1) to (4), Bacillus licheniformis (Baclicus lincheniformis), husky good fortune genus bacillus (Bacillus safensis) proportionally mixes, make in the microbiobacterial agent obtained, the viable count of subtilis (Bacillus subtilis) is 50% of total viable count, the viable count of Bacillus licheniformis (Baclicus lincheniformis) is 25% of total viable count, the viable count of husky good fortune genus bacillus (Bacillus safensis) is 25% of total viable count, obtain microbiobacterial agent A4.
Embodiment 5-8
Microbiobacterial agent A1-A4 obtained in embodiment 1-4 is used to carry out grow seedlings of vegetable respectively, concrete steps are as follows: 2. (1) test materials 1. trial crops watermelon, cucumber, tomato, Flower of Aztec Marigold tests the preparation of microbial nutrition base: microbiobacterial agent A1-A4 is admixed in Nutrient medium raw material according to formula rate mix respectively, then insert in Nutrient medium hydraulic efficiency installation, be pressed into integrated grow seedling nutritional base.(2) test process.5 process (comprising 1 contrast) are established in test, and each process repeats for 3 times, each repetition 50 strain.Survey item: seedling stage (seedling vigorous index, seedling rate, sickness rate), (flowering period, output, quality, sickness rate) (3) test method after field planting.The microbial nutrition base random district group arrangement prepared, put in seedbed, expansion of watering, is sown into vernalization seed in Nutrient medium, covers overburden soil.(4) interpretation of result.Result is as shown in table 1 below.
Comparative example 1
Carry out grow seedlings of vegetable according to the method identical with embodiment 5-8, difference is not use microbiobacterial agent in seedling raising process, and result is as shown in table 1 below.
The statistical study of table 1 watermelon indices
As can be seen from the result in upper table 1, microbiobacterial agent provided by the invention has obvious effect to plant indices after watermelon seedling culturing and field planting.As can be seen from Table 1, seedling rate respectively processed and did not have notable difference with contrasting seedling stage; But seedling vigorous index A4 is up to 0.325, contrasting the poorest is 0.077, each process with contrast between reach difference pole conspicuous level, between respectively processing, A2, A3, A4 and A1 reach conspicuous level, do not have difference between A2, A3, A4; Sickness rate contrast is up to 5%, each process not morbidity; After watermelon seedlings field planting to land for growing field crops, flowering period is A2 the earliest, and being August 1, is secondly A1, A2, and being contrast the latest, is August 10, illustrates that using microbiobacterial agent nursery to improve yields positive results; Ultimate production A3 is up to 1441 kilograms, and minimum is contrast, 1092 kilograms, and each process reaches conspicuous level with contrasting, and does not have difference between process; Quality of watermelon, pol is up to A3 and A4, and minimum is contrast; Watermelon late onset rate is up to contrast, reaches 13.33%, and minimum is A3 and A4, and morbidity Major Diseases is blight; Comprehensively use microbiobacterial agent of the present invention can obtain good stand seedling vigorous index as can be seen from above, thus make the aspects such as final yield and quality all obviously be better than contrast, have broad application prospects in grow seedlings of vegetable field.In addition, owing to using microbiobacterial agent provided by the invention can improve plant rhizospheric microorganism environment, probiotics increases thus suppresses the growth of harmful bacteria, thus can also improve the disease resistance of plant.
Claims (9)
1. a microbiobacterial agent, this microbiobacterial agent comprises substratum and thalline, it is characterized in that, described thalline comprises subtilis (Bacillus subtilis), Bacillus licheniformis (Baclicus lincheniformis) and husky good fortune genus bacillus (Bacillus safensis).
2. microbiobacterial agent according to claim 1, wherein, the total viable count contained by every gram of described microbiobacterial agent is 2-10 × 10
7individual.
3. microbiobacterial agent according to claim 2, wherein, with the total viable count in described microbiobacterial agent for benchmark, the viable count of subtilis (Bacillus subtilis) is the 10-50% of total viable count, the viable count of Bacillus licheniformis (Baclicus lincheniformis) is the 10-50% of total viable count, the viable count of husky good fortune genus bacillus (Bacillus safensis) is the 10-50% of total viable count.
4. microbiobacterial agent according to claim 3, wherein, with the total viable count in described microbiobacterial agent for benchmark, the viable count of subtilis (Bacillus subtilis) is the 40-50% of total viable count, the viable count of Bacillus licheniformis (Baclicus lincheniformis) is the 15-35% of total viable count, the viable count of husky good fortune genus bacillus (Bacillus safensis) is the 15-35% of total viable count.
5. the preparation method of microbiobacterial agent according to claim 1, it is characterized in that, the method comprises: subtilis (Bacillus subtilis), Bacillus licheniformis (Baclicus lincheniformis) and husky good fortune genus bacillus (Bacillus safensis) are inoculated in substratum and are cultivated.
6. preparation method according to claim 5, wherein, the described method of carrying out cultivating that is inoculated in substratum comprises: in respective independently culture system, cultivating subtilis (Bacillus subtilis), Bacillus licheniformis (Baclicus lincheniformis) and husky good fortune genus bacillus (Bacillus safensis) respectively, and proportionally mixing cultivating the microorganism obtained respectively.
7. according to the preparation method that claim 6 is stated, wherein, by the cultivation in respective independently culture system and the mixing after cultivating, make total viable count of the every gram of microbiobacterial agent obtained be 2-10 × 10
7individual, wherein, the viable count of subtilis (Bacillus subtilis) is the 10-50% of total viable count, the viable count of Bacillus licheniformis (Baclicus lincheniformis) is the 10-50% of total viable count, the viable count of husky good fortune genus bacillus (Bacillus safensis) is the 10-50% of total viable count.
8. preparation method according to claim 7, wherein, with the total viable count in described microbiobacterial agent for benchmark, the viable count of subtilis (Bacillus subtilis) is the 40-50% of total viable count, the viable count of Bacillus licheniformis (Baclicus lincheniformis) is the 15-35% of total viable count, the viable count of husky good fortune genus bacillus (Bacillus safensis) is the 15-35% of total viable count.
9. the application of the microbiobacterial agent described in any one in claim 1-4 in grow seedlings of vegetable.
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| CN108728379A (en) * | 2018-06-01 | 2018-11-02 | 甘肃省农业科学院土壤肥料与节水农业研究所 | A kind of composite bacteria agent and preparation method thereof for feces of livestock and poultry quick composting |
| CN114507095A (en) * | 2021-12-15 | 2022-05-17 | 承德宝通矿业有限公司 | Organic fertilizer prepared from phosphorus-containing fine-fraction iron ore tailings and preparation method thereof |
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