CN110408557A - A kind of bacillus subtilis and preparation method thereof for improving Activities of Phosphatase in Soil - Google Patents
A kind of bacillus subtilis and preparation method thereof for improving Activities of Phosphatase in Soil Download PDFInfo
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- CN110408557A CN110408557A CN201910520485.XA CN201910520485A CN110408557A CN 110408557 A CN110408557 A CN 110408557A CN 201910520485 A CN201910520485 A CN 201910520485A CN 110408557 A CN110408557 A CN 110408557A
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
- C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- 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
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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Abstract
The invention discloses a kind of bacillus subtilises and preparation method thereof for improving Activities of Phosphatase in Soil, more particularly to bacillus subtilis preparation method step: (1) being inoculated into Bacillus subtilis strain and adjust in back bevel culture medium PH6.8 ~ 7.2, temperature is controlled at 29 DEG C ~ 31 DEG C, carries out the culture of strain level-one after cultivating by 24 ~ 48 hours；(2) bacillus subtilis is inoculated on seed culture medium after inclined-plane culture, controls seed culture medium PH6.8 ~ 7.2, and temperature regulates and controls at 29 DEG C ~ 31 DEG C, cultivates progress strain second level culture in 12 ~ 24 hours；(3) third stage culture is carried out after high-temperature sterilization in 29 DEG C ~ 31 DEG C of fermentor, after cultivation and fermentation ferment, effective clump count >=5 × 10 of bacillus subtilis9/ ml, the inactivation rate of gemma are 6% ~ 12%；(4) activation of bacillus subtilis.In paddy growth process, the Activities of Phosphatase in Soil compared with conventional fertilizer application processing is respectively increased 53.64%, 49.63%, 18.12% and 12.48% in rice plant of tillering stage, dry field phase, the phase of blossoming and bearing fruit and harvest time, while rice yield improves 9.90%.
Description
Technical field
The present invention relates to microbial-bacterial fertilizer technical fields, and in particular to a kind of withered grass that Activities of Phosphatase in Soil can be improved
Bacillus and preparation method thereof.
Background technique
Grain-production be it is essential in human survival growth course, in recent years, since China is long-term and persistence
Dependent on application chemical fertilizer, cause agricultural occur a series of pollution problem, soil hardening, crops in process of production
And plant is damaged, its yield is caused also constantly to decline, and weakens crop continuous production ability, the environmental structure of soil there is also
Destruction to a certain extent, is seriously had to be solved by the quality security problem caused by soil, environment and agricultural product, and is solved
These problems, then be unable to do without the development and utilization to microbial-bacterial fertilizer, it can make soil have the function that activation, promotes fertility effect
Fruit is obvious, and can guarantee agricultural product quality and safety, potentially acts as analyst and (decomposes the organic matter in soil and degrade poisonous and harmful
Substance), conversion person's (available nutrient in soil can be carried out to the conversion of effective form to supply crop growth), production
(be not only able to convert the nitrogen in air to plant can be absorbed and the ammonia utilized or other itrogenous organic substances person, and can will
Soil with organic matter resolves into humus), it is to push the input that can not be lacked during agricultural sustainable development, function is special
Point complements each other with agricultural sustainable development road.
Bacillus subtilis is one kind of bacteria Bacillus, is mainly widely present in soil and humus, to ring
Border has extremely strong adaptation and resist ability, and not only low to nutritional requirement required for growing but also breeding rapidly, can be attached to
Plant surface；The growth of pathogenic bacteria can be effectively suppressed, the antibacterial material of generation can also survive at high temperature, active duration
It is long；A variety of antibacterial materials can be generated, there is disease resistance, have the function of preventing and treating to plant disease, in crop production
On increasing production of rice can be made to increase income.
Soil enzyme is a kind of bioactive substance, itself can carry out catalytic action, can be used as soil microbiology activity
An and key index of productivity.Soil phosphorus loss is that one kind has catalyzing hydrolysis effect to Organic phosphate compound
Enzyme mainly exists in the form of adsorbed state in the soil, and it is close that phosphatase activity coordinates available phosphorus ability to the growth of crop
Correlation, activity is proportional with fertility level, and is in inverse ratio with soil depth.
This research proposed adoption microbial-bacterial fertilizer, which is used as, is applied to rice crop for examination fertilizer, studies bacillus subtilis to water
The situation of change of rice yield and Activities of Phosphatase in Soil, by measuring in each growth period soil physico-chemical property of rice and soil
The changing rule of phosphatase activity, analysis application bacillus subtilis embody the function influence for improving Activities of Phosphatase in Soil
Influence of the bacillus subtilis to Rice Productivity out.
Summary of the invention
The object of the present invention is to provide a kind of bacillus subtilis, required nutrition in plant growth is sufficiently achieved,
It not only makes moderate progress to soil physical and chemical property but also improves Activities of Phosphatase in Soil to a certain extent, improve soil environment knot
Structure enables the organic matter in soil to resolve into humic acid, supplies needed for crop and plant growth；Can fixed nitrogen, dissolving phosphor and dissolving potassium,
Make increasing production of rice, produce the green agricultural product beneficial to human health, agricultural is pushed to go on the road of sustainable development.
The object of the invention is also to provide a kind of bacillus subtilises that Activities of Phosphatase in Soil can be improved.
The object of the invention is also to provide bacillus subtilis effects played in paddy growth process, improve soil
Earth nutrient and soil enzyme activities make crop and plant reach certain volume increase.
The object of the invention is also to provide a kind of preparation methods of bacillus subtilis.
Effective clump count >=5 × 10 of bacillus subtilis of the invention9A/ml.
The present invention also provides the preparation methods of the bacillus subtilis, and steps are as follows:
Slant medium is mainly used for the culture of strain, strain is connected to culture medium (pH is 6.8 ~ 7.2), temperature is controlled at 29 DEG C
It ~ 31 DEG C, cultivates 24 ~ 48 hours, carries out the culture of strain level-one；
Bacillus subtilis after inclined-plane culture is inoculated on seed culture medium, controls seed culture medium pH6.8 ~ 7.2, temperature control
System is cultivated 12 ~ 24 hours at 29 DEG C ~ 31 DEG C, carries out the culture of strain second level；
Carry out third stage culture after high-temperature sterilization in 29 DEG C ~ 31 DEG C of fermentor, after cultivation and fermentation, bacillus subtilis has
Imitate clump count >=5 × 109A/ml, spore forming rate reach 88% ~ 94%；
Activate bacillus subtilis, Bacillus: water: brown sugar=1g:100ml:5g is blown 12 hours and can be applied under room temperature, placement
Time is no more than 24 ~ 48h.
Effective clump count >=5 × 10 of the bacillus subtilis9A/ml.
The pH value of the bacillus subtilis is 6.8 ~ 7.2.
According to bacillus subtilis described in accordance with the claim 1, it is characterized in that, the bacillus subtilis is solid
Pulvis.
The inclined-plane culture based component of bacillus subtilis according to claim 1 be peptone, yeast extract, agar,
Glucose (C6H12O6) and sodium chloride (NaCl), each composition proportion is (1.6:1:3:1:2)；
Each ingredient of the seed culture medium of bacillus subtilis according to claim 1 is peptone, yeast extract, grape respectively
Sugar (C6H12O6) and sodium chloride (NaCl), each composition proportion is (1.6:1:1:2).
The present invention also provides method of administration of bacillus subtilis during paddy growth, which is characterized in that
Root irrigation will be carried out after bacillus subtilis activation dilution, can promote paddy growth, the organic materials in soil can be had
Disease between effect degradation, the bacillary and fungoid soil for preventing and treating plant itself institute band pass, increases P elements utilization rate, has to crop
Play the role of obvious growth promotion, volume increase, to be reduced using chemical fertilizer, reach improve crop quality, reduce farming at
This.
The positive effect of the present invention is as follows:
The ability of paddy rice anti contravariance can be improved in bacillus subtilis of the invention, and application bacillus subtilis can improve soil
Environmental structure effectively facilitates the soil organism and resolves into humic acid rapidly, makes crop be easier to absorb, increases income crop yield,
Have the function of fixed nitrogen, dissolving phosphor and dissolving potassium, improve fertilizer efficiency utilization rate and quality of agricultural product, ensure food safety, improves Agro-ecology
Environment not only pushes local Recycling Economy Construction but also plays positive effect to the development for accelerating biological industry, as a kind of micro-
Bio-bacterial manure can not only build up fertility, moreover it is possible to improve Activities of Phosphatase in Soil, make crop under the premise of decrement is using chemical fertilizer
Increasing both production and income.
The cost price of its production of bacillus subtilis prepared by the present invention is low, and operating method is simple, survives effective bacterium number living
Height will not endanger environment, be able to suppress pest and disease damage and improve crop yield, have extremely far-reaching social effect and economic valence
Value.
Bacillus subtilis prepared by the present invention is significantly improved compared with the Activities of Phosphatase in Soil that conventional fertilizer application is handled, In
Rice plant of tillering stage, dry field phase, the phase of blossoming and bearing fruit and harvest time, have been respectively increased 53.64%, 49.63%, 18.12% and 12.48%, together
When rice yield improve 9.90%.
Specific embodiment
Technical solution of the present invention can be illustrated by following specific related embodiment.
Case study on implementation one
A kind of bacillus subtilis preparation method improving Activities of Phosphatase in Soil, specific preparation process is as follows:
(1) Bacillus subtilis strain is inoculated into the slant medium of pH6.8 ~ 7.2, temperature controls the culture at 29 DEG C ~ 31 DEG C
24 ~ 48h carries out the culture of strain level-one；Each ingredient of slant medium is peptone, yeast extract, agar, glucose respectively
(C6H12O6) and sodium chloride (NaCl), each composition proportion is (1.6:1:3:1:2)；
(2) bacillus subtilis after inclined-plane culture is inoculated on seed culture medium, controls seed culture medium pH6.8 ~ 7.2, temperature
Degree is controlled at 29 DEG C ~ 31 DEG C, and culture 12 ~ for 24 hours, carry out the culture of strain second level；Each ingredient of seed culture medium is peptone, ferment respectively
Female cream, glucose (C6H12O6) and sodium chloride (NaCl), each composition proportion is (1.6:1:1:2)；
(3) third stage culture is carried out after high-temperature sterilization in 29 DEG C ~ 31 DEG C of fermentor, after cultivation and fermentation, bacillus subtilis
Effective clump count >=5 × 109A/ml, gemma inactivation rate are 6% ~ 12%；
(4) bacillus subtilis is activated.
The application method and effect of each growth cycle of two rice of embodiment
The rice striking root phase, bacillus subtilis using preceding need carry out 50 times dilution after root irrigation is made to crops, to soil
Environment has some improvement, and along with the generation of a variety of antibiotic and enzyme, increases soil fertility；Improve crop disease-resistant evil, drought
The abilities such as calamity, being allowed to volume increase to the continuous growth and development for promoting plant has indispensable remarkable effect.Rice plant of tillering stage,
Foliage-spray is carried out after 100 times of dilutions, the permeability of soil can be effectively improved, grow rice preferably.In Rice Heading
Phase, the phase of blossoming and bearing fruit promote Root Growth of Rice, microbial enzymatic activities and physics and chemistry to carry out foliage-spray after 100 times of dilutions
Character changes and improves, drought resistant cold resistant capability enhancing.
Preparation manipulation method of the present invention is simpler, and the effective bacterium survived number living is high, and price is low, nontoxic, pollution-free, energy
Enough promote nutrition conversion and absorption, improve soil enzyme activities, have the function of prevention resistance disease, by microorganism with it is traditional
Fertilizer or organic fertilizer combination are one of the Main ways that fertilizer industry is delivered in recent years, and microbial-bacterial fertilizer is rationally matched with chemical fertilizer to be applied, and is subtracted
It can increase production again while low applying quantity of chemical fertilizer, result of study can provide ginseng for the further improvement and popularization of microbial-bacterial fertilizer
It examines, is the environmentally friendly microbial-bacterial fertilizer of current China's Developing Efficient Eco-agriculture sustainable development.
Claims (6)
1. a kind of bacillus subtilis and preparation method thereof for improving Activities of Phosphatase in Soil, which is characterized in that including following step
It is rapid:
Its main preparation step of the bacillus subtilis is that Bacillus subtilis strain is seeded to inclined-plane training first by (1)
It supports in base, adjustable inclined surface apparatus culture medium PH6.8 ~ 7.2, temperature is controlled at 29 DEG C ~ 31 DEG C, after culture in 24 ~ 48 hours
Carry out the culture of strain level-one；(2) bacillus subtilis is inoculated on seed culture medium after inclined-plane culture, adjusts control seed
Culture medium PH6.8 ~ 7.2, temperature are controlled at 29 DEG C ~ 31 DEG C, carry out the culture of strain second level after culture 12 ~ 24 hours；(3) high temperature goes out
Carry out third stage culture after bacterium in 29 DEG C ~ 31 DEG C of fermentor, after cultivation and fermentation, effective clump count of bacillus subtilis >=
5×109A/ml, the inactivation rate of gemma is 6% ~ 12%；(4) activation of bacillus subtilis is Bacillus: water: brown sugar=1g:
100ml:5g is blown 12 hours and can be applied under room temperature, and standing time is no more than 24 ~ 48h.
2. bacillus subtilis according to claim 1, which is characterized in that the culture medium described in it includes following components:
Slant medium: peptone, yeast extract, agar, glucose (C6H12O6), each composition proportion of sodium chloride (NaCl)
(1.6:1:3:1:2)；
Seed culture medium: peptone, yeast extract, glucose (C6H12O6), each composition proportion of sodium chloride (NaCl) (1.6:1:
1:2).
3. a kind of bacillus subtilis preparation methods for improving soil enzyme activities described in any item according to claim 1 ~ 2,
Be characterized in that itself the following steps are included:
(1) Bacillus subtilis strain is inoculated into the slant medium that PH is 7.0, for regulating and controlling temperature at 30 DEG C, culture 24 is small
Shi Hou starts the culture of strain level-one；(2) bacillus subtilis is inoculated in the seed culture medium that PH is 7.0 after inclined-plane culture
On, temperature is controlled at 30 DEG C, carries out the culture of strain second level after culture 22 hours；(3) at 30 DEG C of fermentor after high-temperature sterilization
Carry out third stage culture in ± 1 DEG C, after cultivation and fermentation, effective clump count >=5 × 10 of bacillus subtilis9/ ml, gemma
Inactivation rate be 6% ~ 12%；(4) activation of bacillus subtilis.
4. bacillus subtilis according to claim 1, which is characterized in that the bacillus subtilis is solid powder
Agent, effective clump count >=5 × 109/ ml, pH value are 6.8 ~ 7.2.
Promote paddy growth 5. bacillus subtilis described according to claim 1 ~ 3 has, volume increase, Resistant, improve soil
Nutrient and enzymatic activity, by the way that pouring root can simultaneously antagonism be pathogenic in soil-borne disease for effective degrading pesticide residues, pre- anti-bacteria in the soil
Microorganism reduces the generation of disease, and the growth and development of raising and plant to P elements utilization rate, volume increase tool have certain effect
Effect, can effectively prevent crops during the growth process by pest and disease damage erosion, by reducing applying for chemical fertilizer and pesticide
Dosage, the application of plant growth etc. can preferably be promoted by being ensured food safety with this.
6. this bacillus subtilis is added according to ratio to fertilize during rice growth, the effect after use compared with
Obviously, soil nutrient and soil enzyme activities are improved, to meet the needs of people are to grain, reduces applying quantity of chemical fertilizer to the shadow of environment
It rings, there is certain practice significance to promotion microbial-bacterial fertilizer development in an all-round way.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN111647527A (en) * | 2020-05-21 | 2020-09-11 | 浙江工业大学 | Method for improving saline-alkali soil and preserving fertilizer by utilizing bacillus subtilis |
| CN116349803A (en) * | 2023-05-08 | 2023-06-30 | 南京师范大学 | Rice-fish co-cropping specialized composite biological bait and application thereof |
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| CN103333694A (en) * | 2013-06-05 | 2013-10-02 | 华南理工大学 | Microbial agent capable of converting unavailable phosphorus into available phosphorus in soil, and preparation method and applications thereof |
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|---|---|---|---|---|
| CN111647527A (en) * | 2020-05-21 | 2020-09-11 | 浙江工业大学 | Method for improving saline-alkali soil and preserving fertilizer by utilizing bacillus subtilis |
| CN116349803A (en) * | 2023-05-08 | 2023-06-30 | 南京师范大学 | Rice-fish co-cropping specialized composite biological bait and application thereof |
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