CN112457117A - Culture medium composition for synthesizing organic selenium, synthesis method and application - Google Patents
Culture medium composition for synthesizing organic selenium, synthesis method and application Download PDFInfo
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- CN112457117A CN112457117A CN202011402692.4A CN202011402692A CN112457117A CN 112457117 A CN112457117 A CN 112457117A CN 202011402692 A CN202011402692 A CN 202011402692A CN 112457117 A CN112457117 A CN 112457117A
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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
- C05G3/40—Mixtures of one or more fertilisers with additives not having a specially fertilising activity for affecting fertiliser dosage or release rate; for affecting solubility
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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
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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
- C05G3/80—Soil conditioners
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- 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
- C12N1/14—Fungi; Culture media therefor
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- 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
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
- C12N1/18—Baker's yeast; Brewer's yeast
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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
Abstract
The invention discloses a culture medium composition for synthesizing organic selenium and a synthesis method thereof, comprising a solid strain culture medium, a liquid culture medium and a solid culture medium; the solid strain culture medium comprises the following raw materials: fig nanometer powder, tryptophan, valine, brown sugar and inorganic selenium powder; the raw materials of the liquid culture medium comprise: culture medium liquid and bacillus subtilis; the culture medium liquid comprises water, fig nano powder, tryptophan, valine, brown sugar, glucose, inorganic selenium powder, yeast and solid culture medium, and the raw materials of the culture medium liquid comprise: fig nano-particles, tryptophan, valine, brown sugar and glucose; the fructus fici nanometer powder contains organic selenium. In the growth process of plants, the organic selenium fertilizer prepared by the invention is applied, thus being beneficial to improving the resistance of crops, endowing the selenium-rich function and reducing the dependence on chemical pesticides; the quality of the agricultural products is improved, and more competitive products are formed to enter the consumer market.
Description
Technical Field
The invention relates to the technical field of agriculture, in particular to a culture medium composition for synthesizing organic selenium, a synthesis method and application.
Background
The influence of heavy metal pollution on the environment is increasingly aggravated, the range is continuously expanded, and the related fields comprise environmental water, soil, atmosphere and the like. Meanwhile, the social knowledge and the living standard of people are generally improved, the attention to the environment is continuously increased, and the awareness of environmental protection is gradually improved. Heavy metals in the soil can enter human bodies along with food chains through rice, wheat and other foods, and serious threat is brought to the health of the human bodies.
Disclosure of Invention
Based on the technical background, the invention provides a culture medium composition for synthesizing organic selenium and a synthesis method thereof, which solve the problems.
The invention is realized by the following technical scheme:
a culture medium composition for synthesizing organic selenium comprises solid strain culture medium, liquid culture medium and solid culture medium; the solid strain culture medium comprises the following raw materials: 85 to 95 percent of fig nano powder, 0.5 to 3.5 percent of tryptophan, 0.1 to 3.5 percent of valine, 0.5 to 5 percent of brown sugar and 0.3 to 3 percent of inorganic selenium powder; the raw materials of the liquid culture medium comprise: adding 350-550 parts of bacillus subtilis or trichoderma into 30-50 parts of culture medium liquid; the culture medium liquid comprises, by weight, 20-40 parts of water, 1600-2300 parts of fig nano powder, 350-550 parts of tryptophan, 200-300 parts of valine, 350-550 parts of brown sugar, 350-550 parts of glucose, 150-250 parts of inorganic selenium powder and 15-25 parts of yeast powder; the raw materials of the solid culture medium comprise: 89-98% of fig nano powder, 0.1-3.5% of tryptophan, 0.05-3% of valine, 0.1-4% of brown sugar and 0.1-4% of glucose;
the fig nano powder also contains organic selenium, for example, 200-400 mug of organic selenium is contained in every 500 g.
The invention provides a novel synthetic organic selenium culture medium composition, which is characterized in that a solid strain culture medium is used for culturing microbial transformation strains, such as beer yeast; converting inorganic selenium into organic selenium by microbial conversion in a liquid culture medium; and finally, the content of the organic selenium is improved by further converting the solid culture medium, the finally obtained solid culture medium can be directly used as the organic selenium fertilizer and is nano organic selenium (the particle size reaches the nano level, for example, the particle size reaches 1-100nm), and the content of the organic selenium in the organic selenium fertilizer reaches more than 98 percent. The fig containing selenium is used as a starter and is matched with other culture medium components, so that inorganic selenium can be promoted to be rapidly converted into organic selenium, and the yield of the organic selenium is extremely high; by adding bacillus subtilis or trichoderma into the liquid culture medium preferably, the absorption of nutrition by germs can be limited, germs can be effectively killed, and the normal growth of microorganism transformation bacteria (such as beer yeast) and saccharomycetes is not influenced.
Further preferably, the solid strain culture medium comprises the following raw materials: 89-93% of fig nano powder, 0.8-3% of tryptophan, 0.5-3% of valine, 1-4% of brown sugar and 0.5-2.5% of inorganic selenium powder; the raw materials of the liquid culture medium comprise: adding 400-520 parts of bacillus subtilis or trichoderma into 35-45 parts of culture medium liquid; the culture medium liquid comprises, by weight, 25-35 parts of water, 1800-2100 parts of fig nano powder, 450-530 parts of tryptophan, 210-280 parts of valine, 450-530 parts of brown sugar, 450-530 parts of glucose, 180-240 parts of inorganic selenium powder and 17-22 parts of yeast powder; the raw materials of the solid culture medium comprise: 93-97% of fig nano powder, 0.5-2% of tryptophan, 0.1-1.5% of valine, 0.5-2.5% of brown sugar and 0.5-2.5% of glucose;
the fig nano powder also contains organic selenium, for example, 200-400 mug of organic selenium is contained in every 500 g.
Further preferably, the solid strain culture medium comprises the following raw materials: 91% of fig nano powder, 2% of tryptophan, 1.5% of valine, 3.5% of brown sugar and 2% of inorganic selenium powder; the raw materials of the liquid culture medium comprise: adding 500g of bacillus subtilis or trichoderma into every 40L of culture medium liquid; the culture medium liquid comprises, by weight, 30L of water, 2000g of fig nano powder, 500g of tryptophan, 250g of valine, 500g of brown sugar, 500g of glucose, 200g of inorganic selenium powder and 20g of yeast powder; the raw materials of the solid culture medium comprise: 96.5% of fig nano powder, 1% of tryptophan, 0.5% of valine, 1% of brown sugar and 1% of glucose;
the fig nano powder contains 300 mu g of organic selenium per 500 g.
Further preferably, the particle size of the solid strain culture medium and the fig nanopowder in the solid culture medium is 1nm-100nm, such as 10nm-80nm, such as 30-50nm, preferably 15nm, 25nm, 35nm, 45nm, 55nm, 60nm, 70nm, 90nm, etc.
Further preferably, in the solid strain culture medium and the liquid culture medium, the inorganic selenium powder preferably adopts sodium selenite.
Further preferably, the water in the liquid culture medium is nano water.
More preferably, the fig nanometer powder in the solid medium is obtained by adding inorganic selenium during fig germination, and converting the inorganic selenium into organic selenium in fig to obtain fig nanometer powder containing 200 μ g-400 μ g of organic selenium per 500 g.
The method for synthesizing the organic selenium by adopting the culture medium composition for synthesizing the organic selenium comprises the following steps:
s1, inoculating a biotransformation strain on a solid strain culture medium for culture;
s2, adding the solid strain culture medium cultured in the step S1 into a liquid culture medium, and performing fermentation culture to obtain a liquid strain;
s3, inoculating the liquid strain to a solid culture medium, and continuously culturing to obtain the organic selenium fertilizer.
Further preferably, the preparation process of the solid strain culture medium in step S1 is as follows: weighing the raw materials according to the proportion; cooking the inorganic selenium powder and the fig powder; uniformly stirring the cooked inorganic selenium powder and fig powder with tryptophan, valine and brown sugar; bottling and sterilizing; inoculating on a slant, and culturing solid strains;
the preparation process of the liquid culture medium in the step S2 is as follows: weighing the raw materials according to the proportion; cooking the inorganic selenium powder and the fig powder; uniformly stirring the cooked inorganic selenium powder and fig powder with tryptophan, valine, brown sugar, glucose and yeast powder, bottling, and sterilizing to obtain culture medium liquid; then bacillus subtilis or trichoderma in the culture medium liquid to obtain a liquid culture medium.
The preparation process of the solid culture medium in the step S3 is as follows: weighing the raw materials according to the proportion; cooking the fig powder; uniformly stirring the cooked fig powder, tryptophan, valine, brown sugar and glucose; bottling and sterilizing; inoculating on slant, and culturing.
When the fig powder is cooked or the fig powder and the inorganic selenium powder are cooked, the fig powder or the fig powder and the inorganic selenium powder are added into water to be boiled for 0.5h-5h, preferably 2h-3h, for example, at 100 ℃ (determined based on atmospheric pressure).
Further preferably, in step S1, the solid strain culture medium is cultured at a temperature of 40-50 ℃, at a pH value of 7.2, for a period of 3d-4 d;
in step S2, the fermentation culture temperature of the liquid culture medium is 25-30 ℃, the pH value is 6.5, and the culture time is 7d-10 d;
in step S3, the solid culture medium has a culture temperature of 25-30 deg.C, a pH of 7.0, and a culture time of 8d-11 d.
More preferably, in step S1, the inoculated biotransformation strain is preferably Saccharomyces cerevisiae.
The organic selenium fertilizer is prepared by utilizing the culture medium composition for synthesizing organic selenium, or is synthesized by the method for synthesizing organic selenium.
Preferably, the organic selenium fertilizer is matched with secondary elements, wherein the secondary elements comprise magnesium, boron, zinc, calcium, molybdenum and potassium; the weight portions of the components are as follows: 5-30 parts of organic selenium fertilizer, 700 parts of magnesium 300-containing organic selenium fertilizer, 50-350 parts of boron and 700 parts of zinc 300-containing organic selenium fertilizer; 0-300 parts of calcium, 0-300 parts of molybdenum and 0-600 parts of potassium.
In the secondary elements matched with the organic selenium fertilizer, magnesium, boron and zinc are used as essential elements, and calcium, molybdenum and potassium are used as non-essential elements; the dosage of the magnesium, boron and zinc as essential elements is preferably as follows according to the weight proportion: 10-20 parts of organic selenium fertilizer, 600 parts of magnesium 400-; 300 parts of 100-calcium-containing material, 300 parts of 100-molybdenum-containing material and 600 parts of 200-potassium-containing material, and further preferably 15 parts of organic selenium, 200 parts of calcium, 500 parts of zinc, 200 parts of molybdenum, 500 parts of potassium, 200 parts of boron and 500 parts of magnesium. When in specific use, the organic selenium fertilizer and the secondary element can be mixed, and then added into water to prepare liquid for spraying application.
The invention has the following advantages and beneficial effects:
according to the invention, through solid strain culture, liquid culture medium culture and solid culture medium culture, toxic inorganic selenium is converted into nontoxic organic selenium, and the liquid strain is transferred to the solid culture medium for culture to obtain the organic selenium fertilizer, and the organic selenium fertilizer obtained by the invention has the following advantages:
1. antagonize heavy metals in soil and provide soil fertility
Selenium can antagonize heavy metals in soil, and can be combined with the heavy metals to generate insoluble precipitate, so that the activity of the heavy metals in the soil is reduced, and the heavy metals are difficult to be absorbed by plants, thereby achieving the effect of antagonizing the heavy metals.
After the organic selenium fertilizer is applied to soil, the organic substance can effectively improve the physical and chemical conditions and biological characteristics of the soil, ripen the soil, enhance the fertilizer retention and supply capability and buffer capability of the soil and manufacture good soil conditions for the growth of crops. The organic selenium fertilizer contains more nutrients but has low relative content, slow release, high nutrient content per unit of fertilizer, less components and quick release. The organic acid generated by decomposition can also promote the dissolution of mineral nutrients in soil and chemical fertilizers. The fertilizer is mutually promoted with the fertilizer, is beneficial to the absorption of crops, and improves the utilization rate of the fertilizer.
2. Reduce heavy metal in agricultural products and improve the quality of agricultural products
The nano selenium fertilizer prepared by the invention can effectively inhibit the absorption of heavy metals such as arsenic, lead, cadmium, mercury and the like, reduce the content of heavy metals in agricultural products, improve the product quality, and ensure that the content of protein selenium in grains reaches 40-300 micrograms/kg. The selenium-rich agricultural products are obtained after the plants are selenium-rich. The selenium-rich agricultural technology is implemented in selenium-poor areas, inorganic selenium salt is converted into organic selenoamine by utilizing the physiological function of crops, and the organic selenoamine is enriched in seeds, fruits, stems and leaves of plants for people to eat, so that the problem of selenium supplement of people is solved, and the health level of people can be generally improved.
3. Is beneficial to the production increase of agricultural products
The nano selenium fertilizer prepared by the invention is used for crops such as rice, wheat, corn and vegetables, can obviously improve the stress resistance such as disease resistance, lodging resistance, drought resistance and the like, promotes pollination, improves the seed setting rate and increases the dry grain weight. The yield of rice and wheat is increased by 16-26%. The selenium-enriched rice is rich in selenium elements, so that the price is higher than that of common rice by more than 3-5 times, the market price of the common rice is 2.2 yuan per kilogram, and the market price of the selenium-enriched rice is 13-15 yuan per kilogram, so that the income of one mu of the selenium-enriched rice is 3-5 times that of the common rice, the selenium-enriched rice contains the trace element selenium necessary for a human body, and the immunity of the human body can be enhanced by eating the selenium-enriched rice for a long time.
In conclusion, the invention utilizes the modern technology in the growth process of plants to promote the resistance of crops, endow the selenium-rich function and reduce the dependence on chemical pesticides; the quality of agricultural products is improved, and more competitive products are formed to enter the consumer market; thereby being beneficial to realizing a sustainable agriculture mode with ecological friendliness, soil improvement, quality improvement and income increase.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples, and the exemplary embodiments and descriptions thereof are only used for explaining the present invention and are not used as limitations of the present invention.
Example 1
The embodiment provides a method for synthesizing organic selenium, which comprises the following specific operation steps:
step 1, solid strain culture medium culture:
(1) the solid strain culture medium comprises the following raw materials: 91% of fig nano powder, 2% of tryptophan, 1.5% of valine, 3.5% of brown sugar and 2% of inorganic selenium powder; here, the inorganic selenium powder is sodium selenite;
(2) weighing the raw materials according to the proportion; cooking the inorganic selenium powder and the fig powder; mixing the cooked inorganic selenium powder and fig powder with tryptophan, valine and brown sugar, and stirring uniformly;
(3) bottling the mixed materials, and sterilizing; inoculating beer yeast on the inclined plane, and culturing strains; the culture temperature is 45 ℃, the pH value is 7.2, and the culture time is 3 d;
step 2, liquid culture medium culture:
(1) the liquid culture medium comprises the following raw materials: the culture medium liquid and the bacillus subtilis are added, and 500g of the bacillus subtilis is added in each 40L of the culture medium liquid.
Wherein the culture medium liquid comprises the following components in parts by weight: 30L of water, 2000g of fig nano powder, 500g of tryptophan, 250g of valine, 500g of brown sugar, 500g of glucose, 200g of inorganic selenium powder and 20g of yeast powder.
(2) Weighing the raw materials according to the proportion; cooking the inorganic selenium powder and the fig powder; mixing the cooked inorganic selenium powder and fig powder with tryptophan, valine, brown sugar, glucose and yeast powder, and stirring uniformly to obtain a culture medium solution; then bacillus subtilis is added into the culture medium liquid.
Wherein, the inorganic selenium powder adopts sodium selenite, and the water adopts nanometer water.
(3) Putting the liquid culture medium into a liquid fermentation tank, adding the solid strain culture medium cultured in the step (1) into the liquid fermentation tank, and uniformly mixing the solid strain culture medium with the liquid culture medium for fermentation culture; the fermentation culture temperature is 25 ℃, the pH value is 6.5, and the culture time is 8 d;
and 3, culturing in a solid culture medium:
(1) the solid culture medium comprises the following raw materials: 96.5 percent of fig nano powder, 1 percent of tryptophan, 0.5 percent of valine, 1 percent of brown sugar, 1 percent of glucose and 300 mu g of organic selenium contained in each 500g of fig nano powder.
(2) Weighing the raw materials according to the proportion; cooking the fig powder; uniformly stirring the cooked fig powder, tryptophan, valine, brown sugar and glucose; bottling and sterilizing; and (3) inoculating the liquid strain slant cultured in the step (2) to a solid culture medium for solid culture.
(3) Setting the culture temperature of the solid culture medium to be 25 ℃, the pH value to be 7.0 and the culture time to be 10 d; finally obtaining the organic selenium fertilizer.
Wherein, the nanometer powder of fig is prepared by adding inorganic selenium into fig during germination, and converting inorganic selenium into organic selenium in fig to obtain nanometer powder of fig containing 300 μ g of organic selenium per 500 g.
Example 2
The embodiment provides a method for synthesizing organic selenium, which comprises the following specific operation steps:
step 1, solid strain culture medium culture:
(1) the solid strain culture medium comprises the following raw materials: 85% of fig nano powder, 3.5% of tryptophan, 3.5% of valine, 5% of brown sugar and 3% of inorganic selenium powder; here, the inorganic selenium powder is sodium selenite;
(2) weighing the raw materials according to the proportion; cooking the inorganic selenium powder and the fig powder; mixing the cooked inorganic selenium powder and fig powder with tryptophan, valine and brown sugar, and stirring uniformly;
(3) bottling the mixed materials, and sterilizing; inoculating beer yeast on the inclined plane, and culturing strains; the culture temperature is 45 ℃, the pH value is 7.2, and the culture time is 3 d;
step 2, liquid culture medium culture:
(1) the liquid culture medium comprises the following raw materials: the culture medium liquid and the bacillus subtilis are added, and 400g of the bacillus subtilis is added in every 40L of the culture medium liquid.
Wherein the culture medium liquid comprises the following components in parts by weight: 25L of water, 1600g of fig nano powder, 400g of tryptophan, 200g of valine, 350g of brown sugar, 400g of glucose, 150g of inorganic selenium powder and 15g of yeast powder.
(2) Weighing the raw materials according to the proportion; cooking the inorganic selenium powder and the fig powder; mixing the cooked inorganic selenium powder and fig powder with tryptophan, valine, brown sugar, glucose and yeast powder, and stirring uniformly to obtain a culture medium solution; then bacillus subtilis is added into the culture medium liquid.
Wherein, the inorganic selenium powder adopts sodium selenite, and the water adopts nanometer water.
(3) Putting the liquid culture medium into a liquid fermentation tank, adding the solid strain culture medium cultured in the step (1) into the liquid fermentation tank, and uniformly mixing the solid strain culture medium with the liquid culture medium for fermentation culture; the fermentation culture temperature is 25 ℃, the pH value is 6.5, and the culture time is 8 d;
and 3, culturing in a solid culture medium:
(1) the solid culture medium comprises the following raw materials: 91% of fig nano powder, 1.5% of tryptophan, 1.5% of valine, 3% of brown sugar, 3% of glucose, and 300 mug of organic selenium per 500g of fig nano powder.
(2) Weighing the raw materials according to the proportion; cooking the fig powder; uniformly stirring the cooked fig powder, tryptophan, valine, brown sugar and glucose; bottling and sterilizing; and (3) inoculating the liquid strain slant cultured in the step (2) to a solid culture medium for solid culture.
(3) Setting the culture temperature of the solid culture medium to be 25 ℃, the pH value to be 7.0 and the culture time to be 10 d; finally obtaining the organic selenium fertilizer.
Wherein, the nanometer powder of fig is prepared by adding inorganic selenium during the germination of fig, and converting inorganic selenium into organic selenium in fig to obtain 200 μ g of nanometer powder of fig containing organic selenium per 500 g.
Example 3
The embodiment provides a method for synthesizing organic selenium, which comprises the following specific operation steps:
step 1, solid strain culture medium culture:
(1) the solid strain culture medium comprises the following raw materials: 95% of fig nano powder, 1% of tryptophan, 1% of valine, 2% of brown sugar and 1% of inorganic selenium powder; here, the inorganic selenium powder is sodium selenite;
(2) weighing the raw materials according to the proportion; cooking the inorganic selenium powder and the fig powder; mixing the cooked inorganic selenium powder and fig powder with tryptophan, valine and brown sugar, and stirring uniformly;
(3) bottling the mixed materials, and sterilizing; inoculating beer yeast on the inclined plane, and culturing strains; the culture temperature is 45 ℃, the pH value is 7.2, and the culture time is 3 d;
step 2, liquid culture medium culture:
(1) the liquid culture medium comprises the following raw materials: the culture medium liquid and the bacillus subtilis are added, wherein 550g of the bacillus subtilis is added in each 40L of the culture medium liquid.
Wherein the culture medium liquid comprises the following components in parts by weight: 40L of water, 2300g of fig nano powder, 550g of tryptophan, 300g of valine, 500g of brown sugar, 500g of glucose, 250g of inorganic selenium powder and 25g of yeast powder.
(2) Weighing the raw materials according to the proportion; cooking the inorganic selenium powder and the fig powder; mixing the cooked inorganic selenium powder and fig powder with tryptophan, valine, brown sugar, glucose and yeast powder, and stirring uniformly to obtain a culture medium solution; then bacillus subtilis is added into the culture medium liquid.
Wherein, the inorganic selenium powder adopts sodium selenite, and the water adopts nanometer water.
(3) Putting the liquid culture medium into a liquid fermentation tank, adding the solid strain culture medium cultured in the step (1) into the liquid fermentation tank, and uniformly mixing the solid strain culture medium with the liquid culture medium for fermentation culture; the fermentation culture temperature is 25 ℃, the pH value is 6.5, and the culture time is 8 d;
and 3, culturing in a solid culture medium:
(1) the solid culture medium comprises the following raw materials: fig nano powder 98%, tryptophan 0.5%, valine 0.5%, brown sugar 0.5%, glucose 0.5%, and fig nano powder containing 300 μ g of organic selenium per 500 g.
(2) Weighing the raw materials according to the proportion; cooking the fig powder; uniformly stirring the cooked fig powder, tryptophan, valine, brown sugar and glucose; bottling and sterilizing; and (3) inoculating the liquid strain slant cultured in the step (2) to a solid culture medium for solid culture.
(3) Setting the culture temperature of the solid culture medium to be 25 ℃, the pH value to be 7.0 and the culture time to be 10 d; finally obtaining the organic selenium fertilizer.
Wherein, the nanometer powder of fig is prepared by adding inorganic selenium during the germination of fig, and converting inorganic selenium into organic selenium in fig to obtain nanometer powder of fig containing 400 μ g of organic selenium per 500 g.
Example 4
The applicant performs the test on Mongolian Yangzhou Yunqiacun soil improvement heavy metal base located in Pengzhou city of Sichuan province, and the test is performed in one mu of land. The specific test operations are as follows:
first, demonstration test
1. Preparing a test fertilizer:
taking the organic selenium fertilizer prepared in the example 1, preparing a test fertilizer according to the following mixture ratio: 15g of organic selenium, 200g of calcium, 500g of zinc, 200g of molybdenum, 500g of potassium, 200g of boron, 500g of magnesium and 80kg of water.
2. Application method
The test base is required to be flat as a mirror, the height difference is not more than 3cm, the face deviation phenomenon does not exist, and the earth surface needs soft, movable and bulky mud with the mud. The water layer management aspect is mainly to cause the field surface to catch mud. The test fertilizer formula is installed, the materials are mixed and stirred for 5min, the materials are uniformly sprayed on the water surface of the field by a sprayer before the rice is transplanted, and the aligned liquid test fertilizer is completely sprayed on the field surface of one mu.
Rice seedlings are planted within 1 day, the average selenium content in the selenium-rich soil is over 0.44mg/kg after 20 days, the rice is ripe and harvested, the rice and the soil are separately removed, the soil is detected by a designated inspection institution, and the soil and the rice meet the national heavy metal improvement standard.
Second, control test
The difference from the control test is that the fertilizer formula is as follows: 2.5 percent of monopotassium phosphate, 0.5 percent of vitamin, 3 percent of calcium superphosphate, 6 percent of lime and 88 percent of coarse cereals.
Example 5
1. Detection method
(1) The detection method of selenium in the organic selenium fertilizer comprises the following steps:
GB5009.93-2017。
(2) the detection method of selenium in soil comprises the following steps:
NYT 1104-2006。
(3) the detection method of the heavy metals in the soil comprises the following steps:
the detection is carried out based on the soil environment quality agricultural land soil pollution risk control standard (GB 15618-:
the cadmium content detection method comprises the following steps: GB/T17141-1997;
the mercury content detection method comprises the following steps: HJ 680 and 2013;
the arsenic-content detection method comprises the following steps: HJ 680 and 2013;
the lead content detection method comprises the following steps: GB/T17141-1997;
the detection method of the chromium content comprises the following steps: HJ 491-;
and (3) pH detection: NY/T1377-2007.
(4) The detection method of the selenium content in the rice comprises the following steps:
the detection method of the total arsenic content comprises the following steps: GB 5009.11-2014; provision of "when the total arsenic level does not exceed the inorganic arsenic limit value", according to GB 2762-; the standard requirement value of the total arsenic of the solid detection project is derived from the limit of inorganic arsenic in GB 2762-.
The lead content detection method comprises the following steps: GB 5009.12-2017;
the detection method of cadmium content comprises the following steps: GB 5009.15-2014;
the mercury content detection method comprises the following steps: GB 5009.17-2014;
the detection method of the chromium content comprises the following steps: GB 5009.123-2017;
the detection method of the selenium content comprises the following steps: GB 5009.93-2017.
2. The result of the detection
(1) Yield of organic selenium
The embodiment realizes the conversion of the toxic inorganic selenium into the non-toxic organic selenium, and has high conversion efficiency. Taking the example 1 as an example, the organic selenium fertilizer prepared in the example 1 is detected to contain 9000 μ g of organic selenium per gram of organic selenium fertilizer, and the content of the organic selenium reaches 99.8%.
(2) Heavy metal content in soil
TABLE 1 detection results of heavy metal content in soil
(3) The rice contains heavy metals and selenium
TABLE 2 Rice heavy metals and selenium content
As shown in tables 1 and 2, when rice is harvested, 500g of each of rice and soil in a demonstration test is collected and sent to an inspection institution designated by an official part for inspection, the rice reaches the organic selenium content standard, and meanwhile, the heavy metal in the soil meets the standard requirement of green food soil; the effect of reducing heavy metals and improving selenium content is far lower than the effect of the application by collecting 500g of rice and soil in a control test.
(4) Rice yield increase
Compared with a control test, after the selenium fertilizer prepared in the embodiment 1 is applied in a demonstration test, the selenium content of agricultural products can be obviously improved, and the yield is increased by 20%, so that the commodity value is greatly improved (in the market, the selling price of selenium-rich rice is 3-5 times that of common rice). In addition, the embodiment only takes the planting of the selenium-rich rice as an example, and is also applicable to the planting of selenium-rich wheat, selenium-rich corn, selenium-rich peanut, selenium-rich soybean, selenium-rich apple, selenium-rich nectarine, selenium-rich grape, selenium-rich jujube, selenium-rich garlic, selenium-rich watermelon, selenium-rich tea, selenium-rich vegetables, selenium-rich fruits and the like.
The above-mentioned embodiments are intended to illustrate the objects, technical solutions and advantages of the present invention in further detail, and it should be understood that the above-mentioned embodiments are merely exemplary embodiments of the present invention, and are not intended to limit the scope of the present invention, and any modifications, equivalent substitutions, improvements and the like made within the spirit and principle of the present invention should be included in the scope of the present invention.
Claims (10)
1. A culture medium composition for synthesizing organic selenium is characterized by comprising a solid strain culture medium, a liquid culture medium and a solid culture medium;
the solid strain culture medium comprises the following raw materials: 85 to 95 percent of fig nano powder, 0.5 to 3.5 percent of tryptophan, 0.1 to 3.5 percent of valine, 0.5 to 5 percent of brown sugar and 0.3 to 3 percent of inorganic selenium powder;
the raw materials of the liquid culture medium comprise: adding 350-550 parts of bacillus subtilis or trichoderma into 30-50 parts of culture medium liquid; the culture medium liquid comprises, by weight, 20-40 parts of water, 1600-2300 parts of fig nano powder, 350-550 parts of tryptophan, 200-300 parts of valine, 350-550 parts of brown sugar, 350-550 parts of glucose, 150-250 parts of inorganic selenium powder and 15-25 parts of yeast powder;
the raw materials of the solid culture medium comprise: 89-98% of fig nano powder, 0.1-3.5% of tryptophan, 0.05-3% of valine, 0.1-4% of brown sugar and 0.1-4% of glucose;
the fig nano powder also contains organic selenium.
2. The composition of claim 1, wherein the solid culture medium comprises: 89-93% of fig nano powder, 0.8-3% of tryptophan, 0.5-3% of valine, 1-4% of brown sugar and 0.5-2.5% of inorganic selenium powder;
the raw materials of the liquid culture medium comprise: adding 400-520 parts of bacillus subtilis or trichoderma into 35-45 parts of culture medium liquid; the culture medium liquid comprises, by weight, 25-35 parts of water, 1800-2100 parts of fig nano powder, 450-530 parts of tryptophan, 210-280 parts of valine, 450-530 parts of brown sugar, 450-530 parts of glucose, 180-240 parts of inorganic selenium powder and 17-22 parts of yeast powder;
the raw materials of the solid culture medium comprise: 93-97% of fig nano powder, 0.5-2% of tryptophan, 0.1-1.5% of valine, 0.5-2.5% of brown sugar and 0.5-2.5% of glucose;
the fig nano powder also contains organic selenium.
3. The composition as claimed in claim 2, wherein the solid culture medium comprises: 91% of fig nano powder, 2% of tryptophan, 1.5% of valine, 3.5% of brown sugar and 2% of inorganic selenium powder;
the raw materials of the liquid culture medium comprise: adding 500g of bacillus subtilis or trichoderma into every 40L of culture medium liquid; the culture medium liquid comprises, by weight, 30L of water, 2000g of fig nano powder, 500g of tryptophan, 250g of valine, 500g of brown sugar, 500g of glucose, 200g of inorganic selenium powder and 20g of yeast powder;
the raw materials of the solid culture medium comprise: 96.5% of fig nano powder, 1% of tryptophan, 0.5% of valine, 1% of brown sugar and 1% of glucose;
the fig nano powder contains 300 mu g of organic selenium per 500 g.
4. The organic selenium-synthesizing culture medium composition as claimed in claim 1, wherein the nanometer fig powder in the solid culture medium is prepared by adding inorganic selenium during germination of fig, and the inorganic selenium is converted into organic selenium in fig to obtain 200 μ g-400 μ g of nanometer fig powder per 500g of fig containing organic selenium.
5. A method for synthesizing organic selenium by using the culture medium composition for synthesizing organic selenium in any one of claims 1 to 4, comprising the steps of:
s1, inoculating a biotransformation strain on a solid strain culture medium for culture;
s2, adding the solid strain culture medium cultured in the step S1 into a liquid culture medium, and performing fermentation culture to obtain a liquid strain;
s3, inoculating the liquid strain to a solid culture medium, and continuously culturing to obtain the organic selenium fertilizer.
6. The method of claim 5, wherein the step S1 of preparing the solid bacterial culture medium comprises: weighing the raw materials according to the proportion; cooking the inorganic selenium powder and the fig powder; uniformly stirring the cooked inorganic selenium powder and fig powder with tryptophan, valine and brown sugar; bottling and sterilizing; inoculating on a slant, and culturing solid strains;
the preparation process of the liquid culture medium in the step S2 is as follows: weighing the raw materials according to the proportion; cooking the inorganic selenium powder and the fig powder; uniformly stirring the cooked inorganic selenium powder and fig powder with tryptophan, valine, brown sugar, glucose and yeast powder, bottling, and sterilizing to obtain culture medium liquid; then bacillus subtilis or trichoderma in the culture medium liquid to obtain a liquid culture medium.
The preparation process of the solid culture medium in the step S3 is as follows: weighing the raw materials according to the proportion; cooking the fig powder; uniformly stirring the cooked fig powder, tryptophan, valine, brown sugar and glucose; bottling and sterilizing; inoculating on slant, and culturing.
7. The method of claim 5, wherein the organic selenium is obtained from the reaction of selenium,
in step S1, the solid strain culture medium is cultured at 40-50 deg.C, pH 7.2, and culture time 3d-4 d;
in step S2, the fermentation culture temperature of the liquid culture medium is 25-30 ℃, the pH value is 6.5, and the culture time is 7d-10 d;
in step S3, the solid culture medium has a culture temperature of 25-30 deg.C, a pH of 7.0, and a culture time of 8d-11 d.
8. The method of claim 5, wherein in step S1, the inoculated biological transformed strain is Saccharomyces cerevisiae.
9. An application of an organic selenium fertilizer, wherein the organic selenium fertilizer is prepared by using the culture medium composition for synthesizing organic selenium in any one of claims 1 to 4, or the organic selenium fertilizer is synthesized by the method for synthesizing organic selenium in any one of claims 5 to 8.
10. The application of the organic selenium fertilizer as claimed in claim 9, wherein the organic selenium fertilizer is used in combination with secondary elements, wherein the secondary elements comprise magnesium, boron, zinc, calcium, molybdenum and potassium; the weight portions of the components are as follows:
5-30 parts of organic selenium fertilizer, 700 parts of magnesium 300-containing organic selenium fertilizer, 50-350 parts of boron and 700 parts of zinc 300-containing organic selenium fertilizer;
0-300 parts of calcium, 0-300 parts of molybdenum and 0-600 parts of potassium.
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