CN113372147A - Special bacterial fertilizer for selenium-rich vegetables and preparation method thereof - Google Patents
Special bacterial fertilizer for selenium-rich vegetables and preparation method thereof Download PDFInfo
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Abstract
The invention relates to the technical field of microorganisms, in particular to a special bacterial fertilizer for selenium-rich vegetables, which is prepared from the following raw materials in parts by weight: 65-75 parts of crushed crop straw, 10-15 parts of bran, 40-50 parts of bean curd residue, 5-10 parts of rice biochar, 1-5 parts of molasses and 3-5 parts of compound microbial agent. The bacterial fertilizer can be used for applying the special fertilizer for the selenium-rich vegetables without adding exogenous selenium and causing no selenium pollution risk to soil, the selenium content of the vegetables is in a recommended standard range, and the produced selenium-rich vegetables are natural selenium-rich vegetables. The edible natural selenium-rich vegetables are a safe and effective way for supplementing selenium.
Description
[ technical field ] A method for producing a semiconductor device
The invention relates to the technical field of microorganisms, and particularly relates to a special bacterial fertilizer for selenium-rich vegetables and a preparation method thereof.
[ background of the invention ]
Selenium is one of the essential trace elements for the life of an organism, and has the important functions of nourishing, repairing cardiac muscle, clearing free radicals in the body, delaying senility, regulating the microcirculation system of the human body and the like. At present, with the continuous improvement of living standard of people, people are more urgent to pursue health, and the demand for selenium-rich food is more and more increased. Biological selenium enrichment is an effective way for obtaining organic selenium, and the method for taking selenium-enriched foods such as fruits and vegetables is the most important way for obtaining organic selenium for human bodies. However, the natural fruits and vegetables have low selenium content, and most of the natural fruits and vegetables need to apply selenium fertilizer to strengthen and supply crops so as to produce selenium-rich agricultural products. The biological effectiveness of soil selenium is highly influenced by microorganisms in the environment, and the microorganisms can realize the conversion of selenium form and valence state through the self metabolism, so that the effectiveness of the soil selenium is improved.
At present, most selenium fertilizers are prepared by adding selenite or selenium-rich ore into common fertilizers. Although the method has low cost and simple operation, the method has some defects. Selenate and selenite have high toxicity, and if the selenate and selenite are not applied properly, soil is easily polluted and even a human body is easily damaged; the selenium-rich ore has complex components, so the selenium element in the ore is not easy to be absorbed and utilized by plants, and the biological effectiveness is low. Therefore, the development of safe, reliable and high-bioavailability selenium-rich fertilizer is urgently needed, and the method has important significance for promoting the sustainable development of selenium fertilizer industry and developing functional agriculture.
[ summary of the invention ]
In view of the above, there is a need to provide a selenium-rich bacterial fertilizer for vegetables, which can effectively improve the effectiveness of selenium in soil.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
the special bacterial fertilizer for the selenium-rich vegetables is prepared from the following raw materials in parts by weight: 65-75 parts of crushed crop straw, 10-15 parts of bran, 40-50 parts of bean curd residue, 5-10 parts of rice biochar, 1-5 parts of molasses and 3-5 parts of compound microbial agent;
the composite microbial agent is prepared from a Sinomonas atrocerea (Sinomonas atrocerea) strain TXB1-10 freeze-dried powder and a Achromobacter denitrificans (Achromobacter denitirichiana) strain GPB1-5 freeze-dried powder in a mass ratio of 1.0-1.5: 1.0, mixing to obtain a compound microbial agent;
the Chinese unicellular bacterium darkblack (Sinomonas atrocyanea) strain TXB1-10 has a preservation number of CCTCC NO: m2020984; achromobacter denitrificans (Achromobacter denitiricans) strain GPB1-5 with the preservation number of CCTCC NO: m2020344.
Further, the strain TXB1-10 is obtained by separating from the soil of a selenium-rich area in Guangxi county of China, and the Sinomonas atrocerulosa (Sinomonas atrocyanea) strain is obtained; the strain GPB1-5 is separated from the soil in the selenium-rich area of Guangxi Guiping of China.
Further, lyophilized powder of the Sinomonas atrocerivia (Sinomonas atrocyanea) strain TXB1-10 and lyophilized powder of Achromobacter denitrificans (Achromobacter denitirica) strain GPB1-5 were prepared by the following steps:
(1) performing large-scale fermentation culture on a Sinomonas atrocyanea strain TXB1-10 and a denitrifying Achromobacter (Achromobacter denitirica) strain GPB1-5 in an LB culture solution for 2-3 days respectively, and centrifuging to obtain bacterial sludge;
(2) mixing a protective agent with distilled water to obtain a protective agent solution, and then uniformly mixing the protective agent solution with the bacterial sludge to obtain a bacterial sludge-protective agent solution;
(3) pre-freezing the bacteria mud-protective agent solution to obtain frozen bacteria;
(4) then, freeze-drying technology is adopted to prepare the freeze-dried powder microbial inoculum.
Further, in the step (2), the volume ratio of the bacterial sludge to the protective agent solution is 3-4: 1.
Further, the protective agent comprises the following components: 8-12mg/L skimmed milk, 5-8mg/L fructose and 0.05-0.1mg/L poly-gamma-glutamic acid.
The invention also provides a preparation method of the special bacterial fertilizer for the selenium-rich vegetables, which comprises the following steps:
(1) freeze-dried powder of a Chinese unicellular bacterium (Sinomonas atrocyanea) strain TXB1-10 and freeze-dried powder of a colorless denitrifying bacillus (Achromobacter dentifrices) strain GPB1-5 are mixed according to the mass ratio of 1.0-1.5: 1.0, mixing to obtain a compound microbial agent;
(2) preparing a mixed fermentation material of crushed crop straws of 120-150 meshes, bean curd residues, rice biochar and molasses, and mixing and fermenting 65-75 parts of the crushed crop straws, 10-15 parts of bran, 40-50 parts of bean curd residues, 5-10 parts of the rice biochar and 1-5 parts of molasses by weight;
(3) and (3) adding the compound microbial agent in the step (1) to continue fermenting after the temperature of the fermented material is reduced to below 50 ℃, turning the pile once every 24-48 hours until the humidity of the fermented material is about 12%, and finishing the fermentation.
Further, the fermentation temperature of the step (2) is 30-60 ℃, and the fermentation time is 15-20 days.
Compared with the prior art, the invention has the following beneficial effects:
the special bacterial fertilizer for the selenium-rich vegetables can be used for preventing the soil from being polluted by selenium without adding exogenous selenium, the selenium content of the vegetables is in the recommended standard range by applying the special bacterial fertilizer for the selenium-rich vegetables, and the produced selenium-rich vegetables are natural selenium-rich vegetables. The edible natural selenium-rich vegetables are a safe and effective way for supplementing selenium. When the compound microbial agent is prepared, the adopted protective agent comprises the following components: 8-12mg/L skimmed milk, 5-8mg/L fructose and 0.05-0.1mg/L poly-gamma-glutamic acid, the strain TXB1-10 of the Sinomonas atrocerea and the strain GPB1-5 of the Achromobacter denitrificans are prepared into freeze-dried powder, and the survival rate and the selenium-rich effect of the strains can be effectively improved.
[ description of the drawings ]
FIG. 1 is a morphogram of the strain of Chinese Aplysia atrophaea (Sinomonas atrocyanea) TXB1-10 on a plate according to the examples of the present application;
FIG. 2 is a diagram showing the morphology of Achromobacter denitrificans (Achromobacter denitificas) strain GPB1-5 on a plate in examples of the present application.
[ detailed description ] embodiments
All of the features disclosed in this specification, or all of the steps in any method or process so disclosed, may be combined in any combination, except combinations of features and/or steps that are mutually exclusive.
Any feature disclosed in this specification (including any accompanying claims, abstract) is merely an example of a generic series of equivalent or similar features, unless explicitly described as such.
Example 1:
the present example is a strain of Chinese unicellular bacterium melanomonas (Sinomonas atrocyanea) TXB 1-10; screening method for Achromobacter denitrificans (Achromobacter denitirichicans) strain GPB 1-5:
collecting soil samples from multiple selenium-rich areas such as Guangxi Yongfu (sand shale red soil, full selenium of 0.567mg/kg), Yulin (sand shale red soil, full selenium of 1.804mg/kg), Guiping (sand shale red soil, full selenium of 1.188mg/kg), Teng (Teng) (sand shale red soil, full selenium of 0.580mg/kg), and weighing 10g of soil samples, screening selenium-resistant bacteria by using a selenium-added liquid culture medium, separating by using a dilution coating plate method and a plate scribing method, determining 4 selenium-resistant bacteria as candidate strains through a growth curve, respectively naming the strains as YLB1-6 (Yulin), TXB1-10 (Teng county), TXB2-5 (Teng county) and GPB1-5 (Guiping), analyzing by a 16S rDNA sequence and a developmental phylogenetic tree, YLB1-6 is Bacillus, B1-10 is Simonosanocis, Bacillus is 2, and GPB1 is TXylancis 2-1, the selenium-enriched bacteria have the conversion rate of 55.31-74.22% to selenium, and the treatment of each strain can obviously improve the exchangeable selenium content of soil and has stronger activation effect on soil selenium. Has potential application value in soil microorganism selenium reinforcement.
After the strain is activated, the strain is transferred into a triangular flask of 100ml of liquid culture medium according to the inoculation amount of 5 percent, and shaking culture is carried out under the conditions of proper selenium concentration, proper selenium adding time and proper culture time of the strain. Centrifuging the cultured bacterial suspension for 30min at 4000 r/min, taking supernate, performing digestion reaction, measuring a fluorescence intensity value by using an atomic fluorescence photometer, obtaining the selenium concentration of a sample solution from a standard curve, and obtaining the selenium enrichment capacity of the strain by using a selenium conversion rate formula.
Selenium conversion (%) - (total selenium content-residual inorganic selenium content)/total selenium content x 100
After a selenium-rich test, the contents of residual inorganic selenium of YLB1-6, TXB1-10, TXB2-5 and GPB1-5 are 1.547, 1.358, 2.681 and 2.936 mu g/ml respectively, the selenium conversion rates of the strains are 74.22%, 66.05%, 55.31% and 63.30% in sequence, and the selenium conversion rate differences among the strains of YLB1-6, TXB1-10, GPB1-5 and TXB2-5 all reach an extremely significant level. The selenium-enriching capacities of the 4 selenium-resistant bacteria are sorted from large to small as follows: YLB1-6 > TXB1-10 > GPB1-5 > TXB 2-5.
The detailed deletion step and identification scheme refer to a culture method of a bacterial strain for efficiently transforming soil selenium in Chinese patent CN 201811201689.9.
The strain of Chinese unicellular darkling (Sinomonas atrocyanea) TXB1-10 is deposited in the China center for type culture Collection, address: in China, Wuhan university No. 299 in eight roads in Wuchang district, Wuhan city, Hubei province, with the preservation number of CCTCC NO: m2020984, the preservation date is 12/28/2020.
The strain of Achromobacter denitrificans (Achromobacter densitificans) GPB1-5 was deposited in the China center for type culture Collection at the address: in China, Wuhan university No. 299 in eight roads in Wuchang district, Wuhan city, Hubei province, with the preservation number of CCTCC NO: m2020344, the preservation date is 2020 and 07/23 days.
The morphological characteristics of the bacterial strain TXB1-10 of the Chinese unicellular bacterium (Sinomonas atrocyanea) on a plate are shown in figure 1, the bacterial colony of the bacterial strain is circular, the diameter is 0.8 mm-1.2 mm, the bacterial colony is milky white, the surface is smooth and convex, the edge is neat, and the bacterial colony is easy to pick up; the morphological characteristics of the Achromobacter denitrificans (Achromobacter densificans) GPB1-5 strain on a plate are shown in figure 2, the bacterial colony of the strain is small, round, white, transparent, smooth and moist in surface, hemispherical in middle protrusion, neat in edge and easy to pick up.
Example 2:
lyophilized powder of Chinese unicellular bacterium atrocerea (Sinomonas atrocerea) strain TXB1-10 and lyophilized powder of Achromobacter denitrificans (Achromobacter dentifrices) strain GPB1-5 are prepared by the following steps:
(1) performing large-scale fermentation culture on a Sinomonas atrocyanea strain TXB1-10 and a denitrifying Achromobacter (Achromobacter denitirica) strain GPB1-5 in an LB culture solution for 2-3 days respectively, and centrifuging to obtain bacterial sludge;
(2) mixing a protective agent and distilled water to obtain a protective agent solution, and uniformly mixing the bacterial sludge and the protective agent solution according to the volume ratio of 3:1 to obtain a bacterial sludge-protective agent solution; the protective agent comprises the following components: 8mg/L skimmed milk, 5mg/L fructose and 0.05mg/L poly-gamma-glutamic acid;
(3) pre-freezing the bacteria mud-protective agent solution to obtain frozen bacteria;
(4) then, freeze-drying technology is adopted to prepare the freeze-dried powder microbial inoculum.
The formula of the LB culture solution is as follows: 10.0g of tryptone, 5.0g of yeast powder, 10.0g of sodium chloride, 1L of distilled water and pH7.0 +/-0.2 (sterilization).
Example 3:
the preparation method is basically the same as that of the example 2, except that: the protective agent comprises the following components: 10mg/L skimmed milk, 6mg/L fructose and 0.08mg/L poly-gamma-glutamic acid.
Example 4:
the preparation method is basically the same as that of the example 2, except that: the protective agent comprises the following components: 12mg/L skimmed milk, 8mg/L fructose and 0.1mg/L poly-gamma-glutamic acid.
Comparative example 1:
the preparation method is basically the same as that of the example 2, except that: the protective agent comprises the following components: 10mg/L skim milk and 0.08mg/L poly-gamma-glutamic acid.
Comparative example 2:
the preparation method is basically the same as that of the example 2, except that: the protective agent comprises the following components: 10mg/L skim milk and 6mg/L fructose.
Comparative example 3:
the preparation method is basically the same as that of the example 2, except that: the protective agent comprises the following components: 6mg/L fructose and 0.08mg/L poly-gamma-glutamic acid.
The applicant also designed a control group, which was:
(1) performing large-scale fermentation culture on a Sinomonas atrocyanea strain TXB1-10 and a denitrifying Achromobacter (Achromobacter denitirica) strain GPB1-5 in an LB culture solution for 2-3 days respectively, and centrifuging to obtain bacterial sludge;
(2) uniformly mixing the bacterial sludge and the distilled water solution according to the volume ratio of 3:1 to obtain a bacterial sludge solution;
(3) pre-freezing the bacteria mud solution to obtain frozen bacteria;
(4) then, freeze-drying technology is adopted to prepare the freeze-dried powder microbial inoculum.
The freeze-dried powders prepared in the examples 2, 3 and 4 and the comparative examples 1, 2 and 3 and the control group are subjected to the determination of the survival rate and the selenium enrichment capacity, and the determination results are shown in the table 1:
TABLE 1 record of survival rate and selenium enrichment capacity of different lyophilized powder protective strains
As can be seen from Table 1, the survival rate of the control group was lower than that of the strain containing the protective agent, indicating that the survival rate of the strain can be improved by adding the appropriate protective agent during the production of the lyophilized powder. For the TXB1-10 strain, the survival rate is higher and lower, namely, example 3 is larger than example 2 is larger than example 4 is larger than comparative example 2 is larger than comparative example 1 is larger than comparative example 3; for the GPB1-5 strain, the survival rate is high and low, namely, example 3 > example 4 > example 2 > comparative example 3 > comparative example 1 > comparative example 2.
From the viewpoint of selenium conversion, the TXB1-10 strain has the selenium conversion rate of example 3 > example 4 > example 2 > comparative example 1 > comparative example 3; for the GPB1-5 strain, the selenium conversion rate is high and low, namely, example 3 > example 4 > example 2 > comparative example 3 > comparative example 1 > comparative example 2.
Example 5: and (3) selenium-rich vegetable test:
scheme of group 1: a preparation method of a special bacterial fertilizer for selenium-rich vegetables comprises the following steps:
(1) freeze-dried powder of a Chinese unicellular bacterium (Sinomonas atrocyanea) strain TXB1-10 and freeze-dried powder of a colorless denitrifying bacillus (Achromobacter dentifrices) strain GPB1-5 are mixed according to the mass ratio of 1.0: 1.0, mixing to obtain a compound microbial agent;
(2) preparing a mixed fermentation material of crushed crop straws of 120 meshes, bean curd residue, rice biochar and molasses, mixing 65 parts of crushed crop straws, 10 parts of bran, 40 parts of bean curd residue, 5 parts of rice biochar and 1 part of molasses by weight, and fermenting for 20 days while keeping the fermentation temperature at 30 ℃;
(3) and (3) adding 3 parts of the compound microbial agent in the step (1) to continue fermentation when the temperature of the fermented material is reduced to below 50 ℃, turning the pile once every 24 hours until the humidity of the fermented material is about 12%, and finishing the fermentation.
Group 2 protocol: a preparation method of a special bacterial fertilizer for selenium-rich vegetables comprises the following steps:
(1) freeze-dried powder of a Chinese unicellular bacterium (Sinomonas atrocyanea) strain TXB1-10 and freeze-dried powder of a colorless denitrifying bacillus (Achromobacter dentifrices) strain GPB1-5 are mixed according to the mass ratio of 1.5: 1.0, mixing to obtain a compound microbial agent;
(2) preparing a mixed fermentation material of 150-mesh crushed crop straws, bean curd residues, rice biochar and molasses, mixing 75 parts of the crushed crop straws, 15 parts of bran, 50 parts of the bean curd residues, 10 parts of the rice biochar and 5 parts of molasses by weight, and fermenting for 15 days while keeping the fermentation temperature at 60 ℃;
(3) and (3) when the temperature of the fermented material is reduced to below 50 ℃, adding 5 parts of the compound microbial agent in the step (1) for continuous fermentation, and turning the pile once every 48 hours until the humidity of the fermented material is about 12%, so that the fermentation is finished.
Group 3 protocol: a preparation method of a special bacterial fertilizer for selenium-rich vegetables comprises the following steps:
(1) freeze-dried powder of a Chinese unicellular bacterium (Sinomonas atrocyanea) strain TXB1-10 and freeze-dried powder of a colorless denitrifying bacillus (Achromobacter dentifrices) strain GPB1-5 are mixed according to the mass ratio of 1.0: 1.0, mixing to obtain a compound microbial agent;
(2) preparing a mixed fermentation material of crushed crop straws, bean curd residues, rice biochar and molasses which are 120-150 meshes, mixing and fermenting 70 parts of crushed crop straws, 12 parts of bran, 45 parts of bean curd residues, 8 parts of rice biochar and 3 parts of molasses by weight, and keeping the fermentation temperature at 40 ℃ during fermentation for 18 d;
(3) and (3) adding 4 parts of the compound microbial agent in the step (1) to continue fermentation when the temperature of the fermented material is reduced to below 50 ℃, turning the pile once every 36 hours until the humidity of the fermented material is about 12%, and finishing the fermentation.
Group 4 protocol: essentially the same as the protocol of group 3, except that the microbial inoculum comprised only the Sinomonas atrocyanea strain TXB1-10 lyophilized powder.
Group 5 protocol: essentially the same as the protocol of group 3, except that the microbial inoculum comprises only Achromobacter denitrificans (Achromobacter dentifrices) strain GPB1-5 lyophilized powder.
Control group 1 protocol: basically the same as the scheme of group 3, except that no microbial agent is added to the fermentation material.
Control group 2 protocol: essentially the same as the protocol of group 3, except that the microbial agent is prepared by the following steps:
(1) performing large-scale fermentation culture on a Sinomonas atrocyanea strain TXB1-10 and a denitrifying Achromobacter (Achromobacter denitirica) strain GPB1-5 in an LB culture solution for 2-3 days respectively, and centrifuging to obtain bacterial sludge; (2) uniformly mixing the bacterial sludge and the distilled water solution according to the volume ratio of 3:1 to obtain a bacterial sludge solution; (3) pre-freezing the bacteria mud solution to obtain frozen bacteria; (4) then, freeze-drying technology is adopted to prepare the freeze-dried powder microbial inoculum.
The fertilizers prepared by the control group 1, the control group 2, the groups 1, 2, 3, 4 and 5 are prepared for later use. Selecting soil with selenium content of 1.10 mg/kg; planting the crops into beet cores; and (5) carrying out a potted plant planting contrast test. The potted plant is fertilized with 0.15g of nitrogen (per kilogram of soil)N), 0.1g of phosphorus (P)2O5) And 0.15g of potassium (K)2O) is used as a base fertilizer, 10 grains of rapeseeds are sowed in each pot, and the seedlings are fixed to 6 plants after 10 days. After the seedlings of the rapeseeds are settled, 1.00g of fertilizers prepared by a control group 1, a control group 2, groups 1, 2, 3, 4 and 5 are respectively applied to each kilogram of soil, and a proper amount of water is poured. And (4) sampling and analyzing the selenium content of the beet when the beet center is collected.
TABLE 2 sugar beet dried heart sample selenium content
As can be seen from Table 2, the selenium content of the heart, stem and leaf of beet is effectively increased by the treatment of each bacterial manure. The total selenium content of the over-ground part sample (dry sample) of the sugar beet heart of the control group 1 is 0.2237 +/-0.0204 mg/kg, the total selenium content of the over-ground part sample (dry sample) of the sugar beet heart is 0.3673 +/-0.0012 mg/kg, 0.3520 +/-0.0046 mg/kg, 0.3742 +/-0.0085 mg/kg, 0.4314 +/-0.0317 mg/kg and 0.4083 +/-0.0071 mg/kg respectively by treatment of the applied group 4(TXB1-10), the group 5(GPB1-5) and the bacterial manure of the mixed strains (groups 1, 2 and 3), and is respectively increased by 64.2%, 57.4%, 67.3%, 92.9% and 82.5% compared with the control group 1; the total selenium content of the sample (dry sample) of the overground part of the sugar beet heart of the control group 2 is 0.3109 +/-0.0107, and the total selenium content of the sample (dry sample) of the overground part of the sugar beet heart is respectively improved by 18.2 percent, 13.2 percent, 20.3 percent, 38.8 percent and 31.3 percent compared with the control group 2 by applying the treatment of the group 4(TXB1-10), the group 5(GPB1-5) and the bacterial manure of the mixed strain (groups 1, 2 and 3).
From the data of roots, the bacterial manure treatment adopting the method can also effectively improve the selenium content of the roots of the heart of the beet. The total selenium content of the sugar beet core root samples (dry samples) of the control group 1 is 0.4513 +/-0.0125 mg/kg, the treatment of the applied group 4(TXB1-10), the group 5(GPB1-5) and the mixed strain bacterial manure (groups 1, 2 and 3) respectively has the total selenium content of 0.6449 +/-0.0115 mg/kg, 0.5672 +/-0.0479 mg/kg, 0.6796 +/-0.0247 mg/kg, 0.6869 +/-0.0186 mg/kg and 0.6544 +/-0.0175 mg/kg which are respectively 42.9%, 25.7%, 50.6%, 52.2% and 45.0% higher than that of the control group 1; the total selenium content of the sugar beet core root samples (dry samples) of the control group 2 is 0.5393 +/-0.0154 mg/kg, and the total selenium content of the sugar beet core root samples (dry samples) is respectively improved by 19.6%, 5.2%, 26.0%, 27.4% and 21.3% compared with the control group 2 by the treatment of the group 4(TXB1-10), the group 5(GPB1-5) and the bacterial manure of mixed strains (groups 1, 2 and 3).
In conclusion, the selenium-rich effect of each processed cabbage heart is the best in the group 2 technical scheme.
The fertilizers prepared by the control group 1, the control group 2, the groups 1, 2, 3, 4 and 5 are prepared for later use. Selecting soil with total selenium content of 0.51 mg/kg; the planting crops are pakchoi; and performing a planting contrast test in a selenium-rich vegetable garden. And (4) applying base fertilizer and sowing in the vegetable garden according to the conventional method. After the rape seeds emerge, uniformly spreading 150-200 kg/mu of fertilizers prepared by a control group 1, a control group 2 and groups 1, 2, 3, 4 and 5 in 6 test areas divided in the soil of the vegetable garden respectively, and watering with proper amount of water. The selenium content of the pakchoi is sampled and analyzed when the pakchoi is collected.
TABLE 3 selenium content of dried pakchoi
As can be seen from Table 3, the selenium content in the stem and leaf and root of the pakchoi obtained by the planting method of the present invention is significantly increased. The selenium content of the stem and leaf sample (dry sample) of the pakchoi in the control group 1 is 0.0833 +/-0.0027 mg/kg, and the selenium content of the root sample (dry sample) is 0.0968 +/-0.0018 mg/kg; after the bacterial agents of the groups 1, 2, 3, 4 and 5 are applied, the selenium content of stem and leaf samples (dry samples) of the pakchoi is 0.1838 +/-0.0051 mg/kg, 0.1931 +/-0.0037 mg/kg, 0.1750 +/-0.0021 mg/kg, 0.1662 +/-0.0041 mg/kg and 0.1506 +/-0.0075 mg/kg respectively, and the selenium content of root samples (dry samples) is 0.1320 +/-0.0057 mg/kg, 0.1560 +/-0.0042 mg/kg, 0.1475 +/-0.0078 mg/kg, 0.1410 +/-0.0057 mg/kg and 0.1145 +/-0.0035 mg/kg. After the planting method is adopted, the selenium content of the stems and leaves of the pakchoi of the groups 1, 2, 3, 4 and 5 is respectively improved by 120.6%, 131.8%, 110.0%, 99.5% and 80.8% compared with the control 1, and the selenium content of the roots is respectively improved by 36.4%, 61.2%, 52.4%, 45.7% and 18.9% compared with the control 1.
The selenium content of the stem and leaf sample (dry sample) of the pakchoi in the control group 2 is 0.1310 + -0.0070 mg/kg, and the selenium content of the root sample (dry sample) is 0.1046 + -0.0013 mg/kg; after the planting method is adopted, the selenium content of the stems and leaves of the pakchoi of the groups 1, 2, 3, 4 and 5 is respectively increased by 40.3%, 47.4%, 33.6%, 26.9% and 15.0% compared with the control group 2, and the selenium content of the roots of the pakchoi of the groups 1, 2, 3, 4 and 5 is respectively increased by 26.2%, 49.1%, 41%, 34.8% and 9.5% compared with the control group 2.
In conclusion, the selenium enrichment effect of the pakchoi in each treatment is the best in the group 2 technical scheme. The selenium content of the stem and leaf or root samples (dry samples) of pakchoi was slightly lower in control 1 than in control 2. Compared with the microbial inoculum treated by the method, the microbial inoculum added in the control group 2 without treatment is obviously reduced in promoting the selenium enrichment of the pakchoi, and the results of the method disclosed by the invention are just proved to be the same as the results in the table 1.
In conclusion, the method can obviously improve the selenium content of the stems and leaves of the overground parts of the pakchoi under the condition of not applying exogenous selenium, and the selenium content of the pakchoi applied with the bacterial fertilizer meets the selenium content standard (0.10-1.00mg/kg) of vegetables (in dry weight) GH/T1135-2017 which are standard selenium-rich agricultural products in the industry of the people's republic of China.
The group 2 was also tested in the field for best results. The specific scheme is as follows:
the selenium-rich vegetable garden develops a field test for applying the special bacterial fertilizer for the selenium-rich vegetables. The total selenium content in the soil is 0.51mg/kg, and the planted crop is Chinese cabbage. The vegetable garden is conventionally fertilized with base fertilizer, sowed in a direct sowing mode, thinned at proper time and planted at the density of 100 seedlings/m2Left and right. After the seedlings of the rapeseeds emerge, uniformly spreading 2250-3000 kg/ha of the special bacterial fertilizer for the selenium-rich vegetables obtained by the scheme of the group 2 in the invention in the soil of the vegetable garden by taking no bacterial fertilizer as a control, and irrigating a proper amount of water. The experiment was repeated 3 times, each cell having an area of 20m2And sampling and measuring the yield of the Chinese cabbage when the Chinese cabbage is harvested.
TABLE 4 Brassica campestris Small-area yield
| Treatment of | Cell output (kg) | Increase of control (%) |
| Control | 52.62±0.91 | -- |
| Bacterial manure treatment | 64.37±3.05 | 22.3 |
The result of plot experiment shows that the yield of bacterial manure treatment is increased by 22.3% compared with that of contrast treatment.
The above examples are merely illustrative of several embodiments of the present invention, and the description thereof is more specific and detailed, but not to be construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present invention should be subject to the appended claims.
Claims (7)
1. The special bacterial fertilizer for the selenium-rich vegetables is characterized by being prepared from the following raw materials in parts by weight: 65-75 parts of crushed crop straw, 10-15 parts of bran, 40-50 parts of bean curd residue, 5-10 parts of rice biochar, 1-5 parts of molasses and 3-5 parts of compound microbial agent;
the composite microbial agent is prepared from a Sinomonas atrocerea (Sinomonas atrocerea) strain TXB1-10 freeze-dried powder and a Achromobacter denitrificans (Achromobacter denitirichiana) strain GPB1-5 freeze-dried powder in a mass ratio of 1.0-1.5: 1.0, mixing to obtain a compound microbial agent;
the Chinese unicellular bacterium darkblack (Sinomonas atrocyanea) strain TXB1-10 has a preservation number of CCTCC NO: m2020984; achromobacter denitrificans (Achromobacter denitiricans) strain GPB1-5 with the preservation number of CCTCC NO: m2020344.
2. The special bacterial fertilizer for selenium-enriched vegetables as claimed in claim 1, wherein the strain of Chinese unicellular atrocyanea atrox (Sinomonas atrocynaea) TXB1-10 is separated from the soil in selenium-enriched district in Guangxi county of China; the strain GPB1-5 is separated from the soil in the selenium-rich area of Guangxi Guiping of China.
3. The special bacterial fertilizer for selenium-enriched vegetables as claimed in claim 1, wherein the lyophilized powder of Chinese unicellular atrophaea (Sinomonas atrocyanoea) strain TXB1-10 and the lyophilized powder of Achromobacter denitrificans (Achromobacter dentifrices) strain GPB1-5 are prepared by the following steps:
(1) performing large-scale fermentation culture on a Sinomonas atrocyanea strain TXB1-10 and a denitrifying Achromobacter (Achromobacter denitirica) strain GPB1-5 in an LB culture solution for 2-3 days respectively, and centrifuging to obtain bacterial sludge;
(2) mixing a protective agent with distilled water to obtain a protective agent solution, and then uniformly mixing the protective agent solution with the bacterial sludge to obtain a bacterial sludge-protective agent solution;
(3) pre-freezing the bacteria mud-protective agent solution to obtain frozen bacteria;
(4) then, freeze-drying technology is adopted to prepare the freeze-dried powder microbial inoculum.
4. The special bacterial fertilizer for selenium-enriched vegetables as claimed in claim 3, wherein in the step (2), the volume ratio of the bacterial sludge to the protective agent solution is 3-4: 1.
5. The special bacterial fertilizer for selenium-enriched vegetables as claimed in claim 3, wherein the protective agent comprises the following components: 8-12mg/L skimmed milk, 5-8mg/L fructose and 0.05-0.1mg/L poly-gamma-glutamic acid.
6. The preparation method of the special bacterial fertilizer for the selenium-enriched vegetables according to any one of claims 1 to 5, which is characterized by comprising the following steps:
(1) freeze-dried powder of a Chinese unicellular bacterium (Sinomonas atrocyanea) strain TXB1-10 and freeze-dried powder of a colorless denitrifying bacillus (Achromobacter dentifrices) strain GPB1-5 are mixed according to the mass ratio of 1.0-1.5: 1.0, mixing to obtain a compound microbial agent;
(2) preparing a mixed fermentation material of crushed crop straws of 120-150 meshes, bean curd residues, rice biochar and molasses, and mixing and fermenting 65-75 parts of the crushed crop straws, 10-15 parts of bran, 40-50 parts of bean curd residues, 5-10 parts of the rice biochar and 1-5 parts of molasses by weight;
(3) and (3) adding the compound microbial agent in the step (1) to continue fermenting after the temperature of the fermented material is reduced to below 50 ℃, turning the pile once every 24-48 hours until the humidity of the fermented material is about 12%, and finishing the fermentation.
7. The method according to claim 6, wherein the fermentation temperature of the step (2) is 30-60 ℃, and the fermentation time is 15-20 d.
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