CN114982413A - Solid-state fermentation preparation and application of biochar-based bacillus inoculant - Google Patents
Solid-state fermentation preparation and application of biochar-based bacillus inoculant Download PDFInfo
- Publication number
- CN114982413A CN114982413A CN202210679938.5A CN202210679938A CN114982413A CN 114982413 A CN114982413 A CN 114982413A CN 202210679938 A CN202210679938 A CN 202210679938A CN 114982413 A CN114982413 A CN 114982413A
- Authority
- CN
- China
- Prior art keywords
- biochar
- soil
- solid
- bacillus
- jhm03
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 239000002054 inoculum Substances 0.000 title claims abstract description 21
- 241000193830 Bacillus <bacterium> Species 0.000 title claims abstract description 13
- 238000002360 preparation method Methods 0.000 title claims abstract description 5
- 238000010563 solid-state fermentation Methods 0.000 title abstract description 10
- 239000002689 soil Substances 0.000 claims abstract description 49
- 235000013311 vegetables Nutrition 0.000 claims abstract description 38
- 230000004151 fermentation Effects 0.000 claims abstract description 26
- 238000000855 fermentation Methods 0.000 claims abstract description 24
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 17
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 17
- 239000007787 solid Substances 0.000 claims abstract description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 15
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 14
- 241000194103 Bacillus pumilus Species 0.000 claims abstract description 11
- 235000013379 molasses Nutrition 0.000 claims abstract description 8
- 238000004321 preservation Methods 0.000 claims description 7
- 230000001737 promoting effect Effects 0.000 claims description 7
- 235000010469 Glycine max Nutrition 0.000 claims description 4
- 244000068988 Glycine max Species 0.000 claims description 3
- 235000013312 flour Nutrition 0.000 claims description 3
- 239000003124 biologic agent Substances 0.000 claims description 2
- 238000011081 inoculation Methods 0.000 claims 1
- 239000000843 powder Substances 0.000 claims 1
- 239000002068 microbial inoculum Substances 0.000 abstract description 23
- 235000019764 Soybean Meal Nutrition 0.000 abstract description 4
- 230000006872 improvement Effects 0.000 abstract description 4
- 239000004455 soybean meal Substances 0.000 abstract description 4
- 229910052793 cadmium Inorganic materials 0.000 abstract description 3
- BDOSMKKIYDKNTQ-UHFFFAOYSA-N cadmium atom Chemical compound [Cd] BDOSMKKIYDKNTQ-UHFFFAOYSA-N 0.000 abstract description 3
- 244000005700 microbiome Species 0.000 abstract description 2
- 239000002994 raw material Substances 0.000 abstract description 2
- 239000000758 substrate Substances 0.000 abstract description 2
- 238000010521 absorption reaction Methods 0.000 abstract 1
- 229910001385 heavy metal Inorganic materials 0.000 description 11
- 230000000813 microbial effect Effects 0.000 description 11
- 244000000626 Daucus carota Species 0.000 description 6
- 235000002767 Daucus carota Nutrition 0.000 description 6
- 235000003228 Lactuca sativa Nutrition 0.000 description 6
- 235000009337 Spinacia oleracea Nutrition 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 241000208822 Lactuca Species 0.000 description 5
- 241000219315 Spinacia Species 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 244000088415 Raphanus sativus Species 0.000 description 4
- 235000006140 Raphanus sativus var sativus Nutrition 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 230000009467 reduction Effects 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 238000001816 cooling Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 238000005067 remediation Methods 0.000 description 3
- 241000894006 Bacteria Species 0.000 description 2
- 235000011293 Brassica napus Nutrition 0.000 description 2
- 240000008100 Brassica rapa Species 0.000 description 2
- 235000000540 Brassica rapa subsp rapa Nutrition 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000035558 fertility Effects 0.000 description 2
- 238000004382 potting Methods 0.000 description 2
- 241000186361 Actinobacteria <class> Species 0.000 description 1
- 239000002028 Biomass Substances 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 240000008415 Lactuca sativa Species 0.000 description 1
- 239000005708 Sodium hypochlorite Substances 0.000 description 1
- 244000300264 Spinacia oleracea Species 0.000 description 1
- DGEZNRSVGBDHLK-UHFFFAOYSA-N [1,10]phenanthroline Chemical compound C1=CN=C2C3=NC=CC=C3C=CC2=C1 DGEZNRSVGBDHLK-UHFFFAOYSA-N 0.000 description 1
- 230000001133 acceleration Effects 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000012876 carrier material Substances 0.000 description 1
- 239000003610 charcoal Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000009833 condensation Methods 0.000 description 1
- 230000005494 condensation Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 230000000249 desinfective effect Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000004134 energy conservation Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000002354 inductively-coupled plasma atomic emission spectroscopy Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 238000000120 microwave digestion Methods 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910052664 nepheline Inorganic materials 0.000 description 1
- 239000010434 nepheline Substances 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 230000033116 oxidation-reduction process Effects 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 230000009919 sequestration Effects 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 239000003516 soil conditioner Substances 0.000 description 1
- 238000003900 soil pollution Methods 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 239000012086 standard solution Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000003756 stirring Methods 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01B—SOIL WORKING IN AGRICULTURE OR FORESTRY; PARTS, DETAILS, OR ACCESSORIES OF AGRICULTURAL MACHINES OR IMPLEMENTS, IN GENERAL
- A01B79/00—Methods for working soil
- A01B79/02—Methods for working soil combined with other agricultural processing, e.g. fertilising, planting
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Forests & Forestry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Organic Chemistry (AREA)
- Environmental Sciences (AREA)
- Ecology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Soil Sciences (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biomedical Technology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mechanical Engineering (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Botany (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a solid-state fermentation process of a biological carbon-based bacillus preparation and application of a microbial inoculum, belonging to the application fields of microorganisms, soil improvement agents and ecological improvement. The invention provides a solid microbial inoculum JHM03 which takes biochar as a main raw material and contains Bacillus pumilus (Bacillus pumilus) YG35 with the serial number of CCTCC M20211144. The formula of the solid fermentation medium is as follows: biochar is used as a substrate, molasses is 2.5%, soybean meal is 2.0%, water is 48%, the culture condition is that the inoculum size is 13%, the temperature is 30 ℃, and fermentation is carried out for 4 days. The prepared biochar-based bacillus inoculant JHM03 has the effective viable count of more than or equal to 2.5 multiplied by 10 8 CFU/g. The microbial inoculum JHM03 passivates soil cadmium, reduces the absorption of cadmium by vegetables, remarkably promotes the growth of the vegetables,increase the organic carbon of soil.
Description
Technical Field
The invention belongs to the application fields of microorganisms, soil conditioners and ecological improvement, and relates to a process for preparing a microbial agent by bacillus solid fermentation and application of the microbial agent in the fields of soil remediation, soil carbon sink increase and the like.
Background
The heavy metal pollution of soil in China is increasingly serious. The heavy metal content of vegetables growing in the heavy metal contaminated soil generally exceeds the standard, and the lead and cadmium exceed the standard at most. In the three methods for restoring the heavy metal polluted farmland soil, physical restoration consumes manpower and material resources, addresses the symptoms and does not address the root causes, chemical restoration may cause secondary pollution, and microbial restoration in biological restoration restores the polluted soil through a biological adsorption and biological oxidation-reduction mechanism, has the advantages of good effect, investment saving, low cost, easiness in management and operation, no secondary pollution and the like, and increasingly becomes a hotspot of the research on restoring the polluted soil.
The development of the field of Chinese microbial agents is very rapid, and the application of the Chinese microbial agents in the aspect of soil pollution remediation is gradually rising. With the popularization and development of ecological agriculture and organic agriculture in China, the application of the solid-state fermentation technology in agricultural production is increasingly wide, and the solid-state fermentation technology covers a plurality of fields such as soil improvement agents, plant protection, biological feed and the like. Compared with liquid fermentation, the solid fermentation has the advantages of less water consumption, less discharge of three wastes, less energy consumption, less equipment investment, strong raw material applicability and high product concentration. The development and popularization of the solid fermentation technology to replace or partially replace liquid fermentation have important significance in energy conservation and emission reduction. The microbial agents such as bacteria, actinomycetes and fungi can be produced by a solid state fermentation method. However, the technologies for optimizing fermentation media and culture conditions of remediation microbial agents and products meeting national standards of agricultural microbial agents are rare.
The biochar is a high-tech product prepared by pyrolyzing renewable biomass at high temperature under a closed anoxic condition. The biochar has rich carbon content, more pores and strong stability, can improve the soil texture, improve the soil fertility, enhance the soil carbon sequestration capacity and reduce the poison of heavy metals to plants, and is widely applied as a soil conditioner. Biochar can be used as a carrier material for the microbial agent in consideration of its porosity and nutritional effects. Most studies have mixed or applied biochar with the bacterial solution. The application of producing the microbial inoculum by solid state fermentation by using the biochar as a substrate is not reported.
Disclosure of Invention
The invention aims to provide a solid-state fermentation process capable of improving the effective viable count of a microbial agent, aiming at the problem of low viable count of solid-state fermentation.
The purpose of the invention can be realized by the following technical scheme:
application of Bacillus pumilus YG35 with preservation number of CCTCC NO: M20211144 in reducing Cd content in vegetables and farmland soil, promoting vegetable growth and/or increasing organic carbon content in soil.
Application of Bacillus pumilus YG35 with preservation number of CCTCC NO: M20211144 in preparation of biological agent for reducing Cd content in vegetable and farmland soil, promoting vegetable growth and/or increasing organic carbon content in soil.
A biochar-based Bacillus inoculant produced by fermenting the Bacillus pumilus YG35 of claim 1 in a solid fermentation medium containing biochar.
As a preferred mode of the present invention, the solid fermentation medium is composed of biochar, molasses, soybean meal and water.
As a further optimization of the invention, the solid state fermentation medium takes the mass of the biochar as a reference, and the other components are as follows in percentage by weight: 2-3% of molasses, 1-3% of soybean flour and 45-50% of water.
As a further preferred aspect of the present invention, the solid fermentation medium is prepared by sieving biochar with a 20-mesh sieve, placing 100g of biochar in a 500mL conical flask, adding 2.5% molasses (by weight), 2.0% soybean meal (by weight), and 50mL of water, stirring well, sealing, sterilizing at 121 ℃ for 25min, and cooling.
As a preferred embodiment of the present invention, the number of YG35 cells in the solid microbial inoculum reaches 2X 10 9 CFU/g is higher than the standard.
In a preferred embodiment of the present invention, the fermentation is carried out at a fermentation temperature of 30-33 deg.C, preferably 30 deg.C, and an optimal inoculum size of 13% (% represents ml/100g biochar), for 72-96 h.
The biochar-based bacillus agent disclosed by the invention is applied to reducing the Cd content in the soil of vegetables and farmlands, promoting the growth of the vegetables and/or increasing the organic carbon content in the soil.
Advantageous effects
The Bacillus pumilus YG35 strain is classified and named as Bacillus pumilus YG35 and is preserved in China center for type culture collection (CCTCC M20211144) with the preservation date of 2021 year, 9 months and 6 days. The solid fermentation culture medium and conditions are optimized, the effective viable count of the finished microbial inoculum is improved, and the biochar-based microbial inoculum is prepared.
The biochar-based microbial inoculum (JHM03) containing B.puminus YG35 can promote vegetable growth, remarkably reduce the content of Cd in heavy metal contaminated soil and vegetables, and increase the content of organic carbon in soil. Compared with the prior art, the method has the following advantages:
(1) the effective viable count of the B.puminus YG35 biochar-based microbial inoculum (JHM03) can reach 2.38 multiplied by 10 9 CFU/g。
(2) The effective viable count of the biochar-based microbial inoculum (JHM03) can be maintained at 8.7 × 10 after being placed at room temperature for 6 months 8 CFU/g。
(3) The content of the effective Cd in the heavy metal contaminated soil which is sprayed with the biochar-based microbial inoculum (JHM03) is obviously reduced by 37.2 percent (P < 0.05).
(4) The Cd content of edible parts of spinach, lettuce, carrot and white radish treated by applying the biochar-based microbial inoculum (JHM03) is remarkably reduced by 36.7-66.7% (P < 0.05).
(5) Spinach and lettuce edible part treated by the biochar-based inoculant (JHM03) has the dry weight remarkably increased by 74.7% and 92.8% (P <0.05), and carrot and white turnip have the dry weight remarkably increased by 3.6 times and 3.7 times (P < 0.05).
(6) The organic carbon content of the rhizosphere soil of spinach, lettuce, carrot and white radish which are applied with the biochar-based microbial inoculum (JHM03) is obviously increased by 52.4-94.8% (P is less than 0.05).
Detailed Description
The following claims are hereby incorporated into the detailed description of the invention, with the understanding that the invention is not to be limited in any way, as any number of modifications may be made by one within the scope of the claims and the invention is still within the scope of the claims.
The following examples are, unless otherwise indicated, all of the routine experimentation and procedures known in the art.
EXAMPLE 1 solid fermentation culture
Placing Bacillus pumilus YG35 (preservation number is CCTCC NO: M20211144, the strain is disclosed in CN 113913331A) preserved in a glycerol tube of a refrigerator at-80 ℃ in a water bath kettle at 37 ℃ for thawing for 60s, then inoculating into an LB flat plate, culturing in an incubator at 30 ℃ for 24h, and activating strains; the activated strain is inoculated into a 250mL conical flask filled with 150mL LB liquid medium, and is subjected to shaking culture at 30 ℃ and 180r/min for 16h to serve as seed liquid. Adding the seed solution into a solid fermentation culture medium (biochar is sieved by a 20-mesh sieve, 100g of biochar is put into a 500mL conical flask, then 2.5% of molasses (weight ratio), 2.0% of soybean meal (weight ratio) and 50mL of water are added, uniformly stirred, sealed, sterilized at 121 ℃ for 25min and cooled to obtain the solid fermentation culture medium), inoculating the strain with 13%, and standing and culturing for 3d at 30 ℃ to obtain the biochar-based microbial inoculum JHM 03. Sampling, diluting and coating an LB plate according to 10 times of gradient, and calculating the effective viable count. The result shows that the effective viable count can reach 2.38 multiplied by 10 9 CFU/g。
Example 2 biochar-based inoculant JHM03 preservation
The effective viable count of the charcoal-based microbial inoculum JHM03 obtained in example 1 is kept at 8.7 × 10 in the microbial inoculum JHM03 after 6 months of storage at room temperature 8 CFU/g, which meets the national standard of the number of effective viable bacteria in the agricultural microbial agent.
Example 3 Effect of biochar-based microbial inoculum on reduction of Cd content in heavy metal contaminated soil and vegetables
Collecting heavy metal polluted soil in the inhabited nepheline area of Nanjing City. Potting experiments were arranged, each pot containing 1.5kg of soil, and 700g of deionized water was added to the pot to maintain a suitable water content throughout the soil environment. Treatment group 5% of the inoculum JHM03 prepared in example 1 was mixed with potting soil, and no inoculum treatment was appliedAs a control. Lettuce, spinach, carrot and white radish were selected as the test vegetables. The method comprises the steps of disinfecting the surfaces of vegetable seeds for 20min by using a 5% sodium hypochlorite solution, placing the vegetable seeds on sterile gauze, placing the vegetable seeds on a sterile incubator at the temperature of 30 ℃, keeping the humidity at 60% -80% for germination acceleration, and selecting the seeds with the same size and exposed white, and placing the seeds on a water culture bowl for culture. When the bud grows to about 1cm, selecting vegetable seedlings with uniform growth vigor and size for planting. Each treatment was repeated three times, and a conventional management was performed for 45 days. And collecting vegetable rhizosphere soil to determine the content of available Cd in the soil. The samples were baked to constant weight and ground with a tissue grinder. Accurately weighing 0.1g of vegetable sample, performing microwave digestion, and then using 5% HNO 3 And (4) performing constant volume, and determining the content of Cd in the digestion solution by adopting ICP-OES. As can be seen from Table 5, compared with the control, the Cd content of the edible part of the vegetable treated by the microbial inoculum is significantly reduced by 36.7-66.7%, wherein the Cd content reduction rate of carrot is the largest (Table 1). The Cd contents of spinach, lettuce and white radish meet the relevant regulations of the national Ministry of agriculture (GB 2762-.
As can be seen from Table 2, the content of Cd in the effective state of the vegetable rhizosphere soil after being treated by the microbial inoculum is obviously reduced by 28.9-37.5%, the bioavailability is reduced, and the Cd toxicity is relieved.
TABLE 1 biochar-based inoculant JHM03 for reducing Cd content (mg/kg) in edible part of vegetables
TABLE 2 reduction of available Cd content in vegetable rhizosphere soil by biochar-based inoculant JHM03
Example 4 action of biochar-based inoculant JHM03 in heavy metal soil to promote vegetable growth
The vegetables treated in each of examples 3 were collected, and the dry weight of the edible part of the vegetables was measured to evaluate the effect of increasing the yield of the microbial inoculum. As can be seen from Table 3, compared with the control, the dry weight of the edible parts of spinach and lettuce treated by the microbial inoculum is remarkably increased by 74.7% and 92.8% (P <0.05), the weight of carrot and dried white turnip is remarkably increased by 3.6 times and 3.7 times, and the yield of the microbial inoculum on vegetables is remarkably increased.
TABLE 3 biochar-based inoculant JHM03 for promoting vegetable growth
Example 5 biochar-based inoculant JHM03 in heavy metal soil for increasing organic carbon content of vegetable rhizosphere soil
The rhizosphere soil of the vegetables treated in example 3 was collected, dried soil sample (0.1-1 g) of 100 mesh was collected and placed in the bottom of a glass test tube without water, and 1/6K was added precisely by using a pipette 2 Cr 2 O 7 5mL of standard solution is added into 5mL of concentrated sulfuric acid by using a pipette, the mixture is shaken evenly, a pipe opening is covered with a bent neck funnel, and water vapor is evaporated by condensation. Placing the test tube in an iron wire cage, placing the test tube in a 190 ℃ oil bath pot, adjusting the temperature of the oil bath pot to 170-180 ℃, boiling for 5min when bubbles are generated when liquid in the test tube boils, and taking out the test tube and cooling to room temperature. After cooling, transferring the liquid and residue in the tube into a triangular flask without damage, so that the total volume of the solution in the triangular flask is 60-70 mL, and the concentration of the mixed solution is kept at 2-3 mol L -1 And then titrating by using an phenanthroline indicator, and converting the content of organic carbon in the soil sample after titration is finished.
As can be seen from Table 4, compared with the control, the organic carbon content of the vegetable rhizosphere soil treated by the microbial inoculum is remarkably increased by 52.4-94.8% (P <0.05), which is beneficial to the cultivation of soil fertility.
TABLE 4 biochar-based inoculant JHM03 for increasing organic carbon in vegetable rhizosphere soil
Claims (8)
1. Application of Bacillus pumilus YG35 with preservation number of CCTCC NO: M20211144 in reducing Cd content in vegetables and farmland soil, promoting vegetable growth and/or increasing organic carbon content in soil.
2. Application of Bacillus pumilus YG35 with preservation number of CCTCC NO: M20211144 in preparation of biological agent for reducing Cd content in vegetable and farmland soil, promoting vegetable growth and/or increasing organic carbon content in soil.
3. A biochar-based Bacillus inoculant produced by fermenting the Bacillus pumilus YG35 according to claim 1 in a solid fermentation medium containing biochar.
4. The biochar-based bacillus inoculant according to claim 3, wherein the solid fermentation medium consists of biochar, molasses, soy flour and water.
5. The biochar-based bacillus inoculant according to claim 4, wherein the solid fermentation medium is prepared from the following components in percentage by weight based on the mass of biochar: 2-3% of molasses, 1-3% of soybean flour and 45-50% of water.
6. The biochar-based bacillus inoculant according to claim 4, wherein the solid fermentation medium is prepared from the following components in percentage by weight based on the mass of biochar: molasses 2.5%, soybean powder 2.0%, and water 48%.
7. The biochar-based bacillus inoculant according to any one of claims 3 to 6, wherein the fermentation conditions are that the fermentation temperature is 30-33 ℃, the optimal inoculation amount is 13%, and the fermentation lasts for 72-96 h.
8. The use of the biochar-based bacillus agent as claimed in any one of claims 3 to 6 for reducing the Cd content of vegetables and farmland soil, promoting vegetable growth and/or increasing the organic carbon content of soil.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210679938.5A CN114982413B (en) | 2022-06-16 | 2022-06-16 | Solid state fermentation preparation and application of biochar-based bacillus inoculant |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210679938.5A CN114982413B (en) | 2022-06-16 | 2022-06-16 | Solid state fermentation preparation and application of biochar-based bacillus inoculant |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114982413A true CN114982413A (en) | 2022-09-02 |
| CN114982413B CN114982413B (en) | 2023-12-08 |
Family
ID=83036063
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210679938.5A Active CN114982413B (en) | 2022-06-16 | 2022-06-16 | Solid state fermentation preparation and application of biochar-based bacillus inoculant |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114982413B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113913331A (en) * | 2021-10-12 | 2022-01-11 | 南京农业大学 | Saline-alkali-tolerant bacillus pumilus for producing polyglutamic acid and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102399723A (en) * | 2011-11-07 | 2012-04-04 | 南京大学 | Bacillus with electrogenesis characteristic and application thereof to microbiological fuel cell |
| CN102911901A (en) * | 2012-10-25 | 2013-02-06 | 青岛蔚蓝生物集团有限公司 | Bacillus pumilus strain and application thereof |
| CN111172052A (en) * | 2018-12-06 | 2020-05-19 | 东南大学 | Bacillus P75 for improving soil fertility of sandstone and application thereof |
| CN113913331A (en) * | 2021-10-12 | 2022-01-11 | 南京农业大学 | Saline-alkali-tolerant bacillus pumilus for producing polyglutamic acid and application thereof |
-
2022
- 2022-06-16 CN CN202210679938.5A patent/CN114982413B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102399723A (en) * | 2011-11-07 | 2012-04-04 | 南京大学 | Bacillus with electrogenesis characteristic and application thereof to microbiological fuel cell |
| CN102911901A (en) * | 2012-10-25 | 2013-02-06 | 青岛蔚蓝生物集团有限公司 | Bacillus pumilus strain and application thereof |
| CN111172052A (en) * | 2018-12-06 | 2020-05-19 | 东南大学 | Bacillus P75 for improving soil fertility of sandstone and application thereof |
| CN113913331A (en) * | 2021-10-12 | 2022-01-11 | 南京农业大学 | Saline-alkali-tolerant bacillus pumilus for producing polyglutamic acid and application thereof |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113913331A (en) * | 2021-10-12 | 2022-01-11 | 南京农业大学 | Saline-alkali-tolerant bacillus pumilus for producing polyglutamic acid and application thereof |
| CN113913331B (en) * | 2021-10-12 | 2023-02-17 | 南京农业大学 | Saline-alkali-tolerant bacillus pumilus for producing polyglutamic acid and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114982413B (en) | 2023-12-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102432393B (en) | Composite organic soil for raising rice seedlings | |
| CN108929700A (en) | A kind of charcoal base immobilized microbial inoculum soil conditioner and preparation method thereof | |
| CN109652329A (en) | A kind of preparation and application of bacillus solid microbial | |
| CN110591724A (en) | Conditioner for treating heavy metal contaminated soil and preparation method and application thereof | |
| CN111057665B (en) | Cellulose degrading bacterium n3 for producing IAA and application thereof | |
| CN109456915B (en) | Bacillus safensis strain X3 and application thereof | |
| CN116024118B (en) | Saline-alkali tolerant microbial agent SYM-6 and application thereof | |
| CN109928854A (en) | A kind of biological organic fertilizer and preparation method thereof with rehabilitating soil function | |
| CN109868242B (en) | Salt-tolerant acetoin-producing bacillus subtilis and application thereof | |
| CN114982413B (en) | Solid state fermentation preparation and application of biochar-based bacillus inoculant | |
| CN109468343B (en) | Promoter for producing biogas through straw anaerobic fermentation and preparation method and application thereof | |
| CN107467075B (en) | Application of bacillus pumilus as rice growth promoter | |
| CN111172073B (en) | Bacillus subtilis strain and application thereof in plant growth | |
| CN116121147B (en) | Chenopodium ambrosioides seed endophytic Larimol agrobacterium and application thereof | |
| CN111234825A (en) | Biomass charcoal for improving saline-alkali soil, preparation method, and preparation and application of derivatives thereof | |
| CN113897316B (en) | Bacillus licheniformis BLc06, functional melon and fruit seedling biological matrix prepared from same and application | |
| CN115651846A (en) | Trichoderma fungicide, biological organic fertilizer, preparation method and application thereof | |
| CN110615722A (en) | Biochar-based multi-component fertilizer and preparation method thereof | |
| CN113817625B (en) | Flavobacterium acidophilus and application thereof in improvement of saline-alkali soil | |
| CN112063552B (en) | Microbial agent for preventing and treating asparagus wilt, preparation method and application thereof | |
| CN112779187B (en) | Bacillus quinquefolius and application thereof | |
| CN109337844B (en) | Pseudomonas moraxella X2 and application thereof | |
| CN107937299A (en) | A kind of method that thermophilic Pb is passivated Pb into ore deposit bacterium and its in sludge high temperature compost | |
| CN114703081B (en) | Brevundimonas ST3CS3 and application thereof | |
| CN116102382B (en) | Fungus encapsulated fertilizer, preparation method and application |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB02 | Change of applicant information | ||
| CB02 | Change of applicant information |
Address after: 210043 Jiangsu Nanjing Qixia District Bagua Zhou street Jiangsu Qixia modern agriculture industrial park Nanjing Agricultural University modern horticulture industry science and Technology Innovation Center Applicant after: NANJING AGRICULTURAL University Address before: Weigang Xuanwu District of Nanjing Jiangsu province 210095 No. 1 Applicant before: NANJING AGRICULTURAL University |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |