CN116355782A - Serratia rubra and application thereof - Google Patents
Serratia rubra and application thereof Download PDFInfo
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- CN116355782A CN116355782A CN202210836233.XA CN202210836233A CN116355782A CN 116355782 A CN116355782 A CN 116355782A CN 202210836233 A CN202210836233 A CN 202210836233A CN 116355782 A CN116355782 A CN 116355782A
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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
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01C—PLANTING; SOWING; FERTILISING
- A01C21/00—Methods of fertilising, sowing or planting
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05C—NITROGENOUS FERTILISERS
- C05C11/00—Other nitrogenous fertilisers
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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
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
- C05F17/20—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation using specific microorganisms or substances, e.g. enzymes, for activating or stimulating the treatment
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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
- C05G5/00—Fertilisers characterised by their form
- C05G5/20—Liquid fertilisers
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- 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
- C12R2001/425—Serratia
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- 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
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Abstract
The invention relates to application of Serratia rubra (Serratia rubidaea) in preparation of organic fertilizer, wherein the Serratia rubra is preserved in China Center for Type Culture Collection (CCTCC) in Wuhan City of Hubei province in 2022, 7 th month and 6 th day, and the preservation number is CCTCC M20221042. The application not only solves the problem of resource waste of low-value fish, fish meat practical waste and the like, realizes the reutilization of waste resources, but also utilizes the amino acid fertilizer prepared by the serratia rubra of the application to realize high fertilizer efficiency, no pollution and no public nuisance, saves the production cost and can lighten the environmental pollution to a certain extent.
Description
Technical Field
The invention relates to bacteria, in particular to Serratia rubra and application thereof in preparing organic fertilizer.
Background
The deterioration of the soil environment brings the problem of crop yield reduction, and has great significance for increasing crop yield and reasonably and scientifically fertilizing. The fertilizer has been widely used because of its low cost, stable ingredients, easy preservation, etc. Along with the continuous development of green agriculture, the defects of the traditional fertilizer are gradually displayed: the fertilizer has single active ingredients and low utilization rate, and excessive use can cause eutrophication of water body, soil pollution, hardening and the like after application. The biological liquid fertilizer has high fertilizer efficiency, no pollution and no public hazard, and meets the development requirements of green agriculture. At present, the price of the biological liquid fertilizer cultivated by the fermentation of the biological bacterial agent is higher, so that a more economical fertilizer production mode is still a hot spot for current research.
Disclosure of Invention
Aiming at the technical problems in the prior art, the invention provides application of Serratia rubra (Serratia rubidaea) in preparing organic fertilizer, wherein the Serratia rubra is preserved in China center for type culture collection (CCTCC M) of Wuhan City in Hubei province for 7 months and 6 days, and the address is eight paths 299 of Wuchang district in Wuhan City in Hubei province, and the preservation number is CCTCC M20221042.
An organic fertilizer comprising: serratia rubra and medium for decomposing the same; or comprises fermentation liquor or fermentation liquor clear liquor obtained after the medium is decomposed by Serratia rubra; wherein, the Serratia rubra is preserved in China Center for Type Culture Collection (CCTCC) of Wuhan City, hubei province at 7 months and 6 days in 2022, and has the address of eight paths 299 of Wuchang district, hubei province, and the preservation number of CCTCC M20221042.
As described above, the medium is a protein-rich animal utility waste including, but not limited to, fish utility waste, shrimp utility waste, crab utility waste, or shell utility waste.
As the organic fertilizer, the medium is tuna practical waste.
The organic fertilizer is amino acid fertilizer.
The organic fertilizer is an amino acid liquid fertilizer.
The preparation method of the organic fertilizer comprises the following steps: providing a medium for decomposition of Serratia; sterilizing the medium; inoculating Serratia rubra to the sterilized medium; fermenting the serratia rubra; wherein the fermentation temperature is 25-30deg.C, or 30-35deg.C; the fermentation time is 12-16 hours.
The application of the organic fertilizer prepared by Serratia rubra (Serratia rubidaea) in improving the germination rate of plant seeds and promoting plant growth is provided, wherein the Serratia rubra is preserved in China Center for Type Culture Collection (CCTCC) of Wuhan City in Hubei province for 7 months and 6 days, and the address is eight paths 299 of Wuchang district in Wuhan City in Hubei province, and the preservation number is CCTCC M20221042.
A method of increasing germination of plant seeds comprising: obtaining the organic fertilizer according to any one of the above; and mixing the organic fertilizer with plant seeds, a medium for culturing plants or sown soil.
A method of promoting plant growth comprising: obtaining the organic fertilizer according to any one of the above claims; and spraying the organic fertilizer to plant leaf surfaces or applying the organic fertilizer to soil where plants grow.
The application not only solves the problem of resource waste of low-value fish, fish meat practical waste and the like, realizes the reutilization of waste resources, but also utilizes the amino acid fertilizer prepared by the serratia rubra of the application to realize high fertilizer efficiency, no pollution and no public nuisance, saves the production cost and can lighten the environmental pollution to a certain extent.
Drawings
Preferred embodiments of the present invention will be described in further detail below with reference to the attached drawing figures, wherein:
FIG. 1 is a screen identification of enzyme producing strains isolated from soil according to one embodiment of the present application, wherein FIG. 1A is a hydrolysis circle graph generated by R3 strain when screening casein plates according to one embodiment of the present application; FIG. 1B is a gel electrophoresis separation profile of PCR products obtained after amplification of bacterial genes with 16S primers according to one embodiment of the present application; FIG. 1C is a phylogenetic tree of protease producing strains obtained after sequence alignment according to one embodiment of the present application; and
FIG. 2 is an effect of amino acid fertilizer obtained after fermentation of fish utility waste by Serratia rubra on germination and growth of cabbage seeds according to one embodiment of the present application.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention, and it is apparent that the described embodiments are some embodiments of the present invention, but not all embodiments of the present invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
In the following detailed description, reference is made to the accompanying drawings, which form a part hereof, and in which is shown by way of illustration specific embodiments in which the application may be practiced. In the drawings, like reference numerals describe substantially similar components throughout the different views. Various specific embodiments of the present application are described in sufficient detail below to enable those skilled in the art to practice the teachings of the present application. It is to be understood that other embodiments may be utilized and structural and logical changes may be made to the embodiments of the present application.
The relevant terms referred to in this application have the following meanings:
the terms "Serratia rubidaea", "Serratia rubra" refer to a species of Serratia (Serratia) that is a gram-negative Bacillus pumilus that secretes proteases. Serratia is very widely distributed in nature and can be found in water, plant leaves, soil, and human respiratory and intestinal tracts. Early Serratia has been considered non-pathogenic, but recently Serratia has received widespread attention as a opportunistic pathogen. However, until now, there is no report that Serratia can cause plant diseases.
In this application, serratia rubra was isolated from Hainan mangrove soil and was self-isolated and preserved in the laboratory. In the application, serratia rubra (Serratia rubidaea) is preserved in China Center for Type Culture Collection (CCTCC) of Wuhan City, hubei province at 7 th month and 6 th day of 2022, and has a preservation number of CCTCC M20221042, wherein the address is eight 299 of Wuhan City, hubei province.
The term "fermentation" in this application refers to the use of Serratia rubra degradation medium to obtain an amino acid fertilizer. In some embodiments, the medium is protein-rich animal utility waste.
The term "medium" may be a practical waste material of fish, shrimp, crab, shellfish etc. which has not been sterilized. When the medium is sterilized, the medium becomes a culture medium for fermentation and decomposition of Serratia rubra. In some embodiments, the sterilization treatment is to sterilize the medium in a sterilization pot at 121 ℃ for 15-25min, preferably 20min; or sterilizing at 117 deg.C for 20-40min, preferably 30min. In one embodiment, the medium is tuna fish utility waste. In one embodiment, the tuna is homogenized in the medium. In some embodiments, an appropriate amount of pure water may be added to the medium.
It should be understood by those skilled in the art that Serratia rubra can secrete protease, and the meat-based practical waste materials of aquatic products such as fish, shrimp, crab and shellfish all comprise protein as main component, so that the substances such as fish, shrimp, crab and shellfish all comprise protein as main component can be used as the medium for fermentation of Serratia rubra in the application. In the present application, the tuna fish flesh utility waste is only one example of the present application, and the kind of the medium is not limited thereto.
The term "utility waste" refers to the leftovers or fertilizer which are remained and separated after the protein-rich substances such as fish, shrimp, crab, shell and the like are processed, and contains rich proteins so as to facilitate the decomposition of the protein-rich substances by Serratia rubra. In some embodiments, the utility waste is a can, post-food processing fertilizer.
The term "tuna homogenate" refers to biological tissue obtained by grinding, mashing, splitting, crushing, blending, etc. tuna, which has a fine composition and is well mixed.
The term "organic fertilizer" in this application refers to an amino acid fertilizer obtained after degradation of the medium by Serratia rubra. In some embodiments, the organic fertilizer may be a solid fertilizer, a liquid fertilizer, or a solid-liquid mixed fertilizer. The solid and liquid forms of the organic fertilizer are not limited, and a person skilled in the art can select an organic fertilizer with a proper form according to actual needs. In one embodiment, the organic fertilizer of the present application refers to an amino acid liquid fertilizer obtained after degrading the medium with Serratia. Further, the organic fertilizer disclosed by the application refers to an amino acid liquid fertilizer obtained after degradation of tuna by Serratia.
The term "Fu Lin Fenfa (Lowry method)" is one of the sensitive protein assay methods that can be used to determine protease activity. The method utilizes the Fu Lin Fen method to determine the activity of protease secreted by a plurality of strains, so as to screen proper strains.
The terms "protease activity" and "protease activity" have the same meaning and refer to the amount of enzyme capable of converting 1. Mu. Mole of a substrate in 1 minute under specific conditions (25 ℃ C., other conditions being optimal) or the amount of enzyme capable of converting 1. Mu. Mole of the relevant groups in the substrate. In the present application, the enzyme activity unit is defined as 1 protease activity unit expressed as U/mL, corresponding to the amount of enzyme consumed to hydrolyze casein at 40℃in 1mL of enzyme solution per minute to produce 1. Mu.g of tyrosine. According to one embodiment of the present application, protease enzyme activity is calculated according to the following formula:
enzyme activity (U/mL) =od 680 ×K×(4/10)×N
The terms "liquid chromatography-mass spectrometry" and "LC-MS" refer to a technique called liquid chromatography-mass spectrometry, which uses liquid chromatography as a separation system and mass spectrometry as a detection system. The amino acid content change of the protein-rich medium in the offal before and after degradation by Serratia rubra is measured by using a liquid chromatography-mass spectrometry instrument.
With the maturation of the modern water-saving irrigation technology, the production and use of liquid fertilizer are rapidly developed. The amino acid liquid fertilizer is used as one of liquid fertilizers, and has the advantages of quick fertilizer efficiency, cleanness, no pollution, improvement of crop quality, improvement of plant metabolism and stress resistance, improvement of soil physicochemical properties, improvement of water and fertilizer retention capacity and the like. The amino acid liquid fertilizer is prepared by fermenting or hydrolyzing the practical wastes of animals and plants, so that the reutilization of waste protein resources can be realized, the added value of the product is increased, and the prepared liquid fertilizer can also be used for improving the yield and quality of crops. However, the preparation of amino acid liquid fertilizer by Serratia rubra is not reported at present.
The method for preparing the amino acid liquid fertilizer by hydrolysis comprises the following steps: enzymatic hydrolysis, acid hydrolysis and alkaline hydrolysis. The enzymatic hydrolysis needs to be carried out under milder conditions, and the reaction conditions are controlled strictly; the hydrolysate in the acid hydrolysis process has peculiar smell, and the produced acid liquor also causes pollution to the environment; alkali hydrolysis changes the L-form amino acid into D-form and the hydrolysis method has no specificity. Compared with acid-base hydrolysis and enzyme hydrolysis, the method has the advantages that the protease-producing strain is directly added into the practical waste for microbial fermentation, so that the production cost is saved to a great extent, and the method is considered to be the most economical and effective means for treating the animal and plant practical waste.
In the processing process of fish products, fish resources cannot be fully utilized, and more than 60% of practical waste materials are generated, wherein the fish products are rich in fish proteins. Microorganisms are gradually used as the basis for preparing high-efficiency and safe biological liquid fertilizer, and have wide application in modern agricultural production. The liquid fertilizer produced by biodegradation has the following characteristics: the fertilizer is high in utilization efficiency and small in dosage; the nutrient absorption is fast, and the fertilizer efficiency is good; the pertinence is strong; the spraying is convenient, economical and environment-friendly; can supplement the deficiency of nutrient absorption of the root of crops. The fish flesh practical waste is used as a raw material, serratia rubra is utilized for fermentation, the serratia rubra is prepared into amino acid liquid fertilizer, the yield and quality of crops are improved through leaf surface spraying, seed soaking, culture medium or soil mixing and other modes, the reutilization of waste fish protein resources can be realized, the added value of fish products is increased, the secondary utilization of energy sources can be realized, and the environmental pollution can be reduced to a certain extent.
The invention aims to provide Serratia and application thereof, and the Serratia can be used for producing an amino acid liquid fertilizer by degrading fish utility waste. The germination experiment of the seeds proves that the produced liquid fertilizer can promote the germination of the cabbage seeds. It will be appreciated by those skilled in the art that the fertility of amino acid fertilizers is not only reflected in the period of seed germination, but that a large amount of various amino acids are required for each stage of crop growth. The efficacy of the amino acid fertilizer is illustrated by taking seed germination as an example, but the amino acid fertilizer obtained by fermenting salmonella rubrum is used for various periods of plant growth, and the amino acid fertilizer is within the protection scope of the application.
According to one embodiment of the application, the method for preparing the amino acid liquid fertilizer by fermenting fish meat with Serratia rubra comprises the following steps:
homogenizing fish flesh waste, adding appropriate amount of pure water, and sterilizing at 121deg.C for 20min.
Inoculating Serratia rubra into the sterilized fish culture medium, and culturing at 30deg.C and pH of 6 for 12-16 hr to obtain fermentation broth.
Centrifuging the fermentation liquor at 5000rpm, removing thalli, and obtaining the supernatant as the obtained amino acid liquid fertilizer stock solution.
The technical solutions of the present application will be described below by way of examples:
example 1: isolation and purification of soil strain and protease enzyme activity determination
According to one embodiment of the present application, bacteria may be isolated and purified from mangrove soil and tested for protease-producing activity.
1. Preparing a soil dilution. 10g of mangrove soil was weighed and mixed into 100mL of sterile water. The obtained supernatant is the soil stock solution.
Then diluting the soil stock solution by 10 times and 10 times 2 Multiple of 10 3 Multiple of 10 4 Multiple of 10 5 Multiple of 10 6 Multiple, etc. And respectively marking test tubes as 0,1,2,3,4,5 and 6 according to the dilution times of the soil stock solution.
2. And (5) coating a plate. The microorganisms in each test tube in the step 1 are coated on a casein solid culture medium and are encapsulated by a sealing film. According to experimental and statistical requirements, each dilution of solution can be spread on the surface of a plurality of solid media. For example, each dilution of solution was spread on 3 solid medium surfaces.
3. And (5) culturing microorganisms. The solid culture medium coated with bacteria is placed in a constant temperature environment at 30 ℃ for culturing for more than 12 hours.
4. Counting. The number of colonies capable of producing hydrolysis circles on each casein solid medium was counted, the morphological characteristics of the colonies were observed, and the results were recorded.
5. The flat panel is scored and separated. The colonies obtained in the above steps are streaked and separated, and then subjected to a suitable temperature for re-cultivation. By the operation, the monoclonal strain can be separated, propagated and stored, and the subsequent sequencing and classification tests are facilitated.
6. And (5) preserving and identifying strains.
7. Fu Lin Fenfa enzyme activity was measured. Table 1 is a listing of parameters for measuring enzyme activity according to Fu Lin Fenfa of one embodiment of the present application.
FIG. 1 is a screen identification of enzyme producing strains isolated from soil according to one embodiment of the present application. FIG. 1A is a diagram showing hydrolysis circles generated by R3 strain during screening of casein plates, and it can be seen from the diagram that the R3 strain appears red in appearance, which can cause the casein plates to generate hydrolysis circles. FIG. 1B is a gel electrophoresis separation profile of the PCR product obtained after amplification of bacterial genes with 16S primers; FIG. 1C is a phylogenetic tree of protease-producing strains obtained after sequence alignment, which was analyzed by sequence alignment to identify the strain as Serratia rubra (Serratia rubidaea).
TABLE 1 Fu Lin Fenfa measurement of enzyme Activity parameters
Example 2: identification of strain species
According to one embodiment of the present application, the strain species may be initially identified using 16S universal primers. According to one embodiment of the present application, strain identification may be performed directly using the strain as a substrate, or genomic DNA may be extracted first and strain identification may be performed using genomic DNA as a substrate.
1. PCR amplification of 16S rDNA was performed. The universal primers 27F and 1492R were selected for PCR amplification, wherein the DNA sequences of 27F and 1492R were as follows:
27F：5’-AGAGTTTGATCCTGGCTCAG-3’；
1492R：5’-GGTTACCTTGTTACGACTT-3’。
according to the selected different DNA polymerases, a proper PCR reaction system and reaction conditions are selected. The application is not limited to the use of any kind of DNA polymerase, nor to the PCR reaction system and reaction conditions after the use of the same DNA polymerase. For example, when Taq DNA polymerase is selected for PCR amplification, reference may be made to the reaction system as set forth in Table 1:
TABLE 2PCR systems table (50. Mu.L)
The reaction conditions for PCR were: firstly, performing pre-denaturation at 94 ℃ for 5min, then performing a circulation procedure, wherein each circulation procedure comprises denaturation at 94 ℃ for 30s, annealing at 55 ℃ for 30s, extension at 72 ℃ for 1min for 30s, performing 30 circulation, and finally performing heat preservation at 72 ℃ for 7min.
According to one embodiment of the present application, the content and quality of DNA fragments obtained by PCR using the extracted genomic DNA as a template with 16S universal primers were detected using agarose gel electrophoresis. FIG. 1B is an agarose gel electrophoresis of a DNA fragment obtained from a bacterial genomic DNA extracted as a template after PCR with 16S universal primers according to one embodiment of the present application. As shown in FIG. 1B, 3 groups of experiments all obtain PCR products with single bands and concentration meeting the requirement of subsequent sequencing identification.
2. And (5) identifying strain types. According to one embodiment of the present application, the PCR products obtained from the above experiments may be subjected to a sequencing process. Comparing the obtained sequencing result with the existing strain sequence to finally obtain the species of the identified strain. For example, the result of sequencing is input to NCBI website (https:// www.ncbi.nlm.nih.gov) for blast to obtain the species of strain identified. FIG. 1C is a phylogenetic tree obtained by PCR amplification of the isolated strain using the 16S primer pair, and sequence alignment, identifying Serratia rubra (Serratia rubidaea), according to one embodiment of the present application. From the phylogenetic tree shown in FIG. 1C, the evolutionary relationship of Serratia rubra (Serratia rubidaea) and other bacterial species of the present application can be clearly seen.
Example 3: preparation of amino acid liquid fertilizer
1. Preparation of fish culture medium
The tuna utility waste is purchased from the market and crushed into homogenates by a meat grinder, and the tuna homogenates are prepared according to different proportions: the culture medium was prepared with water (1:1, 1:4,1:9, 1:19), and the prepared fish culture was sterilized at 121℃for 20min.
2. Fermentation of amino acid liquid fertilizer
Adding 1% of Serratia rubra strain into the sterilized fish culture medium, and fermenting at 30deg.C and 150rpm for 24 hr.
3. Determination of amino acid content before and after fermentation
And determining the fermentation effect of Serratia rubra by measuring the content change of amino acids in the fish culture medium before and after fermentation through liquid chromatography-mass spectrometry (LC-MS). Wherein the amino acid content before and after fermentation is shown in Table 3.
TABLE 3 amino acid content Table before and after fermentation
Example 4: seed germination experiments
1. Selecting vegetable heart seeds with relatively consistent size and plump grains and without injury of diseases and insects, soaking the vegetable heart seeds in 75% ethanol for 5min, and then cleaning the vegetable heart seeds with ultrapure water.
2. The concentration of the amino acid liquid fertilizer is set to be 50, 100, 150, 200, 300 and 400mg/L in six concentration gradients, the amino acid liquid fertilizer purchased in the market is used as a positive control, and clear water is used as a negative control. Each group of 35 seeds are uniformly sown in a flat plate with the diameter of 10cm and laid with filter paper sheets infiltrated by each treatment liquid, and the seeds are placed in an illumination incubator for 12h of illumination culture every day. Three replicates were set for each treatment.
3. And (3) adding a plurality of drops of liquid which are treated correspondingly into the flat plate every day, recording the germination condition of the seeds every 12 hours, recording the root and bud length of the seeds after germination for three days, and determining the influence of different treatments on the germination process of the seeds.
FIG. 2 is an effect on germination and growth of cabbage seeds after fermentation of fish utility waste by Serratia rubra according to one embodiment of the present application. Fig. 2A and 2B are both results after 3 days of germination. Wherein the vertical axis in FIG. 2A is shown in order of soaking seed treatment with clear water, with 50mg/L of the amino acid fertilizer of the present application, with 100mg/L of the amino acid fertilizer of the present application, with 150mg/L of the amino acid fertilizer of the present application, with 200mg/L of the amino acid fertilizer of the present application, and with commercially available amino acid fertilizers purchased on the market; the horizontal axis represents 10 repetitions of each treatment. In fig. 2B, the ordinate indicates the root length and bud length after germination of the seeds, and the abscissa indicates the above-mentioned fresh water, amino acid fertilizers of different concentrations and commercial liquid fertilizers for seed treatment. Table 4 shows seed germination rates over different time periods. As is obvious from the combination of fig. 2A-2B and table 4, the amino acid liquid fertilizer produced by fermentation can promote germination of vegetable cores, has the same physiological function as the amino acid liquid fertilizer purchased in the market, and shows that the fermentation liquid can replace the amino acid liquid fertilizer in the market to a certain extent. Further, the growth quality of the amino acid liquid fertilizer treated seeds produced by fermentation in the application is better than that of the amino acid liquid fertilizer treated seeds obtained by water treatment and market purchase no matter in root elongation or bud length.
TABLE 4 seed germination rate
The serratia rubra disclosed by the invention is separated from the soil of the mangrove forest in Hainan, and the serratia rubra is used for fermenting the fish practical waste to prepare the amino acid liquid fertilizer, so that the germination rate of seeds is obviously improved in a seed soaking mode. Not only realizes the reutilization of the waste fish protein resource, but also can promote the germination of seeds.
The above embodiments are provided for illustrating the present invention and not for limiting the present invention, and various changes and modifications may be made by one skilled in the relevant art without departing from the scope of the present invention, therefore, all equivalent technical solutions shall fall within the scope of the present disclosure.
Claims (10)
1. The application of Serratia rubra (Serratia rubidaea) in preparing organic fertilizer is characterized in that Serratia rubra is preserved in China Center for Type Culture Collection (CCTCC) in Wuhan City of Hubei province in 2022, 7 months and 6 days, and the preservation number is CCTCC M20221042.
2. An organic fertilizer comprising: serratia rubra and medium for decomposing the same; or comprises fermentation liquor or fermentation liquor clear liquor obtained after the medium is decomposed by Serratia rubra; wherein, the Serratia rubra is preserved in China Center for Type Culture Collection (CCTCC) of Wuhan City of Hubei province in 2022, 7 months and 6 days, and the preservation number is CCTCC M20221042.
3. The organic fertilizer of claim 2, wherein the medium is a protein-rich animal utility waste including, but not limited to, fish utility waste, shrimp utility waste, crab utility waste, or conch utility waste.
4. The organic fertilizer of claim 2, wherein the medium is tuna utility waste.
5. The organic fertilizer of claim 2, which is an amino acid fertilizer.
6. The organic fertilizer of claim 2, which is an amino acid liquid fertilizer.
7. A method for preparing the organic fertilizer according to any one of claims 2 to 6, comprising:
providing a medium for decomposition of Serratia;
sterilizing the medium;
inoculating Serratia rubra to the sterilized medium; and
fermenting the serratia rubra;
wherein the fermentation temperature is 25-30deg.C, or 30-35deg.C; the fermentation time is 12-16 hours.
8. The application of the organic fertilizer prepared by Serratia rubra (Serratia rubidaea) in improving the germination rate of plant seeds and promoting plant growth is provided, wherein the Serratia rubra is preserved in China Center for Type Culture Collection (CCTCC) in Wuhan City of Hubei province in the period of 7 months and 6 days of 2022, and the preservation number is CCTCC M20221042.
9. A method of increasing germination of plant seeds comprising:
obtaining the organic fertilizer of any one of claims 2-6; and
the organic fertilizer is mixed with plant seeds, a culture medium for culturing plants or sown soil.
10. A method of promoting plant growth comprising:
obtaining the organic fertilizer of any one of claims 2-6; and
the organic fertilizer is sprayed on the leaf surfaces of plants or applied to the soil where plants grow.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| KR100825639B1 (en) * | 2007-03-16 | 2008-04-25 | 장창순 | The preparation method of liquid fertilizer containing the amino acid from the high concentration organic wastewater as raw materials |
| CN104371951A (en) * | 2014-09-15 | 2015-02-25 | 河北省科学院生物研究所 | Serratia sp. A5 and uses thereof |
| CN110564644A (en) * | 2019-09-17 | 2019-12-13 | 山东农业大学 | Serratia atropurpurea with growth promoting effect and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100825639B1 (en) * | 2007-03-16 | 2008-04-25 | 장창순 | The preparation method of liquid fertilizer containing the amino acid from the high concentration organic wastewater as raw materials |
| CN104371951A (en) * | 2014-09-15 | 2015-02-25 | 河北省科学院生物研究所 | Serratia sp. A5 and uses thereof |
| CN110564644A (en) * | 2019-09-17 | 2019-12-13 | 山东农业大学 | Serratia atropurpurea with growth promoting effect and application thereof |
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