CN115636704A - Compound composition of stagnant corynebacterium fermentation broth, preparation method and application thereof - Google Patents
Compound composition of stagnant corynebacterium fermentation broth, preparation method and application thereof Download PDFInfo
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- CN115636704A CN115636704A CN202211287052.2A CN202211287052A CN115636704A CN 115636704 A CN115636704 A CN 115636704A CN 202211287052 A CN202211287052 A CN 202211287052A CN 115636704 A CN115636704 A CN 115636704A
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Abstract
The invention discloses a compound composition of corynebacterium parvum fermentation liquor and a preparation method thereof, and simultaneously provides application of the compound composition in the aspects of improving the yield of pepper and promoting the growth of pepper plants. The corynebacterium parvum strain adopted by the invention is separated from natural soil in a natural environment, and the corynebacterium parvum fermentation liquor compound composition obtained by compounding the corynebacterium parvum strain with other compound substances including amino acid can induce plants to generate growth hormone, promote the roots of pepper plants to generate a large number of adventitious roots, ensure that the pepper plants grow rapidly and grow in a large amount, and improve the nutrient absorption efficiency and the growth and development capacity of the pepper.
Description
Technical Field
The invention belongs to the field of plant growth regulation, and relates to a compound composition of corynebacterium parvum fermentation broth and application of the compound composition in the aspect of improving the yield of capsicum.
Background
The pepper has wide planting adaptability, long industrial chain and rich nutritive value, and is not only an important fresh vegetable, but also a raw material of a traditional pepper seasoning product. At present, in pepper cultivation, under the conditions of weak plant growth vigor or limitation of natural environmental factors, and the like, pepper is often subjected to phenomena of growth retardation, resistance reduction, flower formation rate and fruit setting rate reduction, even flower and fruit dropping and the like, with the increase of successive cropping years, pepper diseases and insect pests are aggravated, yield is reduced, quality is deteriorated, pepper yield reduction is caused by 20-50%, and even the yield is up to 70% in serious land.
In order to ensure continuous production and income increase of the peppers, people adopt various prevention and control means. Chemical prevention and control are the main means adopted at present, chemical pesticides are widely applied to prevention and control of pepper diseases and insect pests, but excessive use of the pesticide can cause drug resistance of a prevention and control object, destroy a soil microbial community, cause problems of agricultural product residue, standard exceeding and the like, seriously affect safety of people and livestock, and even destroy the balance of a natural agricultural ecosystem. Biological control is widely regarded by various countries in the world due to the characteristics of high efficiency, no pollution, low cost and the like, wherein the microbial agent can improve the physicochemical and biological shapes of the micro-ecological environment of the rhizosphere of crops, promote the growth of the root system and enhance the resistance of the crops.
But the crop universality of the microbial inoculum is poor, the effect of a single microbial inoculum is not obvious enough, the culture and production of the microbial compound microbial inoculum are difficult, the adaptation environment of different microbial inocula is greatly different, and the requirement on the use environment is high.
The corynebacterium stagnatinum is a naturally-occurring wild corynebacterium, has the capability of directly converting a plurality of purine derivatives to generate corresponding nucleotides, and various auxotrophic mutants thereof can be used for synthesizing or accumulating various nucleotide derivatives in a segmented way, 5' -inosinic acid, adenylic acid succinate synthetase, purine nucleotides and other substances are developed by CJ first sugar manufacturing company, are directly or indirectly used as raw materials of food additives, and have a good degradation effect on 2,4-dichlorophen in polluted water bodies and soil environments. More importantly, the patent CN202010069133.X improves a strain of corynebacterium parvum, a screening method and application thereof, discloses that the corynebacterium parvum has the capability of producing indoleacetic acid, can also increase the fertilizer efficiency of compost products and promote the growth of plants, and also has the capability of promoting the germination of Chinese cabbage seeds, but when the corynebacterium parvum is used, the effect of a pure corynebacterium parvum fermentation broth is limited, the stability is poor, and the influence on different pepper strains cultivated at the same time is not completely the same.
Disclosure of Invention
In order to solve one of the problems, the invention provides a compound composition of Corynebacterium parvum (Corynebacterium parvum) fermentation liquor, which is compounded with raw materials such as compound amino acid and the like, so that the action stability of a Corynebacterium parvum microbial inoculum on pepper is improved, the yield of a single pepper plant can be generally improved, and compared with a Corynebacterium parvum microbial inoculum which is only used for standing or an independent amino acid compound fertilizer, the effect is more obvious, and a good synergistic effect is shown.
In order to achieve the above object, in one aspect, the invention provides a compound composition of corynebacterium parvum fermentation broth, wherein the compound composition comprises corynebacterium parvum bacterial liquid, amino acid, and one or two of humic acid and potassium-containing substance.
Specifically, the number of effective viable bacteria in the finished product of the compound composition is more than or equal to 5 multiplied by 10 8 CFU/ml。
Specifically, the amino acid is compound amino acid powder containing tryptophan, arginine, glutamic acid, glycine, lysine, alanine and phenylalanine.
Specifically, the potassium-containing substance is monopotassium phosphate, and the humic acid is mineral potassium fulvate.
More specifically, in the compound composition, the mass ratio of the corynebacterium parvum liquid to the other composite substances including the amino acid is 1000-10.
More specifically, the other compound substances including amino acid in the invention are tryptophan, arginine, glutamic acid, glycine, lysine, alanine, phenylalanine, monopotassium phosphate and mineral potassium fulvate. Wherein, the mass ratio of tryptophan, arginine, glutamic acid, glycine, lysine, alanine, phenylalanine, potassium dihydrogen phosphate and mineral potassium fulvate is 2.
In another aspect, the present invention provides a method for preparing a complex composition of Corynebacterium parvum (Corynebacterium stauntonii) fermentation broth, specifically comprising:
1. preparation of a Strain of Corynebacterium parvum
Adopting liquid culture method, using Bacillus arrestus strain as zymocyte, fermenting until the indoleacetic acid content is more than or equal to 4000ppm and the number of effective viable bacteria contained in the zymocyte is more than or equal to 1.5 × 10 9 CFU/mL is ready for use.
2. Preparation of mixed prepared microbial inoculum compound composition
And adding a compound substance comprising various amino acids into the bacterial liquid prepared in the step, and uniformly mixing.
In a third aspect, the invention also provides a method for improving the yield of the peppers, which is to irrigate bacterial manure or bacterial manure diluent containing corynebacterium parvum bacterial liquid and amino acid to the pepper root soil in the transplanting period, the seedling period, the bud period and the fruiting period of the peppers respectively.
Furthermore, the compound composition prepared by the steps is applied to pepper cultivation, and the composition is diluted by 500 times and poured into pepper root soil respectively in the transplanting period, the seedling period, the bud period and the fruiting period of peppers.
In a fourth aspect, the compound composition of the fermentation broth of Corynebacterium parvum (Corynebacterium staenitis) provided by the invention is applied to pepper cultivation, and test results show that the growth and development conditions, the root growth condition, the single fruit weight and the yield of pepper plants treated by the compound composition are obviously superior to those of single agents and clear water contrast.
Through the technical scheme, the compound composition of the fermentation broth of the corynebacterium parvum provided by the invention has at least the following beneficial effects or advantages:
1. the compound composition can promote the growth and development of pepper plants, has a promotion effect obviously superior to that of a single agent, and can improve the plant height growth amount by more than 5% and the stem thickness growth amount by more than 3% compared with the single use of corynebacterium parvum fermentation broth.
2. The compound composition disclosed by the invention can promote the growth of pepper root systems.
3. The compounded composition can increase the weight of pepper fruits.
4. The compound composition can increase the yield of the pepper, has a yield increase effect obviously superior to that of a single dose, and can increase the weight of a single red pepper and a single dry pepper by more than 20 percent and increase the weight of a single green pepper by nearly 20 percent compared with the weight of a single red pepper and a single dry pepper which are fermented by Corynebacterium sp (Corynebacterium staphylium) alone.
Drawings
FIG. 1 is a diagram showing the colony shape and color of Corynebacterium parvum ZWCY102A in the medium of example 1 of the present invention.
FIG. 2 is a comparison of the growth of pepper plants after treatment 1 and treatment 2 of example 5 of the present invention.
Fig. 3 is a comparison of pepper yields after treatment 1 and treatment 3 of example 5 of the present invention.
Detailed Description
The following examples are given to illustrate the technical aspects of the present invention, but the present invention is not limited to the following examples.
The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by manufacturers, and are conventional products available on the market.
Example 1
This example describes the screening, isolation and cultivation of the strain ZWCY102A used in the present invention, ZWCY102A being the number of the internal experiment of Corynebacterium parvum used in the present invention.
In a test base used by Vechu rain plant Nutrition Co., ltd, in Henan province of the Yana of the three-gate gorges of Henan, the same test field is sampled from plot soil which has better growth vigor, lighter soil hardening compared with other plots and relatively higher crop yield, and dominant microbial strains are screened and cultured.
The specific screening method adopts a dilution plate method, after a soil sample collected from soil of a test base is dissolved with sterile water indoors, supernatant is sequentially diluted and coated on a plate, single bacteria are selected for plate lineation purification, and primary screening and secondary screening are gradually carried out to select dominant strains. In the screening process of the strain, a beef extract peptone culture medium is adopted for culture at 30 ℃, and the growth condition after 34h of culture is shown in figure 1.
The preparation method of the NA plate culture medium comprises the following steps: dissolving 3g of beef extract, 10g of peptone and 5g of NaCl, adding 18g of agar powder, sterilizing at 121 ℃ for 20min, pouring 10-15ml of culture medium into each culture dish, cooling and solidifying to obtain the NA plate culture medium of the strain to be inoculated.
On the basis of screening the obtained strains, further fermenting the strains to prepare a fermentation solution for application.
The specific fermentation process is as follows: a single colony of the Corynebacterium stasis ZWCY102A preserved and cultured on a solid plate is picked up and inoculated into a seed culture medium in a 500ml triangular flask with the liquid loading volume of 100ml, and the colony is subjected to shaking culture for 34h on a constant temperature shaking table with the temperature of 30 ℃ and the rotating speed of 200r/min to serve as a seed liquid.
The culture medium of the seed liquid is prepared by adding 10g of peptone, 5g of sodium chloride, 3g of beef extract powder and 0.5g/L of tryptophan into 950ml of deionized water, and shaking until the solute is completely dissolved; then adjusting the pH value to 7.0 by using 5mol/L NaOH, and fixing the volume to 1L by using deionized water; finally sterilizing at 121 deg.C for 20min.
Then transferring the seed liquid after shake flask culture into a fermentation culture medium according to the inoculation proportion of 2.5 percent of volume ratio, culturing the seed liquid at the temperature of 30 ℃ and the ventilation ratio of 1:1 under the aerobic condition of 200r/min until the yield of the indoleacetic acid is more than or equal to 4000ppm and the number of effective viable bacteria in the liquid is more than or equal to 1.5 multiplied by 10 9 CFU/mL is ready for use.
The fermentation medium contains the following components in each kilogram of the medium: 10g of tryptone, 5g of yeast extract, 10g of NaCl10g, 2g of tryptophan and 1L of deionized water.
Example 2
This example describes a preparation method of a complex composition of a fermentation broth of corynebacterium parvum, which specifically includes the following steps:
(1) The strain of Corynebacterium parvum ZWCY102A cultured in example 1 was collected.
(2) Mixed preparation of microbial inoculum compound composition
And (2) adding a composite substance including various amino acids into the bacterial liquid prepared in the step (1), uniformly mixing, and reserving for later use.
Example 3
The compound composition of the corynebacterium parvum fermentation liquor obtained by the preparation method of the embodiment 2 comprises corynebacterium parvum bacterial liquid and a plurality of amino acids, and the number of effective viable bacteria in a finished product of the compound composition is more than or equal to 5 multiplied by 10 8 CFU/ml。
In this embodiment, the plurality of amino acids are composite amino acid powder containing tryptophan, arginine, glutamic acid, glycine, lysine, alanine, and phenylalanine.
In this example, the potassium-containing substance was potassium dihydrogen phosphate, and humic acid was mineral potassium fulvate.
In the embodiment, the mass ratio of tryptophan, arginine, glutamic acid, glycine, lysine, alanine, phenylalanine, potassium dihydrogen phosphate and mineral potassium fulvate is 2. And uniformly mixing the tryptophan, the arginine, the glutamic acid, the glycine, the lysine, the alanine, the phenylalanine, the monopotassium phosphate and the mineral source potassium fulvate in the proportion for later use.
The following specifications of reagents were obtained according to the ratio of the bacteria liquid of the dead corynebacterium species to the complex substance in the examples: (1) 1000ml of stagnant Corynebacterium ZWCY102A bacterial liquid and 10g of compound substance. (2) 1000ml of stagnant Corynebacterium ZWCY102A bacterial liquid and 1g of compound substance. (3) 1000ml of stagnant corynebacterium ZWCY102A bacterial liquid and 100g of compound substance.
Example 4
This example describes the method of applying the corynebacterium parvum complex composition of the present invention to pepper planting.
Planting place: the planting place is located in test field of capsicum in Shaanzhou region of Sanxia city, henan province
Planting the variety: selecting new generation Chinese red pepper variety with planting density of 5 plants/m 2 ；
Measuring the average content of organic matters in the soil of the planting land mass to be 10g/kg, the alkaline hydrolysis nitrogen to be 33mg/kg, the quick-acting phosphorus to be 6.5mg/kg, the quick-acting potassium to be 98.5mg/kg and the total amount of microorganisms to be 5.1 multiplied by 10 in the soil environment before planting 5 /g。
In the embodiment, a compound composition of 1000ml of corynebacterium parvum ZWCY102A bacterial liquid and 10g of corynebacterium parvum fermentation liquid of a composite substance is diluted by 500 times and is respectively irrigated in soil at the root of the capsicum in the transplanting period, the seedling period, the bud period and the fruiting period of the capsicum.
The test result of the embodiment shows that the plant growth hormone can be induced to produce the growth hormone, the related physiological and biochemical characteristics and the genetic performance of the plant are changed, the root generates a large amount of adventitious roots, the plant grows rapidly, the growth quantity is increased, and therefore the nutrient absorption efficiency and the growth and development capacity of the plant are improved, and the pepper growth promoting effect is achieved.
Example 5
A comparative experiment is carried out simultaneously in the same planting environment as in example 4, and the specific test steps are as follows:
1. selecting strong pepper seedlings, processing 45 seedlings each, and dividing into 3 cells, wherein the specific processing groups are shown in table 3-1, and each cell is 3m 2 For one-time repetition, 2 peppers are left among all cells to serve as a protection line. The blank group is also 45, and 2 peppers are left between the cells as a protection row.
2. And (4) irrigating 1000mL of each seedling in stages according to the growth period of the pepper on a soil plough layer around the pepper, and performing normal water and fertilizer management until the pepper is harvested.
TABLE 3-1 test treatment groups
In particular, the composite material in treatment 1 in table 3-1 was the same as the composite material in treatment 3 in terms of composition and preparation method.
The embodiment exerts the influence on each stage of the pepper growth, the specific implementation time is shown in the table 3-2, and the growth condition of the pepper after the influence of each stage is recorded and analyzed.
TABLE 3-2 test procedure
After the planting is finished, the growth condition of the pepper root system is observed, and the result shows that the pepper plants treated by the strain fermentation liquor 1 and the strain fermentation liquor 2 have developed underground root systems which are superior to those of a control group which does not adopt bacterial liquid. Statistics were made on the yield of pepper in each test plot and the results are shown in tables 3-3.
TABLE 3-3 growth of peppers under different treatments
The growth conditions of the pepper plants of the treatment groups 1 and 2 are shown in the specification and the attached figure 2, the yield of the pepper of the treatment groups 1 and 3 is compared with that of the pepper of the treatment group 3 and the attached figure 3, and the table 3-3 is combined with the attached figure 2 and the attached figure 3, so that the single agent of the corynebacterium parvum ZWCT102A bacterial liquid prepared in the embodiment 2 of the invention shows a certain yield increase effect no matter the single compound substance or the single agent is used. However, other parallel experiments similar to the embodiment prove that the growth promotion effect and the yield increase effect of the capsicum are not obviously increased by singly and continuously increasing the single agent of the corynebacterium parvum ZWCT102A bacterial liquid or singly and continuously increasing the dosage of the compound substance, but after the two are compounded and applied, the two further show a certain synergistic effect, the yield of the capsicum can be further increased, a better application prospect is shown, and the compound composition obtained in different proportions in the embodiment 3 has approximate yield increase effect.
In conclusion, the compound composition of the corynebacterium parvum fermentation broth provided by the invention can effectively promote the rooting of pepper plants, promote the growth and development of the pepper plants, and simultaneously can effectively increase the weight and yield of pepper fruits.
As described above, the present invention can be preferably implemented, and the above-mentioned embodiments only describe the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various changes and modifications of the technical solution of the present invention made by those skilled in the art without departing from the design spirit of the present invention shall fall within the protection scope defined by the present invention.
Claims (9)
1. The compound composition of the fermentation liquor of the corynebacterium parvum is characterized by comprising the components of corynebacterium parvum bacterial liquid, amino acid, humic acid and/or potassium-containing substances.
2. The compound composition as claimed in claim 1, wherein the number of active bacteria in the finished compound composition is not less than 5 x 10 8 CFU/ml。
3. The compound composition as claimed in claim 1, wherein the amino acid is compound amino acid powder containing tryptophan, arginine, glutamic acid, glycine, lysine, alanine and phenylalanine.
4. The compound composition as claimed in claim 1, wherein the potassium-containing substance is potassium dihydrogen phosphate, and the humic acid is mineral potassium fulvate.
5. The compound composition according to claim 1, wherein the mass ratio of the corynebacterium parvum bacterial liquid to the other composite substances including the amino acid in the compound composition is 1000-10.
6. The compound composition according to claim 5, wherein the other compound substances including amino acids are tryptophan, arginine, glutamic acid, glycine, lysine, alanine, phenylalanine, potassium dihydrogen phosphate and mineral potassium fulvate, and the mass ratio of tryptophan, arginine, glutamic acid, glycine, lysine, alanine, phenylalanine, potassium dihydrogen phosphate and mineral potassium fulvate is 2.
7. The preparation method of the compound composition as claimed in any one of claims 1 to 6, which comprises fermenting in a liquid culture mode with a strain of Corynebacterium parvum as a fermentation strain until the indoleacetic acid content is more than or equal to 4000ppm and the effective viable count is more than or equal to 1.5 x 10 9 CFU/mL zymogen liquid; adding a composite substance including various amino acids into the bacterial liquid, and uniformly mixing.
8. A method for improving the yield of hot peppers is characterized in that bacterial manure or bacterial manure diluent containing corynebacterium parvum bacterial liquid and amino acid is poured into the soil at the root of hot peppers in the transplanting period, the seedling period, the bud period and the fruiting period respectively.
9. The use of the compounded composition of any one of claims 1-6 in pepper cultivation.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103553828A (en) * | 2013-11-12 | 2014-02-05 | 天津北洋百川生物技术有限公司 | Liquid compound biological fertilizer with long shelf life as well as preparation method and application thereof |
| CN109608273A (en) * | 2019-02-15 | 2019-04-12 | 王刚 | Long-acting organic mineralization activity fertilizer of one kind and preparation method thereof |
| CN111363694A (en) * | 2020-01-21 | 2020-07-03 | 中国科学院城市环境研究所 | Stagnant corynebacterium, screening method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103553828A (en) * | 2013-11-12 | 2014-02-05 | 天津北洋百川生物技术有限公司 | Liquid compound biological fertilizer with long shelf life as well as preparation method and application thereof |
| CN109608273A (en) * | 2019-02-15 | 2019-04-12 | 王刚 | Long-acting organic mineralization activity fertilizer of one kind and preparation method thereof |
| CN111363694A (en) * | 2020-01-21 | 2020-07-03 | 中国科学院城市环境研究所 | Stagnant corynebacterium, screening method and application thereof |
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