Chitin biological peptide preparation, preparation method thereof, functional fertilizer and application
Through the synergistic effect of chitin biopeptide preparation and Bacillus melon, combined with humic acid treatment, the problems of soil nutrient loss and root-knot nematode disease in the Hami melon planting area in Hainan Province were solved, the yield and quality of Hami melon were improved, and the root prevention and control was achieved efficiently The effect of nematode.
Patent Information
- Application Number
- CN202411442187.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-10-16
- Publication Date
- 2025-01-07
AI Technical Summary
Under the high temperature, high humidity and high fertilizer conditions in the Hami melon planting area in Hainan Province, carbon mineralization in the soil is fast turnover and the amount of organic matter accumulated is small, resulting in soil nutrient loss, low fertilizer utilization rate, and serious root knot nematode diseases, affecting the yield and quality of Hami melon.
Using chitin biopeptide preparation and Bacillus megaly, the chitin biopeptide fermentation broth and biopeptide fermentation broth are prepared by mixing chitin biopeptide fermentation broth, combining humic acid adsorption and drying to make functional fertilizers, which are used to improve the yield and solubleness of cantaloupe. The content of protein and sugar can reduce the content of organic acids and prevent and treat root knot nematodes.
The yield of cantaloupe was significantly increased by 54.07%, the content of soluble protein and soluble sugar increased by 30.88% and 29.56%, the content of organic acid was reduced by 67.42%, and the incidence rate of root knot nematodes was effectively prevented and treated, with the maximum prevention and control effect of up to 82.75%.
Smart Images
Figure BDA0005086631030000131 
Figure BDA0005086631030000141 
Figure BDA0005086631030000142
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of fertilizers, and more specifically, to a chitin biopeptide preparation and a preparation method thereof, a functional fertilizer and application thereof. Background Art
[0002] Hami melon is one of the important cash crops in my country. It is rich in nutrition and has high economic value. It has become an effective way for farmers to quickly increase their income and become rich. With its unique climate and light conditions, Hainan Province produces sweet and juicy Hami melons with delicate taste and good storage and transportation. It is widely popular and occupies 80% of the national winter Hami melon market. Hami melon is a high nutrient input crop system. Every 100 kg of fresh Hami melon needs to absorb 2.5-3.5 kg of N, 1.4-1.8 kg of P2O5, and 4.4-4.6 kg of K2O from the soil.
[0003] At present, there are many commercial fertilizers on the market, such as general-purpose biological organic fertilizers, microbial agents, organic-inorganic compound fertilizers and amino acid water-soluble fertilizers, but few are designed for the high temperature and high humidity soil environment of Hami melon cultivation in Hainan Province. Hainan Island is located between 108°37′ and 111°03′ east longitude and 18°10′ and 20°10′ north latitude, with an average annual temperature of 22 to 27°C, annual sunshine of 1750 to 2650 hours, and annual precipitation of 1000 mm to 2600 mm. The main Hami melon growing areas in Hainan Province are Sanya, Ledong, Dongfang and Lingshui. The soil involved in the growing areas is all sandy. Although this type of soil is loose and has good water and air permeability, under the high temperature, high humidity and high fertilizer conditions of intensive planting, the carbon mineralization turnover in the soil is fast, the accumulation of organic matter is small, and the ability to retain water and fertilizer is poor, which in turn causes soil nutrient loss, low fertilizer utilization rate, serious root-knot nematode disease and a decline in Hami melon yield and quality, which is not conducive to the sustainable development of the Hami melon planting system. Summary of the invention
[0004] In view of this, the object of the present invention is to provide a chitosan biopeptide preparation and a preparation method thereof, a functional fertilizer and an application thereof. The chitosan biopeptide preparation of the present invention can synergize with Bacillus megaterium to increase the yield of Hami melon, increase the content of soluble protein and soluble sugar in Hami melon, reduce the content of organic acid, and effectively prevent and control the incidence of root-knot nematodes.
[0005] In order to achieve the above object, the present invention adopts the following technical solutions:
[0006] A method for preparing a chitin biopeptide preparation, comprising:
[0007] The chitin fermentation liquid and the biopeptide fermentation liquid are mixed, and dried at 70 to 80° C. to obtain a chitin biopeptide preparation;
[0008] The preparation method of the chitosan fermentation liquid comprises:
[0009] Under the conditions of 50-55°C and 150-170r / min, black soldier fly shells, trichoderma viride and water are mixed to obtain chitosan fermentation liquid;
[0010] The preparation method of the biological peptide fermentation liquid comprises:
[0011] Under the conditions of 50-60°C and 150-170r / min, black soldier fly larvae and compound enzyme are mixed to obtain biological peptide fermentation liquid;
[0012] The complex enzyme comprises papain and lipase.
[0013] Trichoderma viride is a type of Trichoderma, which is widely distributed in nature and often grows on wood, seeds and plant residues. Trichoderma viride can produce a variety of biologically active enzymes, and it is one of the strains with the highest cellulase activity. The cellulase produced has a degrading effect on crops.
[0014] Preferably, the volume ratio of the chitin fermentation broth to the biopeptide fermentation broth is 1:(1-5), preferably 1:1.
[0015] Preferably, in the chitin bio-peptide preparation, the oligosaccharide content is 200-220 mg / g, and the total amount of free amino acids is 220-230 mg / g; among the free amino acids, the content of threonine, methionine, proline, alanine, and phenylalanine accounts for 80%-85% of the total amount of free amino acids; the ratio of threonine, methionine, proline, alanine, and phenylalanine is 1:(0.4-0.6):(1.1-1.3):(1.3-1.4):(1.9-2.1).
[0016] Preferably, the mass ratio of the black soldier fly shell, trichoderma viride and water is (0.5-5):(0.5-5):100, preferably (0.5-2):(1-3):100, and more preferably 1:1.5:100.
[0017] Preferably, the particle size of the black soldier fly shell is 300-500 mesh.
[0018] Preferably, the mixing of the black soldier fly shells, Trichoderma viride and water is carried out at 50-55° C., for 9-50 hours, at a mixing speed of 100-200 r / min.
[0019] Preferably, the content of chitosan oligosaccharide in the chitin fermentation broth is greater than 10 mg / ml.
[0020] Preferably, the content of free amino acids in the bio-peptide fermentation broth is greater than 14 mg / ml.
[0021] Preferably, the mass ratio of the black soldier fly larvae to the composite enzyme is 100:(0.05-0.2); in the composite enzyme, the mass ratio of papain to lipase is (1-2):1, preferably 2:1.
[0022] Preferably, the black soldier fly larvae and the complex enzyme are mixed at 50-60° C., for 9-15 hours, at a mixing speed of 100-200 r / min.
[0023] Preferably, the method for preparing the chitosan biopeptide preparation specifically comprises:
[0024] Chitosan fermentation liquid and biopeptide fermentation liquid are mixed to obtain a mixed liquid, and then the mixed liquid is adsorbed with humic acid, and then dried at 70-80°C to obtain a chitosan biopeptide preparation; the ratio of humic acid to the mixed liquid is 1:(10-15). After the mixed liquid is concentrated and dried with humic acid, it is made into a powder for subsequent use.
[0025] Preferably, the ratio of humic acid to the mixed solution is 1:15 (w / v).
[0026] Preferably, the drying time is 18 to 26 hours.
[0027] The invention also provides a chitin biopeptide preparation prepared by a method for preparing the chitin biopeptide preparation.
[0028] The present invention also provides a functional fertilizer containing a chitin biopeptide preparation, comprising a basic fertilizer and Bacillus megaterium;
[0029] In parts by mass, the basic fertilizer includes 30-50 parts of mature compost, 25-35 parts of earthworm castings, 20-30 parts of peat, 2-4 parts of chitin biopeptide preparation, and 1-3 parts of potassium humate;
[0030] The ratio of Bacillus megaterium to basic fertilizer is (10-20): 1 ml / g; the concentration of Bacillus megaterium is 1×10 8 ~3×10 8 cfu / ml.
[0031] Bacillus magaterium is a large spore-forming, gram-positive bacillus that moves with peritrichous flagella. It is an aerobic bacterium and a beneficial soil bacterium. In order to solve the problem of low available phosphorus content in the soil, Bacillus megaterium is used as a phosphate-solubilizing bacterium in the production of microbial fertilizers. With the deepening of research, the role of Bacillus megaterium has been further explored, and it has begun to be widely used in phosphorus and nitrogen fixation, biological control, animal husbandry, water purification, enzyme industry and other aspects.
[0032] Preferably, the concentration of Bacillus megaterium is 1×10 8 cfu / ml.
[0033] Preferably, the method for preparing the mature compost comprises:
[0034] The harvested Hami melon, cucumber, cherry tomatoes, whole plants of green radish and whole plants of morning glory are pre-crushed to a length of 1-3 cm, mixed with compost, and water is added to control the moisture content to 50%. The system is ventilated for 1 hour every day, the pile temperature is controlled to be continuously ≥55°C, the pile is turned every 2 to 3 days, and the compost is composted for 15 to 21 days to select the mature compost with a particle size of 3 to 5 mm.
[0035] Peat is a brown-black solid fuel that is gradually formed by the natural compression of plant residues over a long period of time. The main components of peat are carbon, hydrogen, oxygen, and a small amount of nitrogen, sulfur, phosphorus and other elements. It is formed by the continuous natural decomposition of residues including sawdust, bark, straw, wheat straw, mushroom straw, reeds and other weeds, dried vegetables, fruits and herbs.
[0036] Preferably, the organic matter content of the peat is 70%-80%, the humic acid content is 40%-50%, and the pH value is 5.0-5.5.
[0037] Preferably, the earthworm castings are the product of earthworms devouring livestock and poultry excrement and / or crop straw; and the storage time of the earthworm castings does not exceed 3 months.
[0038] Preferably, the basic fertilizer includes, by weight, 30 parts of mature compost, 33 parts of earthworm castings, 30 parts of peat, 4 parts of chitin biopeptide preparation, and 3 parts of potassium humate.
[0039] Preferably, the basic fertilizer includes 41 parts of mature compost, 25 parts of earthworm castings, 30 parts of peat, 3 parts of chitin biopeptide preparation, and 1 part of potassium humate, calculated by mass.
[0040] Preferably, the basic fertilizer includes 40 parts of mature compost, 30 parts of earthworm castings, 25 parts of peat, 3 parts of chitin biopeptide preparation, and 2 parts of potassium humate, calculated by mass.
[0041] Preferably, the basic fertilizer includes 50 parts of mature compost, 27 parts of earthworm castings, 20 parts of peat, 2 parts of chitin biopeptide preparation, and 1 part of potassium humate, calculated by mass.
[0042] Preferably, the basic fertilizer includes 34 parts of mature compost, 35 parts of earthworm castings, 25 parts of peat, 4 parts of chitin biopeptide preparation, and 2 parts of potassium humate, calculated by mass.
[0043] The present invention also provides the use of chitin biopeptide preparations and / or functional fertilizers containing the chitin biopeptide preparations in preventing and controlling root-knot nematodes.
[0044] Preferably, the chitosan biopeptide preparation and / or the functional fertilizer containing the chitosan biopeptide preparation can prevent and control root-knot nematodes of Hami melon.
[0045] The present invention also provides the use of chitin biopeptide preparations and / or functional fertilizers containing chitin biopeptide preparations in (1) to (4);
[0046] (1) Increase the yield of Hami melon;
[0047] (2) increasing the soluble protein content in Hami melon;
[0048] (3) increasing the soluble sugar content in Hami melon;
[0049] (4) Reduce the content of organic acids in Hami melon.
[0050] Compared with the prior art, the present invention has the following beneficial effects:
[0051] 1. The functional fertilizer of the present invention uses decomposed compost, earthworm castings and peat as main raw materials, has sufficient organic nutrients, and can effectively promote the growth of Hami melon seedlings.
[0052] 2. The chitosan biopeptide preparation of the present invention can act synergistically with Bacillus megaterium to increase the yield of Hami melon by 54.07%, increase the soluble protein and soluble sugar by 30.88% and 29.56% respectively, and reduce the organic acid content by 67.42%. It can effectively prevent and control the incidence of Hami melon root-knot nematodes, and its prevention and control effect can reach up to 82.75%.
[0053] 3. The free amino acids of the chitin biopeptide preparation of the present invention contain specific contents of threonine, methionine, proline, alanine and phenylalanine, which can effectively improve the synergistic effect between the chitin biopeptide preparation and Bacillus megaterium. DETAILED DESCRIPTION
[0054] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0055] In order to further illustrate the present invention, the following examples are used for detailed description. The raw materials used in the following examples of the present invention are all commercially available commodities; Bacillus megaterium and Bacillus subtilis used in the examples of the present invention are purchased from Hubei Qiming Bioengineering Co., Ltd.; Trichoderma viride is purchased from Guangxi Nongbao Bioengineering Co., Ltd. The peat used in the examples of the present invention has an organic matter content of 70%-80%, a humic acid content of 40%-50%, and a pH value of 5.0-5.5.
[0056] The preparation method of the mature compost used in the embodiment of the present invention includes:
[0057] The harvested cantaloupe, cucumber, cherry tomatoes, whole plants of green radish and whole plants of morning glory are pre-crushed to a length of 1-3 cm, mixed with compost, and water is added to control the moisture content to 50%. The system is ventilated for 1 hour every day, the pile temperature is controlled to be ≥55°C, the pile is turned every 2 days, and the compost is composted for 18 days to select the mature compost with a particle size of 3 to 5 mm.
[0058] The vermicompost used in the embodiment of the present invention is the product of earthworms eating rice straw, and the storage time is 3 months.
[0059] Example 1
[0060] The preparation method of the functional fertilizer containing the chitosan biopeptide preparation of this embodiment comprises the following steps:
[0061] The following materials were mixed in mass percentage: 30% of mature compost, 33% of earthworm castings, 30% of peat, 4% of chitin biopeptide preparation, and 3% of potassium humate; after mixing, Bacillus megaterium was inoculated into the system at an inoculation amount of 10 ml / g and a concentration of 10 8 cfu / ml.
[0062] In the functional fertilizer of this embodiment, the oligosaccharide content in the chitin biopeptide preparation is 200 mg / g, and the total amount of free amino acids is 220 mg / g; the content of threonine, methionine, proline, alanine, and phenylalanine in the free amino acids accounts for 80% of the total amount of free amino acids, and the ratio of threonine, methionine, proline, alanine, and phenylalanine is 1:0.4:1.1:1.3:1.9; the remaining free amino acids are: arginine 2.31%, histidine 0.82%, lysine 0.52%, tyrosine 1.43%, leucine 3.27%, isoleucine 2.66%, valine 3.08%, glycine 1.65%, glutamic acid 2.03%, serine 1.52%, and aspartic acid 0.71%; the preparation method of the chitin biopeptide preparation comprises the following steps:
[0063] (1) Weigh 10 g of black soldier fly shell powder crushed by a colloid mill and sieved through a 300-mesh sieve and 15 g of green Trichoderma, add them to 1 liter of distilled water, and stir evenly; culture at 50°C and 150 r / min for 46 hours, take a sample, and determine the chitosan oligosaccharide content in the solution to be greater than 10 mg / ml, thereby obtaining chitosan fermentation liquid.
[0064] (2) Select clean and disease-free black soldier fly larvae and crush them into a homogenate until the homogenate diameter is less than 100 nm; add papain and lipase into a complex enzyme in a mass ratio of 2:1, and add the complex enzyme to the homogenate. The amount of the complex enzyme added is 0.05 wt% of the mass of the homogenate. Stir the system evenly, oscillate at 150 r / min at 57° C. for 9 h, take a sample, and determine the free amino acid content in the solution to be greater than 14 mg / ml, thereby obtaining a biopeptide fermentation broth.
[0065] (3) The chitosan fermentation liquid obtained in step (1) and the biopeptide fermentation liquid obtained in step (2) are mixed uniformly at a ratio of 1:1 (v / v) to obtain a mixed liquid, and then the mixed liquid is adsorbed with humic acid (the ratio of humic acid to the mixed liquid is 1:15 (w / v)). After the sample is adsorbed, it enters a spray drying tower and is dried at 70° C. for 20 hours to obtain a chitosan biopeptide preparation.
[0066] Example 2
[0067] The preparation method of the functional fertilizer containing the chitosan biopeptide preparation of this embodiment comprises the following steps:
[0068] The following materials were mixed in mass percentage: 41% of mature compost, 25% of earthworm castings, 30% of peat, 3% of chitin biopeptide preparation, and 1% of potassium humate; after mixing, Bacillus megaterium was inoculated into the system at an inoculation amount of 15 ml / g and a concentration of 10 8 cfu / ml; the mature compost included cantaloupe, cucumber, cherry tomato, green radish and morning glory in a mass ratio of 5:2:3:4:1, with a particle size of 5mm.
[0069] In the functional fertilizer of this embodiment, the oligosaccharide content in the chitin biopeptide preparation is 210 mg / g, and the total amount of free amino acids is 225 mg / g; the content of threonine, methionine, proline, alanine, and phenylalanine in the free amino acids accounts for 83% of the total amount of free amino acids, and the ratio of threonine, methionine, proline, alanine, and phenylalanine is 1:0.5:1.2:1.35:2.0; the remaining free amino acids are: arginine 2.06%, histidine 0.61%, lysine 0.54%, tyrosine 1.26%, leucine 2.44%, isoleucine 2.58%, valine 2.54%, glycine 1.63%, glutamic acid 1.54%, serine 1.26%, and aspartic acid 0.54%; the preparation method of the chitin biopeptide preparation comprises the following steps:
[0070] (1) Weigh 10 g of black soldier fly shell powder crushed by a colloid mill and passed through a 300-mesh sieve and 15 g of green Trichoderma, add them to 1 liter of distilled water, and stir evenly; culture at 51°C and 160 r / min for 48 hours, take a sample, and determine the chitosan oligosaccharide content in the solution to be greater than 10 mg / ml, thereby obtaining chitosan fermentation liquid.
[0071] (2) Select clean and disease-free black soldier fly larvae and crush them into a homogenate until the diameter of the homogenate is less than 100 nm; add papain and lipase into a complex enzyme in a mass ratio of 2:1 to the homogenate, and add the complex enzyme to the homogenate in an amount of 0.10 wt% of the mass of the homogenate; stir the system evenly, oscillate at 160 r / min at 55° C. for 10 h, take a sample, and determine the free amino acid content in the solution to be greater than 14 mg / ml, thereby obtaining a biopeptide fermentation broth.
[0072] (3) The chitosan fermentation liquid obtained in step (1) and the biopeptide fermentation liquid obtained in step (2) are mixed evenly at a ratio of 1:1 (v / v), and then adsorbed with humic acid at a ratio of 1:15 (w / v). After the sample is adsorbed, it enters a spray drying tower and is dried at 70°C for 24 hours to obtain a chitosan biopeptide preparation.
[0073] Example 3
[0074] The preparation method of the functional fertilizer containing the chitosan biopeptide preparation of this embodiment comprises the following steps:
[0075] The following materials were mixed in mass percentage: 40% mature compost, 30% earthworm castings, 25% peat, 3% chitin biopeptide preparation, and 2% potassium humate; after mixing, Bacillus megaterium was inoculated into the system at an inoculation amount of 20 ml / g and a concentration of 10 8 cfu / ml; the mature compost included cantaloupe, cucumber, cherry tomato, green radish and morning glory in a mass ratio of 5:2:3:4:1, with a particle size of 5mm.
[0076] In the functional fertilizer of this embodiment, the oligosaccharide content in the chitin biopeptide preparation is 220 mg / g, and the total amount of free amino acids is 230 mg / g. The content of threonine, methionine, proline, alanine, and phenylalanine in the free amino acids accounts for 85% of the total amount of free amino acids, and the ratio of threonine, methionine, proline, alanine, and phenylalanine is 1:0.6:1.3:1.4:2.1; the remaining free amino acids are: arginine 2.06%, histidine 0.61%, lysine 0.54%, tyrosine 1.26%, leucine 2.44%, isoleucine 2.58%, valine 2.54%, glycine 1.63%, glutamic acid 1.54%, serine 1.26%, and aspartic acid 0.54%; the preparation method of the chitin biopeptide preparation includes the following steps:
[0077] (1) Weigh 10 g of black soldier fly shell powder crushed by a colloid mill and passed through a 300-mesh sieve and 15 g of green Trichoderma, add them to 1 liter of distilled water, and stir evenly; culture at 52°C and 170 r / min for 50 h, take a sample, and determine the chitosan oligosaccharide content in the solution to be greater than 10 mg / ml, thereby obtaining chitosan fermentation liquid.
[0078] (2) Select clean and disease-free black soldier fly larvae and crush them into a homogenate until the diameter of the homogenate is less than 100 nm; add papain and lipase into a complex enzyme in a mass ratio of 2:1 to the homogenate, and add the complex enzyme to the homogenate in an amount of 0.15 wt% of the mass of the homogenate; stir the system evenly, oscillate at 170 r / min for 11 h at 57° C., take a sample, and determine the free amino acid content in the solution to be greater than 14 mg / ml, thereby obtaining a biopeptide fermentation broth.
[0079] (3) The chitosan fermentation liquid and the biopeptide fermentation liquid obtained by fermentation were mixed evenly at a ratio of 1:1 (v / v), and then adsorbed with humic acid at a ratio of 1:15 (w / v). After the sample was adsorbed, it entered a spray drying tower and was dried at 70°C for 12 hours to obtain a chitosan biopeptide preparation.
[0080] Example 4
[0081] The preparation method of the functional fertilizer containing the chitosan biopeptide preparation of this embodiment comprises the following steps:
[0082] The following materials in percentage by mass were mixed: 50% mature compost, 27% earthworm castings, 20% peat, 2% chitin biopeptide preparation, and 1% potassium humate; after mixing, Bacillus megaterium was inoculated into the system at an inoculation amount of 10 ml / g and a concentration of 10 8 cfu / ml; the mature compost included cantaloupe, cucumber, cherry tomato, green radish and morning glory in a mass ratio of 5:2:3:4:1, with a particle size of 5mm.
[0083] In the functional fertilizer of this embodiment, the oligosaccharide content in the chitin biopeptide preparation is 200 mg / g, and the total amount of free amino acids is 230 mg / g. The content of threonine, methionine, proline, alanine, and phenylalanine in the free amino acids accounts for 80% of the total amount of free amino acids, and the ratio of threonine, methionine, proline, alanine, and phenylalanine is 1:0.6:1.1:1.3:2.0; the remaining free amino acids are: arginine 2.26%, histidine 0.72%, lysine 0.64%, tyrosine 1.58%, leucine 3.07%, isoleucine 2.61%, valine 3.08%, glycine 1.43%, glutamic acid 2.57%, serine 1.49%, and aspartic acid 0.55%; the preparation method of the chitin biopeptide preparation includes the following steps:
[0084] (1) Weigh 10 g of black soldier fly shell powder crushed by a colloid mill and sieved through a 300-mesh sieve and 15 g of green Trichoderma, add them to 1 liter of distilled water, and stir evenly; culture at 51°C and 160 r / min for 50 h, take a sample, and determine the chitosan oligosaccharide content in the solution to be greater than 10 mg / ml, thereby obtaining chitosan fermentation liquid.
[0085] (2) Select clean and disease-free black soldier fly larvae, crush them into a homogenate until the homogenate diameter is less than 100 nm; add papain and lipase in a mass ratio of 2:1 to form a composite enzyme, and add the composite enzyme to the homogenate, and the amount of the composite enzyme added is 0.12wt% of the mass of the homogenate. Stir the system evenly, oscillate at 170r / min for 10h at 56°C, take a sample, and determine the free amino acid content in the solution to be greater than 14mg / ml, and obtain a biopeptide fermentation liquid.
[0086] (3) The chitosan fermentation liquid obtained in step (1) and the biopeptide fermentation liquid obtained in step (2) are mixed evenly at a ratio of 1:1 (v / v), and then adsorbed with humic acid at a ratio of 1:15 (w / v). After the sample is adsorbed, it enters a spray drying tower and is dried at 70°C for 18 hours to obtain a chitosan biopeptide preparation.
[0087] Example 5
[0088] The preparation method of the functional fertilizer containing the chitosan biopeptide preparation of this embodiment comprises the following steps:
[0089] The following materials were mixed in percentage by mass: 34% of mature compost, 35% of earthworm castings, 25% of peat, 4% of chitin biopeptide preparation, and 2% of potassium humate; after mixing, Bacillus megaterium was inoculated into the system at an inoculation amount of 20 ml / g and a concentration of 10 8 cfu / ml; the mature compost included cantaloupe, cucumber, cherry tomato, green radish and morning glory in a mass ratio of 5:2:3:4:1, with a particle size of 5mm.
[0090] In the functional fertilizer of this embodiment, the oligosaccharide content in the chitin biopeptide preparation is 210 mg / g, and the total amount of free amino acids is 230 mg / g. The content of threonine, methionine, proline, alanine, and phenylalanine in the free amino acids accounts for 82% of the total amount of free amino acids, and the ratio of threonine, methionine, proline, alanine, and phenylalanine is 1:0.5:1.2:1.3:2.1; the remaining free amino acids are: arginine 2.63%, histidine 0.70%, lysine 0.54%, tyrosine 1.34%, leucine 2.23%, isoleucine 2.87%, valine 2.63%, glycine 1.49%, glutamic acid 1.55%, serine 1.37%, and aspartic acid 0.65%; the preparation method of the chitin biopeptide preparation includes the following steps:
[0091] (1) Weigh 10 g of black soldier fly shell powder crushed by a colloid mill and sieved through a 300-mesh sieve and 15 g of green Trichoderma, add them to 1 liter of distilled water, and stir evenly; culture at 52°C and 170 r / min for 46 hours, take a sample, and determine the chitosan oligosaccharide content in the solution to be greater than 10 mg / ml, thereby obtaining chitosan fermentation liquid.
[0092] (2) Select clean and disease-free black soldier fly larvae and crush them into a homogenate until the diameter of the homogenate is less than 100 nm; add papain and lipase into a complex enzyme in a mass ratio of 2:1 to the homogenate, and add the complex enzyme to the homogenate in an amount of 0.15 wt% of the mass of the homogenate; stir the system evenly, oscillate at 150 r / min for 11 h at 55° C., take a sample, and determine the free amino acid content in the solution to be greater than 14 mg / ml, thereby obtaining a biopeptide fermentation broth.
[0093] (3) The chitosan fermentation liquid obtained in step (1) and the biopeptide fermentation liquid obtained in step (2) are mixed evenly at a ratio of 1:1 (v / v), and then adsorbed with humic acid at a ratio of 1:15 (w / v). After the sample is adsorbed, it enters a spray drying tower and is dried at 70°C for 24 hours to obtain a chitosan biopeptide preparation.
[0094] Test Example 1
[0095] In order to further illustrate the promotion and application value of the functional fertilizer of the present invention, the functional fertilizer prepared in the following steps was tested on Hami melon for fertilizer effect and root-knot nematode control.
[0096] Planting site conditions: Coastal area, with an average annual sunshine of 2545.4 hours, annual rainfall of 769.4 mm, and an average annual temperature of 24-25°C. The planting site is sandy, with loose soil, good drainage, and a pH value between 5.7-7.6.
[0097] Test materials: Hami melon, Duzhimi No. 2.
[0098] Experimental design: The experimental design includes 50 treatments, 3 replicates per treatment, and 8 m per plot. 2 , randomized block arrangement. The specific experimental design is as follows.
[0099] Treatment 1: functional fertilizer prepared in Example 1;
[0100] Treatment 2: functional fertilizer prepared in Example 2;
[0101] Treatment 3: functional fertilizer prepared in Example 3;
[0102] Treatment 4: functional fertilizer prepared in Example 4;
[0103] Treatment 5: functional fertilizer prepared in Example 5;
[0104] Control 1-1: The chitin biopeptide preparation in the functional fertilizer of Example 1 was omitted;
[0105] Control 1-2: The chitin biopeptide preparation in the functional fertilizer of Example 2 was omitted;
[0106] Control 1-3: The chitin biopeptide preparation in the functional fertilizer of Example 3 was omitted;
[0107] Control 1-4: The chitin biopeptide preparation in the functional fertilizer of Example 4 was omitted;
[0108] Control 1-5: The chitin biopeptide preparation in the functional fertilizer of Example 5 was omitted;
[0109] Control 2-1: The Bacillus megaterium in the functional fertilizer of Example 1 was omitted;
[0110] Control 2-2: The Bacillus megaterium in the functional fertilizer of Example 2 was omitted;
[0111] Control 2-3: The Bacillus megaterium in the functional fertilizer of Example 3 was omitted;
[0112] Control 2-4: The Bacillus megaterium in the functional fertilizer of Example 4 was omitted;
[0113] Control 2-5: The Bacillus megaterium in the functional fertilizer of Example 5 was omitted;
[0114] Control 3-1: The Bacillus megaterium in Example 1 was replaced with sterilized Bacillus megaterium;
[0115] Control 3-2: The Bacillus megaterium in Example 2 was replaced with sterilized Bacillus megaterium;
[0116] Control 3-3: The Bacillus megaterium in Example 3 was replaced with sterilized Bacillus megaterium;
[0117] Control 3-4: The Bacillus megaterium in Example 4 was replaced with sterilized Bacillus megaterium;
[0118] Control 3-5: The Bacillus megaterium in Example 5 was replaced with sterilized Bacillus megaterium;
[0119] Control 4-1: The chitin biopeptide preparation and Bacillus megaterium in the functional fertilizer of Example 1 were omitted;
[0120] Control 4-2: The chitin biopeptide preparation and Bacillus megaterium in the functional fertilizer of Example 2 were omitted;
[0121] Control 4-3: The chitin biopeptide preparation and Bacillus megaterium in the functional fertilizer of Example 3 were omitted;
[0122] Control 4-4: The chitin biopeptide preparation and Bacillus megaterium in the functional fertilizer of Example 4 were omitted;
[0123] Control 4-5: The chitin biopeptide preparation and Bacillus megaterium in the functional fertilizer of Example 5 were omitted;
[0124] Control 5-1: The Bacillus megaterium in Example 1 was replaced with Bacillus subtilis;
[0125] Control 5-2: The Bacillus megaterium in Example 2 was replaced with Bacillus subtilis;
[0126] Control 5-3: The Bacillus megaterium in Example 3 was replaced with Bacillus subtilis;
[0127] Control 5-4: The Bacillus megaterium in Example 4 was replaced with Bacillus subtilis;
[0128] Control 5-5: The Bacillus megaterium in Example 5 was replaced with Bacillus subtilis;
[0129] Control 6-1: a functional fertilizer prepared in Example 1 in which the content of threonine, methionine, proline, alanine and phenylalanine accounts for 72.4% of the total free amino acids, and the ratio of threonine, methionine, proline, alanine and phenylalanine remains unchanged;
[0130] Control 6-2: The functional fertilizer prepared in Example 2 in which the content of threonine, methionine, proline, alanine and phenylalanine accounts for 72.4% of the total free amino acids, and the ratio of threonine, methionine, proline, alanine and phenylalanine remains unchanged;
[0131] Control 6-3: The functional fertilizer prepared in Example 3 in which the content of threonine, methionine, proline, alanine and phenylalanine accounts for 72.4% of the total free amino acids, and the ratio of threonine, methionine, proline, alanine and phenylalanine remains unchanged;
[0132] Control 6-4: The functional fertilizer prepared in Example 4 in which the content of threonine, methionine, proline, alanine and phenylalanine accounts for 72.4% of the total free amino acids, and the ratio of threonine, methionine, proline, alanine and phenylalanine remains unchanged;
[0133] Control 6-5: The functional fertilizer prepared in Example 5 in which the content of threonine, methionine, proline, alanine and phenylalanine accounts for 72.4% of the total free amino acids, and the ratio of threonine, methionine, proline, alanine and phenylalanine remains unchanged;
[0134] Control 7-1: a functional fertilizer prepared in Example 1 in which the content of threonine, methionine, proline, alanine and phenylalanine accounts for 90.6% of the free amino acids, and the ratio of threonine, methionine, proline, alanine and phenylalanine remains unchanged;
[0135] Control 7-2: The functional fertilizer prepared in Example 2, wherein the content of threonine, methionine, proline, alanine, and phenylalanine accounts for 90.6% of the total free amino acids, and the ratio of threonine, methionine, proline, alanine, and phenylalanine remains unchanged;
[0136] Control 7-3: The functional fertilizer prepared in Example 3 in which the content of threonine, methionine, proline, alanine and phenylalanine accounts for 90.6% of the total free amino acids, and the ratio of threonine, methionine, proline, alanine and phenylalanine remains unchanged;
[0137] Control 7-4: The functional fertilizer prepared in Example 4 in which the content of threonine, methionine, proline, alanine and phenylalanine accounts for 90.6% of the total free amino acids, and the ratio of threonine, methionine, proline, alanine and phenylalanine remains unchanged;
[0138] Control 7-5: The functional fertilizer prepared in Example 5 in which the content of threonine, methionine, proline, alanine and phenylalanine accounts for 90.6% of the total free amino acids, and the ratio of threonine, methionine, proline, alanine and phenylalanine remains unchanged;
[0139] Control 8-1: In the functional fertilizer prepared in Example 1, threonine, methionine, proline, alanine, and phenylalanine are replaced by tryptophan, leucine, arginine, histidine, and glycine in sequence, and the dosage and ratio are consistent with those in Example 1;
[0140] Control 8-2: In the functional fertilizer prepared in Example 2, threonine, methionine, proline, alanine, and phenylalanine are replaced by tryptophan, leucine, arginine, histidine, and glycine in sequence, and the dosage and ratio are consistent with those in Example 2;
[0141] Control 8-3: In the functional fertilizer prepared in Example 3, threonine, methionine, proline, alanine, and phenylalanine are replaced by tryptophan, leucine, arginine, histidine, and glycine in sequence, and the dosage and ratio are consistent with those in Example 3;
[0142] Control 8-4: In the functional fertilizer prepared in Example 4, threonine, methionine, proline, alanine, and phenylalanine are replaced by tryptophan, leucine, arginine, histidine, and glycine in sequence, and the dosage and ratio are consistent with those in Example 4;
[0143] Control 8-5: In the functional fertilizer prepared in Example 5, threonine, methionine, proline, alanine, and phenylalanine are replaced by tryptophan, leucine, arginine, histidine, and glycine in sequence, and the dosage and ratio are consistent with those in Example 5;
[0144] Absolute control 1: no application of the functional fertilizer prepared in Example 1;
[0145] Absolute control 2: no application of the functional fertilizer prepared in Example 2;
[0146] Absolute control 3: no application of the functional fertilizer prepared in Example 3;
[0147] Absolute control 4: no application of the functional fertilizer prepared in Example 4;
[0148] Absolute control 5: The functional fertilizer prepared in Example 5 was not applied.
[0149] According to the experimental design, the functional fertilizer prepared for each treatment (except the absolute control) was applied at an application rate of 120 kg / mu. At the same time, in order to ensure the normal growth of Hami melon, each treatment applied 25 kg / mu of compound fertilizer, 15 kg / mu of potassium sulfate, and 0.5 kg / mu of borax. Rotary tillage, land preparation, and ridge formation. Large ridges were planted alternately in double rows, with a plant spacing of 15 cm, a ridge width of 40 cm, and a row spacing of 35 cm between ridges. Hami melon seedlings with consistent growth were selected for transplanting, and then the water and fertilizer management of each treatment was consistent. On the 10th, 30th, and 60th days after the seedlings were transplanted, the rhizosphere soil and root system of Hami melon were collected respectively, the number of root-knot nematodes in the soil was investigated, and the disease index and control effect were calculated. The specific statistical data are shown in Table 1. When the Hami melon was harvested, the yield was counted, and its quality indicators such as soluble sugar, vitamin C and organic acid were determined. The specific data are shown in Table 2.
[0150] Table 1 Effects of different fertilizer treatments on root-knot nematodes in Hami melon at different stages
[0151]
[0152]
[0153] Note: Different capital letters in the table indicate significant differences among treatments (P<0.05), and different lowercase letters indicate significant differences within treatments (P<0.05), the same below
[0154] As shown in Table 1, compared with no addition of chitin biopeptide preparation (control 1-1 to 1-5), no addition of Bacillus megaterium (control 2-1 to 2-5), inoculation of sterilized Bacillus megaterium (control 2-1 to 2-5), no addition of chitin biopeptide preparation and Bacillus megaterium (control 4-1 to 4-5), Bacillus megaterium replaced by Bacillus subtilis (control 5-1 to 5-5), the content of threonine, methionine, proline, alanine, and phenylalanine in functional fertilizers accounting for less than 80% of the total free amino acids (control 6-1 to 6-5) or large Compared with 85% (control 7-1 to 7-5) and threonine, methionine, proline, alanine and phenylalanine in the functional fertilizer replaced by other amino acids (control 8-1 to 8-5), the Hami melons applied with the functional fertilizer prepared by the present invention (treatments 1 to 5) have significantly increased control effects on root-knot nematodes in the soil, which are specifically manifested as follows: 30 days after the Hami melons are transplanted, the control effects on root-knot nematodes in the soil are 81.98% to 82.75%; 60 days after the Hami melons are transplanted, the control effects on root-knot nematodes in the soil are 80.13% to 80.24%.
[0155] Compared with the control without adding chitin biopeptide preparation (control 1-1 to 1-5), without adding Bacillus megaterium (control 2-1 to 2-5), inoculating sterilized Bacillus megaterium (control 3-1 to 3-5) and replacing Bacillus megaterium with Bacillus subtilis (control 5-1 to 5-5), the control effect of root-knot nematodes in the soil was significantly reduced after applying functional fertilizers prepared without adding chitin biopeptide preparation and Bacillus megaterium (control 4-1 to 4-5) 10 days, 30 days and 60 days after Hami melon transplantation. It can be seen that the use of Bacillus megaterium and chitin biopeptide preparation together has a certain synergistic effect and can improve the control effect of root-knot nematodes.
[0156] Table 2 Effects of different fertilizer treatments on Hami melon yield and quality
[0157]
[0158]
[0159]
[0160] As shown in Table 2, compared with the functional fertilizers prepared in the examples (absolute controls 1 to 5), without adding chitin biopeptide preparations (controls 1-1 to 1-5), without adding Bacillus megaterium (controls 2-1 to 2-5), inoculated with sterilized Bacillus megaterium (controls 2-1 to 2-5), without adding chitin biopeptide preparations and Bacillus megaterium (controls 4-1 to 4-5), Bacillus megaterium replaced by Bacillus subtilis (controls 5-1 to 5-5), the contents of threonine, methionine, proline, alanine, and phenylalanine in the functional fertilizers accounted for 1.3% and 1.5% of the free threonine, methionine, proline, alanine, and phenylalanine in the functional fertilizers. Compared with the total amount of amino acids being less than 80% (control 6-1 to 6-5) or greater than 85% (control 7-1 to 7-5) and the functional fertilizers in which threonine, methionine, proline, alanine and phenylalanine are replaced by other amino acids (control 8-1 to 8-5), application of the functional fertilizers prepared by the present invention (treatments 1 to 5) significantly increased Hami melon yield, soluble protein and soluble sugar contents, with the highest increases of 54.07%, 30.88% and 29.56%, respectively, while the organic acid content was significantly reduced, with a reduction of 67.42%.
[0161] Compared with the case where no chitin biopeptide preparation was added (control 1-1 to 1-5), no Bacillus megaterium was added (control 2-1 to 2-5), sterilized Bacillus megaterium was inoculated (control 3-1 to 3-5), and Bacillus megaterium was replaced by Bacillus subtilis (control 5-1 to 5-5), the yield of Hami melon, soluble protein and soluble sugar content were significantly reduced, while the organic acid content was significantly increased by applying the functional fertilizer prepared by Bacillus megaterium without adding chitin biopeptide preparation (control 4-1 to 4-5). It can be seen that the use of Bacillus megaterium and chitin biopeptide preparation together has a certain synergistic effect, which can increase the yield of Hami melon, increase the content of soluble protein and soluble sugar, and reduce the content of organic acid.
[0162] The above description of the disclosed embodiments enables those skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.
Claims
1. A method for preparing a chitin biopeptide preparation, characterized in that: include: The chitin fermentation liquid and the biopeptide fermentation liquid are mixed, and dried at 70 to 80° C. to obtain a chitin biopeptide preparation; The preparation method of the chitosan fermentation liquid comprises: Under the conditions of 50-55°C and 150-170r / min, black soldier fly shells, trichoderma viride and water are mixed to obtain chitosan fermentation liquid; The preparation method of the biological peptide fermentation liquid comprises: Under the conditions of 50-60°C and 150-170r / min, black soldier fly larvae and compound enzyme are mixed to obtain biological peptide fermentation liquid; The complex enzyme comprises papain and lipase.
2. The preparation method according to claim 1, characterized in that: The mass ratio of the black soldier fly shell, green Trichoderma and water is (0.5-5):(0.5-5):
100.
3. The preparation method according to claim 1, characterized in that: The black soldier fly shells, green Trichoderma and water are mixed at 50-55° C., for 9-50 hours, at a mixing speed of 100-200 r / min.
4. The preparation method according to claim 1, characterized in that: The mass ratio of the black soldier fly larvae to the complex enzyme is 100:(0.05-0.2); In the composite enzyme, the mass ratio of papain to lipase is (1-2):
1.
5. The preparation method according to claim 1, characterized in that: The black soldier fly larvae and the complex enzyme are mixed at 50-60° C., for 9-15 hours, at a rotating speed of 100-200 r / min.
6. The preparation method according to claim 1, characterized in that: The volume ratio of the chitin fermentation liquid to the biological peptide fermentation liquid is 1:(1-5).
7. The chitosan biopeptide preparation obtained by the preparation method of the chitosan biopeptide preparation according to any one of claims 1 to 6.
8. A functional fertilizer containing a chitin biopeptide preparation, characterized in that: Includes basic fertilizer and Bacillus megaterium; In parts by mass, the basic fertilizer comprises 30 to 50 parts of decomposed compost, 25 to 35 parts of earthworm castings, 20 to 30 parts of peat, 2 to 4 parts of the chitin biopeptide preparation prepared by the preparation method of the chitin biopeptide preparation according to any one of claims 1 to 6 or the chitin biopeptide preparation according to claim 7, and 1 to 3 parts of potassium humate; The ratio of the Bacillus megaterium to the basic fertilizer is (10-20): 1 ml / g; The concentration of Bacillus megaterium is 1×10 8 ~3×10 8 cfu / ml.
9. Use of the chitin bio-peptide preparation obtained by the preparation method of the chitin bio-peptide preparation according to any one of claims 1 to 6 or the chitin bio-peptide preparation according to claim 7, and / or the functional fertilizer containing the chitin bio-peptide preparation according to claim 8 in controlling root-knot nematodes.
10. Use of the chitin biopeptide preparation obtained by the preparation method of the chitin biopeptide preparation according to any one of claims 1 to 6 or the chitin biopeptide preparation according to claim 7, and / or the functional fertilizer containing the chitin biopeptide preparation according to claim 8 in (1) to (4); (1) Increase the yield of Hami melon; (2) increasing the soluble protein content in Hami melon; (3) increasing the soluble sugar content in Hami melon; (4) Reduce the content of organic acids in Hami melon.