A special fertilizer for tobacco based on microbial rooting agent

Abstract

The present invention provides a tobacco-specific fertilizer based on a microbial rooting agent, and relates to the technical field of fertilizers. The tobacco-specific fertilizer of the present invention comprises components of Antarctic krill fermentation liquid, a microbial rooting agent, monoammonium phosphate and potassium chloride; the Antarctic krill fermentation liquid is prepared by grinding and washing defatted Antarctic krill, hydrolyzing with a composite protease, and then hydrolyzing the solid residue with chitinase, neutral protease and trypsin for secondary hydrolysis. The microbial rooting agent is prepared by fermenting composite bacteria, and then with a liquid culture medium and a mutagen, and the composite bacteria include components of Alteromonas, Bacillus brevis, Corbettella, Flavobacterium and Bacillus brevis. The tobacco-specific fertilizer of the present invention is used in tobacco cultivation and planting to enhance the absorption efficiency of the root for fertilizer, improve the vitality of the root system and the ability to resist lodging, and at the same time improve the diversity of soil nutrients and bacteria and fungi.

Description

Technical Field

[0001] The invention belongs to the technical field of fertilizers, and in particular relates to a special fertilizer for tobacco based on a microbial rooting agent. Background Art

[0002] Tobacco needs to absorb more nutrients during its entire growth cycle. After ordinary fertilizers are applied to cultivated land, some of the nutrients are lost or fixed and used by the soil and soil microorganisms, and only a small part is absorbed and used by crops. The nutrients such as nitrogen, phosphorus and potassium provided by ordinary fertilizers are insufficient, unevenly distributed and unreasonable, which makes tobacco plants unable to grow and develop normally due to lack of nutrients, weak root growth and development ability, thin plants, and low fertilizer utilization rate. The long-term use of chemical fertilizers will have an adverse effect on the soil and the environment, leading to soil compaction, and a decrease in soil self-regulation and soil self-repair capabilities.

[0003] Organic fertilizer contains rich organic matter and a variety of nutrients required by plants. It is also an excellent soil improvement and soil fertility agent. It has a good effect on increasing soil nutrient capacity, improving the structure of root soil bacteria and fungal communities, regulating soil pH, and improving soil nutrient effectiveness. However, the concentration of quick-acting nutrients in organic fertilizer is low and the fertilizer supply is slow. Studies have shown that adding microorganisms can accelerate the release of nutrients and improve the utilization efficiency of organic fertilizers. However, biological fertilizers in the general sense are composed of live bacteria and have strict requirements on soil growth environment conditions. Moreover, some intermediate metabolites that are unfavorable to crops may be produced during the growth and metabolism of microorganisms. At the same time, in the process of fertilization, due to one-sided topdressing and unreasonable fertilizer types, potassium fertilizers are seriously lacking, causing the quality of tobacco leaves to decline.

[0004] Starting from the diversity of fertilizer raw materials, soil, root soil bacteria and fungal community structure, combined with the fertilizer absorption rules of tobacco, the researchers have conducted a large number of formula fertilization studies, which have enabled tobacco fertilizers to develop from single elements to multi-element compound fertilizers, and from formula fertilization to special tobacco mixed fertilizers.

[0005] Chinese invention patent CN102617238A discloses a special fertilizer for tobacco active biological organic and inorganic fertilizer and its preparation method, the special fertilizer includes an active agent, biological fermentation cake fertilizer, inorganic fertilizer, wherein the preparation method of the biological fermentation cake fertilizer is to select functional microbial strains and add rapeseed meal, sesame meal and soybean meal according to weight percentage, and then solid ferment and enzymatically hydrolyze the crude protein in the cake to form biological fermentation cake fertilizer with biologically active free amino acids and active small peptides. The special fertilizer has the characteristics of high nutrition, high activity and high efficiency, can improve the rhizosphere environment of tobacco plants, enhance root vitality, promote leaf development, and fully meet the mineral nutrition requirements of tobacco plant development, reduce environmental pollution, and improve soil.

[0006] Chinese invention patent CN105601439A discloses a tobacco seedling fertilizer, which includes potassium humate, amino acids, azadirachtin, ammonium nitrate, monoammonium phosphate, sodium nitrophenolate, rooting powder, EDTA chelated magnesium, and boric acid. The tobacco fertilizer can fully utilize plant growth regulators, chelated trace elements, amino acids and other functional elements to promote the growth and development of tobacco plant roots according to the physiological habits of tobacco.

[0007] At present, although there are studies on bio-fermented fertilizers for tobacco fertilizers, there are few studies on the use of products of animal protease hydrolysis and plant rooting agents fermented by fermented strains for the preparation of tobacco fertilizers. In view of this, the present invention uses Antarctic krill for enzymatic fermentation and rooting agents to prepare tobacco fertilizers, which can promote early root growth, improve tobacco seedling survival rate, improve fertilizer utilization, enhance root absorption efficiency of fertilizers, slow down the aging of tobacco roots, and achieve the effect of increasing production and improving quality. Summary of the invention

[0008] In view of the problems existing in the prior art, the present invention provides a tobacco-specific fertilizer based on a microbial rooting agent, wherein a compound fertilizer is prepared by compounding Antarctic krill fermentation products with the rooting agent as the tobacco-specific fertilizer, so as to promote early root growth, improve the survival rate of tobacco seedlings, and improve the utilization rate of fertilizer, thereby achieving the effect of increasing production and improving quality.

[0009] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0010] Firstly, the present invention provides a tobacco-specific fertilizer based on a microbial rooting agent, which comprises the following components: Antarctic krill fermentation liquid, a microbial rooting agent, monoammonium phosphate and potassium chloride.

[0011] Preferably, the tobacco-specific fertilizer comprises the following components in parts by weight: 450-600 parts of Antarctic krill fermentation liquid, 80-120 parts of microbial rooting agent, 200-300 parts of monoammonium phosphate and 3-12 parts of potassium chloride.

[0012] Further preferably, the tobacco-specific fertilizer comprises the following components in parts by weight: 480-550 parts of Antarctic krill fermentation liquid, 90-110 parts of microbial rooting agent, 220-260 parts of monoammonium phosphate and 5-10 parts of potassium chloride.

[0013] More preferably, the tobacco-specific fertilizer comprises the following components in parts by weight: 500 parts of Antarctic krill fermentation liquid, 100 parts of microbial rooting agent, 250 parts of monoammonium phosphate and 5-10 parts of potassium chloride.

[0014] In the present invention, the ratio of nitrogen, phosphorus and potassium elements in the tobacco-specific fertilizer is 10-50:5-10:20-25.

[0015] Preferably, the method for preparing the Antarctic krill fermentation liquid comprises the steps of:

[0016] (1) Washing: Grinding the defatted Antarctic krill into powder, mixing with water and washing, to obtain defatted krill powder;

[0017] (2) Enzymatic hydrolysis: mixing the defatted krill powder with the composite protease, and then mixing with water for enzymatic hydrolysis to obtain an enzymatic hydrolysis mixture;

[0018] (3) Enzyme inactivation: After the enzymatic hydrolysis mixture is inactivated, it is filtered to separate the solid residue and the enzymatic hydrolysis solution A;

[0019] (4) secondary enzymatic hydrolysis: mixing the solid residue obtained in step (3) with 2-15 times the amount of water, adding chitinase, neutral protease and trypsin, and performing secondary enzymatic hydrolysis to obtain a secondary enzymatic hydrolysis mixture;

[0020] (5) Enzyme inactivation: After the secondary enzymatic hydrolysis mixture is inactivated, it is filtered to separate the solid residue and the enzymatic hydrolysis solution B;

[0021] (6) Concentration: The enzymatic hydrolysate A and the enzymatic hydrolysate B are combined and concentrated to obtain the Antarctic krill fermentation liquid.

[0022] Further preferably, in step (1), the powder has a particle size ranging from 200 to 500 mesh.

[0023] Further preferably, in step (2), the added amount of the composite protease is 5-15% of the mass of the defatted krill meal, g / g.

[0024] More preferably, the added amount of the composite protease is 10% of the mass of the defatted krill meal, g / g.

[0025] Further preferably, in step (2), the ratio of water to defatted krill powder is 20-60:1, g / mL.

[0026] More preferably, the ratio of water to defatted krill powder is 40-60:1, g / mL.

[0027] Further preferably, in step (2), the enzymatic hydrolysis is carried out at a temperature of 30-80° C. and a hydrolysis time of 20-180 min.

[0028] Further preferably, the enzymolysis temperature is 40-60° C. and the enzymolysis time is 40-120 min.

[0029] More preferably, in steps (3) and (5), the enzyme inactivation temperature is 80-121° C. and the enzyme inactivation time is 5-30 min.

[0030] More preferably, in steps (3) and (5), the filtration is performed after cooling.

[0031] Further preferably, in step (4), the ratio of water to solid residue is 5-12:1, g / mL.

[0032] More preferably, the ratio of water to solid residue is 10:1, g / mL.

[0033] Further preferably, in step (4), the mass ratio of chitinase, neutral protease and trypsin is 1-5:0.5-1.5:0.5-1.5.

[0034] More preferably, the mass ratio of chitinase, neutral protease and trypsin is 2-4:0.8-1.2:0.8-1.2.

[0035] Most preferably, the mass ratio of chitinase, neutral protease and trypsin is 3:1:1.

[0036] Further preferably, in step (4), the enzyme is added in an amount of 5-20% by mass of the solid residue, g / g.

[0037] More preferably, the enzyme is added in an amount of 8-13% of the mass of the solid residue, g / g.

[0038] Further preferably, in step (4), the enzymatic hydrolysis temperature of the secondary enzymatic hydrolysis is 35-85°C, the enzymatic hydrolysis time is 60-240 min, and the enzymatic hydrolysis pH is 6-7.

[0039] More preferably, the enzymatic hydrolysis temperature of the secondary enzymatic hydrolysis is 38-60°C, the enzymatic hydrolysis time is 90-180 min, and the enzymatic hydrolysis pH is 6.4-6.7.

[0040] Most preferably, the enzymatic hydrolysis temperature of the secondary enzymatic hydrolysis is 40° C., the enzymatic hydrolysis time is 120 min, and the enzymatic hydrolysis pH is 6.5.

[0041] More preferably, the solvent for adjusting the pH of the enzymatic hydrolysis is a 0.015-0.025 mol / L dilute hydrochloric acid solution.

[0042] Further preferably, in step (6), the concentration needs to be concentrated to a volume 5-15 times the mass of the original defatted krill meal, g / mL.

[0043] More preferably, the concentration needs to be concentrated to a volume 10 times the mass of the original defatted krill meal, g / mL.

[0044] Preferably, the microbial rooting agent comprises the following components in parts by weight: 90-111 parts of composite bacterial liquid, 100-200 parts of liquid culture medium and 0.3-0.6 parts of mutagen.

[0045] Further preferably, the composite bacterial liquid is prepared by fermenting composite bacteria, and the composite bacteria comprises the following components in parts by weight: 5-12 parts of Alteromonas, 6-8 parts of Bacillus brevis, 30.0-46 parts of Corbettella, 12-20 parts of Flavobacterium and 30-50 parts of Bacillus brevis.

[0046] Further preferably, the liquid culture medium is the LB liquid culture medium conventionally used in the art, and the preparation method is: dissolving peptone (Tryptone), yeast extract (Yeast extract), and sodium chloride (NaCl) in distilled water in proportion, sterilizing by high pressure, and adjusting the pH to neutral to prepare the LB liquid culture medium; in the LB liquid culture medium, the concentration of peptone is 10 g / L, the concentration of yeast extract is 4-6 g / L, and the concentration of sodium chloride is 8-12 g / L.

[0047] Further preferably, the mutagen is selected from at least one of 5-fluorodeoxyuridine, 2-aminopurine and hydroxylamine; when the mutagen is a mixture of 5-fluorodeoxyuridine and 2-aminopurine, the mass ratio thereof is 1:6-13, g / g.

[0048] Preferably, the preparation method of the microbial rooting agent comprises the steps of:

[0049] (1) mixing the composite bacteria with a liquid culture medium in a weight ratio to prepare a composite bacterial liquid;

[0050] (2) the composite bacterial liquid is subjected to preliminary fermentation culture under a preliminary fermentation process to obtain a preliminary fermentation liquid;

[0051] (3) mixing the preliminary fermentation liquid with the mutagen, and carrying out secondary fermentation culture under the secondary fermentation process to obtain a composite fermentation liquid;

[0052] (4) Decolorizing, concentrating, beating, centrifuging and drying the composite fermentation liquid to obtain a microbial rooting agent.

[0053] Preferably, in step (1), the bacterial content in the composite bacterial solution is 0.8-1.2×10 8 Pieces / mL.

[0054] Preferably, in step (2), the volume ratio of the composite bacterial liquid to the liquid culture medium is 1:1-2, v / v.

[0055] Preferably, in step (3), the preliminary fermentation process is specifically as follows: fermentation temperature 20-26.5°C, fermentation pH 6.5-8.0, fermentation stirring speed 160-200rpm / min; fermentation time 12-18h, the fermentation time is determined by the OD value of the bacterial liquid, and the fermentation is completed when the OD value of the bacterial liquid reaches 0.75-0.82.

[0056] Preferably, in step (3), the amount of the mutagen added is 0.12-0.26%, kg / L, of the initial fermentation broth.

[0057] Preferably, in step (3), the secondary fermentation process is specifically as follows: fermentation temperature 20-26.5°C, fermentation pH 6.5-8.0, fermentation stirring speed 160-200 rpm / min; fermentation time 20-30h.

[0058] Preferably, in step (4), the decolorization material used is selected from at least one of activated carbon, clay and lime; the mass volume ratio of the decolorization material to the composite bacterial liquid is 2.0-2.5:100, kg / L.

[0059] Preferably, in step (4), the centrifugation is carried out at a speed of 5000-10000 rpm / min for 10-120 min.

[0060] Furthermore, the present invention provides a method for preparing the above tobacco-specific fertilizer, comprising the steps of: uniformly mixing Antarctic krill fermentation liquid, microbial rooting agent, monoammonium phosphate and potassium chloride according to a proportion to obtain the fertilizer.

[0061] Finally, the present invention provides the use of the Antarctic krill fermentation liquid in the above tobacco-specific fertilizer in preparing the tobacco-specific fertilizer.

[0062] Compared with the prior art, the present invention has the following beneficial effects:

[0063] 1. The tobacco-specific fertilizer prepared by the present invention by compounding Antarctic krill fermentation liquid with a microbial rooting agent and using monoammonium phosphate and potassium chloride can significantly promote the early growth of roots and improve the survival rate of tobacco seedlings; root irrigation before clustering can stimulate the development of the root system, promote nutrient absorption, improve the vigorous growth ability of tobacco, and promote the opening of flue-cured tobacco; application after vigorous growth is expected to slow down the aging of the tobacco root system, promote the growth of upper tobacco leaves, and improve the utilization rate of upper tobacco leaves.

[0064] 2. The Antarctic krill fermentation liquid in the tobacco-specific fertilizer of the present invention is prepared by subjecting defatted Antarctic krill powder to two-step enzymolysis using a composite protease and chitinase, a neutral protease, and a trypsin, thereby increasing the content of nutrients in the enzymolysis liquid. The N / P / K ratio of the prepared tobacco-specific fertilizer is suitable for plant absorption. After use, the root's absorption efficiency of the fertilizer is greatly enhanced, thereby improving the utilization rate of the fertilizer, thereby achieving the effect of increasing production and improving quality.

[0065] 3. The tobacco-specific fertilizer of the present invention has a component Antarctic krill fermentation liquid that is rich in nitrogen, phosphorus, potassium and the natural compound chitin, which can promote the formation of soil aggregate structure, improve the internal pore space of the soil, coordinate the proportions of solid, liquid and gas in the soil, restore the natural colloid balance lost due to excessive soil burden and chemical pollution, increase soil biological activity, promote the release of fast-acting nutrients, be beneficial to root growth, and improve the stress resistance of crops; the combined component microbial rooting agent can improve the crop root environment, promote root development, improve soil microbial flora, and enhance crop photosynthesis. After application, tobacco grows more evenly and vigorously, with a very obvious yield-increasing effect, thereby increasing the yield of tobacco crops. DETAILED DESCRIPTION

[0066] The following non-limiting examples can make those of ordinary skill in the art understand the present invention more comprehensively, but do not limit the present invention in any way. The following content is merely an exemplary description of the scope of the present invention, and those skilled in the art can make various changes and modifications to the present invention according to the disclosed content, and it should also belong to the scope of the present invention. When the embodiment gives a numerical range, it should be understood that, unless otherwise specified in the present invention, the two endpoints of each numerical range and any numerical value between the two endpoints can be selected. Unless otherwise defined, all technical and scientific terms used in the present invention have the same meanings as those of ordinary skill in the technical field to which the present invention belongs.

[0067] The present invention is further described below by way of specific examples. Unless otherwise specified, the various chemical reagents used in the examples of the present invention are obtained through conventional commercial channels.

[0068] In the following examples, the Alteromonas is Alteromonas australica, from TSTO, numbered ATCC43369; the Bacillus pumilus is Bacillus pumilus Meyer and Gottheil, from TSTO, numbered ATCC31132; the Cobetia is Cobetia marina (Cobet et al.) Arahal et al., from TSTO, numbered ATCC25374; the Flavobacterium is Flavobacterium psychrophilum (Borg) Bernardet et al., from TSTO, numbered ATCC49418; the Bacillus pumilus is Bacillus pumilus, from Taisituo Biotechnology, numbered Nisc-2; the 5-fluorodeoxyuridine is from Sigma, CAS number is 50-91-9; the 2-aminopurine is from Sigma, CAS number is 452-06-2; the hydroxylamine is from Sigma, CAS number is 7803-49-8.

[0069] The components, chemical reagents, enzymes and bacteria in the present invention are not significantly affected by the products of different manufacturers. In some embodiments of the present invention, the preparation method of the microbial rooting agent used comprises the steps of:

[0070] (1) In 100L of the composite bacteria, by weight, Alteromonas australica accounted for 7 parts, Bacillus pumilus Meyer and Gottheil accounted for 6.2 parts, Cobetia marina (Cobet et al.) Arahal et al. accounted for 34.5 parts, Flavobacterium psychrophilum (Borg) Bernardet et al. accounted for 12 parts, and Bacillus pumilus accounted for 40.3 parts. The number of bacteria per milliliter was about 1×10 8 ; placed in a 1000 liter fermenter.

[0071] (2) Then, liquid culture medium was added to 250 liters. The preliminary fermentation culture was carried out under the following preliminary fermentation process parameters: temperature 24.5°C, pH=7.0, rotation speed 185 rpm, culture until the OD value of the bacterial liquid was between 0.75-0.82, fermentation for 15 hours, and obtaining the preliminary fermentation liquid.

[0072] (3) adding mutagen 5-fluorodeoxyuridine 0.05 kg and 2-aminopurine 0.3 kg (mass ratio of 1:6) at a dosage of 0.14%, and continuing culturing for 24 hours; the fermentation process parameters are: 24.5° C., pH=7.0, and rotation speed of 185 rpm; and obtaining a composite fermentation broth.

[0073] (4) 2.5 kg of activated carbon was added to the composite fermentation liquid for decolorization, and then evaporated and concentrated to a slurry state, beaten, precipitated, centrifuged at 8000 rpm / min for 2 h, and finally dried at 60° C. for 8 h to obtain the microbial rooting agent A.

[0074] In other embodiments of the present invention, the preparation method of the microbial rooting agent used comprises the steps of:

[0075] (1) In 100L of the composite bacteria, by weight, Alteromonas australica accounted for 10 parts, Bacillus pumilus Meyer and Gottheil accounted for 7.5 parts, Cobetia marina (Cobet et al.) Arahal et al. accounted for 30.0 parts, Flavobacterium psychrophilum (Borg) Bernardet et al. accounted for 18.5 parts, and Bacillus pumilus accounted for 34 parts; the number of bacteria contained in each milliliter was about 1×10 8 ; placed in a 1000 liter fermenter.

[0076] (2) Then, liquid culture medium was added to 250 liters. The preliminary fermentation culture was carried out under the following preliminary fermentation process parameters: temperature 24°C, pH=6.6, rotation speed 200 rpm, culture until the OD value of the bacterial liquid was between 0.75-0.82, fermentation for 16 hours, and obtaining the preliminary fermentation liquid.

[0077] (3) adding 0.53 kg of mutagen hydroxylamine (H3NO, CAS No. 7803-49-8) at a dosage of 0.21%, and continuing culturing for 24 h; the fermentation process parameters are: temperature 24°C, pH = 6.6, and rotation speed 200 rpm; and obtaining a composite fermentation liquid.

[0078] (4) 2.0 kg of activated carbon was added to the composite fermentation liquid for decolorization, and then evaporated and concentrated to a slurry state, beaten, precipitated, centrifuged at 8000 rpm / min for 2 h, and finally dried at 60° C. for 8 h to obtain the microbial rooting agent B.

[0079] Example 1

[0080] The tobacco-specific fertilizer includes the following components by weight: 500 parts of Antarctic krill fermentation liquid, 100 parts of microbial rooting agent, 250 parts of monoammonium phosphate and 5 parts of potassium chloride. After the above components are evenly mixed according to the proportion, the tobacco-specific fertilizer (N:P2O5:K2O=15:5:25) is obtained.

[0081] The method for preparing the Antarctic krill fermentation liquid comprises the following steps:

[0082] (1) Washing: Grind the defatted Antarctic krill into powder, pass it through a 330-350 mesh sieve, mix it with water, wash it, and dry it at 60° C. to obtain defatted krill powder;

[0083] (2) Enzymatic hydrolysis: defatted krill powder and composite protease were mixed at a mass ratio of 10:1 (g / g), the obtained mixture was mixed evenly with 50 times the amount of water (g / mL), and enzymatic hydrolysis was performed at a temperature of 40° C. for 50 min to obtain an enzymatic hydrolysis mixture;

[0084] (3) Inactivation of enzyme: After the enzymatic mixture is inactivated at 100°C for 20 min, it is filtered to separate the solid residue and the enzymatic solution A;

[0085] (4) secondary enzymolysis: mixing the solid residue obtained in step (3) with 10 times the amount of water (g / mL), adding chitinase, neutral protease and trypsin, wherein the mass ratio of chitinase, neutral protease and trypsin is 3:1:1 (g / g / g), and the amount of enzyme added is 10% (g / g) of the mass of the solid residue; performing secondary enzymolysis, adjusting the pH to 6.5 with a dilute hydrochloric acid solution with a concentration of 0.02 mol / L, the enzymolysis temperature is 40°C, and the enzymolysis time is 120 min to obtain a secondary enzymolysis mixture;

[0086] (5) Inactivation of enzyme: After the enzymatic mixture is inactivated at 100°C for 20 min, it is filtered to separate the solid residue and the enzymatic solution B;

[0087] (6) Concentration: Enzyme hydrolysate A and enzyme hydrolysate B are combined and concentrated to a volume 10 times the mass of the original defatted krill powder (g / mL) to obtain Antarctic krill fermentation liquid.

[0088] The microbial rooting agent is microbial rooting agent A.

[0089] Example 2

[0090] The difference from Example 1 is that the weight parts of the tobacco fertilizer are different: 500 parts of Antarctic krill fermentation liquid, 100 parts of microbial rooting agent, 250 parts of monoammonium phosphate and 10 parts of potassium chloride. After the above ingredients are mixed evenly according to the proportion, the tobacco fertilizer (N:P2O5:K2O=15:5:25) is obtained.

[0091] The microbial rooting agent is microbial rooting agent A.

[0092] Example 3

[0093] The difference from Example 2 is that the tobacco fertilizer has different components by weight: 480 parts of Antarctic krill fermentation liquid, 90 parts of microbial rooting agent, 220 parts of monoammonium phosphate and 10 parts of potassium chloride. After the above components are mixed evenly according to the proportion, the tobacco fertilizer is obtained.

[0094] The microbial rooting agent is microbial rooting agent A.

[0095] Example 4

[0096] The difference from Example 2 is that the tobacco fertilizer has different components by weight: 550 parts of Antarctic krill fermentation liquid, 110 parts of microbial rooting agent, 260 parts of monoammonium phosphate and 10 parts of potassium chloride. After the above components are mixed evenly according to the proportion, the tobacco fertilizer is obtained.

[0097] The microbial rooting agent is microbial rooting agent A.

[0098] Example 5

[0099] The difference from Example 2 is that the tobacco fertilizer has different components by weight: 450 parts of Antarctic krill fermentation liquid, 80 parts of microbial rooting agent, 200 parts of monoammonium phosphate and 3 parts of potassium chloride. After the above components are mixed evenly according to the proportion, the tobacco fertilizer is obtained.

[0100] The microbial rooting agent is microbial rooting agent B.

[0101] Example 6

[0102] The difference from Example 2 is that the tobacco fertilizer has different components by weight: 600 parts of Antarctic krill fermentation liquid, 120 parts of microbial rooting agent, 300 parts of monoammonium phosphate and 12 parts of potassium chloride. After the above components are mixed evenly according to the proportion, the tobacco fertilizer is obtained.

[0103] The microbial rooting agent is microbial rooting agent A.

[0104] Example 7

[0105] Different from Example 2, the preparation method of the Antarctic krill fermentation liquid in the tobacco fertilizer is different, including the steps of:

[0106] (1) Washing: Grinding the defatted Antarctic krill into powder, passing through a 200-300 mesh sieve, mixing with water for washing, and drying at 60° C. to obtain defatted krill powder;

[0107] (2) Enzymatic hydrolysis: defatted krill powder and composite protease were mixed at a mass ratio of 20:1 (g / g), the obtained mixture was mixed evenly with 20 times the amount of water (g / mL), and enzymatic hydrolysis was performed at a temperature of 60° C. for 120 min to obtain an enzymatic hydrolysis mixture;

[0108] (3) Inactivation of enzyme: After the enzymatic mixture is inactivated at 121°C for 5 min, it is filtered to separate the solid residue and the enzymatic solution A;

[0109] (4) secondary enzymolysis: mixing the solid residue obtained in step (3) with 5 times the amount of water (g / mL), adding chitinase, neutral protease and trypsin, wherein the mass ratio of chitinase, neutral protease and trypsin is 2:0.8:0.8 (g / g / g), and the amount of enzyme added is 5% (g / g) of the mass of the solid residue; performing secondary enzymolysis, adjusting the pH to 6 with a dilute hydrochloric acid solution with a concentration of 0.02 mol / L, the enzymolysis temperature is 60° C., and the enzymolysis time is 90 min to obtain a secondary enzymolysis mixture;

[0110] (5) Inactivation of enzyme: After the enzymatic mixture is inactivated at 121°C for 5 min, it is filtered to separate the solid residue and the enzymatic solution B;

[0111] (6) Concentration: Enzyme hydrolysate A and enzyme hydrolysate B are combined and concentrated to a volume 10 times the mass of the original defatted krill powder (g / mL) to obtain Antarctic krill fermentation liquid.

[0112] Example 8

[0113] Different from Example 2, the preparation method of the Antarctic krill fermentation liquid in the tobacco fertilizer is different, including the steps of:

[0114] (1) Washing: Grinding the defatted Antarctic krill into powder, passing through a 400-500 mesh sieve, mixing with water for washing, and drying at 60° C. to obtain defatted krill powder;

[0115] (2) Enzymatic hydrolysis: defatted krill powder and composite protease were mixed in a mass ratio of 20:3 (g / g), and the obtained mixture was mixed evenly with 60 times the amount of water (g / mL), and enzymatic hydrolysis was performed at a temperature of 80° C. for 20 min to obtain an enzymatic hydrolysis mixture;

[0116] (3) Inactivation of enzyme: After the enzymatic mixture is inactivated at 80°C for 30 min, it is filtered to separate the solid residue and the enzymatic solution A;

[0117] (4) secondary enzymolysis: mixing the solid residue obtained in step (3) with 12 times the amount of water (g / mL), adding chitinase, neutral protease and trypsin, wherein the mass ratio of chitinase, neutral protease and trypsin is 4:1.5:1.5 (g / g / g), and the amount of enzyme added is 20% (g / g) of the mass of the solid residue; performing secondary enzymolysis, adjusting the pH to 7.0 with a dilute hydrochloric acid solution with a concentration of 0.02 mol / L, the enzymolysis temperature is 85° C., and the enzymolysis time is 180 min to obtain a secondary enzymolysis mixture;

[0118] (5) Inactivation of enzyme: After the enzymatic mixture is inactivated at 80°C for 30 min, it is filtered to separate the solid residue and the enzymatic solution B;

[0119] (6) Concentration: Enzyme hydrolysate A and enzyme hydrolysate B are combined and concentrated to a volume 10 times the mass of the original defatted krill powder (g / mL) to obtain Antarctic krill fermentation liquid.

[0120] Comparative Example 1

[0121] Traditional commercially available ordinary potassium sulfate compound fertilizer has a total nutrient content of 45%, of which nitrogen, phosphorus and potassium each account for 15%.

[0122] Comparative Example 2

[0123] The difference from Example 2 is that the weight ratio of the tobacco fertilizer is different, specifically, the components are as follows: 500 parts of Antarctic krill fermentation liquid, 100 parts of microbial rooting agent, 10 parts of monoammonium phosphate and 250 parts of potassium chloride. After the above components are mixed evenly according to the proportion, the tobacco fertilizer is obtained.

[0124] Comparative Example 3

[0125] Different from Example 2, the weight ratio of the tobacco fertilizer is different, specifically, the components by weight are: 100 parts of Antarctic krill fermentation liquid, 100 parts of microbial rooting agent, 250 parts of monoammonium phosphate and 10 parts of potassium chloride. After the above ingredients are mixed evenly according to the proportion, the tobacco fertilizer is obtained.

[0126] Comparative Example 4

[0127] The difference from Example 2 is that the Antarctic krill fermentation liquid is replaced by humic acid fermentation liquid.

[0128] That is, humic acid is subjected to the preparation method of fermentation liquid described in Example 2 to prepare humic acid fermentation liquid. The humic acid is humic acid raw powder purchased from Shanxi Xinjinxiang Agricultural Technology Co., Ltd.

[0129] Comparative Example 5

[0130] The difference from Example 2 is that the microbial rooting agent is replaced by 1-naphthylacetic acid (NAA).

[0131] Comparative Example 6

[0132] The difference from Example 2 is that the preparation method of Antarctic krill fermentation liquid is different, specifically, the enzymatic hydrolysis time and enzymatic hydrolysis temperature in step (2) are different: the enzymatic hydrolysis temperature is 20° C., and the enzymatic hydrolysis time is 360 min.

[0133] Comparative Example 7

[0134] The difference from Example 2 is that the preparation method of Antarctic krill fermentation broth is different. Specifically, in the enzymatic hydrolysis process of step (4), the mass ratio of chitinase, neutral protease and trypsin used is different: 1:5:5, g / g / g.

[0135] Comparative Example 8

[0136] The difference from Example 2 is that the preparation method of the Antarctic krill fermentation liquid is different, specifically, in the enzymatic hydrolysis process of step (4), the type of enzyme used is different, neutral protease and trypsin are replaced by pepsin, that is, the mass ratio of chitinase to pepsin is 3:2.

[0137] Comparative Example 9

[0138] Different from Example 2, the preparation method of Antarctic krill fermentation liquid is different, specifically a one-step enzymatic hydrolysis method, comprising the steps of:

[0139] (1) Same as Example 2;

[0140] (2) Enzymatic hydrolysis: defatted krill meal was mixed with composite protease, chitinase, neutral protease and trypsin (3:1:1, g / g / g) in a mass ratio of 10:1 (g / g). The obtained mixture was mixed evenly with 50 times the amount of water (g / mL) and enzymatic hydrolysis was performed at a temperature of 40°C for 170 min to obtain an enzymatic hydrolysis mixture.

[0141] (3) Inactivation of enzyme: After the enzymatic mixture is inactivated at 100°C for 20 min, it is filtered to separate the solid residue and the enzymatic solution;

[0142] (4) Concentration: The volume of the enzymatic hydrolysate is concentrated to 10 times the mass of the original defatted krill powder (g / mL) to obtain Antarctic krill fermentation liquid.

[0143] Comparative test

[0144] 1. Sample preparation: Take 2 g of the fertilizer prepared in the example and the comparative example, respectively, and dissolve it in 1 L of water. Stir until completely dissolved to prepare a tobacco-specific fertilizer nutrient solution for use.

[0145] 2. Experimental tobacco: 5 tobacco seedlings of the same size were grouped together, and each group was treated with tobacco-specific fertilizer and nutrient solution. 1L of tobacco-specific fertilizer and nutrient solution was used for irrigation, and the water content was maintained at 60% according to the field method. 100mL of water was used every 7 days, and the cultivation lasted for 25 days in total.

[0146] In this experiment, the transplanting and irrigation of tobacco seedlings began in mid-April. After 25 days of cultivation, the tobacco plants were measured and the soil was collected for testing.

[0147] 3. Detection

[0148] 3.1 Root system detection

[0149] The test results of the effects of tobacco-specific fertilizers on root volume, lateral root number and root activity are shown in Table 1.

[0150] It can be seen from Table 1 that the tobacco-specific fertilizer of the present invention has the ability to significantly promote root growth and development and enhance root vitality.

[0151] Table 1

[0152]

[0153] 3.2 Leaf surface, plant height and stem girth detection

[0154] The test results are shown in Table 2. It can be seen from Table 2 that the tobacco-specific fertilizer of the present invention can increase the area of ​​tobacco leaves and improve the lodging resistance of tobacco plants.

[0155] Table 2

[0156]

[0157] 3.3 Soil nutrient element detection

[0158] The detection method is: detection according to national standard methods, involving GB7859-1987, GB7137-1987, GB7853-1987, GB7855-1987, GB9834-1988.

[0159] The test results are shown in Table 3. It can be seen from Table 3 that the tobacco-specific fertilizer of the present invention can promote the release of alkaline nitrogen and quick-acting potassium in the soil of the planted tobacco plants.

[0160] Table 3

[0161] pH Alkaline nitrogen (mg / kg) Available phosphorus (mg / kg) Available potassium (mg / kg) Organic matter (g / kg) Example 1 6.22 77.97 38.09 177.13 16.31 Example 2 6.10 80.47 37.81 176.28 16.68 Example 8 6.13 78.64 38.15 177.06 16.48 Comparative Example 1 6.28 64.46 34.06 160.72 12.23 Comparative Example 3 6.26 68.79 35.61 164.29 13.07 Comparative Example 4 6.31 67.43 34.75 162.06 12.84 Comparative Example 5 6.17 66.58 35.26 163.87 12.91 Comparative Example 7 6.22 67.45 35.39 160.25 13.52 Comparative Example 9 6.18 68.46 33.40 164.06 12.73

[0162] 3.4 Diversity detection of soil bacterial and fungal community structure

[0163] The diversity of soil bacterial and fungal community structure was detected according to the national standards for soil testing.

[0164] The test results are shown in Table 4. It can be seen from Table 4 that the tobacco-specific fertilizer of the present invention can improve the diversity of bacterial and fungal community structures and improve the fragrance index of the soil.

[0165] Table 4

[0166]

[0167]

[0168] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A tobacco-specific fertilizer based on a microbial rooting agent, characterized in that: The components include, by weight: 450-600 parts of Antarctic krill fermentation liquid, 80-120 parts of microbial rooting agent, 200-300 parts of monoammonium phosphate and 3-12 parts of potassium chloride; The method for preparing the Antarctic krill fermentation liquid comprises the steps of: (1) Washing: Grinding the defatted Antarctic krill into powder, mixing with water and washing, to obtain defatted krill powder; (2) Enzymatic hydrolysis: mixing the defatted krill powder with the composite protease, and then mixing with water for enzymatic hydrolysis to obtain an enzymatic hydrolysis mixture; wherein the enzymatic hydrolysis temperature is 30-80° C. and the enzymatic hydrolysis time is 20-180 min; (3) Enzyme inactivation: After the enzymatic hydrolysis mixture is inactivated, it is filtered to separate the solid residue and the enzymatic hydrolysis solution A; (4) secondary enzymolysis: mixing the solid residue obtained in step (3) with 2-15 times the amount of water, adding chitinase, neutral protease and trypsin, and performing secondary enzymolysis to obtain a secondary enzymolysis mixture; the mass ratio of the chitinase, neutral protease and trypsin is 1-5g:0.5-1.5g:0.5-1.5g; the enzymolysis temperature of the secondary enzymolysis is 35-85°C, and the enzymolysis time is 60-240min; (5) Enzyme inactivation: After the secondary enzymatic hydrolysis mixture is inactivated, it is filtered to separate the solid residue and the enzymatic hydrolysis solution B; (6) Concentration: combining the enzymatic hydrolysate A and the enzymatic hydrolysate B, and concentrating them to obtain the Antarctic krill fermentation liquid; The microbial rooting agent comprises the following components in parts by weight: 90-111 parts of composite bacteria, 100-200 parts of liquid culture medium and 0.3-0.6 parts of mutagen; the microbial rooting agent is prepared by fermenting the composite bacteria, and the composite bacteria comprises the following components in parts by weight: 5-12 parts of Alteromonas, 6-8 parts of Bacillus brevis, 30.0-46 parts of Corbettella, 12-20 parts of Flavobacterium and 30-50 parts of Bacillus brevis; the mutagen is a mixture of 5-fluorodeoxyuridine and 2-aminopurine in a mass ratio of 1g:6-13g, or is hydroxylamine; The preparation method of the microbial rooting agent comprises the steps of: (1) mixing the composite bacteria with a liquid culture medium in a weight ratio to prepare a composite bacterial liquid; (2) the composite bacterial liquid is subjected to preliminary fermentation culture under a preliminary fermentation process to obtain a preliminary fermentation liquid; (3) mixing the preliminary fermentation liquid with the mutagen, and carrying out secondary fermentation culture under the secondary fermentation process to obtain a composite fermentation liquid; (4) Decolorizing, concentrating, beating, centrifuging and drying the composite fermentation liquid to obtain a microbial rooting agent.

2. The tobacco-specific fertilizer according to claim 1, characterized in that: The invention comprises the following components in parts by weight: 480-550 parts of Antarctic krill fermentation liquid, 90-110 parts of microbial rooting agent, 220-260 parts of monoammonium phosphate and 5-10 parts of potassium chloride.

3. The tobacco-specific fertilizer according to claim 1, characterized in that: In the method for preparing Antarctic krill fermentation liquid, in step (1), the powder is ground to a particle size ranging from 200 to 500 mesh; in step (2), the added amount of the composite protease is 5-15% of the mass of the defatted krill powder, g / g; the ratio of water to defatted krill powder is 20-60g:1mL; in steps (3) and (5), the enzyme inactivation temperature is 80-121°C and the enzyme inactivation time is 5-30min.

4. The tobacco-specific fertilizer according to claim 3, characterized in that: In step (2) of the method for preparing Antarctic krill fermentation broth, the added amount of the composite protease is 10% of the mass of the defatted krill powder; and the ratio of water to defatted krill powder is 40-60g:1mL.

5. The tobacco-specific fertilizer according to claim 1, characterized in that: In the step (4) of the method for preparing Antarctic krill fermentation liquid, the ratio of water to solid residue is 5-12:1, g / mL; the amount of enzyme added is 5-20% of the mass of the solid residue, g / g; the pH of the secondary enzymolysis is 6-7; the solvent for adjusting the pH of the enzymolysis is a 0.015-0.025 mol / L dilute hydrochloric acid solution; in the step (6), the volume of the concentrated liquid needs to be concentrated to 5-15 times the mass of the original defatted krill powder, g / mL.

6. The tobacco-specific fertilizer according to claim 5, characterized in that: In step (4) of the preparation method, the ratio of water to solid residue is 10:1, g / mL; the mass ratio of chitinase, neutral protease and trypsin is 2-4g:0.8-1.2g:0.8-1.2g; the amount of enzyme added is 8-13% of the mass of the solid residue, g / g; the enzymatic hydrolysis temperature of the secondary enzymatic hydrolysis is 38-60°C, the enzymatic hydrolysis time is 90-180min, and the enzymatic hydrolysis pH is 6.4-6.7; in step (6), the volume of the concentrated krill meal needs to be concentrated to 10 times the mass of the original defatted krill meal, g / mL.

7. The method for preparing the tobacco-specific fertilizer according to any one of claims 1 to 6, characterized in that: The method comprises the following steps: uniformly mixing Antarctic krill fermentation liquid, microbial rooting agent, monoammonium phosphate and potassium chloride according to a certain proportion to obtain the product.

8. Use of the Antarctic krill fermentation liquid in the tobacco-specific fertilizer according to any one of claims 1 to 6 in preparing tobacco-specific fertilizer.

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