CN104072241A - Method for preparing organic leaf fertilizer special for herbal fruits - Google Patents
Method for preparing organic leaf fertilizer special for herbal fruits Download PDFInfo
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- 238000000034 method Methods 0.000 title abstract description 11
- 241000235058 Komagataella pastoris Species 0.000 claims abstract description 42
- 241000196324 Embryophyta Species 0.000 claims abstract description 40
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 33
- 239000007788 liquid Substances 0.000 claims abstract description 31
- 238000002360 preparation method Methods 0.000 claims abstract description 29
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- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 25
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 claims abstract description 22
- 238000000855 fermentation Methods 0.000 claims abstract description 22
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- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 56
- 102000004169 proteins and genes Human genes 0.000 claims description 37
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- 238000003756 stirring Methods 0.000 claims description 26
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- -1 1 part Substances 0.000 claims description 12
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- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 7
- 244000061456 Solanum tuberosum Species 0.000 claims description 7
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 7
- 239000008103 glucose Substances 0.000 claims description 7
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 6
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- 230000004913 activation Effects 0.000 claims description 3
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- AIUDWMLXCFRVDR-UHFFFAOYSA-N dimethyl 2-(3-ethyl-3-methylpentyl)propanedioate Chemical class CCC(C)(CC)CCC(C(=O)OC)C(=O)OC AIUDWMLXCFRVDR-UHFFFAOYSA-N 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 3
- 235000019319 peptone Nutrition 0.000 claims description 3
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- 239000000126 substance Substances 0.000 claims description 3
- 235000017060 Arachis glabrata Nutrition 0.000 claims description 2
- 244000105624 Arachis hypogaea Species 0.000 claims description 2
- 235000010777 Arachis hypogaea Nutrition 0.000 claims description 2
- 235000018262 Arachis monticola Nutrition 0.000 claims description 2
- 244000068988 Glycine max Species 0.000 claims description 2
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- 238000004519 manufacturing process Methods 0.000 abstract description 6
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- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 abstract description 5
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- 239000000203 mixture Substances 0.000 description 11
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 9
- 239000011591 potassium Substances 0.000 description 9
- 229910052700 potassium Inorganic materials 0.000 description 9
- 241000219109 Citrullus Species 0.000 description 8
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- 239000000047 product Substances 0.000 description 7
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 6
- 229910052757 nitrogen Inorganic materials 0.000 description 6
- 239000011574 phosphorus Substances 0.000 description 6
- 229910052698 phosphorus Inorganic materials 0.000 description 6
- 239000001965 potato dextrose agar Substances 0.000 description 5
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- 150000001413 amino acids Chemical class 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- GNTDGMZSJNCJKK-UHFFFAOYSA-N divanadium pentaoxide Chemical compound O=[V](=O)O[V](=O)=O GNTDGMZSJNCJKK-UHFFFAOYSA-N 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
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- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 description 1
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- 241000700605 Viruses Species 0.000 description 1
- FPWJLQXCGHQXLL-UHFFFAOYSA-N [P].OP(O)(O)=O Chemical compound [P].OP(O)(O)=O FPWJLQXCGHQXLL-UHFFFAOYSA-N 0.000 description 1
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- JEIPFZHSYJVQDO-UHFFFAOYSA-N ferric oxide Chemical compound O=[Fe]O[Fe]=O JEIPFZHSYJVQDO-UHFFFAOYSA-N 0.000 description 1
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- 239000011707 mineral Substances 0.000 description 1
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- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 150000002903 organophosphorus compounds Chemical class 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 description 1
- 230000008635 plant growth Effects 0.000 description 1
- CHWRSCGUEQEHOH-UHFFFAOYSA-N potassium oxide Chemical compound [O-2].[K+].[K+] CHWRSCGUEQEHOH-UHFFFAOYSA-N 0.000 description 1
- 229910001950 potassium oxide Inorganic materials 0.000 description 1
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Abstract
The invention relates to a method for preparing organic leaf fertilizer special for herbal fruits. The method comprises the following steps: (1) using pichia pastoris as an initial strain to prepare a pichia pastoris fermentation liquor, and then adding water and maize germ to prepare a phosphatase dissolving liquid; (2) adding water and a neutral protease preparation agent to prepare a protease dissolving liquid; (3) extracting plant ash through water so as to obtain a plant ash extracting solution; (4) taking the phosphatase dissolving liquid, the protease dissolving liquid and the plant ash extracting solution to be mixed according to ratio, and adjusting the pH value to be 6.0-6.5 so as to obtain the leaf fertilizer special for the herbal fruits. The leaf fertilizer is used for herbal fruit organic agricultural product production, and can provide reasonable available nutrients for the herbal fruits; shown by the field experiment, the yield-increasing effect on watermelon, melon and the like is good.
Description
Technical field
The present invention relates to a kind of preparation method of foliage fertilizer, particularly the preparation method of the special-purpose organic foliar fertilizer of a kind of herbaceous fruit, belongs to fertilizer production technology field.
Background technology
Herbaceous fruit, refers to fruit crop annual, biennial or perennial, that stem is herbaceous stem.Compare with general crop, herbaceous fruit generally needs more potassium nutrition.According to the study, in the time of infertility, the ratio of strawberry absorbed nitrogen, phosphorus, potassium is 1:0.39:1.81, and the ratio of watermelon absorbed nitrogen, phosphorus, potassium is 3:1:4, and the ratio of muskmelon absorbed nitrogen, phosphorus, potassium is 6:3:11.
Foliage fertilizer is a kind of fertilizer, and crop yield is played an important role.Organic foliar fertilizer is rich in it and promotes the active substance of plant growth, containing the fertilizer of chemosynthesis, is not widely used in organic farm products and produces.But the production technology of organic foliar fertilizer is difficult to accomplish that the nutrient demand according to herbaceous fruit provides nutrition at present.For example, with the composite foliage fertilizer of organic composition, mainly with amino acid, be directly main, only can provide nitrogen nutrition; Another kind of is the organic foliar fertilizer obtaining by organic substance decomposing, as patent document CN101250066 provides a kind of method of utilizing waste molasses of sugar plant to produce biological foliage fertilizer, it utilizes waste molasses boiling sterilization, microbe inoculation bacterial classification when its temperature drops to below 35 ℃, comprise aerobic bacteria, yeast and actinomycetes, carry out cultivation and fermentation and obtain.But the kind of this foliage fertilizer nutritive element and quantity are difficult for determining.The quantity of nutritive element and suitable ratio are the important factors that promotes herbaceous fruit increase output, improves quality.
Summary of the invention
For the deficiencies in the prior art, the invention provides the preparation method of the special-purpose organic foliar fertilizer of a kind of herbaceous fruit.
Term explanation:
1, herbaceous fruit, refers to fruit crop annual, biennial or perennial, that stem is herbaceous stem, and the over-ground part of this stem of plant is at the end withered in vegetative period.Herbaceous fruit of the present invention mainly comprises strawberry, watermelon, muskmelon, hami melon etc.
2, PDA slant medium, is the abbreviation to potato dextrose agar, formula: 200 grams of potatos, 20 grams of glucose, 15～20 grams, agar, 1000 milliliters, tap water.
Technical scheme of the present invention is as follows:
A preparation method for the special-purpose organic foliar fertilizer of herbaceous fruit, comprises that step is as follows:
(1) prepare phosphoric acid enzymolysis solution
A, take pichia pastoris phaff as initial bacterial classification, with PDA slant medium, Pichia Pastoris kind is activated;
B, by mass parts, remove 200 parts of skin potatos, add 1000 parts, water, boil 30 minutes, filtered through gauze, filtrate keeps the skin wet 1000 parts, then adds 10 parts of glucose, 4 parts of peptones, 1 part, magnesium sulfate, is heated to all the components and dissolves; Under 0.11 MPa pressure, sterilizing is 30 minutes, makes liquid nutrient medium; By in the Pichia Pastoris access liquid nutrient medium after step a, activation, in the shaking flask of 28 ℃, 120 revs/min, cultivate 45～48 hours; Obtain pichia pastoris phaff seed liquor;
C, by step b, the pichia pastoris phaff seed liquor that the makes inoculum size access fermentation tank culture medium with 3～5wt%, this fermentation tank culture medium mass parts formula is: 200 parts of potatos, 8 parts of glucose, 1 part of methyl alcohol, 1000 parts, water; Under 30 ℃ of conditions, cultivate 35～45 hours, obtain Fermentation in Pichia Pastoris liquid;
D, by mass parts, 1～3 part of the Fermentation in Pichia Pastoris liquid that get step c, makes, add 10～30 parts of 40～60 parts, water and maize germs, fully mix, under 45～55 ℃ of conditions, place 30～60 minutes, during every 15～20 minutes, stir once, each 1 minute, press filtration after finishing, filtrate is phosphoric acid enzymolysis solution; Standby;
(2) prepare protein enzymatic hydrolyzate
By mass parts, get 20～40 parts of maize germs, add 40～60 parts, water, boil 10～20 minutes, adjust pH value to 6.5～7.5, add 0.03～0.06 part of neutral protein zymin, stir 2～5 minutes, under 45～55 ℃ of conditions, place 2～3 hours, every 0.5 hour, stir once during this time, each 2～3 minutes, finish rear press filtration, filtrate is protein enzymatic hydrolyzate; Standby;
(3) prepare plant ash extracting solution
By mass parts, get 20～40 parts of plant ash, add 40～80 parts, water, stir, room temperature is placed 20～30 minutes, filters, and filtrate is plant ash extracting solution; Standby;
(4) press proportion by weight, get 10～30 parts of the phosphoric acid enzymolysis solutions of step (1), 40～60 parts of 20～40 parts of the protein enzymatic hydrolyzates of step (2), the plant ash extracting solutions of step (3), mix, adjust pH value to 6.0～6.5, filter, sterilizing, obtains the special-purpose organic foliar fertilizer of herbaceous fruit of the present invention.
The preparation method of the special-purpose organic foliar fertilizer of herbaceous fruit according to the present invention, preferred, the proportion by weight in above-mentioned steps (4) is as follows: 15 parts of phosphoric acid enzymolysis solutions, 35 parts of protein enzymatic hydrolyzates, 50 parts of plant ash extracting solutions.
According to the present invention, pichia pastoris phaff described in step (1), is preferably pichia pastoris phaff (Pichia pastoris) ACCC21071, ACCC21072, ACCC21073ACCC21074 or the ACCC21075 of Chinese agriculture microbial strains preservation administrative center (ACCC).
Preferred according to the present invention, step (1) c, in, the inoculum size of Pichia Pastoris is 4wt%, cultivates 40 hours; Steps d, in, 2 parts of Fermentation in Pichia Pastoris liquid, 50 parts, water, 20 parts of maize germs, temperature 50 C, places 40 minutes; In step (2), 30 parts of maize germs, add 50 parts, water, boil 15 minutes, adjust pH value to 7.0, and 0.04 part of neutral protein zymin, stirs 4 minutes, and temperature 50 C is placed 2.5 hours; In step (3), 30 parts of plant ash, add 75 parts, water.
Preferred according to the present invention, the neutral protein zymin used of step (2) is selected two kinds of specifications, NE PW30 (enzyme activity is 30,000U/g) and/or NE PW50 (enzyme activity is 50,000U/g); Can select one of them, also use capable of being combined; Further optimization protein zymin SUKAPro NEPW50 and SUKAPro NEPW30 are with the combination of 1: 1 mass ratio.
Preferred according to the present invention, to be crop stalk utilize mode and make the waste residue under residual after organic substance decomposing through burning or other plant ash described in step (3), it mainly utilizes composition is mineral nutrient element contained in crop straw, especially take potassium element and various trace elements as main.Crop stalk is selected from one of corn stalk, wheat straw, soybean straws, paddy stalk, peanut straw or vegetables straw or combination.
The present invention carries out liquid fermenting by pichia pastoris phaff and obtains the fermented liquid that contains Phosphoric acid esterase, and in described Fermentation in Pichia Pastoris liquid, Phosphatase Activity is 600U/g～1000U/g.
Preferred according to the present invention, in step (1) and step (2), maize germ used is first pulverized, and crosses 60～80 mesh sieves.
Preferred according to the present invention, in preparation process, all filtrations use mesh size at 80～100 orders.
Pichia Pastoris kind is activated by prior art with PDA slant medium in step of the present invention (1); In the methods of the invention, do not have be particularly limited all can be by prior art.
The application of the special-purpose organic foliar fertilizer of herbaceous fruit prepared by method of the present invention, produces for herbaceous fruit organic farm products.
The application process of the special-purpose organic foliar fertilizer of herbaceous fruit prepared by method of the present invention is, by organic foliar fertilizer thin up 10～15 quality of the present invention doubly, at herbaceous fruit, in vegetative period, is sprayed on vacuum side of blade, connects and sprays 3～5 times, 7～15 days, interval.
The composition of the special-purpose organic foliar fertilizer of herbaceous fruit of the present invention is phosphoric acid enzymolysis solution, protein enzymatic hydrolyzate and plant ash extracting solution, wherein said phosphoric acid enzymolysis solution, to utilize the Phosphoric acid esterase of secreting in Fermentation in Pichia Pastoris process to decompose the organo phosphorous compounds in maize germ, the liquid that separation obtains is rich in the available phosphorus that crop can absorb, and is the available phosphorus source of fertilizer of the present invention; Described protein enzymatic hydrolyzate, is to utilize protease preparation to decompose the protein in maize germ, and the liquid that separation obtains, contains amino acid and active substance of plant, is effective nitrogenous source of fertilizer of the present invention; Described plant ash extracting solution, is the liquid obtaining with flooding plant ash, is effective potassium and the trace element source of fertilizer of the present invention.
Excellent results of the present invention:
1. for herbaceous fruit provides rational available nutrient
Herbaceous fruit is the crop that needs potassium amount relatively many, also needs available nitrogen and available phosphorus and the trace element of rational proportion simultaneously.Existing organic foliar fertilizer take nitrogenous as main or its nutritive element kind and quantity uncertain, can not adapt to its nutritional needs of herbaceous fruit.The present invention take height containing potassium element and micro-plant ash extracting solution, containing available nitrogen as main protein enzymatic hydrolyzate be master containing available phosphorus phosphoric acid enzymolysis solution is raw material reasonable coordination, make each nutrient balance in fertilizer meet herbaceous fruit nutritional needs.
2. organic foliar fertilizer of the present invention is suitable for the production of herbaceous fruit organic farm products
The nutrient that in foliage fertilizer product of the present invention, all crops can absorb all comes from organism, meets organic farm products and produces the requirement of not using chemosynthesis fertilizer, is suitable for herbaceous fruit organic farm products and produces.
3. effect of increasing production is good
Product comprehensive nutrition provided by the invention, reasonable ratio.Through field experiment, watermelon, muskmelon have all been obtained good effect of increasing production, than conventional foliage fertilizer volume increase 5.8～10.5%.
Embodiment
Below in conjunction with embodiment, the present invention will be further described, but be not limited to this.
Raw material consumption umber in embodiment is mass parts.Per-cent in embodiment and comparative example (%) is mass percent.Raw material is described as follows:
Maize germ derives from Shandong wide life grease limited-liability company, pulverizes 80 mesh sieves; Pichia pastoris phaff (Pichia pastoris) ACCC21071 and ACCC21073, Chinese agriculture microbial strains preservation administrative center (ACCC) buys; Neutral protein zymin is SUKAPro NE, and (enzyme activity is respectively 50,000U/g and 30, and 000U/g), Su Kehan (Weifang) biotechnology company limited produces for model NE PW50 and NE PW30; Plant ash, Heze city of Shandong province Tong Li biotechnology company limited byproduct, is corn stalk ash.
Embodiment 1: the preparation method of the special-purpose organic foliar fertilizer of a kind of herbaceous fruit, and mass parts is composed as follows:
10 kilograms of phosphoric acid enzymolysis solutions, 30 kilograms of protein enzymatic hydrolyzates, 60 kilograms of plant ash extracting solutions;
Preparation process is as follows:
(1) prepare phosphoric acid enzymolysis solution
A, by prior art, with PDA slant medium, Pichia Pastoris kind ACCC21071 is activated;
B, remove 200 grams of skin potatos, add 1000 grams, water, boil filtered through gauze 30 minutes, filtrate keeps the skin wet 1000 grams, then adds 10 grams of glucose, 4 grams of peptones, 1 gram, magnesium sulfate, be heated to all the components and dissolve, under 0.11 MPa pressure, sterilizing is 30 minutes, is prepared into liquid nutrient medium; By in the Pichia Pastoris access liquid nutrient medium after step a, activation, in the shaking flask of 28 ℃, 120 revs/min, cultivate 46 hours; Obtain pichia pastoris phaff seed liquor;
C, get 200 grams of potatos, 8 grams of glucose, 1 gram of methyl alcohol, 1000 grams, water, by prior art, prepare liquid nutrient medium, pack in fermentor tank, inoculum size with 4wt% accesses step b, pichia pastoris phaff seed liquor, under 30 ℃ of conditions, cultivate 40 hours, obtain Fermentation in Pichia Pastoris liquid;
D, by mass parts, get 1 kilogram of step c, Fermentation in Pichia Pastoris liquid, 40 kilograms, water, 10 kilograms of maize germs, fully mix, under 55 ℃ of conditions, place 30 minutes, during stir once, 1 minute time, finishes rear press filtration, and filtrate is phosphoric acid enzymolysis solution; Standby;
(2) get 40 kilograms of maize germs, add 60 kilograms, water, boil 10 minutes, adjust pH value to 7.5, add 0.06 kilogram of neutral protein zymin SUKAPro NEPW50, stir 3 minutes, under 45 ℃ of conditions, place 2 hours, during every 0.5 hour, stir once, each 2 minutes, press filtration after finishing, obtains protein enzymatic hydrolyzate; Standby;
(3) get 40 kilograms of plant ash, add 80 kilograms, water, stir, place 20 minutes, filter, obtain plant ash extracting solution; Standby;
(4), by proportioning, get the phosphoric acid enzymolysis solution of step (1), the plant ash extracting solution of the protein enzymatic hydrolyzate of step (2), step (3) mixes, adjust pH value to 6.0～6.5, filter, by prior art autoclaving, obtain the special-purpose organic foliar fertilizer of herbaceous fruit of the present invention.
Embodiment 2: the preparation method of the special-purpose organic foliar fertilizer of a kind of herbaceous fruit, and mass parts is composed as follows:
15 kilograms of phosphoric acid enzymolysis solutions, 35 kilograms of protein enzymatic hydrolyzates, 50 kilograms of plant ash extracting solutions;
Preparation process:
(1) prepare phosphoric acid enzymolysis solution
The preparation of Fermentation in Pichia Pastoris liquid is with embodiment 1, and difference is bacterial classification ACCC21073;
By mass parts, get 2 kilograms of above-mentioned Fermentation in Pichia Pastoris liquid, 50 kilograms, water, 20 kilograms of maize germs, fully mix, under 50 ℃ of conditions, place 40 minutes, during stir once, 1 minute time, finish rear press filtration, filtrate is phosphoric acid enzymolysis solution; Standby;
(2) get 30 kilograms of maize germs, add 50 kilograms, water, boil 15 minutes, adjust pH value to 7.0, add 0.04 kilogram of neutral protein zymin SUKAPro NEPW50 and SUKAPro NEPW30 with the composition of 1: 1 mass ratio, stir 4 minutes, under 50 ℃ of conditions, place 2.5 hours, during every 0.5 hour, stir once, each 2 minutes, press filtration after finishing, obtains protein enzymatic hydrolyzate; Standby;
(3) get 30 kilograms of plant ash, add 75 kilograms, water, stir, place 25 minutes, filter, obtain plant ash extracting solution; Standby;
(4), by proportioning, get the phosphoric acid enzymolysis solution of step (1), the plant ash extracting solution of the protein enzymatic hydrolyzate of step (2), step (3) mixes, adjust pH value to 6.0～6.5, filter, by prior art autoclaving, obtain organic foliar fertilizer of the present invention.
Embodiment 3: the preparation method of the special-purpose organic foliar fertilizer of a kind of herbaceous fruit, and mass parts is composed as follows:
30 kilograms of phosphoric acid enzymolysis solutions, 25 kilograms of protein enzymatic hydrolyzates, 45 kilograms of plant ash extracting solutions;
Preparation process is as follows:
(1) prepare phosphoric acid enzymolysis solution
The preparation of Fermentation in Pichia Pastoris liquid is with embodiment 1, bacterial classification ACCC21071;
By mass parts, get 3 kilograms of the Fermentation in Pichia Pastoris liquid that make, 45 kilograms, water, 30 kilograms of maize germs, fully mix, under 45 ℃ of conditions, place 60 minutes, during stir twice, each 1 minute, finish rear press filtration, filtrate is phosphoric acid enzymolysis solution; Standby;
(2) get 25 kilograms of maize germs, add 40 kilograms, water, boil 12 minutes, adjust pH value to 6.8, add 0.03 kilogram of neutral protein zymin SUKAPro NEPW50, stir 3 minutes, under 47 ℃ of conditions, place 3 hours, during every 0.5 hour, stir once, each 2 minutes, press filtration after finishing, obtains protein enzymatic hydrolyzate; Standby;
(3) get 25 kilograms of plant ash, add 40 kilograms, water, stir, place 28 minutes, filter, obtain plant ash extracting solution; Standby;
(4) by proportioning, get the phosphoric acid enzymolysis solution of step (1), the protein enzymatic hydrolyzate of step (2), step (3) plant ash extracting solution mix, adjust pH value to 6.0～6.5, filtration, autoclaving, obtains organic foliar fertilizer of the present invention.
Embodiment 4: the preparation method of the special-purpose organic foliar fertilizer of a kind of herbaceous fruit, and mass parts is composed as follows:
25 kilograms of phosphoric acid enzymolysis solutions, 20 kilograms of protein enzymatic hydrolyzates, 55 kilograms of plant ash extracting solutions;
Preparation process is as follows:
(1) prepare phosphoric acid enzymolysis solution
The preparation of Fermentation in Pichia Pastoris liquid is with embodiment 1, bacterial classification ACCC21073;
By mass parts, get 1.5 kilograms of the Fermentation in Pichia Pastoris liquid that make, 60 kilograms, water, 25 kilograms of maize germs, fully mix, under 48 ℃ of conditions, place 50 minutes, during stir twice, each 1 minute, finish rear press filtration, filtrate is phosphoric acid enzymolysis solution; Standby;
(2) get 20 kilograms of maize germs, add 45 kilograms, water, boil 18 minutes, adjust pH value to 6.9, add 0.05 kilogram of neutral protein zymin SUKAPro NEPW30, stir 5 minutes, under 55 ℃ of conditions, place 2 hours, during every 0.5 hour, stir once, each 2 minutes, press filtration after finishing, obtains protein enzymatic hydrolyzate; Standby;
(3) get 35 kilograms of plant ash, add 60 kilograms, water, stir, place 22 minutes, filter, obtain plant ash extracting solution; Standby;
(4) by proportioning, get the phosphoric acid enzymolysis solution of step (1), the plant ash extracting solution of the protein enzymatic hydrolyzate of step (2), step (3) mixes, adjust pH value to 6.0～6.5, filter, autoclaving, obtains organic foliar fertilizer of the present invention.
Herbaceous fruit virus disease of tobacco
The aqueous solution of urea of comparative example 1:1%, common urea, nitrogenous 46%, commercially available prod;
Comparative example 2:0.5% potassium dihydrogen phosphate aqueous solution, common potassium primary phosphate, containing Vanadium Pentoxide in FLAKES 52%, potassium oxide 34%, commercially available prod.
1, for studying thing watermelon, experimental cultivar capital is glad.Test site: Changyue County, Shandong Province.Test period: year June in March, 2014 to 2014.
The special-purpose organic foliar fertilizer of herbaceous fruit of embodiment of the present invention 1-4 dilutes 13 quality doubly, is sprayed on the water melon leaf back side, and comparative example 1, comparative example 2 need not be diluted and be sprayed directly on to the water melon leaf back side.
If comparative example 1, comparative example 2 and embodiment 1-4 be totally 6 processing, 30 square metres of community areas, repeat 3 times; Foliage fertilizer sprays when watermelon starts to sit melon, and 10 days, interval connects and sprays 3 times.Other control measures are identical.Effect is in Table 1.
Table 1. is compared the watermelon effect of embodiment 1～4 with comparative example
| The inventive method | Than the stimulation ratio % of comparative example 1 | Than the stimulation ratio % of comparative example 2 |
| Embodiment 1 | 9.3 | 8.1 |
| Embodiment 2 | 8.4 | 8.3 |
| Embodiment 3 | 5.8 | 6.2 |
| Embodiment 4 | 7.2 | 7.6 |
2. supply to study thing muskmelon, experimental cultivar Qingzhou silver melon.Test site: Shandong Province's Qingzhou City.Test period: year June in March, 2014 to 2014.
The special-purpose organic foliar fertilizer of herbaceous fruit of embodiment of the present invention 1-4 dilutes 10 quality doubly, is sprayed on muskmelon vacuum side of blade, and comparative example 1, comparative example 2 need not be diluted and be sprayed directly on to muskmelon vacuum side of blade.
If comparative example 1, comparative example 2 and embodiment 1-4 be totally 6 processing, 21 square metres of community areas, repeat 3 times; Foliage fertilizer sprays when muskmelon is sat first batch of melon, and 15 days, interval connects and sprays 4 times.Other control measures are identical.Effect is in Table 2.
Table 2. is compared the muskmelon effect of embodiment 1～4 with comparative example
| The inventive method | Than the stimulation ratio % of comparative example 1 | Than the stimulation ratio % of comparative example 2 |
| Embodiment 1 | 7.9 | 7.6 |
| Embodiment 2 | 9.7 | 10.5 |
| Embodiment 3 | 8.2 | 9.2 |
| Embodiment 4 | 6.5 | 8.5 |
Claims (8)
1. a preparation method for the special-purpose organic foliar fertilizer of herbaceous fruit, comprises that step is as follows:
(1) prepare phosphoric acid enzymolysis solution
A, take pichia pastoris phaff as initial bacterial classification, with PDA slant medium, Pichia Pastoris kind is activated;
B, by mass parts, remove 200 parts of skin potatos, add 1000 parts, water, boil 30 minutes, filtered through gauze, filtrate keeps the skin wet 1000 parts, then adds 10 parts of glucose, 4 parts of peptones, 1 part, magnesium sulfate, is heated to all the components and dissolves; Under 0.11 MPa pressure, sterilizing is 30 minutes, makes liquid nutrient medium; By in the Pichia Pastoris access liquid nutrient medium after step a, activation, in the shaking flask of 28 ℃, 120 revs/min, cultivate 45～48 hours; Obtain pichia pastoris phaff seed liquor;
C, by step b, the pichia pastoris phaff seed liquor that the makes inoculum size access fermentation tank culture medium with 3～5wt%, this fermentation tank culture medium mass parts formula is: 200 parts of potatos, 8 parts of glucose, 1 part of methyl alcohol, 1000 parts, water; Under 30 ℃ of conditions, cultivate 35～45 hours, obtain Fermentation in Pichia Pastoris liquid;
D, by mass parts, 1～3 part of the Fermentation in Pichia Pastoris liquid that get step c, makes, add 10～30 parts of 40～60 parts, water and maize germs, fully mix, under 45～55 ℃ of conditions, place 30～60 minutes, during every 15～20 minutes, stir once, each 1 minute, press filtration after finishing, filtrate is phosphoric acid enzymolysis solution; Standby;
(2) prepare protein enzymatic hydrolyzate
By mass parts, get 20～40 parts of maize germs, add 40～60 parts, water, boil 10～20 minutes, adjust pH value to 6.5～7.5, add 0.03～0.06 part of neutral protein zymin, stir 2～5 minutes, under 45～55 ℃ of conditions, place 2～3 hours, every 0.5 hour, stir once during this time, each 2～3 minutes, finish rear press filtration, filtrate is protein enzymatic hydrolyzate; Standby;
(3) prepare plant ash extracting solution
By mass parts, get 20～40 parts of plant ash, add 40～80 parts, water, stir, room temperature is placed 20～30 minutes, filters, and filtrate is plant ash extracting solution; Standby;
(4) press proportion by weight, get 10～30 parts of the phosphoric acid enzymolysis solutions of step (1), 40～60 parts of 20～40 parts of the protein enzymatic hydrolyzates of step (2), the plant ash extracting solutions of step (3), mix, adjust pH value to 6.0～6.5, filter, sterilizing, obtains the special-purpose organic foliar fertilizer of herbaceous fruit.
2. the preparation method of the special-purpose organic foliar fertilizer of herbaceous fruit as claimed in claim 1, is characterized in that the proportion by weight in step (4) is as follows: 15 parts of phosphoric acid enzymolysis solutions, 35 parts of protein enzymatic hydrolyzates, 50 parts of plant ash extracting solutions.
3. the preparation method of the special-purpose organic foliar fertilizer of herbaceous fruit as claimed in claim 1, is characterized in that the pichia pastoris phaff described in step (1) is ACCC21071, ACCC21072, ACCC21073, ACCC21074 or ACCC21075.
4. the preparation method of the special-purpose organic foliar fertilizer of herbaceous fruit as claimed in claim 1, it is characterized in that step (1) c, in, the inoculum size of Pichia Pastoris is 4wt%, cultivates 40 hours; Steps d, in, 2 parts of Fermentation in Pichia Pastoris liquid, 50 parts, water, 20 parts of maize germs, temperature 50 C, places 40 minutes; In step (2), 30 parts of maize germs, add 50 parts, water, boil 15 minutes, adjust pH value to 7.0, and 0.04 part of neutral protein zymin, stirs 4 minutes, and temperature 50 C is placed 2.5 hours; In step (3), 30 parts of plant ash, add 75 parts, water.
5. the preparation method of the special-purpose organic foliar fertilizer of herbaceous fruit as claimed in claim 1, the neutral protein zymin used that it is characterized in that step (2) is selected two kinds of specifications, be NE PW30 (enzyme activity is 30,000U/g) and/or NE PW50 (enzyme activity is 50,000U/g); Select one of them or be used in combination.
6. the preparation method of the special-purpose organic foliar fertilizer of the herbaceous fruit as described in claim 1 or 5, the neutral protein zymin used that it is characterized in that step (2) is that protease preparation SUKAPro NEPW50 and SUKAPro NEPW30 are with the combination of 1: 1 mass ratio.
7. the preparation method of the special-purpose organic foliar fertilizer of herbaceous fruit as claimed in claim 1, it is characterized in that the plant ash described in step (3) is crop stalk and utilizes mode and make residual lower waste residue after organic substance decomposing through burning or other, described crop stalk is selected from one of corn stalk, wheat straw, soybean straws, paddy stalk, peanut straw or vegetables straw or combines.
8. the preparation method of the special-purpose organic foliar fertilizer of herbaceous fruit as claimed in claim 1, is characterized in that in step (1) and step (2), maize germ used is first pulverized, and crosses 60～80 mesh sieves.
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| CN106348848A (en) * | 2016-08-23 | 2017-01-25 | 山东蓝驼农业技术开发有限公司 | Method for compounding biological fertilizer capable of improving fruit taste |
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