CN104072241B - The preparation method of the special organic foliar fertilizer of a kind of herbaceous fruit - Google Patents
The preparation method of the special organic foliar fertilizer of a kind of herbaceous fruit Download PDFInfo
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- 238000002360 preparation method Methods 0.000 title claims abstract description 30
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims abstract description 62
- 241000235058 Komagataella pastoris Species 0.000 claims abstract description 42
- 241000196324 Embryophyta Species 0.000 claims abstract description 40
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- 238000000855 fermentation Methods 0.000 claims abstract description 22
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- 230000000694 effects Effects 0.000 claims abstract description 13
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- 238000003756 stirring Methods 0.000 claims description 18
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- 239000010902 straw Substances 0.000 claims description 9
- 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
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- 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 4
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- 108010080698 Peptones Proteins 0.000 claims description 3
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 3
- 235000005822 corn Nutrition 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 3
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- 241000209140 Triticum Species 0.000 claims 1
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- 238000000034 method Methods 0.000 abstract description 11
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- 230000000052 comparative effect Effects 0.000 description 17
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 12
- 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
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 7
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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 description 4
- 239000000203 mixture Substances 0.000 description 4
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- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen group Chemical group [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 3
- 230000000050 nutritive effect Effects 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 235000013619 trace mineral Nutrition 0.000 description 3
- 239000011573 trace mineral Substances 0.000 description 3
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- 235000016623 Fragaria vesca Nutrition 0.000 description 2
- 240000009088 Fragaria x ananassa Species 0.000 description 2
- 235000011363 Fragaria x ananassa Nutrition 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 239000013543 active substance Substances 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
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- GNTDGMZSJNCJKK-UHFFFAOYSA-N divanadium pentaoxide Chemical compound O=[V](=O)O[V](=O)=O GNTDGMZSJNCJKK-UHFFFAOYSA-N 0.000 description 2
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- 244000061176 Nicotiana tabacum Species 0.000 description 1
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 1
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 1
- 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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- 238000011081 inoculation Methods 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 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 present invention relates to the preparation method of the special organic foliar fertilizer of a kind of herbaceous fruit.The method comprises: (1) is the obtained Fermentation in Pichia Pastoris liquid of initial bacterial classification with pichia pastoris phaff, then adds water and maize germ obtains phosphoric acid enzymolysis solution; (2) maize germ add water and neutral protein zymin prepare protein enzymatic hydrolyzate; (3) plant ash extracting solution is obtained with flooding plant ash; (4) mixing of phosphoric acid enzymolysis solution, protein enzymatic hydrolyzate and plant ash extracting solution is got by proportioning, adjusted to ph to 6.0 ~ 6.5, obtain the special organic foliar fertilizer of herbaceous fruit, produce for herbaceous fruit organic farm products, rational available nutrient can be provided for herbaceous fruit; Effect of increasing production such as field experiment display watermelon, muskmelon etc. 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 organic foliar fertilizer of a kind of herbaceous fruit, belongs to technical field of fertilizer production.
Background technology
Herbaceous fruit, refers to that annual, biennial or perennial, stem is the fruit crop of herbaceous stem.Compared 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, plays an important role to crop yield.Organic foliar fertilizer with its be rich in promote plant growth active substance, not containing chemosynthesis fertilizer and be widely used in organic farm products produce.But the production technology of current organic foliar fertilizer is difficult to accomplish that the nutrient demand according to herbaceous fruit provides nutrition.Such as, the composite foliage fertilizer of direct organic composition is main mainly with amino acid, only can provide nitrogen nutrition; Another kind of is the organic foliar fertilizer obtained by organic substance decomposing, as patent document CN101250066 provides a kind of method 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 DEG C, comprise aerobic bacteria, yeast and actinomycetes, carry out cultivation and fermentation and obtain.But kind and the quantity of this foliage fertilizer nutritive element are not easily determined.The quantity of nutritive element and suitable ratio are the important factors promoting herbaceous fruit increase yield, oil recovery enhancement.
Summary of the invention
For the deficiencies in the prior art, the invention provides the preparation method of the special organic foliar fertilizer of a kind of herbaceous fruit.
Term illustrates:
1, herbaceous fruit, refer to that annual, biennial or perennial, stem is the fruit crop of herbaceous stem, 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: potato 200 grams, glucose 20 grams, 15 ~ 20 grams, agar, 1000 milliliters, tap water.
Technical scheme of the present invention is as follows:
A preparation method for the special organic foliar fertilizer of herbaceous fruit, comprises step as follows:
(1) phosphoric acid enzymolysis solution is prepared
A, be initial bacterial classification with pichia pastoris phaff, with PDA slant medium, Pichia Pastoris kind activated;
B, by mass parts, get peeled potatoes 200 parts, add water 1000 parts, boil 30 minutes, filtered through gauze, filtrate keeps the skin wet 1000 parts, then adds glucose 10 parts, peptone 4 parts, and 1 part, magnesium sulfate, is heated to all the components and dissolves; Under 0.11 MPa, sterilizing 30 minutes, makes liquid nutrient medium; By in the Pichia Pastoris access liquid nutrient medium after step a, activation, cultivate 45 ~ 48 hours in the shaking flask of 28 DEG C, 120 revs/min; Obtain pichia pastoris phaff seed liquor;
C, by step b, obtained pichia pastoris phaff seed liquor with the inoculum size of 3 ~ 5wt% access fermentation tank culture medium in, this fermentation tank culture medium mass parts formula is: potato 200 parts, glucose 8 parts, methyl alcohol 1 part, 1000 parts, water; Cultivate 35 ~ 45 hours under 30 DEG C of conditions, obtain Fermentation in Pichia Pastoris liquid;
D, by mass parts, get step c, obtained Fermentation in Pichia Pastoris liquid 1 ~ 3 part, add water 40 ~ 60 parts and maize germ 10 ~ 30 parts, abundant mixing, place 30 ~ 60 minutes under 45 ~ 55 DEG C of conditions, period stirred once every 15 ~ 20 minutes, each 1 minute, press filtration after terminating, filtrate is phosphoric acid enzymolysis solution; For subsequent use;
(2) protein enzymatic hydrolyzate is prepared
By mass parts, get maize germ 20 ~ 40 parts, add water 40 ~ 60 parts, boil 10 ~ 20 minutes, adjusted to ph to 6.5 ~ 7.5, add 0.03 ~ 0.06 part of neutral protein zymin, stir 2 ~ 5 minutes, place 2 ~ 3 hours under 45 ~ 55 DEG C of conditions, period stirred once every 0.5 hour, each 2 ~ 3 minutes, terminate rear press filtration, filtrate was protein enzymatic hydrolyzate; For subsequent use;
(3) plant ash extracting solution is prepared
By mass parts, get plant ash 20 ~ 40 parts, add water 40 ~ 80 parts, stir, room temperature is placed 20 ~ 30 minutes, and filter, filtrate is plant ash extracting solution; For subsequent use;
(4) by proportion by weight, get the phosphoric acid enzymolysis solution 10 ~ 30 parts of step (1), the protein enzymatic hydrolyzate 20 ~ 40 parts of step (2), the plant ash extracting solution 40 ~ 60 parts of step (3), mixing, adjusted to ph to 6.0 ~ 6.5, filter, sterilizing, obtains the special organic foliar fertilizer of herbaceous fruit of the present invention.
According to the preparation method of the special organic foliar fertilizer of herbaceous fruit of the present invention, preferably, the proportion by weight in above-mentioned steps (4) is as follows: phosphoric acid enzymolysis solution 15 parts, protein enzymatic hydrolyzate 35 parts, plant ash extracting solution 50 parts.
According to the present invention, pichia pastoris phaff described in step (1), is preferably pichia pastoris phaff (Pichiapastoris) ACCC21071, ACCC21072, ACCC21073ACCC21074 or ACCC21075 of Chinese agriculture Microbiological Culture Collection 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, Fermentation in Pichia Pastoris liquid 2 parts, 50 parts, water, maize germ 20 parts, temperature 50 C, places 40 minutes; In step (2), maize germ 30 parts, adds water 50 parts, boils 15 minutes, adjusted to ph to 7.0, neutral protein zymin 0.04 part, stirs 4 minutes, temperature 50 C, places 2.5 hours; In step (3), plant ash 30 parts, adds water 75 parts.
Preferred according to the present invention, the neutral protein zymin used of step (2) selects two kinds of specifications, i.e. NEPW30 (enzyme activity is 30,000U/g) and/or NEPW50 (enzyme activity is 50,000U/g); One of them can be selected, also use capable of being combined; Further optimization protein zymin SUKAProNEPW50 and SUKAProNEPW30 is with the combination of 1: 1 mass ratio.
Preferred according to the present invention, plant ash described in step (3) is the waste residue under remaining crop stalk makes organic substance decomposing through burning or other utilization mode after, it mainly utilizes composition to be mineral nutrient element contained in crop straw, especially based on potassium element and various trace element.Crop stalk is selected from corn stalk, wheat straw, soybean straws, paddy stalk, one of peanut straw or vegetables straw or combination.
Pichia pastoris phaff is carried out the fermented liquid that liquid fermenting obtains containing Phosphoric acid esterase by the present invention, 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, mistake 60 ~ 80 mesh sieves.
Preferred according to the present invention, in preparation process, all filtration mesh sizes are at 80 ~ 100 orders.
The activation of Pichia Pastoris kind is pressed prior art by the PDA slant medium of using in step of the present invention (1); In the methods of the invention, do not have be particularly limited to all can by prior art.
The application of the special 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 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, is sprayed on vacuum side of blade, connects spray 3 ~ 5 times, 7 ~ 15 days, interval within herbaceous fruit vegetative period.
The composition of the special 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, utilize the Phosphoric acid esterase secreted in Fermentation in Pichia Pastoris process to decompose 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, be utilize the protein in protease preparation decomposition maize germ, being separated the liquid obtained, containing 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 obtained 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 needing potassium amount relatively many, also needs the available nitrogen of rational proportion and available phosphorus and trace element simultaneously.Existing organic foliar fertilizer based on nitrogenous or its nutritive element kind and quantity uncertain, its nutritional needs of herbaceous fruit can not be adapted to.The present invention contains the plain and micro-plant ash extracting solution of potassium, the protein enzymatic hydrolyzate containing available nitrogen and the phosphoric acid enzymolysis solution containing available phosphorus for raw material reasonable coordination based on height, makes 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 all achieve 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 thereto.
Raw material dosage number 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 (Pichiapastoris) ACCC21071 and ACCC21073, Chinese agriculture Microbiological Culture Collection administrative center (ACCC) buys; Neutral protein zymin is SUKAProNE, model NEPW50 and NEPW30 (enzyme activity is respectively 50,000U/g and 30,000U/g), and Su Kehan (Weifang) biotechnology company limited produces; Plant ash, Heze city of Shandong province, with sharp biotechnology company limited byproduct, is corn stalk ash.
Embodiment 1: the preparation method of the special organic foliar fertilizer of a kind of herbaceous fruit, mass parts is composed as follows:
Phosphoric acid enzymolysis solution 10 kilograms, protein enzymatic hydrolyzate 30 kilograms, plant ash extracting solution 60 kilograms;
Preparation process is as follows:
(1) phosphoric acid enzymolysis solution is prepared
A, by prior art PDA slant medium, Pichia Pastoris kind ACCC21071 to be activated;
B, get peeled potatoes 200 grams, add water 1000 grams, boil 30 minutes, filtered through gauze, filtrate keeps the skin wet 1000 grams, then adds glucose 10 grams, peptone 4 grams, 1 gram, magnesium sulfate, be heated to all the components dissolve, under 0.11 MPa, sterilizing 30 minutes, is prepared into liquid nutrient medium; By in the Pichia Pastoris access liquid nutrient medium after step a, activation, cultivate 46 hours in the shaking flask of 28 DEG C, 120 revs/min; Obtain pichia pastoris phaff seed liquor;
C, get potato 200 grams, glucose 8 grams, methyl alcohol 1 gram, 1000 grams, water, liquid nutrient medium is prepared by prior art, load in fermentor tank, with the inoculum size of 4wt% by step b, the access of pichia pastoris phaff seed liquor, cultivate 40 hours under 30 DEG C of conditions, obtain Fermentation in Pichia Pastoris liquid;
D, by mass parts, get step c, Fermentation in Pichia Pastoris liquid 1 kilogram, 40 kilograms, water, maize germ 10 kilograms, abundant mixing, places 30 minutes under 55 DEG C of conditions, and period stirs once, 1 minute time, terminate rear press filtration, filtrate is phosphoric acid enzymolysis solution; For subsequent use;
(2) maize germ 40 kilograms is got, add water 60 kilograms, boil 10 minutes, adjusted to ph to 7.5, add 0.06 kilogram of neutral protein zymin SUKAProNEPW50, stir 3 minutes, place 2 hours under 45 DEG C of conditions, period stirred once every 0.5 hour, each 2 minutes, press filtration after terminating, obtains protein enzymatic hydrolyzate; For subsequent use;
(3) get plant ash 40 kilograms, add water 80 kilograms, stir, place 20 minutes, filter, obtain plant ash extracting solution; For subsequent use;
(4) by proportioning, get the phosphoric acid enzymolysis solution of step (1), the protein enzymatic hydrolyzate of step (2), the plant ash extracting solution of step (3), mixing, adjusted to ph to 6.0 ~ 6.5, filter, by prior art autoclaving, obtain the special organic foliar fertilizer of herbaceous fruit of the present invention.
Embodiment 2: the preparation method of the special organic foliar fertilizer of a kind of herbaceous fruit, mass parts is composed as follows:
Phosphoric acid enzymolysis solution 15 kilograms, protein enzymatic hydrolyzate 35 kilograms, plant ash extracting solution 50 kilograms;
Preparation process:
(1) phosphoric acid enzymolysis solution is prepared
The preparation of Fermentation in Pichia Pastoris liquid is with embodiment 1, and difference is bacterial classification ACCC21073;
By mass parts, get above-mentioned Fermentation in Pichia Pastoris liquid 2 kilograms, 50 kilograms, water, maize germ 20 kilograms, fully mixes, and places 40 minutes under 50 DEG C of conditions, and period stirs once, and 1 minute time, terminate rear press filtration, filtrate is phosphoric acid enzymolysis solution; For subsequent use;
(2) maize germ 30 kilograms is got, add water 50 kilograms, boil 15 minutes, adjusted to ph to 7.0, add 0.04 kilogram of neutral protein zymin SUKAProNEPW50 and SUKAProNEPW30 with the composition of 1: 1 mass ratio, stir 4 minutes, place 2.5 hours under 50 DEG C of conditions, period stirred once every 0.5 hour, each 2 minutes, press filtration after terminating, obtains protein enzymatic hydrolyzate; For subsequent use;
(3) get plant ash 30 kilograms, add water 75 kilograms, stir, place 25 minutes, filter, obtain plant ash extracting solution; For subsequent use;
(4) by proportioning, the phosphoric acid enzymolysis solution of step (1), the protein enzymatic hydrolyzate of step (2), the plant ash extracting solution of step (3) is got, mixing, adjusted to ph 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 organic foliar fertilizer of a kind of herbaceous fruit, mass parts is composed as follows:
Phosphoric acid enzymolysis solution 30 kilograms, protein enzymatic hydrolyzate 25 kilograms, plant ash extracting solution 45 kilograms;
Preparation process is as follows:
(1) phosphoric acid enzymolysis solution is prepared
The preparation of Fermentation in Pichia Pastoris liquid with embodiment 1, bacterial classification ACCC21071;
By mass parts, get obtained Fermentation in Pichia Pastoris liquid 3 kilograms, 45 kilograms, water, maize germ 30 kilograms, fully mixes, and places 60 minutes under 45 DEG C of conditions, and period stirs twice, and each 1 minute, terminate rear press filtration, filtrate is phosphoric acid enzymolysis solution; For subsequent use;
(2) maize germ 25 kilograms is got, add water 40 kilograms, boil 12 minutes, adjusted to ph to 6.8, add 0.03 kilogram of neutral protein zymin SUKAProNEPW50, stir 3 minutes, place 3 hours under 47 DEG C of conditions, period stirred once every 0.5 hour, each 2 minutes, press filtration after terminating, obtains protein enzymatic hydrolyzate; For subsequent use;
(3) get plant ash 25 kilograms, add water 40 kilograms, stir, place 28 minutes, filter, obtain plant ash extracting solution; For subsequent use;
(4) by proportioning, the phosphoric acid enzymolysis solution of step (1), the protein enzymatic hydrolyzate of step (2), step (3) plant ash extracting solution is got, mixing, adjusted to ph to 6.0 ~ 6.5, filter, autoclaving, obtains organic foliar fertilizer of the present invention.
Embodiment 4: the preparation method of the special organic foliar fertilizer of a kind of herbaceous fruit, mass parts is composed as follows:
Phosphoric acid enzymolysis solution 25 kilograms, protein enzymatic hydrolyzate 20 kilograms, plant ash extracting solution 55 kilograms;
Preparation process is as follows:
(1) phosphoric acid enzymolysis solution is prepared
The preparation of Fermentation in Pichia Pastoris liquid with embodiment 1, bacterial classification ACCC21073;
By mass parts, get obtained Fermentation in Pichia Pastoris liquid 1.5 kilograms, 60 kilograms, water, maize germ 25 kilograms, fully mixes, and places 50 minutes under 48 DEG C of conditions, and period stirs twice, and each 1 minute, terminate rear press filtration, filtrate is phosphoric acid enzymolysis solution; For subsequent use;
(2) maize germ 20 kilograms is got, add water 45 kilograms, boil 18 minutes, adjusted to ph to 6.9, add 0.05 kilogram of neutral protein zymin SUKAProNEPW30, stir 5 minutes, place 2 hours under 55 DEG C of conditions, period stirred once every 0.5 hour, each 2 minutes, press filtration after terminating, obtains protein enzymatic hydrolyzate; For subsequent use;
(3) get plant ash 35 kilograms, add water 60 kilograms, stir, place 22 minutes, filter, obtain plant ash extracting solution; For subsequent use;
(4) by proportioning, the phosphoric acid enzymolysis solution of step (1), the protein enzymatic hydrolyzate of step (2), the plant ash extracting solution of step (3) is got, mixing, adjusted to ph 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 of Shandong Province.Test period: year June in March, 2014 to 2014.
The special organic foliar fertilizer of herbaceous fruit of embodiment of the present invention 1-4 dilutes 13 quality doubly, and be sprayed on the water melon leaf back side, 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 process, plot area 30 square metres, repeat 3 times; Foliage fertilizer sprays when watermelon starts to sit melon, 10 days, interval, connects spray 3 times.Other control measures are identical.Effect is in table 1.
The watermelon effect of table 1. embodiment 1 ~ 4 compared 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. for studying 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 organic foliar fertilizer of herbaceous fruit of embodiment of the present invention 1-4 dilutes 10 quality doubly, and be sprayed on muskmelon vacuum side of blade, 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 process, plot area 21 square metres, repeat 3 times; Foliage fertilizer sprays when first batch of melon sat by muskmelon, 15 days, interval, connects spray 4 times.Other control measures are identical.Effect is in table 2.
The muskmelon effect of table 2. embodiment 1 ~ 4 compared 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 organic foliar fertilizer of herbaceous fruit, comprises step as follows:
(1) phosphoric acid enzymolysis solution is prepared
A, be initial bacterial classification with pichia pastoris phaff, with PDA slant medium, Pichia Pastoris kind activated;
B, by mass parts, get peeled potatoes 200 parts, add water 1000 parts, boil 30 minutes, filtered through gauze, filtrate keeps the skin wet 1000 parts, then adds glucose 10 parts, peptone 4 parts, and 1 part, magnesium sulfate, is heated to all the components and dissolves; Under 0.11 MPa, sterilizing 30 minutes, makes liquid nutrient medium; By in the Pichia Pastoris access liquid nutrient medium after step a, activation, cultivate 45 ~ 48 hours in the shaking flask of 28 DEG C, 120 revs/min; Obtain pichia pastoris phaff seed liquor;
C, by step b, obtained pichia pastoris phaff seed liquor with the inoculum size of 3 ~ 5wt% access fermentation tank culture medium in, this fermentation tank culture medium mass parts formula is: potato 200 parts, glucose 8 parts, methyl alcohol 1 part, 1000 parts, water; In 30
0cultivate 35 ~ 45 hours under C condition, obtain Fermentation in Pichia Pastoris liquid;
D, by mass parts, get step c, obtained Fermentation in Pichia Pastoris liquid 1 ~ 3 part, add water 40 ~ 60 parts and maize germ 10 ~ 30 parts, abundant mixing, place 30 ~ 60 minutes under 45 ~ 55 DEG C of conditions, period stirred once every 15 ~ 20 minutes, each 1 minute, press filtration after terminating, filtrate is phosphoric acid enzymolysis solution; For subsequent use;
(2) protein enzymatic hydrolyzate is prepared
By mass parts, get maize germ 20 ~ 40 parts, add water 40 ~ 60 parts, boil 10 ~ 20 minutes, adjusted to ph to 6.5 ~ 7.5, add 0.03 ~ 0.06 part of neutral protein zymin, stir 2 ~ 5 minutes, place 2 ~ 3 hours under 45 ~ 55 DEG C of conditions, period stirred once every 0.5 hour, each 2 ~ 3 minutes, terminate rear press filtration, filtrate was protein enzymatic hydrolyzate; For subsequent use;
(3) plant ash extracting solution is prepared
By mass parts, get plant ash 20 ~ 40 parts, add water 40 ~ 80 parts, stir, room temperature is placed 20 ~ 30 minutes, and filter, filtrate is plant ash extracting solution; For subsequent use;
(4) by proportion by weight, the phosphoric acid enzymolysis solution 10 ~ 30 parts of step (1) is got, the protein enzymatic hydrolyzate 20 ~ 40 parts of step (2), the plant ash extracting solution 40 ~ 60 parts of step (3), mixing, adjusted to ph to 6.0 ~ 6.5, filter, sterilizing, obtains the special organic foliar fertilizer of herbaceous fruit.
2. the preparation method of the special 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: phosphoric acid enzymolysis solution 15 parts, protein enzymatic hydrolyzate 35 parts, plant ash extracting solution 50 parts.
3. the preparation method of the special 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 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, Fermentation in Pichia Pastoris liquid 2 parts, 50 parts, water, maize germ 20 parts, temperature 50 C, places 40 minutes; In step (2), maize germ 30 parts, adds water 50 parts, boils 15 minutes, adjusted to ph to 7.0, neutral protein zymin 0.04 part, stirs 4 minutes, temperature 50 C, places 2.5 hours; In step (3), plant ash 30 parts, adds water 75 parts.
5. the preparation method of the special organic foliar fertilizer of herbaceous fruit as claimed in claim 1, it is characterized in that the neutral protein zymin used of step (2) selects two kinds of specifications, namely NEPW30(enzyme activity is 30,000U/g) and/or NEPW50(enzyme activity be 50,000U/g); Select one of them or combinationally use.
6. the preparation method of the special organic foliar fertilizer of the herbaceous fruit as described in claim 1 or 5, is characterized in that the neutral protein zymin used of step (2) is that protease preparation SUKAProNEPW50 and SUKAProNEPW30 is with the combination of 1: 1 mass ratio.
7. the preparation method of the special organic foliar fertilizer of herbaceous fruit as claimed in claim 1, it is characterized in that the plant ash described in step (3) is the waste residue under crop stalk remains after burning, described crop stalk is selected from corn stalk, wheat straw, soybean straws, paddy stalk, one of peanut straw or vegetables straw or combination.
8. the preparation method of the special 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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| CN101973799A (en) * | 2010-09-15 | 2011-02-16 | 山东光大肥业科技有限公司 | Organic-inorganic fertilizer special for Xinjiang grapes |
| CN102020497A (en) * | 2010-10-25 | 2011-04-20 | 山东光大肥业科技有限公司 | Special organic-inorganic fertilizer for Jinxiang garlic and preparation and application thereof |
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| CN101955390A (en) * | 2010-09-17 | 2011-01-26 | 山东光大肥业科技有限公司 | Special fertilizer for Zhanhua winter jujubes, preparation method and application thereof |
| CN102020497A (en) * | 2010-10-25 | 2011-04-20 | 山东光大肥业科技有限公司 | Special organic-inorganic fertilizer for Jinxiang garlic and preparation and application thereof |
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