CN103497050A - Method for preparing salt tolerant phosphate solubilizing seed coating - Google Patents
Method for preparing salt tolerant phosphate solubilizing seed coating Download PDFInfo
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- CN103497050A CN103497050A CN201310444658.7A CN201310444658A CN103497050A CN 103497050 A CN103497050 A CN 103497050A CN 201310444658 A CN201310444658 A CN 201310444658A CN 103497050 A CN103497050 A CN 103497050A
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- phosphate solubilizing
- salt tolerant
- solubilizing bacteria
- proline
- salt
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Abstract
The invention discloses a method for preparing a salt tolerant phosphate solubilizing seed coating. The method belongs to the field of biotechnology and can be applied in such fields as agriculture and forestry, gardening, etc. The seed coating contains salt tolerant phosphate solubilizing bacteria which is capable of solubilizing insoluble phosphorus in soil to provide available phosphorus for plants. The method comprises the following steps: screening salt tolerant strains from saline alkali soil, and determining the salt tolerant phosphate solubilizing bacteria after verifying the phosphate solubilizing ability, salt tolerance and physiological and biochemical properties; mutating and screening the salt tolerant phosphate solubilizing bacteria by ultraviolet rays and the like, and verifying to obtain a proline deletion mutant; optimizing the fermentation process conditions of a proline mutant of the salt tolerant phosphate solubilizing bacteria; fermenting the proline deletion mutant, stopping the fermentation at the later logarithmic phase and before the stable phase, concentrating the fermentation broth or preparing freeze-dried powder so as to separate the salt tolerant phosphate solubilizing bacteria; and preparing a solution through 3-7% by mass of polyvinyl alcohol and starch (2:1), 0.001-3.0% of sorbitol, 0.001-3.0% of mannitol and 0.001-3.0% of proline, and preparing the seed coating through the solution and 10-20% of phosphate solubilizing bacteria concentrated fermentation broth or fermentation broth freeze-dried powder containing 1*10<5>-1*10<9> CFU, and the like.
Description
Technical field
A kind of preparation method of salt tolerant phosphorus decomposing dressing.The method belongs to biotechnology and agriculture field.Can be widely used in the fields such as agricultural, forestry, gardening.
Background technology
Phosphorus is the main macronutrient of denitrogenating outer restriction crop yield.Phosphorus in soil mainly exists with the form of insoluble phosphorus, yet the phosphorus of this form is the absorption that is not easy to plant.In the saline-alkali soil of high salt, low nutrition, strong surge capability, the available phosphorus phosphorus content in soil is lower especially.Phosphate solubilizing bacteria is that the insoluble phosphorus that a class can be dissolved in soil provides the microorganism that can utilize phosphorus for plant, and it extensively exists and be considered to and play Main Function in the circulation of soil phosphorus in soil.
Summary of the invention
The screening of salt tolerant phosphate solubilizing bacteria; The evaluation of salt tolerant phosphate solubilizing bacteria; The mutagenic obtained proline(Pro) deletion mutant of salt tolerant phosphate solubilizing bacteria; The fermentation technology optimization of salt tolerant phosphate solubilizing bacteria; Separation of fermentative broth salt tolerant phosphate solubilizing bacteria; Adopt membrane-forming agent, plant anti-salt agent (polyvalent alcohol, proline(Pro)), salt tolerant phosphate solubilizing bacteria Fermented Condensed liquid or salt tolerant phosphate solubilizing bacteria lyophilized powder etc. to prepare the kind clothing of seed.
Embodiment
The screening of 1 salt tolerant phosphate solubilizing bacteria: sample in saltings, through enrichment culture, primary dcreening operation salt tolerant bacterial strain; Through multiple sieve screening high yield phosphorus decomposing bacterial strain.
The evaluation of 2 salt tolerant phosphate solubilizing bacterias: the nucleotide sequencing of the Physiology and biochemistry of bacterial strain, 16s or 18s rRNA gene is identified phosphate solubilizing bacteria than reciprocity means.
The mutagenic obtained proline(Pro) deletion mutant of 3 salt tolerant phosphate solubilizing bacteria: phosphate solubilizing bacteria, through factor mutagenesis such as ultraviolet rays, by print contrasting, screens the proline synthesis deletion mutant in perfect medium and proline(Pro) disappearance substratum; Further such mutant is carried out the mensuration of the characteristics such as salt tolerant, phosphorus decomposing; Optimize the kind clothing moiety of phosphorus decomposing in salt affected soil, growth.
The fermentation technology optimization of 4 salt tolerant phosphate solubilizing bacterias: selected mutant is carried out to the optimization of phosphorus decomposing and growth fermention medium and condition;
5 separation of fermentative broth salt tolerant phosphate solubilizing bacterias: enter the logarithmic phase later stage in fermentation, stationary phase early stage is the fermented liquid precipitation, or filter and obtain thalline and ferment filtrate.
6 adopt membrane-forming agent, plant anti-salt agent (polyvalent alcohol), concentrated broth preparation kind of clothing.
6.1 the selection of membrane-forming agent
Seed coat agent is a kind of for the crop seed dressing, has the material of film forming characteristics, can directly or after dilution be coated on seed-coat, forms the preparation of the protective membrane with some strength and permeability, and film-forming properties is the key characteristic of seed coat agent.Good film forming properties makes seed coat agent wrap in equably the rapid film-forming of seed-coat, does not come off, non-caked.China studies at present and applies film forming matter preferably Xylo-Mucine, polystyrene and sodium alginate are arranged.Tested the film-forming properties of 4 kinds of membrane-forming agents in this test, the membrane-forming agent film-forming properties best (table 1) that wherein polyvinyl alcohol and polyvinyl alcohol+starch are made.The characteristics such as starch has wide material sources as a kind of natural matter, and cost is low.And, the biotype fertilizer based on microorganism, starch is suitable as carrier more.Because starch not only can be used as the support substance of activeconstituents, can also provide necessary carbon source for microorganism.Other characteristics such as film-forming properties of comprehensive membrane-forming agent again, the membrane-forming agent of salt tolerant phosphorus decomposing bacterial classification clothing has been selected polyvinyl alcohol+starch component.
The selection of table 1 membrane-forming agent
I be illustrated in water soak the 0.5h caudacoria can be from the glass plate complete scraping; Can not be from the glass plate completely scrape the level into II; Membrane-forming agent can not film forming be the III level at the glass plate.
6.2 the ratio of polyvinyl alcohol and starch is selected
Along with the increase of starch content, swelling property and the solvability of film increase gradually, and shorten time of drying, and film-forming properties descends to some extent, and the mechanical property of film descends.The increase of starch proportion can reinforcing membrane water suction and water-retentivity, reduce the dry time used.But also can reduce the mechanical property (table 2) of film.Considering cost and on the impact of microorganism, finally selected polyvinyl alcohol: starch is 2: 1.
The characteristic of the polyvinyl alcohol of table 2 different ratios+starch membrane-forming agent
I be illustrated in water soak the 0.5h caudacoria can be from the glass plate complete scraping; Can not be from the glass plate completely scrape the level into II; Membrane-forming agent can not film forming be the III level at the glass plate.
6.3 determining of other compositions of seed coat agent
The chemical composition of film seed coat agent is divided into active ingredient and inactive ingredients.Active ingredient comprises the effective constituents such as sterilant, sterilant, growth regulator, micro-fertilizer and microorganism.Inactive ingredients is exactly the auxiliary agent that matches with it, comprises membrane-forming agent, permeate agent, diffusant, floatation agent, emulsifying agent, frostproofer, sanitas, pH adjusting agent, defoamer, warning colorant etc.This paper seed coat agent composition is as follows:
Table 3 seed coat agent composition
6.4 preparation flow
(1) get a certain amount of polyvinyl alcohol and starch (2: 1) in large beaker, adding distilled water to regulate solid content is 5%.Stirring and evenly mixing, at room temperature place 2 hours, makes the abundant water absorption and swelling of polyvinyl alcohol and starch.
(2) 90 ℃ of lower stirring heating, solution is fully dissolved solution, mix, until solution becomes translucent thick liquid.Add the CMC of 0.5% solid content as thickening material, 0.5% glycerine, as plasticising, continues heating 10h again.
(3) be cooled to 50 ℃ of left and right to add trace element, microorganism growth buffering salt, main nutrient composition and anti-salt agent, add biological stain (lemon yellow and leaf mustard are red).
(4) add microorganisms cultures (microorganism powder or Fermented Condensed liquid), make the biomass of microorganism in seed coat agent 1 * 10
5-1 * 10
9between.
(5) seed coat agent is mixed with seed by the solids content ratio of 1: 40, film forming, dry, pack.
6.5 the quality control index of seed coat agent is measured
PH value, viscosity, coating uniformity, dressing expulsion rate etc. are that the important quality of seed coat agent is controlled index, are directly determining stability and dressing and the effect of seed coat agent.As shown in Table 4, the pH value of the seed coat agent of this preparation is 6.84, is slightly acidic, and slightly acidic more is conducive to the sprouting of seed.The viscosity height directly affects dressing fastness and product stability.Viscosity is too little, and seed coat agent is difficult for adhering to, and the clothing film easily comes off; Viscosity is too large, is difficult to dressing even, easily causes the seed adhesion.The about 240mPas of seed coat agent viscosity prepared by this paper
-1, meet the viscosity requirement of seed coat agent.The degree of uniformity that when height of coating uniformity means dressing, the clothing film distributes at seed-coat.Coating uniformity is higher, and the distribution of clothing film is more even, and activeconstituents is evenly distributed on seed, is difficult for producing poisoning.The coating uniformity of the seed coat agent of this test preparation has all reached more than 90%.The height of dressing expulsion rate means that clothing film after dressing is attached to the firm degree of seed-coat.The seed coat agent expulsion rate of preparation is 3%.Illustrate that its ready-made clothes film adheres at seed-coat more firm, be difficult for coming off because of operations such as storings.
The quality index of table 4 seed coat agent
Claims (3)
1. Selection of Salt-Tolerant bacterial strain from saltings is defined as the salt tolerant phosphate solubilizing bacteria after phosphorus decomposing ability, salt resistance ability and Physiology and biochemistry are identified, with ultraviolet ray etc., the salt tolerant phosphate solubilizing bacteria is carried out to mutagenesis, screening and the deletion mutant of acquisition proline(Pro) after identifying.
2. optimize the technological condition for fermentation of salt tolerant phosphate solubilizing bacteria proline(Pro) mutant, carry out the fermentation of proline(Pro) deletion mutant, before entering logarithmic phase later stage and stationary phase, stop fermentation concentrated broth or lyophilized powder processed to separate the salt tolerant phosphate solubilizing bacteria.
3. adopting mass percent is that 3-7% polyvinyl alcohol+starch (2:1), 0.001-3.0% sorbyl alcohol, 0.001-3.0% N.F,USP MANNITOL, 0.001-3.0% proline(Pro) are made solution, with 10-20% containing 1 * 10
5-1 * 10
9cFU phosphate solubilizing bacteria concentrated broth or fermented liquid lyophilized powder etc. prepare seed pelleting.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201310444658.7A CN103497050A (en) | 2013-09-26 | 2013-09-26 | Method for preparing salt tolerant phosphate solubilizing seed coating |
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| CN201310444658.7A CN103497050A (en) | 2013-09-26 | 2013-09-26 | Method for preparing salt tolerant phosphate solubilizing seed coating |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104431355A (en) * | 2014-11-24 | 2015-03-25 | 青岛科技大学 | Method for preparing lactic acid bacterium granule capsule |
| CN109134055A (en) * | 2017-06-19 | 2019-01-04 | 许传高 | A kind of compound fertilizer and its preparation process comprising humic acid |
| CN109134054A (en) * | 2017-06-19 | 2019-01-04 | 许传高 | A kind of biodegradable microballoon coat fertilizer |
| CN109134128A (en) * | 2017-06-28 | 2019-01-04 | 许传高 | The technique for preparing soil conditioning fertilizer using waste gourmet powder fermenting liquor |
-
2013
- 2013-09-26 CN CN201310444658.7A patent/CN103497050A/en active Pending
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104431355A (en) * | 2014-11-24 | 2015-03-25 | 青岛科技大学 | Method for preparing lactic acid bacterium granule capsule |
| CN104431355B (en) * | 2014-11-24 | 2017-05-31 | 青岛科技大学 | A kind of preparation method of lactic acid bacterium particle capsule |
| CN109134055A (en) * | 2017-06-19 | 2019-01-04 | 许传高 | A kind of compound fertilizer and its preparation process comprising humic acid |
| CN109134054A (en) * | 2017-06-19 | 2019-01-04 | 许传高 | A kind of biodegradable microballoon coat fertilizer |
| CN109134128A (en) * | 2017-06-28 | 2019-01-04 | 许传高 | The technique for preparing soil conditioning fertilizer using waste gourmet powder fermenting liquor |
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Application publication date: 20140108 |