CN115925491A - Urease inhibitor for acid soil, preparation method, use method and application - Google Patents
Urease inhibitor for acid soil, preparation method, use method and application Download PDFInfo
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- CN115925491A CN115925491A CN202211661486.4A CN202211661486A CN115925491A CN 115925491 A CN115925491 A CN 115925491A CN 202211661486 A CN202211661486 A CN 202211661486A CN 115925491 A CN115925491 A CN 115925491A
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- 239000002689 soil Substances 0.000 title claims abstract description 75
- 239000002601 urease inhibitor Substances 0.000 title claims abstract description 27
- 239000002253 acid Substances 0.000 title claims abstract description 26
- 229940090496 Urease inhibitor Drugs 0.000 title claims abstract description 23
- 238000000034 method Methods 0.000 title claims abstract description 16
- 238000002360 preparation method Methods 0.000 title abstract description 6
- 108010046334 Urease Proteins 0.000 claims abstract description 39
- 230000000694 effects Effects 0.000 claims abstract description 21
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 12
- 239000004220 glutamic acid Substances 0.000 claims description 31
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 claims description 29
- 239000004202 carbamide Substances 0.000 claims description 29
- 101150073660 glmM gene Proteins 0.000 claims description 13
- 101150014144 ureC gene Proteins 0.000 claims description 13
- 108090000623 proteins and genes Proteins 0.000 claims description 9
- 230000000813 microbial effect Effects 0.000 claims description 8
- 238000000354 decomposition reaction Methods 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 230000015572 biosynthetic process Effects 0.000 claims description 2
- 238000000855 fermentation Methods 0.000 claims description 2
- 230000004151 fermentation Effects 0.000 claims description 2
- 230000005764 inhibitory process Effects 0.000 claims description 2
- 230000007246 mechanism Effects 0.000 claims description 2
- 238000000746 purification Methods 0.000 claims description 2
- 238000003786 synthesis reaction Methods 0.000 claims description 2
- 238000012360 testing method Methods 0.000 description 28
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 18
- 238000012795 verification Methods 0.000 description 10
- 229910052757 nitrogen Inorganic materials 0.000 description 9
- 239000003337 fertilizer Substances 0.000 description 8
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 7
- 230000004720 fertilization Effects 0.000 description 7
- HEPPIYNOUFWEPP-UHFFFAOYSA-N n-diaminophosphinothioylbutan-1-amine Chemical compound CCCCNP(N)(N)=S HEPPIYNOUFWEPP-UHFFFAOYSA-N 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 5
- 239000000203 mixture Substances 0.000 description 5
- 239000000618 nitrogen fertilizer Substances 0.000 description 5
- 238000004382 potting Methods 0.000 description 5
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 4
- OTYBMLCTZGSZBG-UHFFFAOYSA-L potassium sulfate Chemical group [K+].[K+].[O-]S([O-])(=O)=O OTYBMLCTZGSZBG-UHFFFAOYSA-L 0.000 description 4
- 229910052939 potassium sulfate Inorganic materials 0.000 description 4
- 235000011151 potassium sulphates Nutrition 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 241000894007 species Species 0.000 description 4
- -1 superphosphate monohydrate Chemical class 0.000 description 4
- 241000589173 Bradyrhizobium Species 0.000 description 3
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 3
- 241000190932 Rhodopseudomonas Species 0.000 description 3
- 229910021529 ammonia Inorganic materials 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 229910000389 calcium phosphate Inorganic materials 0.000 description 3
- 238000012165 high-throughput sequencing Methods 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 235000019691 monocalcium phosphate Nutrition 0.000 description 3
- 230000007935 neutral effect Effects 0.000 description 3
- 235000015097 nutrients Nutrition 0.000 description 3
- 239000011591 potassium Substances 0.000 description 3
- 229910052700 potassium Inorganic materials 0.000 description 3
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 241000208125 Nicotiana Species 0.000 description 2
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 2
- 240000008042 Zea mays Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000014759 maintenance of location Effects 0.000 description 2
- 239000002686 phosphate fertilizer Substances 0.000 description 2
- 239000001103 potassium chloride Substances 0.000 description 2
- 235000011164 potassium chloride Nutrition 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 239000002426 superphosphate Substances 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 1
- 241001678070 Caballeronia Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000970829 Mesorhizobium Species 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 241000862969 Stella Species 0.000 description 1
- PNNCWTXUWKENPE-UHFFFAOYSA-N [N].NC(N)=O Chemical compound [N].NC(N)=O PNNCWTXUWKENPE-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000004178 amaranth Substances 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- YYRMJZQKEFZXMX-UHFFFAOYSA-N calcium;phosphoric acid Chemical compound [Ca+2].OP(O)(O)=O.OP(O)(O)=O YYRMJZQKEFZXMX-UHFFFAOYSA-N 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 238000003421 catalytic decomposition reaction Methods 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000001310 location test Methods 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000010197 meta-analysis Methods 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
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- Soil Conditioners And Soil-Stabilizing Materials (AREA)
- Fertilizers (AREA)
Abstract
The invention relates to the technical field of urease inhibitors, and particularly provides a urease inhibitor for acid soil, a preparation method, a use method and application thereof. The urease inhibitor provided by the invention has stable urease inhibiting effect on acid soil and compensates the problem of poor effect of NBPT on acid soil.
Description
Technical Field
The invention relates to the technical field of urease inhibitors, and particularly provides a urease inhibitor for acid soil, and a preparation method, a use method and application thereof.
Background
Urea is a nitrogen fertilizer with the largest proportion, the total amount of applied global nitrogen fertilizer in 2018 reaches 1.07 hundred million tons, the urea accounts for about 55 percent, the annual capacity of the Chinese nitrogen fertilizer in recent years is about 4000 million tons, and the urea accounts for about 68 percent. Urease can be present at 10 14 The rate of catalytic decomposition of urea into carbon dioxide and ammonia is doubled, and urease is an inducible enzyme and has high degree of activitySpecificity, and only has catalytic action on urea. Under the action of soil urease, urea is rapidly hydrolyzed, the decomposition rate in 15 days reaches 98%, so that the retention time of the urea in the soil is greatly shortened, the utilization rate of urea nitrogen is obviously reduced, the volatilization of soil ammonia is obviously increased, and serious environmental problems are caused.
The urease inhibitor is a substance capable of effectively reducing the activity of urease, and can effectively delay the hydrolysis of urea and prolong the retention time and fertilizer efficiency of urea in soil, thereby improving the utilization efficiency of the urea. The N-butyl thiophosphoric triamide (NBPT) can generate high-efficiency urease inhibiting effect due to low addition amount, is distinguished from a plurality of urease inhibitors, and is the current mainstream urease inhibitor. Results of a global scale Meta-analysis study show that NBPT addition to urea can increase crop yield by 6% on average and reduce ammonia emission by 53%. However, a number of literature studies have shown that NBPT has no significant urease inhibiting effect in acidic soils (pH < 5.5) and no ammonia gas abatement effect. Therefore, it is important to screen substances that can produce a high urease inhibiting effect in acidic soil.
Disclosure of Invention
In order to solve the technical problems, the invention provides a urease inhibitor for acid soil, a preparation method, a use method and application thereof, and solves the problems that a main stream urease inhibitor NBPT only has obvious effect on neutral and alkaline soil and has poor effect on acid soil.
The invention is realized by providing a urease inhibitor for acid soil, which comprises poly-gamma-glutamic acid.
The invention provides a preparation method of the urease inhibitor for the acid soil, which is obtained by microbial fermentation synthesis and purification.
The invention provides a use method of the urease inhibitor for the acid soil, which is characterized in that a certain amount of poly-gamma-glutamic acid and urea are mixed and completely dissolved in water, and the mixture is uniformly poured on the soil surface or directly spread on the soil surface after being mixed.
Preferably, in the above method of use, the effective amount of poly-gamma-glutamic acid added is 0.5 to 1.5% by mass of urea.
The invention provides application of the urease inhibitor for acid soil, which is used for inhibiting urease activity in acid soil so as to inhibit urea decomposition.
Preferably, in the above application, the urease activity inhibiting mechanism of poly-gamma-glutamic acid is: the method has the advantages of reducing the absolute abundance of the urease functional gene ureC, changing the functional gene microbial diversity of the urease functional gene ureC and reducing the relative abundance of dominant species in the urease functional gene ureC microbial community.
Compared with the prior art, the invention has the advantages that:
the urease inhibitor specially used for the acid soil is provided, the main component of the urease inhibitor comprises poly-gamma-glutamic acid, the urease inhibiting effect of the urease inhibitor on the acid soil is stable, and the problem of poor effect of NBPT on the acid soil is solved.
Drawings
The invention is described in further detail below with reference to the following figures and embodiments:
FIG. 1 shows soil urease activity in example 1;
FIG. 2 is a principal coordinate analysis (PCoA, genus level) of ureC functional genes in soil of example 1;
FIG. 3 is the species relative abundance (genus level) of the ureC functional genes in the soil of example 1.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention will be described in further detail with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1 inhibition of urease by poly-gamma-glutamic acid in acid brown soil:
1. and (3) verification test:
the greenhouse pot experiment is carried out at the research institute of the tobacco terrace coastal zone of the Chinese academy of sciences. The potting test soil was representative dry land cultivation layer 0-20cm brown soil (N41 '49', E123 '34' 4 ") collected from a typical brown soil long term location test station in northeast of Shenyang agricultural university, soil pH (water to soil ratio of 1. The fertilization level (N-P2O 5-K2O) of the potting soil is 360-150-300, the addition amount of poly-gamma-glutamic acid is 30mg/kg of soil, the nitrogen fertilizer is urea, the phosphate fertilizer is superphosphate monohydrate, and the potassium fertilizer is potassium sulfate. Uniformly mixing poly-gamma-glutamic acid, urea, calcium superphosphate monohydrate and potassium sulfate, dissolving the mixture in an aqueous solution, uniformly pouring the aqueous solution on the soil surface, and not additionally applying other fertilizers in the later period. Soil was harvested at 1,3,7, 15, 25, 40 and 55 days after fertilization for urease activity, while the absolute abundance (copy number) of the ureC functional gene encoding urease was determined by real-time fluorescent quantitative PCR and the community structure change of the ureC functional gene was determined by high throughput sequencing molecular biology.
2. Comparative experiment 1:
the method is characterized in that the method is carried out in a research institute of a tobacco terrace coastal zone of China academy of sciences, the experimental conditions are consistent with the verification test, only poly-gamma-glutamic acid is not added into the urea, and urea containing nitrogen and other nitrogen in the poly-gamma-glutamic acid is correspondingly supplemented to the soil of the verification test in order to eliminate the influence of nitrogen nutrients of the poly-gamma-glutamic acid.
3. As a result, the
The Urease content in the test soil (UP) is remarkably smaller than the Urease (Urease) activity (13.00% -16.39%) in the soil (U) of the comparative test 1 from day 1 to day 45 (p <0.05, figure 1), and the reduction amplitudes at days 1,3,7, 15, 25 and 45 are respectively 13.04%, 14.01%, 14.47%, 13.83%, 13.00% and 16.39%, which shows that the poly-gamma-glutamic acid is remarkably inhibited and reduced in the Urease activity in the acid brown soil and has remarkable Urease inhibiting effect.
Meanwhile, the determination result of the real-time fluorescent quantitative PCR method shows that the absolute abundance (copy number) of the ureC of the functional gene encoding urease in the soil in the verification test is reduced by 34.50-175.18%, and the addition of poly-gamma-glutamic acid is proved to obviously inhibit the activity of the urease. The results of high throughput sequencing showed unclassified _ d _ Bacteria (19.95%), bradyrhizobium (11.11%), mesorhizobium (5.10%), rhodopseudomonas (2.99%) were the dominant species in soil, PCoA Analysis (ANOSIM) found to verify that the ureC functional gene microbial communities in the test soil (UP) and the comparative test soil (U) could be clearly distinguished (R =1.00, p =0.098, fig. 2), verifying that the relative abundance of Bradyrhizobium, rhodopseudomonas, leptrix R _ ror, pseudomonas brot, caballeronia, stella, variovax in the test soil was significantly reduced (p <0.01, fig. 3). Thus, the assay results of high throughput sequencing indicate that poly-gamma-glutamate addition causes significant changes in the composition of the ureC functional gene microbial community and significantly reduces the relative abundance (genus level) of the dominant species of Bradyrhizobium and Rhodopseudomonas.
Example 2 inhibitory effect of poly-gamma-glutamic acid on urease in neutral brown soil:
1. and (3) verification test:
taking example 1 as an example, the greenhouse potting test was carried out at Shenyang applied ecology research institute of Chinese academy of sciences. The soil for potting test was farmland soil collected from ten miles of towns in Shenyang city, and had a soil pH (1. The fertilization level (N-P2O 5-K2O) of the potting soil is 360-150-300, the addition amount of the poly-gamma-glutamic acid is 300mg/kg of soil, the nitrogen fertilizer is urea, the phosphate fertilizer is calcium superphosphate monohydrate, and the potassium fertilizer is potassium sulfate. Uniformly mixing poly-gamma-glutamic acid with urea, calcium superphosphate monohydrate and potassium sulfate, dissolving the mixture in an aqueous solution, and injecting the solution into soil without applying other fertilizers in the later stage. Soil was collected on days 1,3,7, 15, 30, 45 and 60 after fertilization for urease activity.
2. And (3) comparison test:
the method is applied to ecological research institute of Shenyang of Chinese academy of sciences, and the experimental conditions are consistent with those of verification tests, except that poly-gamma-glutamic acid is not added into urea. In order to eliminate the influence of nitrogen nutrients of the poly-gamma-glutamic acid, the urea with nitrogen content and the like added by the poly-gamma-glutamic acid in the test soil is correspondingly supplemented and verified.
3. Results
The urease activity in the test soil is verified to be significantly higher than that (4.60% -29.67%) in the soil of the comparative test 1 (p is less than 0.05), and the increases at 1 st, 3 rd, 7 th, 15 th, 30 th, 45 th and 60 th days are respectively 12.88%, 27.55%, 9.09%, 29.67% and 4.60%, 20.03% and 23.29%, which shows that the poly-gamma-glutamic acid can significantly increase the urease activity in neutral brown soil and does not have urease inhibition effect.
Example 3 inhibitory Effect of Poly-gamma-glutamic acid on urease in Red soil
1. And (3) verification test:
taking example 1 as an example, a field cell verification test was carried out at the site of an agricultural test base of Yichun institute of Yichun, jiangxi, and 3.6m wide by 6m long cell with an area of 21.6m 2 In the plot, 4 replicates were set, the crop planted was corn, the soil pH (water-soil ratio of 1. Fertilization level (N-P) 2 O 5 -K 2 O) is 225-90-120, and the nitrogen, phosphorus and potassium fertilizers are urea and heavy calcium (45% 2 O 5 ) And potassium chloride, wherein the addition amount of the poly-gamma-glutamic acid is 1 percent of the mass of the urea. And in 2021, 5 and 6 days, uniformly mixing poly-gamma-glutamic acid with urea, triple superphosphate and potassium chloride, then spreading the mixture in furrows, sowing corn after fertilization, and no additional fertilizer is applied in later period. Soil was harvested at 7, 15, 30, 45 days after fertilization (harvest time) for urease activity.
2. And (3) comparison test:
the field test conditions are consistent with the verification test in the agricultural test base of Yichun academy of Yichun city in Yichun, jiangxi, except that poly-gamma-glutamic acid is not added into the fertilizer, and urea with nitrogen content such as nitrogen content and the like added into the poly-gamma-glutamic acid is correspondingly supplemented to the soil of the verification test in order to eliminate the influence of nitrogen nutrients of the poly-gamma-glutamic acid.
3. Results
The content of urease in the test soil is verified to be obviously less than the urease activity (3.97% -31.61%) in the soil of the comparative test 1 from day 1 to day 30, and the reduction amplitudes at days 7, 15, 30 and 40 are respectively 31.61%, 14.50%, 16.22% and 3.97%, which shows that poly-gamma-glutamic acid is effectively inhibited and reduced in the acid red soil, and has obvious urease inhibiting effect.
The embodiments of the present invention have been described in detail with reference to the drawings, but the present invention is not limited to the above embodiments, and various changes can be made within the knowledge of those skilled in the art without departing from the gist of the present invention.
Claims (6)
1. A urease inhibitor for acid soil is characterized by that its component includes poly-gamma-glutamic acid.
2. The method for preparing a urease inhibitor for acid soil according to claim 1, wherein the urease inhibitor is obtained by microbial fermentation synthesis and purification.
3. The method of using urease inhibitor for acid soil according to claim 1, wherein the amount of poly-gamma-glutamic acid and urea are mixed, dissolved completely in water, and either poured homogeneously or mixed and then directly spread on the soil surface.
4. The method of using urease inhibitor for acid soil according to claim 3, wherein the effective amount of poly-gamma-glutamic acid added is 0.5-1.5% of the mass of urea.
5. Use of the urease inhibitor for acid soil according to claim 1 for inhibiting urease activity and thereby urea decomposition in acid soil.
6. The use of the urease inhibitor for acid soil according to claim 5, wherein the urease activity inhibition mechanism of poly-gamma-glutamic acid is: the method has the advantages of reducing the absolute abundance of the urease functional gene ureC, changing the functional gene microbial diversity of the urease functional gene ureC and reducing the relative abundance of dominant species in the urease functional gene ureC microbial community.
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CN101117303A (en) * | 2007-07-09 | 2008-02-06 | 华中农业大学 | Poly-gama-glutamic acid building fertilizer |
CN101157588A (en) * | 2007-09-21 | 2008-04-09 | 石家庄开发区德赛化工有限公司 | Highly effective carbamide and preparation method and application thereof |
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CN109400371A (en) * | 2018-12-05 | 2019-03-01 | 中国科学院沈阳应用生态研究所 | A kind of carbon coupled mode stability compound fertilizer's fertilizer and preparation method |
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Patent Citations (6)
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CN101117303A (en) * | 2007-07-09 | 2008-02-06 | 华中农业大学 | Poly-gama-glutamic acid building fertilizer |
CN101157588A (en) * | 2007-09-21 | 2008-04-09 | 石家庄开发区德赛化工有限公司 | Highly effective carbamide and preparation method and application thereof |
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