CN104276882B - Combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria, and preparation method and application thereof - Google Patents
Combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria, and preparation method and application thereof Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
- C05G3/80—Soil conditioners
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Abstract
The invention discloses a combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria, which comprises the following components in parts by weight: the weight ratio of the phosphate solubilizing bacteria to the potassium solubilizing bacteria to the polyglutamic acid is 0.1-1: 0.001-0.1. The invention also discloses a preparation method and application of the fertilizer synergist. The phosphate solubilizing bacteria can effectively release phosphate radical ions (PO) in soil or fertilizer4 3+) The potassium bacteria can effectively release potassium ions (K) in soil or fertilizer+) After the polyglutamic acid is compounded with the phosphate solubilizing bacteria and the potassium solubilizing bacteria, a better growth environment can be provided for the survival of the phosphate solubilizing bacteria and the potassium solubilizing bacteria, the nutrient elements released by the phosphate solubilizing bacteria and the potassium solubilizing bacteria can be fully chelated, and the synergistic effect can make the phosphate solubilizing effect and the potassium solubilizing effect more prominent. The combined fertilizer synergist can be used in combination with a fertilizer, so that the utilization rate of the fertilizer is improved, and the combined fertilizer synergist can also be directly applied as a bacterial fertilizer.
Description
Technical Field
The invention relates to a combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria, and a preparation method and application thereof, and belongs to the technical field of biological preparations.
Background
Fertilizers play an increasingly important role in modern agricultural production. China is a big country for using chemical fertilizers, the annual dosage is more than 5000 ten thousand tons, and important contribution is made to agricultural development and grain yield increase. However, the excessive use of chemical fertilizer not only causes a great amount of economic loss, but also brings many adverse consequences to our living environment, such as soil hardening, agricultural product pollution, water quality pollution and greenhouse effect. How to effectively improve the utilization rate of the fertilizer and reduce the using amount of the fertilizer becomes a key subject which needs to be solved urgently in agricultural development in China.
The fertilizer synergist is a novel agrochemical preparation which can adjust the physical and chemical properties of soil and improve the absorption and utilization of fertilizer nutrients by crops, and has the long-acting property of biological fertilizer, the stable effect of organic fertilizer and the synergistic property of trace fertilizer. In recent years, polymers have been increasingly studied as fertilizer synergists. The high molecular polymer fertilizer synergist has the advantages of small addition amount, low use cost, obvious growth promoting effect and the like.
The phosphate solubilizing bacteria can decompose insoluble phosphate minerals in soil, release phosphorus in the insoluble phosphate minerals or decompose organic matters in the soil, and change organic phosphorus into inorganic phosphorus which can be absorbed and utilized by plants, thereby providing effective phosphorus for crops.
The potassium-dissolving bacteria are bacteria which are separated from soil and can differentiate aluminosilicate and apatite minerals, can be used as microbial fertilizers, can decompose insoluble aluminosilicate inorganic minerals such as potassium feldspar and apatite, promote nutrient elements such as insoluble potassium, phosphorus, silicon and magnesium to be converted into soluble nutrients, increase the content of quick-acting nutrients in soil, promote crop growth and development and improve yield.
Polyglutamic acids contain a terminal amine and multiple COOs-NH of radical, nitrogen fertilizers4 +PO in phosphate fertilizer4 3+K in potash fertilizer+Ca of elemental fertilizers2+、Mg2+、Zn2+The polyglutamic acid can be directly bonded with the carboxyl site and the amido site of the polyglutamic acid to form water-soluble and stable polyglutamate, and the water-soluble and stable polyglutamate can slowly provide nutrition for the growth of crops. The phosphate solubilizing bacteria can effectively release PO in soil or fertilizer4 3+The potassium bacteria can effectively release K in soil or fertilizer+The polyglutamic acid can generate a synergistic interaction effect after being compounded with the phosphate solubilizing bacteria and the potassium solubilizing bacteria, not only can provide a better growth environment for the survival of the phosphate solubilizing bacteria and the potassium solubilizing bacteria, but also can fully chelate the phosphate solubilizing bacteria and the potassium solubilizing bacteria to releaseThe nutrient elements make the effect of dissolving phosphorus and potassium more prominent.
Disclosure of Invention
The invention aims to solve the technical problem of providing a combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria.
The technical problem to be solved by the invention is to provide a preparation method of the combined fertilizer synergist.
The invention finally aims to solve the technical problem of providing the application of the combined fertilizer synergist in fertilizer technology and agricultural planting.
In order to solve the technical problems, the technical scheme adopted by the invention is as follows:
a combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria comprises the following components in parts by weight: phosphate solubilizing bacteria, potassium solubilizing bacteria and polyglutamic acid, wherein the weight ratio of the phosphate solubilizing bacteria to the potassium solubilizing bacteria to the polyglutamic acid is 0.1-1: 0.001-0.1;
the phosphate solubilizing microbial inoculum is classified and named as Bacillus subtilis JT-1, is preserved in China Center for Type Culture Collection (CCTCC) and has a preservation address as follows: wuhan university, zip code: 430072. the strain preservation number is CCTCC M2014367, and the preservation date is 2014, 8 and 5 days. The strain is a strain bred by an inventor in a phosphorite soil sample in a Yichang bealock area in Hubei province. The high-efficiency phosphate solubilizing bacteria of the invention can be cultured for 7 days at 30 ℃ and 150rpm by shaking, and the concentration of the phosphate solubilizing bacteria can be 5.0g/LCa3(PO4)2Dissolving phosphorus, wherein the content of water-soluble phosphorus is 682.43-725.28 mg/L.
Strain JT-1 has the following properties:
1. morphological characteristics of colonies:
after culturing for 2-3 days at 30 ℃ on a peptone agar culture medium, the vegetative cells are observed to be unicellular and rod-shaped with the size of 0.5-3 multiplied by 0.5-3 mu m and have elliptic spores under a microscope. The cells were cultured in the above medium at 30 ℃ for 12 hours to allow mass growth. The bacterial colony is white, round, smooth in surface, sticky and convex in the middle.
2. Physiological and biochemical properties:
(1) the culture temperature is as follows: 25-38 ℃, and the optimal temperature is 30-32 ℃;
(2) growing at pH5.0-9.0;
(3) gram staining: positive;
(4) and (3) VP reaction: positive;
(5) starch hydrolysis test: positive;
(6) nitrate reduction reaction: positive;
(7) citric acid utilization: positive;
(8) liquefaction of gelatin: positive;
(9) assay for Phenylpropanoic acid deaminase: negative;
(10) M.R.: negative;
(11) urease test: negative;
(12) propionate utilization: negative;
3. 16S rDNA sequence analysis:
the 16S rDNA most sequence is 1418bp, as shown in SEQ ID No: 1 is shown. And (3) comparing the tested sequences with related species in a GeneBank database to construct a phylogenetic tree based on the 16S rDNA full sequence. The results show that: the strain and the bacillus subtilis reach 100 percent of homology. Therefore, the Bacillus subtilis (Bacillus subtilis) is considered to be used in the invention, in particular to the Bacillus subtilis JT-1.
The potassium-solubilizing bacterium is classified and named as Bacillus subtilis JT-2, and is preserved in China Center for Type Culture Collection (CCTCC) at a preservation address: wuhan university, zip code: 430072. the preservation number of the strain is CCTCCM 2014368, and the preservation date is 2014, 8 and 5 days. The strain is a strain bred by an inventor in mineral soil of Shanxi Lingxian county purple Jinshan area rich in potassium ore. The potassium bacteria of the invention treat potassium ore powder, and the content of decomposed water-soluble potassium ions is 118-132 mg/L after the potassium ore powder is cultured for 25 days at 37 ℃.
Bacillus subtilis JT-2 has the following properties:
1. morphological characteristics of colonies:
after culturing for 2-3 days at 30 ℃ on a peptone agar culture medium, the vegetative cells are observed to be unicellular and rod-shaped with the size of 0.5-3 multiplied by 0.5-3 mu m and have elliptic spores under a microscope. The cells were cultured in the above medium at 30 ℃ for 12 hours to allow mass growth. The bacterial colony is white, round, smooth in surface, sticky and convex in the middle.
2. Physiological and biochemical properties:
(1) the culture temperature is as follows: 25-38 ℃, and the optimal temperature is 30-32 ℃;
(2) growing at pH5.0-9.0;
(3) gram staining: positive;
(4) and (3) VP reaction: positive;
(5) starch hydrolysis test: positive;
(6) nitrate reduction reaction: positive;
(7) citric acid utilization: positive;
(8) liquefaction of gelatin: positive;
(9) assay for Phenylpropanoic acid deaminase: negative;
(10) M.R.: negative;
(11) urease test: negative;
(12) propionate utilization: negative;
3. 16S rDNA sequence analysis:
the 16S rDNA most sequence is 1413bp, as shown in SEQ ID No: 2, respectively. And (3) comparing the tested sequences with related species in a GeneBank database to construct a phylogenetic tree based on the 16S rDNA full sequence. The results show that: the strain and the bacillus subtilis reach 100 percent of homology. Therefore, the Bacillus subtilis (Bacillus subtilis) is considered to be used in the invention, in particular to the Bacillus subtilis JT-2.
Wherein the polyglutamic acid is a pure polyglutamic acid product, a pure polyglutamate product, a polyglutamic acid fermentation liquid or a polyglutamic acid fermentation liquid dried product, and the average molecular weight of the polyglutamic acid is 5-3000 KDa. The adding amount of the polyglutamic acid is calculated by the polyglutamic acid contained in a pure polyglutamic acid product, a pure polyglutamate product, a polyglutamic acid fermentation liquid or a polyglutamic acid fermentation liquid dried product.
The pure polyglutamate product comprises polyglutamate chelated with one or more elements of Na, K, Ca, Mn, Cu, Zn, Fe and the like.
The polyglutamic acid solution comprises polyglutamic acid gel aqueous solution, and polyglutamic acid or polyglutamate pure aqueous solution.
The powder containing polyglutamic acid comprises a powdery dry product of polyglutamic acid fermentation liquor, a pure polyglutamic acid product or a combined powder prepared by the pure polyglutamic acid product and other auxiliary material components.
The fermentation culture containing polyglutamic acid comprises polyglutamic acid fermentation liquor containing a polyglutamic acid production strain and a fermentation medium, and concentrated liquor or dried product of the polyglutamic acid fermentation liquor.
The relevant products of the above-mentioned polyglutamic acid are commercially available or can be prepared according to the prior art.
The polyglutamic acid of the present invention is preferably gamma-polyglutamic acid.
The combined fertilizer synergist is characterized in that auxiliary materials are added, the auxiliary materials are any one or more of bentonite, fulvic acid, dolomite, potassium feldspar powder, calcium phosphate and calcium carbonate powder, and the weight of the auxiliary materials accounts for 5% -40% of the total weight of the combined fertilizer synergist.
Wherein the combined fertilizer synergist is in the form of paste, water or powder.
The preparation method of the combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria comprises the following steps:
(1) the phosphate solubilizing bacteria are prepared into the microbial inoculum by the following method:
activating the phosphate solubilizing bacteria preserved on the inclined plane, inoculating the phosphate solubilizing bacteria on an activation culture medium, and culturing for 18-24 h; selecting strains on an activation culture medium, inoculating the strains in a fermentation culture medium, and culturing at the temperature of 30-35 ℃ for 12-24 hours to obtain fermentation liquor or drying the fermentation liquor to obtain powder;
(2) the potassium bacteria are prepared into a microbial inoculum by the following method:
activating the potassium bacteria preserved on the inclined plane, inoculating the activated potassium bacteria on an activation culture medium, and culturing for 18-24 hours; selecting strains on an activation culture medium, inoculating the strains in a fermentation culture medium, and culturing at the temperature of 30-35 ℃ for 12-24 hours to obtain fermentation liquor or drying the fermentation liquor to obtain powder;
(3) and (3) uniformly mixing the microbial inoculum obtained in the step (1), the microbial inoculum obtained in the step (2) and polyglutamic acid according to the weight ratio of 0.1-1: 0.001-0.1.
In the steps (1) and (2), the activation culture medium is an LB culture medium, and the fermentation culture medium comprises the following components: 10-100g/L of carbon source, 1-40g/L of nitrogen source, 0.01-50g/L of inorganic salt, water as solvent and pH of 5.0-9.0.
Wherein the carbon source is any one or combination of more of glucose, sucrose, maltose, lactose, xylose, fructose, lactic acid, citric acid, glycerol, starch and molasses; the nitrogen source is yeast powder, beef extract, peptone, yeast extract, corn steep liquor, bean cake powder, cottonseed cake powder, urea, (NH)4)2SO4、NH4Cl、(NH4)2HPO4And NH4NO3Any one or a combination of several of them; the inorganic salt is one or a combination of more of sulfate, phosphate, dihydrogen phosphate and hydrochloride.
The number of thalli in the fermentation liquor obtained in the steps (1) and (2) can reach 1 multiplied by 1010-9×1012CFU/ml。
In the step (3), auxiliary materials can be added, wherein the auxiliary materials are one or more of bentonite, fulvic acid, dolomite, potassium feldspar powder, calcium phosphate and calcium carbonate powder, and the weight of the auxiliary materials accounts for 5-40% of the total weight of the combined fertilizer synergist.
The application of the compound fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria in crop growth is also within the protection scope of the invention.
The combined fertilizer synergist is used in combination with a nitrogen fertilizer, a potassium fertilizer, a phosphate fertilizer or the like, or the combined fertilizer synergist is directly applied as a bacterial fertilizer.
Wherein, the crops are grain crops, vegetable crops or melon and fruit crops.
Wherein the implementation mode is water culture, foliage spray, root irrigation or hole application.
Polyglutamic acids contain a terminal amine and multiple COOs-NH of radical, nitrogen fertilizers4 +PO in phosphate fertilizer4 3+K in potash fertilizer+Ca of elemental fertilizers2+、Mg2+、Zn2+The polyglutamic acid can be directly bonded with the carboxyl site and the amido site of the polyglutamic acid to form water-soluble and stable polyglutamate, and the water-soluble and stable polyglutamate can slowly provide nutrition for the growth of crops. The phosphate solubilizing bacteria JT-1 can effectively release PO in soil or fertilizer4 3+The potassium bacteria JT-2 can effectively release K in soil or fertilizer+The polyglutamic acid can generate a synergistic interaction effect after being compounded with the phosphate solubilizing bacteria JT-1 and the potassium solubilizing bacteria JT-2, so that a better growth environment can be provided for the survival of the phosphate solubilizing bacteria JT-1 and the potassium solubilizing bacteria JT-2, the nutrient elements released by the phosphate solubilizing bacteria JT-1 and the potassium solubilizing bacteria JT-2 can be fully chelated, and the phosphate solubilizing and potassium solubilizing effects are more prominent.
Has the advantages that: compared with the prior art, the invention has the following advantages:
1. the phosphate solubilizing bacteria JT-1 has strong inorganic phosphorus solubilizing capability, can improve the bioavailability of indissolvable phosphorus in soil and the utilization efficiency of phosphate fertilizer, saves fertilizer, increases yield and improves soil structure.
2. The potassium bacteria JT-2 has strong potassium dissolving capacity, has strong potassium dissolving activity, and can improve the soil microenvironment and promote the growth of plants.
3. The polyglutamic acid is produced by microbial fermentation, is formed by homopolymerization of D-glutamic acid and L-glutamic acid monomers through glutamine bonds, has the advantages of no toxicity, biodegradability, environmental friendliness and the like, and has the effects of improving the absorption of plants to nutrients and the like.
4. The phosphate solubilizing bacteria, the potassium solubilizing bacteria and the polyglutamic acid can generate a synergistic interaction effect after being compounded. The combined fertilizer synergist can be used together with a fertilizer to improve the utilization rate of the fertilizer, and can also be directly applied as a bacterial fertilizer.
5. The combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria can obviously improve the yield of grain crops such as wheat and rice, leaf vegetables such as rape, melons, grapes and other melons and fruits and economic crops such as autumn tea by 25-65%. Under the condition of ensuring that the crop yield is not reduced, the application of 20-40% of phosphate fertilizer and potash fertilizer by mass can be reduced. The method can also obviously improve the content of protein and starch in grains of grain crops and the content of Vc in leaf vegetables, reduce the content of nitrate, improve the content of various sugars in melon and fruit crops, change the content of flavor substances in tea leaves and improve the quality of crops. Meanwhile, the expression level of antioxidant enzyme and stress resistance related genes of the plant is improved, and the stress resistance of the plant is enhanced, so that the growth of the plant is promoted.
Detailed Description
The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the description of the embodiments is only for illustrating the present invention and should not be taken as limiting the invention as detailed in the claims.
Example 1:
the phosphate solubilizing bactericide is prepared by the following steps:
activating Bacillus subtilis JT-1 stored at 4 ℃ on a slant, inoculating the activated Bacillus subtilis JT-1 on an eggplant-shaped culture flask culture medium (namely an activation culture medium), and culturing for 18-24 h. Transferring the grown strain to fermentation medium, culturing at 30-35 deg.C for 12-24 hr to obtain fermentation liquid or dried product of fermentation liquid, wherein the thallus number in the microbial inoculum can reach 1 × 1010-9×1012CFU/g。
The culture medium in the eggplant-shaped culture bottle is an LB culture medium, and the fermentation culture medium comprises the following components: glucose 10g/L, (NH)4)2SO40.5g/L，NaCl 0.3g/L，MnSO4·4H2O 0.03g/L，KH2PO41g/L, and the pH value is 6.0-8.0.
The aeration quantity of the sterile air in the fermentation culture process of the production tank is 0.6-0.8vvm, and the stirring speed is 180-200 rpm.
Example 2: and (4) measuring the phosphate solubilizing capability of the strain.
Respectively adding 100mL of inorganic phosphorus liquid culture medium into 500mL triangular flasks, wherein the culture medium comprises 10g/L (NH) of glucose4)2SO41g/L，MgSO4·7H2O 0.3g/L，NaCl 0.3g/L，FeSO4·7H2O 0.03g/L，MnSO4·4H2O0.03g/L，Ca3(PO4)25.0 g/L. Then inoculate 1mL of bacterial suspension Bacillus subtilis JT-1, and at the same time, do not inoculate the control (1 mL of sterile water is added into the culture medium), all processes are repeated for 3 times. Shaking and culturing at 30 deg.C and 150rpm for 7d, centrifuging the culture solution at 6000rpm for 20min, and determining phosphorus dissolving amount (water soluble phosphorus content) of 682.43-725.28mg/L by molybdenum-antimony colorimetry, and phosphorus dissolving rate of 68.2% -72.5%. The phosphate solubilizing bacterium JT-1 has higher phosphate solubilizing capability.
Example 3:
the potassium-decomposing microbial inoculum is prepared by the following steps:
activating Bacillus subtilis JT-2 stored at a 4 ℃ inclined plane, inoculating the activated Bacillus subtilis JT-2 to an eggplant bottle culture medium (namely an activation culture medium), and culturing for 24 hours. Selecting a large amount of strains from eggplant bottle, inoculating into fermentation culture medium, culturing at 30 deg.C for 16 hr to obtain fermentation broth or dried product of fermentation broth, wherein the thallus number in the strain can reach 1 × 1010-9×1012CFU/g。
The culture medium in the eggplant-shaped culture bottle is an LB culture medium, and the fermentation culture medium comprises the following components: glucose 8-12g/L, (NH)4)2SO40.05-1g/L，NaCl 0.1-1g/L，MnSO4·4H2O 0.1-0.5g/L，KH2PO40.5-2.0g/L and pH 6.0-8.0.
The ventilation quantity of sterile air in the fermentation culture process of the production tank is 0.6-0.8vvm, the stirring speed is 180-200rpm, the culture temperature is 30-32 ℃, and the culture time is 12-16 h.
Example 4: and (4) measuring the potassium-dissolving capability of the strain.
Preparing a potassium dissolving culture medium (without water-soluble potassium ions), subpackaging 95mL of the potassium dissolving culture medium, adding 0.5g of accurately weighed potassium ore powder into a 250mL triangular flask, sterilizing the flask for 30min at 121 ℃, inoculating 5% of inoculum size of bacterial suspension to be detected Bacillus subtilis JT-2 into shake flask strains, and setting no-inoculation control while making three strains in parallel. And (3) finishing the culture after the culture is carried out for 25 days by using a water-bath shaking table at 37 ℃, centrifuging the culture solution for 5min at 1000r/min at 4 ℃, sucking 4mL of centrifuged supernatant by using a pipette, fully and uniformly mixing the centrifuged supernatant with 4mL of 6mmoL/LiCl, and measuring potassium by using a flame spectrophotometer method to obtain the water-soluble potassium ion content of 118-132 mg/L, wherein the potassium-dissolving rate reaches 45.4-50.8%, and the potassium-dissolving capacity is strong.
Example 5:
the composite fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria is prepared according to the following method:
1) the fermentation liquor prepared in the example 1 is dried to prepare the phosphate-solubilizing microbial inoculum.
2) The fermentation broth obtained in example 2 is dried to obtain the potassium-solubilizing bacterium agent.
3) The polyglutamic acid is wettable powder containing polyglutamic acid, and is prepared by drying polyglutamic acid fermentation liquor, wherein the content of the polyglutamic acid is 25-30%, and the molecular weight is 500-2000 kDa.
4) The polyglutamic acid (calculated by pure polyglutamic acid), the phosphate solubilizing bacteria agent, the potassium solubilizing bacteria agent and the auxiliary material (formed by mixing dolomite and calcium carbonate powder according to the weight ratio of 1: 1) are prepared into the compound fertilizer synergist containing the polyglutamic acid, the phosphate solubilizing bacteria and the potassium solubilizing bacteria according to the weight ratio of 0.05:1:1: 0.8.
Example 6: the combined fertilizer synergist has better effect
In this example, the preparation of the phosphate solubilizing agent, the potassium solubilizing agent and the polyglutamic acid is the same as that in example 5. The rice is used as an experimental object to carry out a cell test, the test is provided with 4 treatments, each treatment is repeated for 3 times, the cell length is 8m, the width is 3m, and the cell area is 24m2Protection rows are arranged around the cell, and the specific test treatment is shown in table 1. All the treatments are applied as base fertilizers, and the dosage of the common compound fertilizer is 450kg/hm2The synergist (one or more of phosphate solubilizing bacteria, potassium solubilizing bacteria and polyglutamic acid) is mixed with the compound fertilizer and then applied, and the addition amount of the synergist is shown in table 1.
TABLE 1 design of rice trials
TABLE 2 Effect of the treatments on the yield of Rice and its constituents
Note: values are expressed as mean ± sd, with different lower case letters indicating significant difference after the same column of data (P <0.05)
As can be seen from Table 2, the effective spike number, thousand grain weight and rice yield of the rice can be improved by applying the polyglutamic acid and the common fertilizer together and applying the potassium solubilizing bactericide and the common fertilizer together, and the yield is increased by 20.0% and 22.2% respectively compared with the case of applying the common fertilizer only. When the combined fertilizer synergist is applied, the yield of rice can be obviously improved by 55.1%. This demonstrates that the combined fertilizer synergist has a more pronounced additive yield increasing effect than the additive yield increasing effect of all synergistic components therein applied separately.
Example 7: the combined fertilizer synergist can reduce applied fertilizers.
In this example, the preparation of the compound fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria was the same as that in example 5. The method is characterized in that potted pakchoi is taken as an experimental object, and 3 treatments are set for experiments, wherein the treatment 1 is an experiment contrast common fertilizer, the treatment 2 is a common fertilizer plus a combined fertilizer synergist, and the treatment 3 is a common fertilizer plus a combined fertilizer synergist with the application amount of 30% by mass. The fertilizer applied was 150mg per kg of soil.
TABLE 3 Effect of the Combined Fertilizer synergists on growth characteristics and yield of potted pakchoi
The combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria is applied in a mixing manner with the fertilizer, can promote the growth of the roots of the pakchoi, thicken and enlarge the roots, has obvious promotion effects on the plant height, the fresh weight of the underground part and the fresh weight of the overground part of the pakchoi, and can increase the yield by 46.8%. When 30% of fertilizer is applied in a reduced amount, the yield of the cabbage can still be improved by 14.9%, which shows that the combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria can effectively reduce the use amount of the fertilizer.
Example 8: the combined fertilizer synergist obviously improves the utilization rate of the fertilizer.
In this example, the preparation of the compound fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria was the same as that in example 5.
And (3) taking corn as an experimental object to perform a cell corn test. The experiment designs 3 treatments, wherein the treatment 1 is no fertilization, the treatment 2 is the application of a common compound fertilizer, and the treatment 3 is a common fertilizer plus a combined fertilizer synergist. The length of the cell is 8m, the width is 2.5m, and the area of the cell is 20m2And protective rows are arranged around the cell. The applied fertilizer was 1050kg/hm2The dosage of the combined fertilizer synergist containing polyglutamic acid, phosphate-solubilizing bacteria and potassium-solubilizing bacteria is 80kg/hm2。
The utilization rate of the combined fertilizer synergist of the invention to the fertilizer is calculated by comparing the nitrogen, phosphorus and potassium absorption amount of the harvested corn plants with the application amount of the nitrogen fertilizer, the phosphorus fertilizer and the potassium fertilizer before sowing, and the test result is as follows:
TABLE 4 Effect of the combination Fertilizer synergist on the utilization of the fertilizer
As can be seen from Table 4, compared with the common compound fertilizer, the combined fertilizer synergist of the invention respectively improves the utilization rate of corn N, P, K by 41.8%, 29.3% and 41.2%. Therefore, the combined fertilizer synergist can obviously promote the yield increase of grain crops and improve the utilization rate of the fertilizer.
Example 9: the combined fertilizer synergist can obviously improve the quality of crops.
In this example, the preparation of the compound fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria was the same as that in example 5.
The melon was used as the test subject, and the test was performed 3 times in total. Treatment 1 is the application of a common compound fertilizer (450 kg/ha)2) And the treatment 2 is to apply a common compound fertilizer (450 kg/ha)2) + combined fertilizer synergist and treatment3, reducing the application of 30 percent of common compound fertilizer and combined fertilizer synergist (315 kg/ha)2) The melon management is daily management.
TABLE 5 Effect of the combination Fertilizer synergist on the Vc and nitrate content of melon
Compared with treatment 1, the combined fertilizer synergist can improve the Vc content by 46.3 percent and reduce the nitrate content by 22.5 percent. When 30% of compound fertilizer is reduced, the Vc content of the combined fertilizer synergist can be increased by 26.8%, the nitrate content can be reduced by 31.3%, and the melon quality can be improved. Test results show that the combined fertilizer synergist not only can reduce the use amount of chemical fertilizers, but also can obviously improve the quality of melons.
Example 10: the combined fertilizer synergist can raise the resistance of plant
In this example, the preparation of the compound fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria was the same as that in example 5.
The small green vegetables are used as experimental objects to perform pot culture, and the experiment is carried out for 3 times of treatment in total. The water control method adopts a Hsiao (1973) mesogenic plant water gradient division method, wherein the treatment 1 is 75-80% of a control (accounting for the percentage of the maximum water holding capacity of the soil), the treatment 2 is 40-45% of moderate stress, and the treatment 3 is 30-35% of severe stress. And (3) normally managing the potted small green vegetables, controlling the moisture before experimental observation, and adding a combined fertilizer synergist into the treatment 2 and the treatment 3 when the moisture gradient reaches 3 moisture gradients, wherein the application amount is 200mg per kilogram of soil. During the period, the volume water content of the soil is measured every day, and the water lost in the day is supplemented, so that each treatment level reaches the set water content.
The results are shown in Table 6:
TABLE 6 Effect of the combination Fertilizer synergist on chlorophyll and fresh weight of Brassica chinensis
The results show that: under the environment condition of moderate drought, the yield of the small green vegetables can be increased by 18.7 percent compared with a control group by applying the combined fertilizer synergist, and under the environment condition of severe drought, the yield of the small green vegetables has no obvious difference compared with the control group by applying the combined fertilizer synergist. The experiment result shows that the combined fertilizer synergist can improve the drought resistance of plants, thereby promoting the growth of the plants.
Claims (10)
1. A combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria is characterized by comprising the following components in parts by weight: phosphate solubilizing bacteria, potassium solubilizing bacteria and polyglutamic acid, wherein the weight ratio of the phosphate solubilizing bacteria to the potassium solubilizing bacteria to the polyglutamic acid is 0.1-1: 0.001-0.1;
wherein the phosphate solubilizing bactericide is classified and named as bacillus subtilis (A), (B)Bacillus subtilis) The strain number is JT-1, the strain is preserved in China center for type culture Collection, the strain preservation number is CCTCC M2014367, and the preservation date is 2014, 8 and 5 days.
Wherein the potassium bacteria are classified and named as bacillus subtilis (A), (B)Bacillus subtilis) The strain number is JT-2, and is preserved in China center for type culture Collection with the strain preservation number of CCTCC M2014368The collection date is 2014, 8 and 5.
2. The combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria according to claim 1, wherein the polyglutamic acid is a pure polyglutamic acid product, a pure polyglutamate product, a polyglutamic acid fermentation broth or a dried polyglutamic acid fermentation broth, and the average molecular weight of the polyglutamic acid is 5-3000 KDa.
3. The combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria according to claim 1, characterized in that an auxiliary material is added to the combined fertilizer synergist, wherein the auxiliary material is any one or more of bentonite, fulvic acid, dolomite, potassium feldspar powder, calcium phosphate and calcium carbonate powder, and the weight of the auxiliary material accounts for 5-40% of the total weight of the combined fertilizer synergist.
4. The combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria as claimed in claim 1, wherein the combined fertilizer synergist is in the form of paste, water or powder.
5. The preparation method of the combined fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria of claim 1, characterized by comprising the following steps:
(1) the phosphate solubilizing bacteria are prepared into the microbial inoculum by the following method:
activating the phosphate solubilizing bacteria preserved on the inclined plane, inoculating the phosphate solubilizing bacteria on an activation culture medium, and culturing for 18-24 h; selecting strains on an activation culture medium, inoculating the strains in a fermentation culture medium, and culturing at the temperature of 30-35 ℃ for 12-24 hours to obtain fermentation liquor or drying the fermentation liquor to obtain powder;
(2) the potassium bacteria are prepared into a microbial inoculum by the following method:
activating the potassium bacteria preserved on the inclined plane, inoculating the activated potassium bacteria on an activation culture medium, and culturing for 18-24 hours; selecting strains on an activation culture medium, inoculating the strains in a fermentation culture medium, and culturing at the temperature of 30-35 ℃ for 12-24 hours to obtain fermentation liquor or drying the fermentation liquor to obtain powder;
(3) and (3) uniformly mixing the microbial inoculum obtained in the step (1), the microbial inoculum obtained in the step (2) and polyglutamic acid according to the weight ratio of 0.1-1: 0.001-0.1.
6. The preparation method according to claim 5, characterized in that an auxiliary material is added in the step (3), the auxiliary material is any one or more of bentonite, fulvic acid, dolomite, potassium feldspar powder, calcium phosphate and calcium carbonate powder, and the weight of the auxiliary material accounts for 5-40% of the total weight of the combined fertilizer synergist.
7. The use of the compound fertilizer synergist containing polyglutamic acid, phosphate solubilizing bacteria and potassium solubilizing bacteria of claim 1 in crop growth.
8. The use according to claim 7, characterized in that the combination fertilizer synergist is used in combination with a nitrogen fertilizer, and or a potassium fertilizer, and or a phosphate fertilizer, or the combination fertilizer synergist is applied directly as a bacterial fertilizer.
9. The use of claim 7, wherein the crop is a food crop, a vegetable crop, or a melon and fruit crop.
10. Use according to claim 7, characterized in that embodiments are hydroponic, foliar spray, root irrigation, or hole application.
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