CN113788722B

CN113788722B - Biological stimulator and preparation method thereof

Info

Publication number: CN113788722B
Application number: CN202111117374.8A
Authority: CN
Inventors: 韩国涛; 班宜民
Original assignee: Shandong Aiguozhi Biotechnology Co ltd
Current assignee: Shandong Aiguozhi Biotechnology Co ltd
Priority date: 2021-09-23
Filing date: 2021-09-23
Publication date: 2022-09-02
Anticipated expiration: 2041-09-23
Also published as: CN113788722A

Abstract

The invention relates to a biological stimulator and a preparation method thereof, which are characterized by comprising tea saponin and a seaweed extract; the seaweed extracting solution is prepared according to the following steps: adding seaweed and enzyme into water, and performing enzymolysis to obtain seaweed extract; the enzyme at least comprises lignin peroxidase and oxidase, the oxidase obtains hydrogen peroxide by catalyzing a substrate, and the substrate is organic matter contained in seaweed itself or organic matter added artificially; the substrate accounts for 4-30% of the seaweed by mass; the mass ratio of the seaweed, the enzyme and the water is 20-35: 0.01-0.5: 65-80.

Description

Biological stimulator and preparation method thereof

Technical Field

The invention relates to a biological stimulator and a preparation method thereof.

Background

In the growth process of crops, the biological stimulin has great influence on cell division, chlorophyll transformation, cell wall formation and stress resistance of the crops, can assist the growth of the crops, can also improve the quality of the crops and increase the yield of the crops, and is always concerned by the market. However, the development of the effect of the biostimulant is related to the extraction process, and the great difference of the use effect can be brought by the difference of the extraction process of the same substance, so that the exploration of an extraction method of the biostimulant is particularly important.

Meanwhile, in order to make the bio-stimulin have better functions, the synergistic effect of several bio-stimulants is usually exerted by compounding the bio-stimulants, and the tea saponin is usually used as a pesticide wetting agent in agriculture, so that the physical property of the pesticide is improved, the adhesive force of the liquid medicine on the body surface of organisms or plants is improved, and the synergistic effect on the pesticide is achieved. The insecticidal composition has a certain insecticidal effect, but when the insecticidal composition is applied to insect killing, only the insect killing effect can be achieved, and nutrition supplement cannot be carried out on plants damaged by the insects, so that the insecticidal composition needs to be compounded with other biological stimulants, and the yield reduction of crops is avoided while the insecticidal effect is achieved.

The seaweed extract has the functions of promoting the growth of crops and improving the stress resistance of the crops, but the yield can be increased by only promoting the growth of the crops, and the invasion of diseases and pests is also avoided, so that the combination of the tea saponin with the insecticidal effect and the seaweed extract with the function of promoting the growth of the crops is the main direction of the research at present.

However, the extraction methods of the seaweed extract are various, the components of the seaweed extract obtained by different preparation processes are different, and the using effect of the seaweed extract is different after the seaweed extract is used with tea saponin, so that the screening of the seaweed extract which can be matched with the tea saponin to further improve the using effect becomes a difficult point in the industry.

Disclosure of Invention

The invention provides a biological stimulator and a preparation method thereof, and aims to solve the technical problem of screening a seaweed extract matched with tea saponin, wherein the extract is matched with the tea saponin and has a better using effect.

In order to solve the technical problems, the invention adopts the following technical scheme:

a biostimulant comprises tea saponin and seaweed extract;

the seaweed extract comprises seaweed extract and seaweed powder;

the seaweed extract is prepared according to the following steps:

adding seaweed and enzyme into water, and performing enzymolysis to obtain seaweed extract;

drying the seaweed extract to obtain seaweed powder;

the enzyme at least comprises cellulase, lignin peroxidase and oxidase, the oxidase obtains hydrogen peroxide by catalyzing a substrate, and the substrate is an organic matter contained in the seaweed or an artificially added organic matter; the substrate accounts for 4-30% of the seaweed mass; the mass ratio of the seaweed to the enzyme to the water is 20-35: 0.01-0.5: 65-80;

the mass ratio of the tea saponin to the seaweed extract is 1-99: 1-99.

The enzymolysis is to add lignin peroxidase and oxidase at the same time for enzymolysis; the enzymolysis condition is that the enzymolysis temperature is 30-62 ℃, and the enzymolysis time is 6-96 h.

And after enzymolysis, enzyme deactivation is also included, wherein the enzyme deactivation temperature is 90-120 ℃, and the enzyme deactivation time is 2-5 min.

And (3) drying, wherein the drying temperature is less than 85 ℃.

Mixing tea saponin and seaweed powder to obtain biological stimulator;

adding tea saponin into the seaweed extract, and dissolving to obtain the biological stimulator.

The invention has the following beneficial technical effects:

1. the cellulose is hydrolyzed by adding the cellulase to obtain the polysaccharide and the glucose, the obtained polysaccharide and the glucose can avoid the hydrolysis of the tea saponin, and the drug effect of the tea saponin is prolonged, because the structure of the tea saponin is ester saponin, namely, hydroxyl on aglucon forms ester with organic acid, the structure of the tea saponin consists of three parts, namely a ligand, an aglucon and the organic acid, wherein the aglucon part comprises glucuronic acid, arabinose, xylose and galactose, so that the hydrolysis of the tea saponin can be inhibited and the drug effect of the tea saponin is prolonged by increasing the concentration of a hydrolysate of the tea saponin; the product of the seaweed enzymolyzed by the cellulase is mainly saccharides, so that favorable conditions are provided for the growth of microorganisms, the microorganisms comprise harmful bacteria and beneficial bacteria, potential safety hazards are brought to the growth of plants, the normal growth of crops can be further ensured through the bacteriostatic action of the tea saponin, and the seaweed extract and the tea saponin obtained through the hydrolysis of the cellulase play complementary roles.

2. This application is through adding lignin peroxidase and oxidase, and the H2O2 that lignin peroxidase produced through catalytic oxidation enzyme oxidation substrate carries out the decoloration to melanin to the marine alga extract of preparation, the colour is light, has better light transmissivity, spouts can reduce the influence of leaf fertilizer to illumination in the leaf surface, reduces the influence to plant leaf photosynthesis, can restore fast after the tea saponin insecticidal because of the damage of insect pest to the plant, makes the crop resume the growth as early as possible.

3. The enzymatic hydrolysis is to add lignin peroxidase and oxidase at the same time, because the lignin is degraded by H2O2 generated by oxidizing a substrate by the lignin peroxidase, and simultaneously, the lignin peroxidase is catalyzed by H2O2 to decolor melanin to obtain a seaweed extract with a lighter color; in the preparation method, if the lignin peroxidase is added firstly, the lignin peroxidase cannot carry out an enzymolysis reaction or slowly progresses because no H2O2 is used for catalyzing, and meanwhile, the enzyme activity of the lignin peroxidase is lost, so that the enzymolysis effect and the enzymolysis time after the addition of the oxidase are influenced; and when the oxidase is added firstly, the oxidase catalyzes to generate H2O2, H2O2 is unstable and easy to decompose, and the enzymolysis effect after the lignin peroxidase is added is influenced due to the reduction of H2O2 after the lignin peroxidase is added.

Detailed Description

The present invention is further illustrated by the following specific examples.

Example 1

A biological stimulator comprises tea saponin and Sargassum extractive solution;

the mass ratio of the tea saponin to the seaweed extract is 5: 95.

A preparation method of seaweed extract comprises the following steps: adding seaweed and enzyme 1 into water, performing enzymolysis for 72h at the temperature of 58 ℃ +/-2 ℃, inactivating enzyme for 2min at the temperature of 100 ℃ to obtain seaweed liquid, adding glucose and enzyme 2 into the seaweed liquid, performing enzymolysis for 24h at the temperature of 58 ℃ +/-2 ℃, and inactivating enzyme for 2min at the temperature of 100 ℃ to obtain seaweed extract;

the enzyme 1 is a composition of cellulase and acid protease according to the mass ratio of 2: 1;

the enzyme 2 is a composition of lignin peroxidase and glucose oxidase according to a mass ratio of 1: 1;

the mass ratio of enzyme 1 to enzyme 2 is 1:1.

The mass ratio of the seaweed, the glucose, the enzyme 1, the enzyme 2 and the water is 30:6:0.03:0.03: 63.94.

Adding tea saponin into the seaweed extract to obtain biological stimulator;

the seaweed is herba Zosterae Marinae, and the tea saponin is obtained from Sabina biological science and technology, water soluble tea saponin, brown powder.

Example 2

A biological stimulator comprises tea saponin and seaweed powder;

the mass ratio of the tea saponin to the seaweed meal is 90: 10.

A preparation method of seaweed powder comprises the following steps: adding seaweed and enzyme 1 into water, performing enzymolysis for 72h at the temperature of 58 ℃ +/-2 ℃, inactivating enzyme for 2min at the temperature of 100 ℃ to obtain seaweed liquid, adding glucose and enzyme 2 into the seaweed liquid, performing enzymolysis for 24h at the temperature of 58 ℃ +/-2 ℃, and inactivating enzyme for 2min at the temperature of 100 ℃ to obtain seaweed extract;

spray drying the seaweed extract at 80 deg.C to obtain seaweed powder;

the mass ratio of enzyme 1 to enzyme 2 is 1:1.

Mixing tea saponin and Sargassum powder to obtain biological stimulator;

the seaweed is herba Zosterae Marinae, and the tea saponin is obtained from Saiangzi Biotech limited, water soluble tea saponin, and brown powder.

Example 3

A biological stimulator comprises tea saponin and seaweed powder;

the mass ratio of the tea saponin to the seaweed meal is 40: 60.

A preparation method of seaweed powder comprises the following steps: adding Sargassum and enzyme into water, performing enzymolysis at 45 + -2 deg.C for 90 hr, and inactivating enzyme at 101 deg.C for 2min to obtain Sargassum extract;

spray drying the seaweed extract at 80 deg.C to obtain seaweed powder;

the enzyme is a composition of cellulase, lignin peroxidase and L-amino acid oxidase according to the mass ratio of 3:1: 1;

the mass ratio of the seaweed, the enzyme and the water is 30:0.05: 69.95.

Mixing tea saponin and seaweed powder to obtain biological stimulator;

the tea saponin is obtained from Saiangtze biotechnology, water-soluble tea saponin, and brown powder.

The seaweed is Sargassum.

Example 4

A biological stimulator comprises tea saponin and seaweed powder;

the mass ratio of the tea saponin to the seaweed meal is 70: 30.

A preparation method of seaweed powder comprises the following steps: adding Sargassum and enzyme into water, performing enzymolysis at 57 + -2 deg.C for 85 hr, and inactivating enzyme at 100 deg.C for 2min to obtain Sargassum extract;

spray drying the seaweed extract at 75 deg.C to obtain seaweed powder;

the enzyme is a composition of pectinase, cellulase, lignin peroxidase and L-amino acid oxidase according to a mass ratio of 1:1:1: 1;

the mass ratio of the seaweed, the enzyme and the water is 27:0.2: 72.8.

Mixing tea saponin and seaweed powder to obtain biological stimulator

The seaweed is Porphyra haitanensis.

Example 5

the mass ratio of the tea saponin to the seaweed extract is 10: 90.

A preparation method of seaweed extract comprises the following steps: adding Sargassum and enzyme into water, performing enzymolysis at 57 + -2 deg.C for 85 hr, and inactivating enzyme at 100 deg.C for 2min to obtain Sargassum extract;

the enzyme is a composition of cellulase, lignin peroxidase and L-amino acid oxidase according to the mass ratio of 2:1: 1;

the mass ratio of the seaweed, the enzyme and the water is 15:0.02: 84.98.

Adding tea saponin into the seaweed extract to obtain biological stimulator;

The seaweed is Eucheuma Gelatinosum.

Example 6

A method for preparing Sargassum extract comprises adding enzyme and glucose into Sargassum solution, performing enzymolysis at 57 + -2 deg.C for 24 hr, and inactivating enzyme at 100 deg.C for 2min to obtain Sargassum extract;

the enzyme is a composition of lignin peroxidase and glucose oxidase according to a mass ratio of 1: 1;

the mass ratio of the seaweed liquid to the enzyme to the glucose is 98.79:0.01: 1.2;

the seaweed liquid is purchased from Futaijiate biotechnology limited company, the preparation method is a chemical method, the color is black, and the whiteness is 2.23 after the seaweed liquid is dried in a vacuum oven at 50 ℃.

Adding tea saponin into the seaweed extract to obtain biological stimulator;

The seaweed is Eucheuma Gelatinosum.

Example 7

the enzyme is a composition of cellulase, lignin peroxidase and L-amino acid oxidase according to the mass ratio of 1:2: 1;

the mass ratio of the seaweed, the enzyme and the water is 32:0.4: 67.6.

Adding tea saponin into the seaweed extract to obtain biostimulant;

The seaweed is Eucheuma Gelatinosum.

Cellulase, glucose oxidase and acid protease used in the application are purchased from Tan Xindeli bioengineering limited company, the enzyme activities are 200000u/g, 10000u/g and 200000u/g respectively, lignin peroxidase is purchased from Shandong Nuojie biotechnology limited company, and the enzyme activity is 10000 u/g.

The beneficial effects of the present invention are further illustrated below in conjunction with experimental data:

experiment one

1.1 test site: shandong Eiguo Biotech Co., Ltd.

1.2 test detection: solid content, seaweed extract color and seaweed powder whiteness;

preparing seaweed powder: drying the seaweed extract in a vacuum oven at 50 deg.C until the water content is less than 0.4 to obtain seaweed powder.

1.3 test materials: comparative 1 (except that lignin peroxidase, glucose oxidase and glucose were not added, and other preparation methods were all the same as those in example 1) was performed by adding cellulase, acid protease and lignin peroxidase at 58 ℃. + -. 2 ℃ for 72 hours, after enzyme deactivation, glucose oxidase and glucose were added, and at 58. + -. 2 ℃ for 24 hours, except that the preparation method of the seaweed extract in example 1 was performed by adding cellulase, acid protease and lignin peroxidase first, and the other preparation methods were the same as those in example 1), comparative 2 was performed by adding cellulase, acid protease, glucose oxidase and glucose first, at 58. + -. 2 ℃ for 72 hours, after enzyme deactivation, lignin peroxidase was added, and after enzyme deactivation, after 24 hours, enzymatic hydrolysis was performed at 58. + -. 2 ℃ for 24 hours, the other seaweed extracts, which are identical to those of example 1), were prepared as comparative example 3, comparative example 4 (seaweed liquid available from Futai Jiate Biotechnology Co., Ltd. by chemical method), comparative example 4, and the seaweed extract prepared in example 1 was prepared as example 1, and the seaweed liquid prepared in example 6 was prepared as example 6.

1.4 Experimental implementation: the solid content is that the prepared seaweed extract is kept stand for 24h, the upper solution is taken, the color is observed by naked eyes according to the coating solid content measuring method GB1725-79, and the whiteness is detected by a whiteness meter.

The present application is consistent in other implementations except for differences in the respective processes.

2 results and analysis

The solid content, the color of the seaweed extract and the whiteness of the seaweed powder are shown in Table 1

TABLE 1

As can be seen from Table 1, lignin peroxidase and oxidase can reduce the color depth of the seaweed extract. The influence on the whiteness is different due to different addition sequences of lignin peroxidase and oxidase.

Experiment two

1.1 test site: shou guang sun collects the street, Shimen, Dong village.

1.2 test detection: chlorophyll content (SPAD).

1.3 test materials: a seaweed extract prepared in comparative example 1 (except that no lignin peroxidase, glucose oxidase and glucose were added, and the other preparation methods were all the same as in example 1), a seaweed extract prepared in comparative example 2 (except that the seaweed extract prepared in example 1 was prepared by adding cellulase, acid protease and lignin peroxidase first, carrying out enzymatic hydrolysis at 58 ℃. + -. 2 ℃ for 72 hours, inactivating the enzyme, adding glucose oxidase and glucose, and carrying out enzymatic hydrolysis at 58 ℃. + -. 2 ℃ for 24 hours, and the other preparation methods were all the same as in example 1), a seaweed extract prepared in comparative example 3 (except that cellulase, acid protease, glucose oxidase and glucose were added first, carrying out enzymatic hydrolysis at 58 ℃. + -. 2 ℃ for 72 hours, inactivating the enzyme, adding lignin peroxidase, and carrying out enzymatic hydrolysis at 58 ℃. + -. 2 ℃ for 24 hours, and the other preparation methods were all the same as in example 1), comparative 4 (seaweed liquid purchased from santa yunnanensis limited biotechnology, by chemical method), the seaweed extract prepared in example 1 and the seaweed extract prepared in example 6 were added to a liquid water-soluble fertilizer containing urea, potassium dihydrogen phosphate and water in a mass ratio of 30:10:60, respectively, wherein the mass ratio of the seaweed extract to the liquid water-soluble fertilizer is 5:95, and the liquid water-soluble fertilizer is respectively referred to as a comparative 1 fertilizer, a comparative 2 fertilizer, a comparative 3 fertilizer, a comparative 4 fertilizer, an example 1 fertilizer and an example 6 fertilizer.

1.4 Experimental implementation: dividing a 50-day-transplanted cauldron mountain 88 test field into 6 cells, 100 square meters per cell, respectively matching a comparative fertilizer 1, a comparative fertilizer 2, a comparative fertilizer 3, a comparative fertilizer 4, a fertilizer of example 1 and a fertilizer of example 6, respectively weighing 30g of the comparative fertilizer 1, the comparative fertilizer 2, the comparative fertilizer 3, the comparative fertilizer 4, the fertilizer of example 1 and the fertilizer of example 6, respectively diluting to 25kg by water to obtain a diluent, and spraying the diluent on leaf surfaces in a spraying manner.

Wherein the chlorophyll is measured by an ultraviolet-visible spectrophotometer. 100 leaves are picked from each experimental cell, the picking part of the leaves is the middle part of the 3 rd branch counted from the top to the bottom, the part receives sufficient sunlight, and the chlorophyll changes obviously.

Note: at most one leaf is picked from each plant.

2 results and analysis

Chlorophyll content (SPAD), see Table 2

TABLE 2

	Chlorophyll before treatment (SPAD)	Treated chlorophyll (SPAD)
			Comparative fertilizer 1	48.6	54.6
Contrast 2 fertilizer	48.3	58.7
			Contrast 3 fertilizer	48.9	56.4
Contrast 4 fertilizer	48.7	52.5
			Example 1 Fertilizer	48.5	60.6
Example 6 Fertilizer	48.6	56.7

As can be seen from the data in tables 1 and 2, the whiteness of the seaweed extract is affected by the addition of lignin peroxidase and oxidase, the solid content of the seaweed extract is not greatly affected, the whiteness affects the use effect of the seaweed extract, the substance composition of the product is changed after the treatment of the oxidase and the oxidase, the method is limited by the research and development efforts of the applicant, the change data of various components cannot be made, the effect after the use can only be verified, the whiteness change shows that the simultaneous addition of the lignin peroxidase and the oxidase has the greatest effect on the whiteness, and the use effect is the best.

Meanwhile, by comparing the data of the fertilizer of comparative example 4 and the fertilizer of example 6, it can be seen that the fertilizer of example 6 treated by lignin peroxidase and oxidase can obviously improve the chlorophyll content in the leaves compared with the untreated fertilizer of comparative example 4, i.e. has better use effect.

Experiment three

1.1 test site: shandong fruit lovers Biotech Co., Ltd. laboratory.

1.2 test detection: height (cm) of corn plants, stem thickness (mm), root length (cm), fresh weight above ground (g) and fresh weight below ground (g).

1.3 test materials: maize, variety Longping 206, date of experiment 5.15 to 6.10.

1.4 Experimental implementation: dividing 25 flowerpots into 5 groups, dividing each group into 5, respectively comparing the biostimulant prepared in 5 (except that no lignin peroxidase, glucose oxidase and glucose are added in the preparation of the seaweed extract, and the other preparation methods are all consistent with those in example 1) with reference of 5, comparing 5 with reference of 6 (except that the preparation method of the seaweed extract in example 1 is that cellulase, acid protease and lignin peroxidase are added firstly, enzymolysis is carried out for 72h at a temperature of 58 ℃ +/-2 ℃, then glucose oxidase and glucose are added, and the other preparation methods are all consistent with those in example 1 with reference of 24h at a temperature of 58 ℃ +/-2 ℃), comparing 6 with reference of 7 (except that the preparation method of the seaweed extract is that cellulase, acid protease, glucose oxidase and glucose are added firstly, and the temperature is 58 ℃ +/-2 ℃), enzymolysis for 72h, then adding lignin peroxidase, and performing enzymolysis for 24h at 58 ℃ +/-2 ℃ with the other being the same as that in example 1) to obtain biostimulant, which is designated as comparative 7, and comparative 8 (except that the seaweed extract is chemically prepared from seaweed extract purchased from santa sieboldii biotechnology limited, the preparation method is the same as that in example 2), which is designated as comparative 8, and the biostimulant prepared in example 1 is designated as example 1.

2 results and analysis

The height (cm) of the corn plants, the stem thickness (mm), the root length (cm), the fresh weight (g) on the ground and the fresh weight (g) under the ground are shown in Table 3

TABLE 3

	Plant height (cm)	Stem diameter (mm)	Root length (cm)	Fresh weight on ground (g)	Fresh weight underground (g)
						Comparative example 5	72.27	8.43	44.31	12.86	4.51
Comparative example 6	72.16	8.41	44.26	12.79	4.48
						Comparative example 7	71.23	8.37	44.15	12.56	4.47
Comparative example 8	67.15	8.15	41.76	11.67	4.05
						Example 1	74.52	8.57	45.13	13.02	4.62

As can be seen from table 2, the seaweed extract obtained by different preparation processes is compounded with tea saponin to obtain different planting effects, which are the worst effect compared with the comparison 4 (seaweed extract prepared by chemical method purchased from tai jiate biotechnology limited), the effect of the application in example 1 (adding lignin peroxidase and glucose oxidase simultaneously) is the best, while the effects of the comparison 2 and the comparison 3 in which lignin peroxidase and glucose oxidase are added separately are almost the same as the effect of the comparison 1 in which lignin peroxidase, glucose oxidase and glucose are not added, i.e. the use effect of compounding the seaweed extract and the tea saponin is directly influenced by the change of the preparation process.

Experiment four

1.1 test site: laboratory of biotechnology limited for eastern lovers.

1.2 test detection: the activity condition of the slug after the drug is used is observed, the death rate of the slug is counted after 24h, the experimental rape is weighed, the slug is extremely strong in tolerance and is not frequently moved for a long time, and whether the slug dies or not is difficult to observe by naked eyes, so that the death rate is uniformly counted after 24h, and the judgment method is as follows: a slug is dropped into a beaker containing 200ml of water, observed for movement and immobility for 10min, counted as dead, and counted as non-dead if there is movement within 10 min.

1.3 test materials: comparison 9 (except that no lignin peroxidase, glucose oxidase and glucose were added in the preparation of the seaweed extract, and other preparation methods were all the same as those in example 2) of the prepared biostimulant, denoted as comparison 9, comparison 10 (except that the preparation method of the seaweed extract in example 2 was that cellulase, acid protease and lignin peroxidase were added first, enzymolysis was carried out at 58 ℃. + -. 2 ℃ for 72 hours, then glucose oxidase and glucose were added, and enzymolysis was carried out at 58 ℃. + -. 2 ℃ for 24 hours, and other methods were all the same as those in example 2) of the prepared biostimulant, denoted as comparison 10, comparison 11 (except that the preparation method of the seaweed extract was that cellulase, acid protease, glucose oxidase and glucose were added first, enzymolysis was carried out at 58 ℃. + -. 2 ℃ for 72 hours, then lignin peroxidase was added, the biostimulant prepared in example 2 was enzymatically hydrolyzed at 58 ℃. + -. 2 ℃ for 24 hours, and the others were identical to those of example 2) in comparative example 11 and comparative example 12 (except that the seaweed extract was chemically prepared from seaweed extract purchased from Futai Jiate Biotechnology Ltd., and the preparation method thereof was identical to that of example 2), and the biostimulant prepared in example 2 in comparative example 12 was denoted as example 2.

1.4 Experimental implementation: preparing 5 culture dishes, placing 5g of rape green leaves in each culture dish, then placing 30 adult slugs with the body length of 32-35 mm, respectively correspondingly comparing 9 (except that no lignin peroxidase, glucose oxidase and glucose are added in the preparation of the seaweed extract, and other preparation methods are all consistent with that of the example 2) prepared biostimulant with reference 9, marking as comparison 9, comparing 10 (except that the preparation method of the seaweed extract of the example 2 is that cellulase, acid protease and lignin peroxidase are added firstly, enzymolysis is carried out for 72h at the temperature of 58 +/-2 ℃, then glucose oxidase and glucose are added, enzymolysis is carried out for 24h at the temperature of 58 +/-2 ℃, and other biostimulant is marked as comparison 10), and comparing 11 (except that the preparation method of the seaweed extract comprises adding cellulase, glucose, and the like firstly, Acid protease, glucose oxidase and glucose are subjected to enzymolysis for 72 hours at the temperature of 58 ℃ plus or minus 2 ℃, then lignin peroxidase is added, and the mixture is subjected to enzymolysis for 24 hours at the temperature of 58 ℃ plus or minus 2 ℃, wherein the rest is the same as that of the biostimulant prepared in example 2), and the biostimulant is marked as comparison 11, and the biostimulant prepared in comparison 12 (except that the seaweed extract is prepared by a chemical method from a seaweed extract purchased from Futai Jiate Biotechnology Co., Ltd., and the other preparation method is the same as that of example 2) is marked as comparison 12, and the biostimulant prepared in example 2 is marked as example 2. 2. + -. 0.2g of a 1000-fold diluted treatment solution was sprayed into each of the petri dishes, and the treated petri dishes were placed in the dark.

2 results and analysis

The death rate of the land slug, the weighing and activity conditions of the experimental rapes before and after the experiment are shown in Table 4

TABLE 4

	Rape weight (g) before experiment	Rape weight after experiment (g)	Weight difference (g) of rape	Situation of activity	Death number (only)	Mortality (%)
							Comparative example 9	5.233	2.918	2.315	Reduction of activity	27	90
Comparative example 10	5.176	2.907	2.269	Reduction of activity	27	90
							Comparative example 11	5.154	2.800	2.354	Reduction of activity	26	86.7
Comparative example 12	5.152	2.385	2.767	Reduction of activity	24	80
							Example 2	5.146	3.874	1.272	Activity is reduced significantly	29	96.7

Note: the activity condition is observed by naked eyes, and because the activity condition is not observed for 24h and is observed by people, subjective judgment can exist, and the activity condition only has reference value.

It can be seen from the mortality of slug in table 3 that the seaweed extract solution prepared by different preparation methods is applied to disinsection, and has different effects, wherein the bio-stimulin prepared in embodiment 2 of the present application has better effect of killing slug, and by the activity and difference of rape before and after experiment, it can be seen that contrast 9, contrast 10, contrast 11, contrast 12 and embodiment 2 all have toxic effect to slug, can reduce the activity of slug and have killing ability, but embodiment 2 of the present application can reduce the activity of slug and reduce its ingestion better, and killing effect is better.

Market feedback

Be applied to market with this application embodiment 2, obtained market feedback, have better effect of killing slug and snail, the Weifang city embankment village town plant family lie a certain use after, very approved the effect of this application, simultaneously, it is fast that he still reacts to use this article to kill after slug and the snail, the plant recovery speed is fast, the leaf color is dark green.

Claims

1. A biostimulant is characterized by comprising tea saponin and seaweed extract; the seaweed extract comprises seaweed extract or seaweed powder; the seaweed extract is prepared according to the following steps:

drying the seaweed extract to obtain seaweed powder;

the enzyme at least comprises cellulase, lignin peroxidase and oxidase, the oxidase obtains hydrogen peroxide by catalyzing a substrate, and the substrate is an organic matter contained in the seaweed and/or an artificially added organic matter; the substrate accounts for 4-30% of the seaweed by mass; the mass ratio of the seaweed to the enzyme to the water is 20-35: 0.01-0.5: 65-80; the mass ratio of the tea saponin to the seaweed extract is 1-99: 1-99;

in the enzymolysis process, lignin peroxidase and oxidase are added simultaneously for enzymolysis; the enzymolysis condition is that the enzymolysis temperature is 30-62 ℃, and the enzymolysis time is 6-96 h.

2. The biostimulant of claim 1, further comprising inactivating the enzyme after the enzymatic hydrolysis, wherein the temperature of inactivating the enzyme is 90-120 ℃ and the time of inactivating the enzyme is 2-5 min.

3. The biostimulant of claim 1 or 2, wherein the drying is at a temperature of less than 85 ℃.

4. The method for preparing biostimulant according to claim 1 or 2, wherein the biostimulant is obtained by mixing tea saponin and seaweed powder; adding tea saponin into the seaweed extract, and dissolving to obtain the biological stimulator.