CN111848298A

CN111848298A - Humic acid insect-prevention antibacterial silicon fertilizer containing alkaloid and preparation method thereof

Info

Publication number: CN111848298A
Application number: CN202010762991.2A
Authority: CN
Inventors: 宋洁; 窦玉芳; 牛育华; 李胜港
Original assignee: Shaanxi University of Science and Technology
Current assignee: Shaanxi University of Science and Technology
Priority date: 2020-07-31
Filing date: 2020-07-31
Publication date: 2020-10-30

Abstract

The invention discloses a humic acid insect-proof antibacterial silicon fertilizer containing alkaloid and a preparation method thereof, relating to the field of agricultural fertilizer application. The preparation raw materials comprise: 100 parts of humate, 1-20 parts of silicon-containing compound and 1-20 parts of alkaloid. Wherein: the alkaloid is extracted and separated from natural plant containing alkaloid, and is obtained by silica gel column chromatography, elution, dissolution and recrystallization. The obtained humic acid insect-proof antibacterial silicon fertilizer containing alkaloid combines the fertilizer and the botanical pesticide, not only meets the nutrient absorption of crops in the growth process, but also can effectively inhibit plant diseases and insect pests in the crops, the product can directly promote the photosynthesis of crop leaves, form silicified cells in the crop bodies and promote the accumulation of organic matters of the crops, and finally achieves the purposes of improving soil, improving the quality of the crops and increasing the yield of the crops, thereby conforming to the development concept of green agriculture.

Description

Humic acid insect-prevention antibacterial silicon fertilizer containing alkaloid and preparation method thereof

Technical Field

The invention belongs to the field of agricultural fertilizer application, and particularly relates to a humic acid insect-proof antibacterial silicon fertilizer containing alkaloid and a preparation method thereof.

Background

The fertilizer is an essential production material in agricultural production, and China is the largest fertilizer market in the world. With the increasing of the application amount of the chemical fertilizer in the farmland, the soil quality of the land applying the chemical fertilizer in successive years is low as a whole, the soil degradation phenomenon is serious, and the phenomena of water and soil loss, soil desertification, acidification, salinization and the like are expanded continuously. Meanwhile, the field has many problems in preventing and treating plant diseases and insect pests, about 90% of the traditional chemical pesticide remains in the environment, not only causes environmental pollution, but also can be transferred into the human body through a food chain to cause harm to the human body. Currently, the series of problems seriously affect the livelihood of people. Therefore, it is very important to develop a fertilizer which can improve the physical and chemical properties of soil and has an insecticidal effect.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention aims to provide a humic acid insect-proof antibacterial silicon fertilizer containing alkaloid and a preparation method thereof, so as to solve the problems of low soil quality and field diseases and insect pests caused by excessive fertilizer.

In order to achieve the purpose, the invention adopts the following technical scheme to realize the purpose:

the invention discloses an insect-proof and antibacterial silicon fertilizer containing alkaloid, which comprises, by mass, 100 parts of humate, 1-20 parts of a silicon-containing compound and 1-20 parts of alkaloid.

Preferably, the humate is one or more of ammonium humate, potassium humate and sodium humate.

Preferably, the alkaloid is extracted and separated from natural plants containing alkaloid, and the natural plants containing alkaloid are any one of coptis chinensis, pyrola, phellodendron amurense, barberry root and fritillaria.

Further preferably, the alkaloid is extracted by:

cleaning natural plant containing alkaloid, drying, pulverizing, mixing with ethanol at a ratio of 1 g: (1-50) ultrasonically stirring the solid-liquid ratio of ml for 30min, refluxing the obtained mixed solution for not less than 4 times, combining the refluxed solutions, then concentrating the refluxed solution under reduced pressure, and freeze-drying to obtain a crude alkaloid product;

dissolving the crude alkaloid product with methanol solution, filtering, subjecting the filtrate to silica gel column chromatography, eluting with methanol-water eluent, collecting sample solution, drying, dissolving with 90% methanol solution, recrystallizing, filtering, and drying to obtain alkaloid.

Preferably, the time of each reflux for extracting the alkaloid is not less than 1.5 h.

Preferably, the volume ratio of methanol to water is 3:1 when the alkaloid is extracted.

Preferably, the silicon-containing compound is one or both of sodium silicate and calcium silicate.

The invention also discloses a preparation method of the humic acid insect-proof antibacterial silicon fertilizer containing alkaloid, which comprises the following steps:

1) weighing humate, a silicon-containing compound and alkaloid according to a proportion;

2) preparing a silicon-containing compound into an aqueous solution with the concentration of 2-10%;

3) dissolving humate in water to obtain a humate solution, blending the silicon-containing compound aqueous solution obtained in the step 2) with the humate solution, controlling the temperature to be 40-70 ℃, and fully stirring to uniformly mix the humate silicon fertilizer solution;

4) adding the humic acid salt silicon fertilizer mixed solution into the humic acid salt silicon fertilizer mixed solution, stirring uniformly, and then adjusting the pH value to be neutral to prepare the humic acid insect-proof antibacterial silicon fertilizer containing alkaloid.

9. The method for preparing humic acid insect-proof antibacterial silicon fertilizer containing alkaloid as claimed in claim 8, wherein in step 4) hydrochloric acid is used to adjust pH to neutral.

Compared with the prior art, the invention has the following beneficial effects:

the molecular chain of the humate in the formula of the insect-proof antibacterial organic silicon fertilizer containing the alkaloid disclosed by the invention has various active functional groups, can interact with a plurality of inorganic and organic compounds, and can play roles in improving the physical and chemical properties of soil, enhancing the fertilizer efficiency and the like. The alkaloid is widely present in plants, is also a widely used pesticide, has good insecticidal effect, is easy to degrade, is pollution-free, and kills pests mainly through the actions of stomach toxicity, contact poisoning and the like. The silicon fertilizer is a good quality fertilizer, a health-care fertilizer and a plant regulating fertilizer, and can be used as a fertilizer for providing nutrients, a soil conditioner and soil improvement. The invention skillfully combines fertilizers (low-rank coal-based humate and silicon fertilizer) and plant-derived pesticides (alkaloids), thereby not only improving the utilization rate of the fertilizers and the soil quality, but also reducing the use of chemical pesticides, not only meeting the nutrient problem of crops in the growth process, but also effectively inhibiting plant diseases and insect pests in the crops to a certain extent, directly promoting the photosynthesis of crop leaves, forming silicified cells in the crops, thickening the cell walls and cuticles on the surface layers of stems and leaves, effectively inhibiting the water loss of the leaves, promoting the accumulation of organic matters in the crops, finally achieving the purposes of improving the soil, improving the crop quality and increasing the crop yield, really realizing scientific fertilization and scientific prevention and control of the plant diseases and insect pests, and conforming to the development concept of green agriculture. Compared with the common humic acid fertilizer, the humic acid insect-proof antibacterial silicon fertilizer containing the alkaloid has more obvious increase on the chlorophyll content, the vitamin C content and the peroxidase activity in each growth stage, and has more obvious reduction on the malonaldehyde content in each growth stage. Meanwhile, the humic acid insect-proof antibacterial silicon fertilizer containing the alkaloid has good antibacterial and insecticidal effects.

The method provides the preparation method of the humic acid insect-proof antibacterial silicon fertilizer containing the alkaloid, the method is simple to operate and high in practicability, the fertilizer prepared by the method is safe and harmless, and the sustainable development of agriculture and the development direction of green agriculture are met.

Detailed Description

The present invention will now be described in further detail with reference to specific examples, which are intended to be illustrative, but not limiting, of the invention.

Example 1

A humic acid insect-proof antibacterial organic silicon fertilizer containing alkaloids comprises 100 parts of ammonium humate, 10 parts of sodium silicate and 10 parts of coptis alkaloids, and the preparation method comprises the following steps of:

1) cleaning Coptidis rhizoma, drying, pulverizing, adding into ultrasonic stirrer, mixing with ethanol at a ratio of 1: 15, stirring for 30min, transferring the obtained mixed solution to a reflux device, refluxing for not less than 4 hours, wherein the refluxing time is not less than 1.5h each time, combining the refluxed solutions, then concentrating the refluxed solutions under reduced pressure, and freeze-drying to obtain crude alkaloid; dissolving the crude alkaloid product with methanol solution, filtering, subjecting the filtrate to silica gel column chromatography, eluting with eluent of methanol and water at a volume ratio of 3:1, collecting sample solution, drying, dissolving with 90% methanol solution, recrystallizing, filtering, and drying to obtain Coptidis rhizoma alkaloid;

2) respectively weighing 100 parts of ammonium humate, 10 parts of sodium silicate and 10 parts of coptis alkaloid according to the mass parts;

3) preparing the weighed sodium silicate into 6% aqueous solution;

4) adding a proper amount of water into ammonium humate for dissolving to obtain an ammonium humate solution, blending a sodium silicate-containing aqueous solution and the ammonium humate solution, controlling the temperature at 40 ℃, and fully stirring to uniformly mix the ammonium humate silicon fertilizer solution;

5) adding the coptis alkaloid into the humic acid ammonium silicon fertilizer solution, uniformly stirring, and then adjusting the pH value to be neutral by using hydrochloric acid to obtain the humic acid insect-proof antibacterial silicon fertilizer containing the alkaloid.

Example 2

The humic acid insect-prevention antibacterial organic silicon fertilizer containing alkaloid comprises 100 parts of potassium humate, 15 parts of calcium silicate and 5 parts of deer tail alkaloid, and the preparation method comprises the following steps of:

1) cleaning, drying and crushing the rhaponticum carthamoides, adding the crushed rhaponticum carthamoides into an ultrasonic stirrer, mixing the crushed rhaponticum carthamoides with ethanol in a proportion of 1: stirring for 30min at a solid-to-liquid ratio of 10, transferring the obtained mixed solution to a reflux device, refluxing for not less than 1.5h for not less than 4 times, combining the refluxed solutions, concentrating the refluxed solution under reduced pressure, and freeze-drying to obtain crude alkaloid; dissolving the crude alkaloid product with methanol solution, filtering, subjecting the filtrate to silica gel column chromatography, eluting with eluent of methanol and water at a volume ratio of 3:1, collecting sample solution, drying, dissolving with 90% methanol solution, recrystallizing, filtering, and drying to obtain rhaponticum carthamoides alkaloid;

2) respectively weighing 100 parts of potassium humate, 15 parts of calcium silicate and 5 parts of deer tail alkaloid according to the mass parts;

3) preparing the weighed calcium silicate into 2% aqueous solution;

4) adding a proper amount of water into potassium humate for dissolving to obtain a potassium humate solution, blending the calcium silicate solution and the potassium humate solution, controlling the temperature at 50 ℃, and fully stirring to uniformly mix the potassium humate and the silicon fertilizer solution;

5) adding rhaponticum carthamoides alkaloid into the potassium silicon humate fertilizer solution, stirring uniformly, and then adjusting the pH to be neutral by using hydrochloric acid to obtain the humic acid insect-proof antibacterial silicon fertilizer containing alkaloid.

Example 3

The humic acid insect-proof antibacterial organic silicon fertilizer containing alkaloid comprises 100 parts of sodium humate, 8 parts of sodium silicate and 12 parts of phellodendron alkaloid, and the preparation method comprises the following steps of:

1) cleaning cortex Phellodendri, drying, pulverizing, adding into ultrasonic stirrer, mixing with ethanol at a ratio of 1: 25 of solid-to-liquid ratio, stirring for 30min, transferring the obtained mixed solution to a reflux device, refluxing for not less than 4 times, wherein the refluxing time is not less than 1.5h each time, combining the refluxed solutions, then concentrating the refluxed solutions under reduced pressure, and freeze-drying to obtain crude alkaloid; dissolving the crude alkaloid product with methanol solution, filtering, subjecting the filtrate to silica gel column chromatography, eluting with eluent of methanol and water at a volume ratio of 3:1, collecting sample solution, drying, dissolving with 90% methanol solution, recrystallizing, filtering, and drying to obtain cortex Phellodendri alkaloid;

2) respectively weighing 100 parts of sodium humate, 8 parts of sodium silicate and 12 parts of phellodendron alkaloid according to the mass parts;

3) preparing the weighed sodium silicate into 6% aqueous solution;

4) adding a proper amount of water into sodium humate for dissolving to obtain a sodium humate solution, blending a sodium silicate aqueous solution and the sodium humate solution, controlling the temperature at 60 ℃, and fully stirring to uniformly mix the sodium humate and the silicon fertilizer solution;

5) adding the phellodendron alkaloid into the sodium humate silicon fertilizer solution, uniformly stirring, and then adjusting the pH to be neutral by using hydrochloric acid to obtain the humic acid insect-proof antibacterial silicon fertilizer containing the alkaloid.

Example 4

The humic acid insect-prevention antibacterial organic silicon fertilizer containing alkaloid comprises 100 parts of potassium humate, 10 parts of sodium silicate and 8 parts of barberry root alkaloid, and the preparation method comprises the following steps of:

1) cleaning radix Berberidis, drying, pulverizing, adding into ultrasonic stirrer, mixing with ethanol at a ratio of 1: 30min, transferring the obtained mixed solution to a reflux device, refluxing for not less than 4 times, wherein the refluxing time is not less than 1.5h each time, combining the refluxed solutions, then concentrating the refluxed solutions under reduced pressure, and freeze-drying to obtain crude alkaloid; dissolving the crude alkaloid product with a methanol solution, filtering, performing silica gel column chromatography on the filtrate, eluting with an eluent of methanol and water in a volume ratio of 3:1, collecting a sample solution, drying, dissolving with a 90% methanol solution, recrystallizing, filtering, and drying to obtain the barberry alkaloid;

2) respectively weighing 100 parts of potassium humate, 10 parts of sodium silicate and 8 parts of barberry alkaloid according to the mass parts;

3) preparing the weighed sodium silicate into a 4% aqueous solution;

4) adding a proper amount of water into potassium humate for dissolving to obtain a potassium humate solution, blending a sodium silicate aqueous solution and the potassium humate solution, controlling the temperature at 70 ℃, and fully stirring to uniformly mix the potassium humate and the silicon fertilizer solution;

5) adding barberry root alkaloid into the potassium humate silicon fertilizer solution, stirring uniformly, and then adjusting the pH to be neutral by using hydrochloric acid to obtain the humic acid insect-proof antibacterial silicon fertilizer containing alkaloid.

Example 5

A humic acid insect-proof antibacterial organic silicon fertilizer containing alkaloid comprises 100 parts of humic acid ammonium, 20 parts of potassium silicate and 20 parts of fritillaria alkaloid, and the preparation method comprises the following steps of:

1) cleaning Bulbus Fritillariae Cirrhosae, drying, pulverizing, adding into ultrasonic stirrer, mixing with ethanol at a ratio of 1: 40, stirring for 30min, transferring the obtained mixed solution to a reflux device, refluxing for not less than 4 times, wherein the refluxing time is not less than 1.5h each time, combining the refluxed solutions, then concentrating the refluxed solutions under reduced pressure, and freeze-drying to obtain crude alkaloid; dissolving the crude alkaloid product with methanol solution, filtering, subjecting the filtrate to silica gel column chromatography, eluting with eluent of methanol and water at a volume ratio of 3:1, collecting sample solution, drying, dissolving with 90% methanol solution, recrystallizing, filtering, and drying to obtain Bulbus Fritillariae Cirrhosae alkaloid;

2) respectively weighing 100 parts of ammonium humate, 20 parts of potassium silicate and 20 parts of fritillaria alkaloid according to the mass parts;

3) preparing the weighed potassium silicate into a 10% aqueous solution;

4) adding a proper amount of water into ammonium humate for dissolving to obtain an ammonium humate solution, blending a potassium silicate aqueous solution and the ammonium humate solution, controlling the temperature at 55 ℃, and fully stirring to uniformly mix the ammonium humate silicon fertilizer solution;

5) adding fritillaria alkaloid into the humic acid ammonium silicon fertilizer solution, stirring uniformly, and then adjusting the pH value to be neutral by using hydrochloric acid to obtain the humic acid insect-proof antibacterial silicon fertilizer containing alkaloid.

Example 6

A humic acid insect-proof antibacterial organic silicon fertilizer containing alkaloid comprises 100 parts of sodium humate, 5 parts of calcium silicate and 15 parts of phellodendron alkaloid, and the preparation method comprises the following steps of:

1) cleaning natural cortex Phellodendri, drying, pulverizing, adding into ultrasonic stirrer, mixing with ethanol at a ratio of 1: stirring for 30min at a solid-to-liquid ratio of 50, transferring the obtained mixed solution to a reflux device, refluxing for not less than 1.5h for not less than 4 times, combining the refluxed solutions, concentrating the refluxed solution under reduced pressure, and freeze-drying to obtain crude alkaloid; dissolving the crude alkaloid product with methanol solution, filtering, subjecting the filtrate to silica gel column chromatography, eluting with eluent of methanol and water at a volume ratio of 3:1, collecting sample solution, drying, dissolving with 90% methanol solution, recrystallizing, filtering, and drying to obtain cortex Phellodendri alkaloid;

2) respectively weighing 100 parts of sodium humate, 5 parts of calcium silicate and 15 parts of phellodendron alkaloid according to the mass parts;

3) preparing the weighed calcium silicate into an aqueous solution with the concentration of 8%;

4) adding a proper amount of water into sodium humate for dissolving to obtain a sodium humate solution, blending the calcium silicate aqueous solution and the sodium humate solution, controlling the temperature at 65 ℃, and fully stirring to uniformly mix the sodium humate and the silicon fertilizer solution;

Ordinary humic acid fertilizer:

the common humic acid fertilizer without added silicon-containing compound and alkaloid is used as a comparison sample.

Plant fertilization verification is performed on the humic acid insect-proof and antibacterial silicon fertilizer and the humic acid fertilizer containing the alkaloids prepared in examples 1-6, and the crop quality and the antibacterial and insecticidal detection results are as follows:

(1) influence of humic acid insect-proof antibacterial silicon fertilizer containing alkaloid on chlorophyll content of green vegetables

Chlorophyll is a substance essential for photosynthesis in plants. The method for testing and calculating the chlorophyll content in the application comprises the following steps:

weighing 0.2g of green vegetable leaves, placing the green vegetable leaves in a mortar, adding a small amount of calcium carbonate, adding 3mL of acetone, uniformly grinding, adding 2mL of acetone, grinding until the leaves are whitish, standing for 3-5 minutes, and filtering the homogenate into a 25mL brown volumetric flask through a funnel. The mortar residue was washed with 5mL of acetone and transferred to a funnel for filtration. Discarding leaves in filter residues, washing chlorophyll remained on filter paper into a brown volumetric flask by using acetone, metering the volume to 25mL by using the acetone, and shaking up. Pure acetone solution is used as a control group, then the absorbance of the control group is respectively measured under the wavelengths of 665nm and 645nm, and the total content of chlorophyll is calculated.

Wherein: v is the total volume (mL) of the extracting solution; w is the fresh weight (g) of the plant sample.

The influence of the humic acid insect-proof and antibacterial silicon fertilizer containing alkaloid and the common humic acid fertilizer prepared in the examples 1 to 6 on the chlorophyll content of the green vegetables is shown in table 1. As can be seen from table 1, the chlorophyll content of the different fertilizer samples showed a tendency of increasing first and then decreasing with the growth of the plants, which is caused by the fact that the chlorophyll content of the plants increases with the increase of the growth period, the plant metabolism is enhanced, the chlorophyll content increases, and the chlorophyll content decreases slowly after reaching the peak growth point. In the same period, after the humic acid insect-proof antibacterial silicon fertilizer containing alkaloid in the examples 1-6 is applied to the green vegetables, the chlorophyll content of the humic acid insect-proof antibacterial silicon fertilizer is obviously higher than that of two groups of control groups, which shows that the synergistic cooperation of the low-order coal-based humate, the silicon-containing compound and the alkaloid has a great promotion effect on the synthesis of chlorophyll.

TABLE 1 Effect of the different examples on the chlorophyll content of green vegetables

(2) Influence of humic acid insect-proof antibacterial silicon fertilizer containing alkaloid on vitamin C content of green vegetables

Vitamin C is an indispensable organic compound for the human body and also an essential element for life. The method for testing and calculating the content of the vitamin C in the application comprises the following steps:

the determination of the vitamin C content adopts the titration determination of the oxidized 2, 6-dichlorophenol indophenol under the acidic condition, and the titration end point is the moment when the solution is changed from colorless to pale red. Firstly, titrating a certain volume of Vc standard solution (0.1mg/mL) with 2, 6-dichlorophenol indophenol until the solution is pink and no color disappears within 15s, then taking 2% oxalic acid with the same amount as the Vc standard solution as a blank control, titrating according to the method, finally weighing 1g of vegetable leaves, shredding, adding 5mL of 2% oxalic acid solution, grinding into slurry, transferring the slurry into a centrifuge tube, simultaneously cleaning a mortar with a little 2% oxalic acid solution, combining the slurry with liquid, centrifuging at a rotating speed of 8000r/min for 5min, transferring supernatant into a volumetric flask for constant volume, quickly titrating a certain amount of liquid with 2, 6-dichlorophenol indophenol until the liquid is pink, and determining V_cAnd (4) content.

In the formula:

v is the amount (mL) of 2, 6-dichlorophenol indophenol used for titrating the sample;

V₁the total volume (mL) of the sample after constant volume;

V₂is the volume (mL) of the sample to be measured;

l is the content of Vc oxidized per 1mL of 2, 6-dichlorophenol indophenol solution (mg/mL), which is determined experimentally.

The influence of the humic acid insect-proof and antibacterial silicon fertilizer containing alkaloid and the common humic acid fertilizer prepared in the examples 1-6 on the vitamin C content of the green vegetables is shown in table 2. As can be seen from table 2, the different fertilizer applications showed approximately the same change in vitamin C content as the change in chlorophyll content. Also affected by the plant growth cycle, the difference in data also indicates: the humic acid insect-proof antibacterial silicon fertilizer containing alkaloid prepared in the examples 1-6 is more beneficial to the synthesis of the plant vitamin C.

TABLE 2 Effect of the different examples on the vitamin C content of green vegetables

(3) Influence of humic acid insect-proof antibacterial silicon fertilizer containing alkaloid on malondialdehyde content

Malondialdehyde is one of the most important products of membrane lipid peroxidation, and its production can exacerbate membrane damage. The method for testing and calculating the content of the malonaldehyde in the application comprises the following steps:

weighing 10g of trichloroacetic acid (TCA), and fixing the volume to a 100mL volumetric flask to prepare a 10% (TCA) solution. Weighing 0.125g of thiobarbituric acid (TBA), and making a constant volume into a 25mL volumetric flask to prepare a 0.6% (TBA) solution. Weighing 0.5g of green vegetable leaves, putting the green vegetable leaves into a mortar, adding 3mL of 10% TCA, grinding and homogenizing, adding 2mL of 10% TCA, continuing grinding, centrifuging at the rotating speed of 4000r/min for 15min (4 ℃), and obtaining the supernatant which is the total volume of the MDA extracting solution. Weighing 2mL of extracting solution (which is the volume of the extracting solution for determination) into a test tube with a plug, adding 2mL of 0.6% TBA (the sum of the volumes of the extracting solution and the TBA is the volume of the reaction solution), uniformly mixing, reacting in a boiling water bath for 20min, immediately transferring into ice water, rapidly cooling, centrifuging under the condition of 4000r/min, detecting the absorbance values of obtained supernatant at 532nm, 600nm and 450nm, and calculating the content of MDA. The control group was 2mL of water plus 2mL of 0.6% TBA.

Wherein: v is the volume (ml) of the reaction solution; v_sVolume (ml) of the extract for measurement; v_tThe total volume (ml) of the extract; w: fresh weight (g) of plant sample.

The influence of the humic acid insect-proof and antibacterial silicon fertilizer containing alkaloid and the common humic acid fertilizer prepared in the examples 1 to 6 on the content of malondialdehyde in green vegetables is shown in table 3. As can be seen from table 3, when the humic acid insect-proofing and antibacterial silicon fertilizers containing alkaloids prepared in examples 1 to 6 are applied, the malondialdehyde content is obviously reduced in the same period, and again, the humic acid insect-proofing and antibacterial silicon fertilizers containing alkaloids can inhibit the production of malondialdehyde of plants to a certain extent, which is beneficial to the growth of plants.

TABLE 3 Effect of the different examples on the malondialdehyde content of the vegetables

(4) Influence of humic acid insect-proof antibacterial silicon fertilizer containing alkaloid on peroxidase activity

Peroxidase is an important oxidoreductase in plants, is related to photosynthesis, respiration, auxin oxidation and the like, and can ensure the permeability of cell membranes so that the plants can be prevented from being damaged by active oxygen and grow vigorously. The methods for testing and calculating peroxidase activity (POD) in the present application are:

the 100mmol/L phosphate buffer solution with pH of 6.0 is prepared by blending sodium dihydrogen phosphate and disodium hydrogen phosphate. 50mL of the above solution was placed in a beaker, 28. mu.L of guaiac wood powder was added, 19. mu.L of 30% hydrogen peroxide solution was added, and the mixture was mixed well and stored in a refrigerator as a reaction mixture for further use. Weighing 0.25g of well-grown green vegetable leaves, putting the green vegetable leaves into a mortar, adding 5mL of phosphate buffer solution with the pH value of 6.0, grinding the mixture into homogenate, centrifuging the homogenate for 15min (4 ℃) at the rotating speed of 7000r/min, transferring the supernatant into a 25mL volumetric flask, fixing the volume to 25mL by using the phosphate buffer solution, and storing the supernatant at low temperature for later use. 3mL of the reaction mixture was added to 1mL of the above extract, and the mixture was placed in a cuvette, and a stopwatch was immediately started to record the reaction time, and the absorbance at 470nm was measured and read every one minute to calculate the POD activity. The control group consisted of 3mL reaction mixture and 1mL phosphate buffer.

Wherein: delta A₄₇₀Is the difference in absorbance; Δ t is the time (min) corresponding to the absorbance difference; v₁: total volume (ml) of the extractive solution; v₂Volume of extract solution for measurement (ml): w: fresh weight (g) of plant sample.

The influence of the humic acid insect-proof and antibacterial silicon fertilizer containing alkaloid and the common humic acid fertilizer prepared in the examples 1 to 6 on the activity of the peroxidase of the green vegetables is shown in table 4. As can be seen from Table 4, the humic acid insect-proofing and antibacterial silicon fertilizer containing alkaloid prepared in examples 1-6 is applied in the same period, the peroxidase activity in the green vegetables is obviously higher, which shows that examples 1-6 can promote the synthesis of the plant peroxidase activity, ensure the permeability of cell membranes and promote the growth of crops.

TABLE 4 Effect of the different examples on peroxidase Activity

(5) Alkaloid-containing humic acid insect-proof antibacterial silicon fertilizer antibacterial experiment test data

The antibacterial property testing and calculating method comprises the following steps:

circular Xinhua filter paper with the diameter of 6mm is subjected to dry heat sterilization (160 ℃ for 2 hours) by adopting a filter paper sheet method for later use. The sterilized paper sheets are put into flavonoid insect-proof antibacterial humic acid selenium fertilizer or common humic acid fertilizer to be fully soaked, taken out and spread in a sterile plate, and naturally volatilized to be used.

The slant preservation strain is transferred to a beef extract peptone agar culture medium, and the medium is placed in a constant temperature incubator at 37 ℃ for 24 hours. Taking activated strain, and preparing into 10 with sterile normal saline⁵cfu/mL of bacterial suspension.

0.2mL of the prepared bacterial suspension is taken, dripped into the surface of a solid culture medium, uniformly coated on the surface of the solid culture medium by using a sterile coater, and a filter paper sheet soaked with the pigment solution and volatilized from the flavonoid insect-proof antibacterial humic acid selenium fertilizer or the common humic acid fertilizer is clamped by using a sterile forceps and is put into a flat dish. A blank was prepared by evaporating the solution with distilled water. 3 replicates were made for each sample. And (3) putting the flat plate into a bacteria incubator, culturing at 37 ℃ for 18h, taking out, observing, measuring the diameter of the inhibition zone, and taking an average value.

The antibacterial properties of the humic acid insect-proofing antibacterial silicon fertilizer containing alkaloid and the common humic acid fertilizer prepared in the examples 1-6 are shown in table 5. As can be seen from Table 5, the humic acid fertilizer has no antibacterial ability in the comparative blank case and the common humic acid fertilizer, and the examples 1 to 6 have certain inhibition on staphylococcus aureus and escherichia coli, which shows that the addition of the natural alkaloid has an obvious antibacterial effect.

Table 5 different examples of the antibacterial tests

(6) Insect killing experimental data of humic acid insect-proof antibacterial silicon fertilizer containing alkaloid

The method for testing and calculating the lethality rate in the above embodiment of the present invention:

dissecting root system of herba Apii Graveolentis infected with root-knot nematode, extracting polypide in laboratory by Bellman funnel method, picking oocysts from the polypide, sterilizing the oocysts with 0.2% sodium hypochlorite solution for 0.5min, and washing with sterile water. Placing the oocysts in sterile water for hatching, and taking the nematodes hatched after 2 days for determination. The fertilizer liquid and the insect sample liquid in each embodiment are mixed for direct contact killing, and the concentration of the insect sample is 1000-1200 per mL; continuously observing for 72 hours under the condition of 100-400 times of microscopic observation, and recording the change condition of the number of the worms to calculate the lethality of the fertilizer to the root-knot nematodes.

The thread killing activity of the humic acid insect-preventing and antibacterial silicon fertilizer containing the alkaloids and the common humic acid fertilizer on the root-knot nematodes prepared in the examples 1 to 6 are shown in Table 6. As can be seen from Table 6, the humic acid insect-repellent antibacterial silicon fertilizers containing alkaloids prepared in examples 1-6 have significant lethal effect on Meloidogyne incognita compared with the blank example and the common humic acid fertilizers.

TABLE 6 nematicidal activity of the different examples against root-knot nematodes

Note: mortality rating was AA: 100 percent; a: 90% -100%; BB: 70% -90%; b: 50% -70%; CC: 30% -50%; c: 15% -30%; DD: 5% -15%; d: < 5%; "-" indicates no dead bodies were found.

In conclusion, the humic acid insect-proof and antibacterial silicon fertilizer containing the alkaloid has great stimulation on the synthesis of chlorophyll, vitamin C and peroxidase activities in the leaves of the green vegetables, plays a role in inhibiting the generation of malondialdehyde, delays the damage and aging degree of the green vegetables, and accelerates the metabolic activity in the bodies of the green vegetables. During the use of the fertilizer, humic acid, silicon fertilizer and plant alkaloid are continuously released, and the three synergistically act on plants, so that the antibacterial capacity of the plants and the lethality to root nematodes are enhanced, the disaster resistance of the plants is improved, and the influence of external adverse environments on the plant growth is obviously reduced.

The above-mentioned contents are only for illustrating the technical idea of the present invention, and the protection scope of the present invention is not limited thereby, and any modification made on the basis of the technical idea of the present invention falls within the protection scope of the claims of the present invention.

Claims

1. The humic acid insect-proof antibacterial silicon fertilizer containing alkaloid is characterized by comprising, by mass, 100 parts of humate, 1-20 parts of a silicon-containing compound and 1-20 parts of alkaloid.

2. The humic acid insect-proof antibacterial silicon fertilizer containing alkaloid as claimed in claim 1, wherein the humate is one or more of ammonium humate, potassium humate and sodium humate.

3. The humic acid insect-prevention antibacterial silicon fertilizer containing alkaloid as claimed in claim 1, wherein the alkaloid is extracted and separated from natural plant containing alkaloid, and the natural plant containing alkaloid is any one of coptis chinensis, pyrola, phellodendron amurense, root of Chinese barberry and fritillaria.

4. The humic acid insect-proof antibacterial silicon fertilizer containing alkaloid as claimed in claim 3, wherein the alkaloid is extracted by the following method:

5. The humic acid insect-proof antibacterial silicon fertilizer containing alkaloid as claimed in claim 4, wherein the reflux time of each time when the alkaloid is extracted is not less than 1.5 h.

6. The humic acid insect-proof antibacterial silicon fertilizer containing alkaloid as claimed in claim 4, wherein the volume ratio of methanol to water is 3:1 when alkaloid is extracted.

7. The humic acid insect-proof antibacterial silicon fertilizer containing alkaloid as claimed in claim 1, wherein the silicon-containing compound is one or two of sodium silicate and calcium silicate.

8. The preparation method of the humic acid insect-proof antibacterial silicon fertilizer containing the alkaloid as claimed in any one of claims 1 to 7, which is characterized by comprising the following steps: