Mixed biological leaching liquor as well as preparation method and application thereof

Abstract

The invention belongs to the technical field of biological control, and particularly relates to a mixed biological leaching solution and a preparation method and application thereof.The preparation method comprises the steps that S1, raw material treatment is conducted, specifically, solidago canadensis, ageratum conyzoides, scandent hop and black walnut roots are selected as plant raw materials, the plant raw materials are cut into pieces and then subjected to high-pressure sterilization, water is added, and mixing is conducted to obtain a mixture; s2, the mixture is soaked for 48-72 h under the environment condition of 20-30 DEG C, and an extraction stock solution is obtained; and S3, centrifuging the leaching stock solution, taking supernate, and filtering to obtain the mixed biological leaching solution. The mixed biological extract prepared by the invention can be used for preventing and treating seed germination, seedling growth and flowering processes of solidago canadensis, and can also be used for preparing a product for preventing and treating solidago canadensis. The problem that a biological medicament for preventing and treating solidago canadensis is lacked at present is solved.

Description

Technical Field

[0001] The present invention belongs to the technical field of biological control, and particularly relates to a mixed biological extract, a preparation method thereof, and an application thereof. Background Art

[0002] For many years, the control of Solidago virgaurea in China has mainly adopted manual control and chemical control measures. In addition, a large number of studies have been conducted on biological control measures such as using fungi isolated from plants, parasitic plant Cuscuta japonica, and insect feeding to control Solidago virgaurea. However, so far, the biological control technology is not yet mature, and the effect is greatly affected by the environment, and it cannot be prepared and used on a large scale in industrial production. Summary of the Invention

[0003] Aiming at the problem of the lack of biological control technology for Solidago virgaurea, the present invention provides a mixed biological extract, a preparation method thereof, and an application thereof. Using Solidago virgaurea, Ageratum conyzoides, Humulus scandens, and Juglans nigra as plant raw materials, a mixed biological extract that can be applied to Solidago virgaurea is prepared through a simple extraction technology. The mixed biological extract in the present invention has stable properties and can also be applied to other products for controlling Solidago virgaurea, solving the problem that the current biological control technology for Solidago virgaurea cannot be prepared and used on a large scale in industrial production.

[0004] The specific technical solutions are as follows:

[0005] A mixed biological extract, the raw materials of the mixed biological extract include Solidago virgaurea, Ageratum conyzoides, Humulus scandens, and Juglans nigra roots, and the mass ratio of Solidago virgaurea, Ageratum conyzoides, Humulus scandens, and Juglans nigra roots in the raw materials is 4:1:2:3 or 1:2:3:4.

[0006] A preparation method of a mixed biological extract, comprising the following steps:

[0007] Weigh Solidago virgaurea, Ageratum conyzoides, Humulus scandens, and Juglans nigra roots according to the ratio, crush them, add water to the crushed raw materials for soaking, and filter after the soaking is completed to obtain the mixed biological extract.

[0008] In any of the above possible implementation manners, a further implementation manner is provided, and the mass ratio of water to the raw materials is 3-8:1.

[0009] In any of the above possible implementation manners, a further implementation manner is provided, the soaking temperature is 20-30 °C, and the soaking time is 48-72 h.

[0010] The present invention also provides an application of the mixed biological extract in controlling Solidago virgaurea.

[0011] For any of the possible implementation manners described above, a further implementation manner is provided. The application includes directly using a mixed biological leaching solution and products prepared by using the mixed biological leaching solution. The products include any one of biological control herbicides, biological enzymes, and biological organic fertilizers.

[0012] For any of the possible implementation manners described above, a further implementation manner is provided. The mixed biological leaching solution and the products prepared by using the mixed biological leaching solution are applied to the seed germination, and / or seedling growth, and / or flowering stage of Solidago virgaurea.

[0013] The beneficial effects of the present invention are as follows:

[0014] 1. By using the leaching method, according to the types and functional differences of allelochemicals released by different plants, Solidago virgaurea, Ageratum conyzoides, Humulus scandens, and Juglans nigra leaching materials are mixed. Through the synergistic effect of multiple components, a complementary inhibitory effect is formed to replace a single allelochemical, which becomes an effective means for controlling Solidago virgaurea.

[0015] 2. Selecting and mixing different plant residues in the present invention can improve the resource utilization rate and reduce the dependence on a single raw material. The present invention uses a simple leaching method to extract effective substances. The method is simple and pollution-free, and it is a completely biological control agent. Moreover, the raw materials of the present invention need to use Solidago virgaurea, which not only consumes Solidago virgaurea but also solves the problem of raw material sources, and the production cost is low. The leaching solution prepared by the present invention has stable properties, is not restricted by the use environment, has no chemical pollution, and is suitable for large-scale industrial production and popularization and use. Description of the Drawings

[0016] Figure 1 The figure shows the experimental results of the inhibition of Solidago virgaurea seed germination by the experimental group and the control group in Experimental Example 1 of the present invention;

[0017] Figure 2 The figure shows the experimental results of the inhibition of Solidago virgaurea seedling growth by the experimental group and the control group in Experimental Example 2 of the present invention;

[0018] Figure 3 The figure shows the experimental results of the inhibition of Solidago virgaurea development by the experimental group and the control group in Experimental Example 3 of the present invention. Detailed Embodiments

[0019] The specific embodiments of the present invention will be described in detail below with reference to the specific drawings. It should be noted that the technical features described in the following embodiments or the combination of technical features should not be considered as isolated, and they can be combined with each other to achieve better technical effects.

[0020] Example 1

[0021] This embodiment provides a method for preparing a mixed biological extract, comprising the following steps:

[0022] Select the upper parts of Solidago virgaurea, Ageratum conyzoides, Humulus scandens and the root bark of Juglans nigra. The dosage ratio of the four materials is Solidago virgaurea: Ageratum conyzoides: Humulus scandens: Juglans nigra = 4:1:2:3, and the cumulative dosage is 100 g. After crushing, add 500 mL of distilled water, soak at room temperature for 48 hours, and filter to obtain the mixed biological extract.

[0023] A mixed biological extract prepared in this embodiment is applied to the control of Solidago virgaurea, including inhibiting the seed germination of Solidago virgaurea, and / or the growth of seedlings, and / or the flowering process.

[0024] A mixed biological extract prepared in this embodiment can also be applied to the preparation of products for Solidago virgaurea, including but not limited to biological control herbicides, biological enzymes, and biological organic fertilizers. Specifically, for example, for a biological control herbicide, take 500 ml of the mixed biological extract, add 1 ml of a surfactant such as Tween 20, mix and stir evenly to prepare the biological control herbicide, and store it at 4°C for later use.

[0025] When in use, spray before Solidago virgaurea blooms. The spraying amount is 50 ml / plant, spray continuously for 3 days, and repeat spraying once after an interval of 7 days.

[0026] Example 2

[0027] The difference between this embodiment and Example 1 lies in the different ratios of plant raw materials. Specifically, in this embodiment, the dosage ratio of the four materials is Solidago virgaurea: Ageratum conyzoides: Humulus scandens: Juglans nigra = 1:2:3:4. Other preparation steps and parameters are the same as those in Example 1.

[0028] Comparative Example 1

[0029] The difference between this comparative example and Example 1 lies in the different ratios of plant raw materials. Specifically, in this comparative example, the dosage ratio of the four materials is Solidago virgaurea: Ageratum conyzoides: Humulus scandens: Juglans nigra = 2:3:4:1. Other preparation steps and parameters are the same as those in Example 1.

[0030] Comparative Example 2

[0031] The difference between this comparative example and Example 1 lies in the different ratios of plant raw materials. Specifically, in this comparative example, the dosage ratio of the four materials is Solidago virgaurea: Ageratum conyzoides: Humulus scandens: Juglans nigra = 3:4:1:2. Other preparation steps and parameters are the same as those in Example 1.

[0032] Experimental Example 1: Seed Germination Inhibition Experiment

[0033] Experimental method: The biocontrol herbicide prepared from the mixed biological leaching solution prepared in the above Examples 1 and 2 (added with 0.5% v / v Tween 20) was used as the experimental group (TR1 and TR4), and the biocontrol herbicide prepared from the mixed biological leaching solution prepared in Comparative Example 1 and Comparative Example 2 (added with 0.5% v / v Tween 20) was used as the control group (TR2 and TR3), and a blank control group and a tribenuron-methyl control group at 4 mg / L were set up. Among them, tribenuron-methyl was from a commercially available product. During the experiment, the seeds of Solidago virgaurea were placed in a filter paper culture dish (5 mL / dish) containing different concentrations of the experimental group or the control group, 50 seeds per dish, with 3 replicates, and cultured at 25 °C. 5 mL of deionized water and 5 mL of the corresponding medicament were added to the culture dishes in the experimental group and the control group respectively. On the 7th day, the germination rate and radicle length of the seeds of Solidago virgaurea under different treatments were counted, and 10 seeds were measured for the radicle length of each treatment.

[0034] Experimental results: As shown in the appendix Figure 1 , the four biocontrol herbicides had obvious inhibitory effects on the seed germination and radicle length of Solidago virgaurea (P < 0.05), but the inhibitory effects were quite different, reducing the seed germination rate to 75.67% - 20.33% respectively (the seed germination rate of the blank control was 96.67%) ( Figure 1 A), and the radicle length decreased by 20.53% - 83.44% respectively compared with the blank control ( Figure 1 B). The inhibitory effects of the four treatment conditions on the seeds of Solidago virgaurea were shown as: TR4 > TR1 > TR3 > TR2. The biocontrol herbicide of TR4 had a significant inhibitory effect on the seed germination and radicle length of Solidago virgaurea, and its inhibitory effect on seed germination was higher than that of the chemical herbicide tribenuron-methyl, and its inhibitory effect on radicle length was equivalent to that of tribenuron-methyl.

[0035] Experimental Example 2: Seedling growth inhibition experiment

[0036] Experimental method: The experimental group and the control group were the same as those in Experimental Example 1. During the experiment, the seedlings of Solidago virgaurea were transplanted from the field to a spray pot containing a 1:1 mixture of soil and humus. The diameter of the pot was 10 cm, and 3 seedlings were transplanted into each pot. When the seedling height reached 10 cm, watering treatment was carried out. The seedlings of Solidago virgaurea were watered with 100 mL of biocontrol herbicides or control agents at different concentrations, twice a week for 4 consecutive weeks. After 4 weeks of treatment, the plant height, aboveground biomass (fresh / dry weight), chlorophyll content (SPAD), and MDA content of Solidago virgaurea under different treatment conditions were measured. For each treatment, the plant height of 3 seedlings was measured; after collecting and weighing the aboveground stems and leaves, they were placed in an oven at 50 °C and dried for 7 days to measure the aboveground dry weight. The SPAD value was measured using a handheld chlorophyll meter SPAD-502Plus. For each treatment, the SPAD values of 6 leaves at the same leaf position were measured, and the measurement was repeated 3 times; the determination of MDA content was carried out using a lipid oxidation (MDA) detection kit (Biosharp), and the specific steps were referred to the instruction manual.

[0037] Experimental results: As shown in Figure 2 the figure, the biocontrol herbicides under different treatments all showed a significant inhibitory effect on the growth of Solidago virgaurea seedlings (P < 0.05), manifested as a significant decrease in plant height, aboveground fresh weight and dry weight, chlorophyll content, and a significant increase in MDA content. The inhibitory effects of the four treatment conditions on the seedlings of Solidago virgaurea were similar to those on the seeds, also showing: TR4 > TR1 > TR3 > TR2. Under the TR2 treatment, the biocontrol herbicide had already shown a significant inhibitory effect on the seedlings of Solidago virgaurea. Compared with the blank control, the plant height, aboveground fresh weight and dry weight of the seedlings decreased by 15.14%, 13.55%, and 16.49% respectively, while the chlorophyll content increased by 3.05 times. Under the TR4 treatment, the biocontrol extract and tribenuron-methyl control showed the strongest inhibitory effect, and the seedlings of Solidago virgaurea basically stopped growing. Compared with the blank control, the plant height, aboveground fresh weight and dry weight, and chlorophyll content of the seedlings decreased by 39.51%, 40.47%, 41.59%, and 27.11% respectively, and the MDA content increased sharply by 8.94 times, indicating that the degree of membrane lipid peroxidation of the cell membrane of Solidago virgaurea seedlings increased.

[0038] Experimental Example 3: Flowering inhibition experiment

[0039] Experimental method: The experimental group and the control group were the same as in Experimental Example 1. During the experiment, the extract or the control agent was sprayed on the leaf surface of the Solidago virgaurea plants before flowering. The spraying volume was 50 mL / plant, and it was sprayed continuously for 3 days. After a 7-day interval, it was sprayed again. The flowering rate, inflorescence length, and pollen viability (TTC staining method) of Solidago virgaurea were counted. The flowering rate was the proportion of the opened flowers on the inflorescences at specific positions of a single plant to the total flower buds. Three inflorescences were counted for each treatment, and their inflorescence lengths were also counted. The pollen viability was measured using a pollen viability detection kit (Biosharp), and the specific steps were referred to the instruction manual. Each treatment was counted 3 times.

[0040] Experimental results: As shown in the Figure 3 appendix, the biocontrol herbicides of the four treatments all had an obvious abortion effect on Solidago virgaurea. After the treatment, the flowering rate, inflorescence length, and pollen viability of Solidago virgaurea were all significantly reduced (P < 0.05). Under the TR4 treatment, the biocontrol extract significantly reduced the flowering rate of Solidago virgaurea and shortened its inflorescence. The flowering rate and inflorescence length decreased by 85.91% and 74.57% respectively compared with the blank control, and the pollen viability was less than 1 / 4 of the blank control. The abortion effect was equivalent to that of the tribenuron-methyl treatment.

[0041] Although several embodiments of the present invention have been given in this article, those skilled in the art should understand that the embodiments in this article can be changed without departing from the spirit of the present invention. The above embodiments are only exemplary and should not be used as the limitation of the scope of the rights of the present invention.

Claims

1. A mixed biological leaching solution, characterized in that, The raw material of the mixed biological leaching solution includes Solidago virgaurea, Ageratum conyzoides, Humulus scandens, and Juglans nigra roots. The mass ratio of Solidago virgaurea, Ageratum conyzoides, Humulus scandens, and Juglans nigra roots in the raw material is 4:1:2:3 or 1:2:3:

4.

2. The preparation method of a mixed biological leaching solution according to claim 1, characterized in that, It includes the following steps: Weigh Solidago virgaurea, Ageratum conyzoides, Humulus scandens, and Juglans nigra roots in proportion and then crush them. Add water to the crushed raw materials for soaking. After the soaking is completed, filter to obtain the mixed biological leaching solution.

3. The preparation method of a mixed biological leaching solution according to claim 2, characterized in that, The mass ratio of the water to the raw materials is 3-8:

1.

4. The preparation method of a mixed biological leaching solution according to claim 2, wherein The soaking temperature is 20-30 °C, and the soaking time is 48-72 h.

5. Application of a mixed biological leaching solution prepared by the method described in claim 1 or any one of claims 2-4 in controlling Solidago virgaurea.

6. The application of a mixed biological leaching solution in controlling Solidago canadensis as claimed in claim 5, characterized in that, The application includes directly using a mixed biological leaching solution and products prepared from the mixed biological leaching solution. The products include any one of biological control herbicides, biological enzymes, and biological organic fertilizers.

7. Use of a mixed biological leaching solution in preventing and controlling Solidago canadensis as described in claim 6, characterized in that, The said mixed biological leaching solution and the products prepared from the mixed biological leaching solution are applied to the seed germination, and / or seedling growth, and / or flowering stage of Solidago virgaurea.

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