CN115363055A - Preparation method and application of bidens bipinnata leaching liquor capable of preventing and treating crop bacterial wilt - Google Patents
Preparation method and application of bidens bipinnata leaching liquor capable of preventing and treating crop bacterial wilt Download PDFInfo
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- CN115363055A CN115363055A CN202210946453.8A CN202210946453A CN115363055A CN 115363055 A CN115363055 A CN 115363055A CN 202210946453 A CN202210946453 A CN 202210946453A CN 115363055 A CN115363055 A CN 115363055A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
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- A—HUMAN NECESSITIES
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Abstract
The invention discloses a preparation method and application of a spanishneedles herb leaching liquor capable of preventing and treating crop bacterial wilt, relates to the technical field of biological prevention and treatment of plant diseases, and can be used for preventing and treating crop bacterial wilt. The method comprises the steps of mixing the bidens bipinnata with a phosphate buffer solution according to the proportion of 200g to 1L, and leaching for 2d at normal temperature to obtain a bidens bipinnata leaching solution for preventing and treating crop bacterial wilt. The pot control effect is measured by adopting a leaching liquor root irrigation inoculation method, and the control effect of the bidens bipinnata leaching liquor treatment on the tobacco bacterial wilt is 50.7% 14d after inoculation. The preparation method of the bidens pilosa leaching liquor provided by the invention has a good prevention and treatment effect on crop bacterial wilt without adding any organic solvent.
Description
Technical Field
The invention belongs to the technical field of biological control of plant diseases, and particularly relates to a preparation method and application of a spanishneedles herb leaching liquor capable of controlling crop bacterial wilt.
Background
The crop bacterial wilt is a typical bacterial vascular bundle disease of plants, and the pathogen of the disease is Ralstonia solanacearum (Ralstonia solanacearum)Ralstonia solanacearum) The disease is an important disease affecting the yield and quality of crops such as eggplants, tomatoes, peppers, potatoes, peanuts and the like in the planting process, and can cause the roots, stems and leaves of the plants to have typical wilting symptoms and seriously affect the growth and development of the plants.
For the prevention and treatment research of crop greensickness, planting resistant varieties, agricultural prevention and treatment measures such as crop rotation, grafting and the like, chemical prevention and biological prevention and treatment and the like are mainly adopted in fields, wherein the chemical prevention and treatment is a relatively common prevention and treatment method, and some prevention and treatment effects are also obtained. Biological control has become a focus of research in recent years. Plants are rich in active substances and beneficial microorganisms, and have natural defense effects on plant diseases and insect pests, so that the prevention and control of diseases by using plant-derived extracts is one of the main methods for biological prevention and control.
Bidens bipinnata (Bidens bipinnata) ((B))Bidens pilosaL.) annual herbaceous plants, produced in the provinces of china. Can grow in villages, roadside and wasteland, is a common Chinese herbal medicine, and has effects of clearing away heat and toxic materials, removing blood stasis and promoting blood circulation. The ethanol extract and the methanol extract of the bidens bipinnata are reported to be used for inhibiting plant pathogenic fungi such as anthracnose of melons, fruits and vegetables and inhibiting human pathogenic bacteria such as pseudomonas aeruginosa, staphylococcus aureus and the like, but do not relate to the plant pathogenic bacteria, and the ethanol extract and the methanol extract are extracted by adopting an organic solvent, so that the environmental pollution is easily caused.
Disclosure of Invention
The invention aims to provide a preparation method and application of a spanishneedles herb leaching liquor with the function of preventing and treating crop bacterial wilt.
In order to achieve the purpose, the invention adopts the following technical scheme:
a method for preparing herba Bidentis Bipinnatae leaching solution for preventing and treating crop bacterial wilt comprises cutting fresh herba Bidentis Bipinnatae into 2-5cm segments, adding phosphate buffer, leaching at room temperature for 2d, and filtering with sterile gauze to obtain herba Bidentis Bipinnatae leaching solution. Wherein the material-liquid ratio of the bidens bipinnata to the phosphate buffer solution is 0.2g/mL.
Further, the concentration of the phosphate buffer was 0.2moL/mL.
The application of the bidens bipinnata leaching liquor in preventing and treating crop bacterial wilt.
Researches show that the leaching concentration (the material-liquid ratio of the bidens bipinnata to the leaching medium phosphate buffer solution) of the bidens bipinnata phosphate buffer solution leaching liquor is 0.2g/mL, and the plate bacteriostasis result is the best when the leaching time is 2d.
The prevention and treatment effect of the spanishneedles herb leaching liquor on the tobacco bacterial wilt is as follows: the result of pot control effect measurement by using a leaching liquor root irrigation inoculation method shows that 14 days after inoculation, the control effect of the bidens bipinnata phosphate buffer solution on tobacco bacterial wilt is 50.7%.
The invention has the advantages that:
(1) the herba Bidentis Bipinnatae leaching liquor has an inhibiting effect on common crop ralstonia solanacearum, and a pot experiment also has a good control effect, so that a new strategy is provided for controlling crop ralstonia solanacearum.
(2) The herba Bidentis Bipinnatae leaching liquor has a lasting effect of more than 20 days on the tobacco bacterial wilt pathogens, has no pollution, residue and toxicity compared with chemical pesticides, has higher safety on the environment, is a green prevention and control measure, and has important significance on the prevention and control of the crop bacterial wilt pathogens.
(3) The herba Bidentis Bipinnatae leaching liquor contains abundant chemical substances which mainly comprise flavone, polyacetylene, terpenoids, volatile oil and the like, and the herba Bidentis Bipinnatae plants contain abundant microorganisms, so that the herba Bidentis Bipinnatae leaching liquor contains abundant microorganisms and active substances and can be applied to the control of crop bacterial wilt.
Drawings
FIG. 1 is a graph showing the bacteriostatic effects of Bidens pilosa leaching solutions of different concentrations on tobacco withering in example 1;
FIG. 2 is a graph showing the bacteriostatic effects of Bidens pilosa leaching solution on tobacco withering at different leaching times in example 2;
FIG. 3 is a graph of plate inhibition results (concentration 0.20g/mL, leaching time 2 d) for the Bidens bipinnata leaching solution of example 2, water leaching solution A, phosphate leaching solution B;
FIG. 4 is a graph showing the control effect of the Bidens pilosa leaching solution on tobacco bacterial wilt in example 4;
FIG. 5 is a 14d schematic representation of the Bidens bipinnata leaching solution of example 4 for controlling tobacco bacterial wilt, phosphate leaching solution A + tobacco ralstonia solanacearum, zhongshengmycin B + tobacco ralstonia solanacearum, and C tobacco ralstonia solanacearum.
Detailed Description
In order to make the aforementioned and other features and advantages of the invention more comprehensible, embodiments accompanied with figures are described in detail below. The method of the present invention is a method conventionally used in the art unless otherwise specified.
The preparation method of the phosphate buffer solution comprises the following steps: respectively adding 11.856 g of NaH 2 PO 4 ·2H 2 O and 44.392 g of Na 2 HPO 4 ·12H 2 Dissolving O in 1L distilled water to obtain 0.2moL/mL phosphate buffer solution for leaching herba Bidentis Bipinnatae.
EXAMPLE 1 plate inhibition Effect of Bidens bipinnata leach liquor of different concentrations on Ralstonia solanacearum
Obtaining a bidens pilosa leaching liquor: collecting fresh plants of herba Bidentis Bipinnatae, weighing 20g, 40g, 60g, 80g and 200g herba Bidentis Bipinnatae respectively, cutting into 2-5cm segments, placing the cut herba Bidentis Bipinnatae in a sterile container, adding 1L water or phosphate buffer solution respectively, standing at room temperature for leaching for 2d, filtering with sterile gauze to remove residue in herba Bidentis Bipinnatae leaching solution, and placing the filtrate in a refrigerator at 4 deg.C for use to obtain leaching solutions with different concentrations.
And (3) determining the bacteriostatic effect of the bidens bipinnata leaching liquor on crop ralstonia solanacearum: selecting ralstonia solanacearum to be tested in an NB culture medium by an inhibition zone method, carrying out constant-temperature shaking culture (28 ℃,180 r/min) for 24h, uniformly mixing a bacterial suspension and a CPG solid culture medium which is melted at a proper temperature according to a ratio of 1; respectively sucking herba Bidentis Bipinnatae leaching solution (5 μ L) with different concentrations on the plate, air drying, culturing in 28 deg.C incubator, and repeating each treatment for 3 times.
As shown in the results of fig. 1: the herba Bidentis Bipinnatae water extractive solution and herba Bidentis Bipinnatae phosphate extractive solution have the best antibacterial effect at a concentration of 0.20g/mL, the average radius of the antibacterial zone is 2.67mm and 4.67mm, respectively, and the corresponding plate antibacterial result is shown in FIG. 3.
Example 2 plate bacteriostatic effect of Bidens pilosa Leaching liquid on tobacco ralstonia solanacearum in different leaching time
Obtaining a bidens pilosa leaching liquor: collecting fresh plants of herba Bidentis Bipinnatae, weighing 200g herba Bidentis Bipinnatae respectively, cutting into 2-5cm segments, placing the cut herba Bidentis Bipinnatae in sterile container, adding 1L water or phosphate buffer solution, standing at room temperature, leaching for different time, filtering with sterile gauze to remove residue in herba Bidentis Bipinnatae leaching solution, and placing the filtrate in refrigerator at 4 deg.C for use to obtain leaching solutions with different concentrations.
The leaching solutions of the sticktight of 1d, 2d, 3d, 4d, 5d, 6d and 7d are respectively leached, and the plate bacteriostasis effect of the sticktight on the tobacco ralstonia solanacearum is determined by adopting a bacteriostasis ring method (the specific operation is the same as that in example 1).
As shown in the results of FIG. 2, the Bidens pilosa water extract and Bidens pilosa phosphate extract both showed the best bacteriostatic effect after 2d leaching, and the radius of the zone of inhibition was 2.67mm and 4.67mm, respectively, and the corresponding plate bacteriostatic results are shown in FIG. 3.
Example 3 plate bacteriostatic effect of Bidens pilosa Leaching liquid on crop pathogenic bacteria
The method for determining the content of Bacillus subtilis in the leaching solution of herba Bidentis Bipinnatae is performed by inhibiting bacteria in the manner of circle (the same operation as in example 1)Ralstonia solanacearium) And ralstonia solanacearum (C. Solani)R. solanacearium) And Ralstonia solanacearum (sweet potato)R. solanacearium) Hot pepper Ralstonia solanacearum (ii) (R. solanacearium) Bacterial wilt of potato (R. solanacearium) Tobacco cavity pathogen: (Pectobacterium carotovorum) Tobacco wild fire pathogen: (Pseudomonas amygdali) Bacterial spot of tomato (A), (B)P. syringae) Pseudomonas chicory (A)P. cichorii) Chinese cabbage Soft rot fungus (B)P. carotovorum) Bacteria of common scab of Capsicum (A), (B), (C)Xanthomonas euvesicatoria) And the results are shown in table 1. Wherein the concentration of the herba Bidentis Bipinnatae leaching liquor is 0.20g/mL, and the leaching time is 2d.
TABLE 1 plate bacteriostatic effect of herba Bidentis Bipinnatae leaching solution on crop pathogenic bacteria
The results in table 1 show that the bidens bipinnata leach liquor has a strong bacteriostatic effect on ralstonia solanacearum of different crops, but has no bacteriostatic effect on plant pathogenic bacteria such as tobacco cavity bacteria, tobacco wildfire bacteria, tomato bacterial spot bacteria, chicory pseudomonas, pakchoi soft rot bacteria, capsicum scab bacteria and the like, which proves that the bidens bipinnata leach liquor obtained by the invention has specificity on the inhibitory effect on ralstonia solanacearum. Wherein, the bacteriostatic effect of the bidens bipinnata phosphate leaching solution is better than that of the bidens bipinnata water leaching solution.
Example 4 measurement of Effect of Bidens bipinnata leaching solution on controlling tobacco bacterial wilt
Selecting healthy tobacco seedlings with consistent growth vigor, respectively irrigating 30mL of bidens bipinnata phosphate leaching liquor with the concentration of 0.20g/mL and the leaching time of 2d and zhongshengmycin (the effective content of the zhongshengmycin is 3%) diluted by 750 times, repeating the treatment for 6 tobacco plants for 3 times, irrigating roots after 24 hours, and inoculating 30mL of tobacco ralstonia solanacearum suspension (OD 600= 0.8).
As shown in the results of FIG. 4 and FIG. 5, after inoculation for 14d, the control effect of the Bidens pilosa phosphate leaching liquor on tobacco ralstonia solanacearum is 50.7%; after 21d of inoculation, the control effect of the composition is still 40.0 percent, which is higher than the control effect (16.4 percent) of the zhongshengmycin diluted by 750 times. The control effect is shown in fig. 5.
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
Claims (4)
1. A preparation method of a spanishneedles herb leaching liquor with the function of preventing and treating crop bacterial wilt is characterized in that: cutting fresh herba Bidentis Bipinnatae into 2-5cm pieces, adding phosphate buffer solution according to the material-to-liquid ratio of 0.2g/mL, leaching at room temperature for 2d, and filtering with sterile gauze to obtain herba Bidentis Bipinnatae leaching solution.
2. The method for preparing the leaching liquor for controlling the crop bacterial wilt and the sponished needles herb according to claim 1, wherein the feed-liquor ratio of the sponished needles herb to the phosphate buffer is 0.2g/mL.
3. The production method according to claim 1, characterized in that: the concentration of the phosphate buffer was 0.20moL/mL, pH =7.
4. Use of the Bidens pilosa Leaching solution prepared by the method of claim 1 for controlling crop bacterial wilt.
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| CN104490970A (en) * | 2014-12-25 | 2015-04-08 | 李健 | Medicament composition for treating tinea corporis and preparation method of medicament composition |
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