CN115363055B - Preparation method and application of spanishneedles herb extract liquid for preventing and treating crop bacterial wilt - Google Patents
Preparation method and application of spanishneedles herb extract liquid for preventing and treating crop bacterial wilt Download PDFInfo
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- CN115363055B CN115363055B CN202210946453.8A CN202210946453A CN115363055B CN 115363055 B CN115363055 B CN 115363055B CN 202210946453 A CN202210946453 A CN 202210946453A CN 115363055 B CN115363055 B CN 115363055B
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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
- 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
- A01N65/08—Magnoliopsida [dicotyledons]
- A01N65/12—Asteraceae or Compositae [Aster or Sunflower family], e.g. daisy, pyrethrum, artichoke, lettuce, sunflower, wormwood or tarragon
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P3/00—Fungicides
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- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
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Abstract
The invention discloses a preparation method and application of a spanishneedles herb leaching solution for 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 sticktight is mixed with phosphate buffer solution according to the proportion of 200g to 1L, and is leached for 2d at normal temperature to obtain the sticktight leaching solution which is used for preventing and treating crop bacterial wilt. The pot control effect is measured by adopting a leaching solution root filling inoculation method, and the control effect of the spanishneedles herb leaching solution treatment on tobacco bacterial wilt is 50.7% after inoculation of 14. 14 d. The preparation method of the spanishneedles herb extract provided by the invention has better control 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 in particular relates to a preparation method and application of a spanishneedles herb leaching solution for controlling crop bacterial wilt.
Background
Crop bacterial wilt is a typical plant bacterial vascular bundle disease, and the pathogen of the disease is ralstonia solanacearum @ or the likeRalstonia solanacearum) Is an important disease affecting the yield and quality of eggplants, tomatoes, peppers, potatoes, peanuts and the like in the planting process of the crops, and can cause the roots, stems and leaves of the plants to have typical wilting symptoms so as to seriously influence the growth and development of the plants.
For the prevention and treatment research of crop bacterial wilt, planting resistant varieties, agricultural prevention and treatment measures such as rotation, grafting and the like, chemical prevention and treatment, 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 obtained, but due to the problems of environmental pollution, pathogen resistance and the like, safer and pollution-free biological prevention and treatment are needed to control the occurrence and development of the diseases. Biological control has therefore become a research hotspot in recent years. Plants themselves are rich in abundant active substances and some beneficial microorganisms, and can have natural defense effects on plant diseases and insect pests, so that prevention and control of diseases by using plant source extracts is one of the main methods of biological control.
Herba Bidentis BipinnataeBidens pilosaL.), annual herbaceous plants, produced in the multiple provinces of china. Can grow in villages, roadsides and barren lands, is a common herbal medicine for Chinese folks, and has the effects of clearing heat, detoxicating, removing stasis and promoting blood circulation. The ethanol extract and the methanol extract of sticktight have been reported to be used for inhibiting plant pathogenic fungi such as melon, fruit and vegetable anthracnose and human pathogenic bacteria such as pseudomonas aeruginosa, staphylococcus aureus and the like, but the ethanol extract and the methanol extract of sticktight are not related to plant pathogenic bacteria, and the ethanol extract and the methanol extract are extracted by adopting an organic solvent, so that environmental pollution is easily caused.
Disclosure of Invention
The invention aims to provide a preparation method and application of a spanishneedles herb extract for preventing and treating crop bacterial wilt, which are characterized in that the spanishneedles herb extract is prepared and applied to preventing and treating crop bacterial wilt, phosphate buffer solution is used as a leaching medium, the environment is not polluted, and the obtained spanishneedles herb extract has a specific inhibition effect on bacterial wilt.
In order to achieve the above purpose, the invention adopts the following technical scheme:
a herba Bidentis Bipinnatae leaching solution for preventing and treating crop bacterial wilt is prepared by cutting fresh herba Bidentis Bipinnatae into 2-5cm pieces, adding phosphate buffer solution, leaching at room temperature for 2d, and filtering with sterile gauze to obtain herba Bidentis Bipinnatae leaching solution. Wherein, the feed liquid ratio of the spanishneedles herb to the phosphate buffer solution is 0.2g/mL.
Further, the concentration of the phosphate buffer was 0.2moL/L.
The application of the sticktight leaching solution in preventing and treating crop bacterial wilt.
The research shows that the leaching solution of the spanishneedles herb phosphate buffer has the best plate bacteriostasis result when the leaching concentration (the feed liquid ratio of spanishneedles herb to leaching medium phosphate buffer) is 0.2g/mL and the leaching time is 2d.
The control effect of the sticktight leaching solution on tobacco bacterial wilt is as follows: the result of measuring the pot control effect by adopting the leaching solution root filling inoculation method shows that 14. 14d percent after inoculation, the control effect of the spanishneedles herb phosphate buffer solution on tobacco bacterial wilt is 50.7 percent.
The invention has the advantages that:
(1) the sticktight leaching solution has an inhibiting effect on common crop bacterial wilt, and a potting experiment also has a good control effect, so that a new strategy for controlling crop bacterial wilt is provided.
(2) The sticktight leaching solution can have a continuous effect of over 20 days on tobacco bacterial wilt, has no pollution, no residue and no 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 crop bacterial wilt.
(3) The herba Bidentis Bipinnatae leaching solution contains abundant chemical substances, mainly including flavone, polyacetylene, terpenes, volatile oil and the like, and contains abundant microorganisms in herba Bidentis Bipinnatae plants, so that the herba Bidentis Bipinnatae leaching solution contains abundant microorganisms and active substances, can be applied to the prevention and treatment of crop bacterial wilt, and has the advantages of being easily decomposed by natural microorganisms, not polluting the environment, not easily generating drug resistance, having relatively strong specificity, having no toxic or harmful effects on beneficial organism groups and the like compared with chemical agents.
Drawings
FIG. 1 shows the bacteriostatic effect of the leaching solution of Bidens pilosa at different concentrations on tobacco bacterial wilt in example 1;
FIG. 2 shows the bacteriostatic effect of the spanishneedles herb extract on tobacco bacterial wilt at different extraction times of example 2;
FIG. 3 is a graph showing the results of bacteriostasis on a flat plate of the extract of Bidens pilosa of example 2 (concentration 0.20g/mL, extraction time 2 d), A water extract, B phosphate extract;
FIG. 4 shows the control effect of the sticktight extract of example 4 on tobacco bacterial wilt;
fig. 5 is a physical diagram of 14 th d of the sticktight extract of example 4 for controlling tobacco bacterial wilt, phosphate extract a+tobacco bacterial wilt, biotin in b+tobacco bacterial wilt, C tobacco bacterial wilt.
Detailed Description
The following examples are provided to illustrate the above features and advantages of the present invention. The method of the invention is a conventional method in the art unless specifically stated otherwise.
The preparation method of the phosphate buffer solution comprises the following steps: respectively NaH of 11.856 g 2 PO 4 ·2H 2 Na of O and 44.392 g 2 HPO 4 ·12H 2 O is dissolved in 1L distilled water, namely 0.2moL/L phosphate buffer solution, and is used for leaching sticktight.
EXAMPLE 1 Flat plate inhibitory Effect of Bidens Bipinnata extract at different concentrations on tobacco bacterial wilt
Obtaining sticktight leaching solution: collecting fresh plants of herba Bidentis Bipinnatae, respectively weighing 20g, 40g, 60g, 80g, and 200g herba Bidentis Bipinnatae, cutting into 2-5cm pieces, placing the cut herba Bidentis Bipinnatae in sterile container, respectively adding 1L water or phosphate buffer solution, 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 refrigerator at 4deg.C for use to obtain leaching solutions with different concentrations.
Determination of antibacterial effect of sticktight leaching solution on crop bacterial wilt: selecting bacterial wilt of tobacco to be tested in NB culture medium by adopting a bacteriostasis circle method, shaking culture at constant temperature (28 ℃ and 180 r/min) for 24 hours, uniformly mixing bacterial suspension with CPG solid culture medium which is melted at proper temperature in a ratio of 1:100, and pouring into a plate; respectively sucking herba Bidentis Bipinnatae leaching solutions (5 μl) with different concentrations, air drying, and culturing in 28 deg.C incubator, and repeating each treatment for 3 times.
The results show in fig. 1: the herba Bidentis Bipinnatae water extract and herba Bidentis Bipinnatae phosphate extract have the best antibacterial effect at concentration of 0.20g/mL, and average radius of antibacterial zone is 2.67mm and 4.67mm respectively, and the corresponding graph of the antibacterial results of the flat plate is shown in figure 3.
EXAMPLE 2 Flat bacteriostatic Effect of Bidens Bipinnata extract with different leaching times on tobacco bacterial wilt
Obtaining sticktight leaching solution: collecting fresh plants of herba Bidentis Bipinnatae, respectively weighing 200g of herba Bidentis Bipinnatae, cutting into 2-5cm pieces, placing the cut herba Bidentis Bipinnatae in a sterile container, respectively adding 1L of water or phosphate buffer solution, standing at room temperature for leaching for different times, filtering with sterile gauze to remove residue in herba Bidentis Bipinnatae leaching solution, and placing the filtrate in a refrigerator at 4deg.C for use to obtain leaching solutions with different concentrations.
The spanishneedles herb extracts of 1d, 2d, 3d, 4d, 5d, 6d and 7d were extracted respectively, and the plate inhibition effect on tobacco bacterial wilt was measured by the inhibition zone method (the specific operation is the same as in example 1).
As shown in the results of figure 2, the herba Bidentis Bipinnatae water extract and herba Bidentis Bipinnatae phosphate extract have the best antibacterial effect when leached for 2d, and the radius of the antibacterial zone is 2.67mm and 4.67mm respectively, and the corresponding plate antibacterial results are shown in figure 3.
Example 3 Flat-plate antibacterial Effect of Bidens Bipinnata extract on crop pathogenic bacteria
The method of bacteriostasis circle (the specific operation is the same as that in example 1) is adopted to determine the leaching solution of sticktight on tobacco bacterial wiltRalstonia solanacearium) Tomato bacterial wiltR. solanacearium) Sweet potato bacterial wiltR. solanacearium) Rhizoctonia solani of capsicumR. solanacearium) Bacterial wilt of potatoR. solanacearium) Tobacco cavity germPectobacterium carotovorum) Tobacco wild fire germPseudomonas amygdali) Bacterial spot pathogen of tomatoP. syringae) Pseudomonas Cichorii (Cichorium intybus)P. cichorii) Chinese cabbage soft rot fungiP. carotovorum) Bacteria of capsicum scabXanthomonas euvesicatoria) And the like, and the results are shown in table 1. Wherein the concentration of herba Bidentis Bipinnatae leaching solution is 0.20g/mL, and the leaching time is 2d.
Table 1 Flat-panel bacteriostatic effects of Bidens pilosa extract on crop pathogenic bacteria
The results in Table 1 show that the sticktight extract has strong antibacterial effect on bacterial wilt of different crops, but has no antibacterial effect on plant pathogenic bacteria such as tobacco cavity bacteria, tobacco wild fire bacteria, tomato bacterial spot bacteria, chicory pseudomonas, cabbage soft rot bacteria, capsicum scab bacteria and the like, which proves that the sticktight extract obtained by the invention has specificity on the inhibition effect on the bacterial wilt. Wherein, the antibacterial effect of the spanishneedles herb phosphate extract is better than that of the spanishneedles herb extract.
Example 4 determination of the control Effect of Bidens Bipinnata extract on tobacco bacterial wilt
Healthy tobacco seedlings with consistent growth vigor are respectively irrigated with 30mL of spanishneedles herb phosphate leaching solution with concentration of 0.20g/mL and leaching time of 2d and Zhongshengmycin diluted by 750 times (the effective content of Zhongshengmycin is 3%), 6 plants are treated each time, each treatment is repeated for 3 times, and 30mL of tobacco bacterial suspension is inoculated after 24 hours, and the tobacco bacterial suspension is irrigated with roots (OD 600 = 0.8).
As shown in fig. 4 and 5, after 14d inoculation, the control effect of the spanishneedles herb phosphate leaching solution on tobacco bacterial wilt is 50.7%; after 21d inoculation, the control effect is 40.0 percent, which is higher than that of the Zhongshengmycin diluted by 750 times (16.4 percent). The control effect is shown in figure 5.
The foregoing description is only of the preferred embodiments of the invention, and all changes and modifications that come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Claims (2)
1. The application of the spanishneedles herb extract in preventing and treating crop bacterial wilt is characterized in that: the preparation method of the spanishneedles herb extract comprises the following steps: cutting fresh herba Bidentis Bipinnatae into 2-5cm pieces, adding phosphate buffer solution according to a feed 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 use according to claim 1, characterized in that: the phosphate buffer concentration was 0.20moL/L, ph=7.
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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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| Publication number | Priority date | Publication date | Assignee | Title |
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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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