CN115644181A - Chrysanthemum white rust control agent and use method thereof - Google Patents
Chrysanthemum white rust control agent and use method thereof Download PDFInfo
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Abstract
The invention relates to the technical field of chrysanthemum white rust prevention and control, and particularly discloses a chrysanthemum white rust prevention and control agent and a using method thereof. The control agent is added with Tween according to the final concentration of 0.2g to 1.0g/L, and after uniform mixing, 2~3 times/day is uniformly sprayed on chrysanthemum plant leaves, so that the occurrence and the propagation of horiba horikoshii can be effectively inhibited, the inhibition rate of horiba horikoshii can reach 100% within 7 to 12 days at the fastest, after spraying, the leaf germs of the chrysanthemum plant infected with the horiba horikoshii are dead, the new leaves are not infected with germs any more, and the plant recovers normal growth. The invention can effectively reduce the use of toxic chemical pesticides, reduce the environmental pollution and improve the yield and the quality of the chrysanthemum.
Description
Technical Field
The invention relates to the technical field of chrysanthemum white rust prevention and control, and particularly relates to a chrysanthemum white rust prevention and control agent and a using method thereof.
Background
Chrysanthemum (Dendronthema moriforlium) Is one of ten traditional famous flowers in China and one of main fresh cut flowers in the world, and has remarkable economic, humanistic and social values. Chrysanthemum White Rust (CWR) is one of the main diseases damaging the production of cultivated chrysanthemum, is an important worldwide chrysanthemum disease, and is listed as a worldwide quarantine disease at present. The white rust of chrysanthemum is caused by Horikoshi's Puccinia (Horikoshi's)Puccinia horiana) Due to the characteristics of fast development speed, wide spread range, high damage degree and the like of the white rust of the chrysanthemum, once the chrysanthemum almost completely loses ornamental function and commercial value after being infected with diseases, destructive damage is generated.
Scholars Wang Shunli, wang Desheng and Huang Jianghua disclose academic articles related to the prevention and treatment of chrysanthemum white rust, and specifically disclose a prevention and treatment technology for chrysanthemum white rust, wherein various chemical pesticides are sprayed to prevent and treat chrysanthemum white rust when spraying medicines, and the chemical pesticides comprise myclobutanil, holy-li, amicida, bayanan, green werai, clindamycin and epoxiconazole. However, the chemical pesticide has a positive effect on agricultural production increase and pest control, and also brings serious side effects, pesticide residues seriously affect the commercial value of tea or medicine prepared from chrysanthemum, and are not beneficial to the sustainable development of the chrysanthemum industry chain.
At present, the agricultural resource environment is increasingly tightened, the reduction and the efficiency of crop pest control are required to be realized to realize high-quality ecology, and a green control way is developed vigorously, so that the method is a requirement on plant protection technology in a new period and is also a direction for agricultural production development. Aiming at the white rust of the chrysanthemum, a prevention and control agent meeting the green prevention and control needs to be developed urgently, a green prevention and control method is constructed, and the sustainable development of the chrysanthemum industry chain is promoted.
Disclosure of Invention
In order to solve the technical problems, the invention provides a chrysanthemum white rust prevention and control agent and a using method thereof.
In order to achieve the purpose, the invention adopts the technical scheme that:
a chrysanthemum white rust control agent comprises a plant extract and a solvent, wherein the final concentration of the chrysanthemum white rust control agent contains 10-250mg/L chlorogenic acid, 20-200mg/L3,5-O-dicaffeoylquinic acid and 6-50mg/L luteolin.
Preferably, the plant extract comprises jerusalem artichoke extract, artemisia pigweed extract and herba ixeris denticulatae extract according to the mass ratio of 1.
Further, the extraction method of the plant extract comprises the following steps:
(1) Respectively crushing the leaves of the jerusalem artichoke, the stems and leaves of the artemisia scoparia and the whole herb of the ixeris denticulata into powder;
(2) Adding 20-25 times of 50-80% methanol or ethanol solution into the powder obtained in the step (1), performing reflux extraction for 2~3 times, 30-45min each time, filtering, and mixing uniformly to obtain an extracting solution;
(3) And (3) concentrating and drying the extracting solution obtained in the step (2) to obtain the plant extract.
Preferably, the plant extract consists of the extract of broadleaf holly leaf and the extract of glossy privet fruit according to the mass ratio of 1.
Further, the extraction method of the plant extract comprises the following steps:
(1) Respectively grinding the broadleaf holly leaf and the glossy privet fruit into powder;
(2) Adding 20-40 times of 70-80% methanol solution into the powder obtained in the step (1), setting ultrasonic waves at 400W, 30 ℃ and 30min, manually shaking for 2 times in the ultrasonic process, placing 2h, further carrying out ultrasonic treatment for 30min, and filtering to obtain an extracting solution;
(3) And (3) concentrating and drying the extracting solution obtained in the step (2) to obtain the plant extract.
Preferably, the drying is one of spray drying, freeze drying, constant temperature drying and microwave drying.
Preferably, the solvent is water or an aqueous solution of pine needles.
Further, the pine needle water solution is prepared by taking needle leaves of Pinus plants as raw materials, adding 10-50 times of water, decocting for 1-3 h, and concentrating to 6 times, wherein the Pinus plants are at least one of Chinese red pine, chinese pine, cedar, korean pine, black pine, chinese pine, slash pine and Yunnan pine.
The invention also provides a use method of the chrysanthemum white rust control agent, which comprises the following steps: adding tween into the chrysanthemum white rust control agent according to the proportion of 200-1000mg/L, uniformly mixing, and uniformly spraying 2~3 times/day on the leaves of chrysanthemum plants.
Preferably, the Tween is at least one of Tween-20, tween-21, tween-40, tween-60, tween-61, tween-80, tween-81 and Tween-85.
The raw materials for preparing the preventing and treating agent do not belong to chemical pesticides, and are used for preventing and treating the white rust of the chrysanthemum, the preparing raw materials are easy to obtain and non-toxic, the preparation method is simple, the industrial production can be realized, and the application of toxic chemical pesticides is effectively reduced, so that the environmental pollution is reduced, meanwhile, the white rust of the chrysanthemum can be effectively prevented and treated, the leaf germs of the chrysanthemum infected with the horiba horikoshii rust fungus die, the new leaves are not further infected, the chrysanthemum plant can recover to grow normally, the yield and the quality of the chrysanthemum are further improved, and the sustainable development of the chrysanthemum industry chain is promoted.
Drawings
FIG. 1 is a graph of the standard curve of chlorogenic acid;
FIG. 2 is a standard curve of luteolin;
FIG. 3 is a standard graph of 3,5-O-dicaffeoylquinic acid;
FIG. 4 is an HPLC chromatogram of chlorogenic acid, luteolin and 3,5-O-dicaffeoylquinic acid;
FIG. 5 is an HPLC chromatogram of example 1;
FIG. 6 is an HPLC chromatogram of example 2;
FIG. 7 is an HPLC chromatogram of example 3;
FIG. 8 is an HPLC chromatogram of example 4;
FIG. 9 is an HPLC chromatogram of example 5;
FIG. 10 is an HPLC chromatogram of example 6;
FIG. 11 is an HPLC chromatogram of example 7;
FIG. 12 is a diagram of the effect of controlling white rust of chrysanthemum.
Detailed Description
The raw materials used in the following examples were purchased from the market or harvested in natural parks or in the field unless otherwise specified.
Example 1
The jerusalem artichoke extract, the artemisia pigweed extract and the ixeris sonchifolia extract are respectively prepared according to the following steps:
(1) Respectively crushing the leaves of jerusalem artichoke, the stems and leaves of artemisia scoparia and the whole plant of ixeris sonchifolia into powder;
(2) Adding 25 times of 50% methanol solution into the powder obtained in step (1), extracting under reflux in 80 deg.C water bath for 3 times, each time for 30min, filtering, and mixing the filtrates to obtain extractive solution;
(3) And (3) concentrating and spray-drying the extracting solution obtained in the step (2) to obtain the plant extract.
And (2) taking the jerusalem artichoke extract, the artemisia pigweed extract and the ixeris sonchifolia extract according to the mass ratio of 1.
Example 2
The jerusalem artichoke extract, the artemisia pigweed extract and the ixeris sonchifolia extract are respectively prepared according to the following steps:
(1) Respectively crushing the leaves of jerusalem artichoke, the stems and leaves of artemisia scoparia and the whole plant of ixeris sonchifolia into powder;
(2) Adding 25 times of 80% methanol solution into the powder obtained in step (1), extracting in 80 deg.C water bath under reflux for 3 times, each time for 40min, filtering, and mixing the filtrates to obtain extractive solution;
(3) And (3) concentrating the extracting solution obtained in the step (2), and freeze-drying to obtain the plant extract.
Picking folium Pini from at least one of Pinus massoniana, pinus armandii, cedar, korean pine, black pine, chinese pine, slash pine, and Yunnan pine, decocting with 10 times of water for 1 hr, concentrating to 6 times, and making into aqueous solution.
And (2) taking the jerusalem artichoke extract, the artemisia pigweed extract and the ixeris sonchifolia extract according to the mass ratio of 1.
Example 3
The jerusalem artichoke extract, the artemisia pigweed extract and the ixeris sonchifolia extract are respectively prepared according to the following steps:
(1) Respectively crushing the leaves of jerusalem artichoke, the stems and leaves of artemisia scoparia and the whole plant of ixeris sonchifolia into powder;
(2) Adding 20 times of 70% ethanol solution into the powder obtained in step (1), extracting in 80 deg.C water bath under reflux for 3 times, each for 30min, filtering, and mixing the filtrates to obtain extractive solution;
(3) And (3) concentrating the extracting solution obtained in the step (2), and drying at constant temperature to obtain the plant extract.
And (2) taking the jerusalem artichoke extract, the artemisia pigweed extract and the ixeris sonchifolia extract according to the mass ratio of 1.
Example 4
Preparing the broadleaf holly leaf extract and the glossy privet fruit extract respectively according to the following steps:
(1) Grinding folium Ilicis and fructus Ligustri Lucidi respectively to obtain powder;
(2) Adding 40 times of 70% methanol solution into the powder obtained in step (1), respectively, performing ultrasonic setting at 400W, 30 deg.C and 30min, shaking up for 2 times during ultrasonic processing, standing for 2h, performing ultrasonic processing for 30min, and filtering to obtain extractive solution;
(3) And (3) concentrating the extracting solution obtained in the step (2) and drying by microwave to obtain the broadleaf holly leaf extract and the glossy privet fruit extract.
And (2) taking the broadleaf holly leaf extract and the glossy privet fruit extract according to the mass ratio of 1.
Example 5
Preparing the broadleaf holly leaf extract and the glossy privet fruit extract respectively according to the following steps:
(1) Grinding folium Ilicis and fructus Ligustri Lucidi respectively to obtain powder;
(2) Adding 20 times of 80% methanol solution into the powder obtained in step (1), respectively, performing ultrasonic setting at 400W, 30 deg.C and 30min, shaking up for 2 times during ultrasonic processing, standing for 2h, performing ultrasonic processing for 30min, and filtering to obtain extractive solution;
(3) Concentrating the extracting solution obtained in the step (2), and drying at constant temperature to obtain the broadleaf holly leaf extract and the glossy privet fruit extract.
And (2) taking the broadleaf holly leaf extract and the glossy privet fruit extract according to the mass ratio of 1.
Example 6
Preparing the broadleaf holly leaf extract and the glossy privet fruit extract respectively according to the following steps:
(1) Grinding folium Ilicis and fructus Ligustri Lucidi respectively to obtain powder;
(2) Adding 20 times of 80% methanol solution into the powder obtained in step (1), respectively, performing ultrasonic setting at 400W, 30 deg.C and 30min, shaking up for 2 times during ultrasonic processing, standing for 2h, performing ultrasonic processing for 30min, and filtering to obtain extractive solution;
(3) And (3) concentrating the extracting solution obtained in the step (2) and drying by microwave to obtain the broadleaf holly leaf extract and the glossy privet fruit extract.
Picking folium Pini from at least one of Pinus massoniana, pinus armandii, cedar, pinus koraiensis, pinus thunbergii, pinus sylvestris, pinus elliottii and Pinus yunnanensis of Pinus, adding 10 times of water, decocting for 1 hr, concentrating to 6 times, and making into aqueous solution of folium Pini.
And (2) taking the broadleaf holly leaf extract and the glossy privet fruit extract according to the mass ratio of 1.
Example 7
Preparing the broadleaf holly leaf extract and the glossy privet fruit extract respectively according to the following steps:
(1) Grinding folium Ilicis and fructus Ligustri Lucidi respectively to obtain powder;
(2) Adding 30 times of 75% methanol solution into the powder obtained in step (1), respectively, performing ultrasonic setting at 400W, 30 deg.C and 30min, shaking up for 2 times during ultrasonic processing, standing for 2h, performing ultrasonic processing for 30min, and filtering to obtain extractive solution;
(3) Concentrating the extracting solution obtained in the step (2), and freeze-drying to obtain the broadleaf holly leaf extract and the glossy privet fruit extract.
Picking folium Pini from at least one of Pinus massoniana, pinus armandii, cedar, korean pine, black pine, chinese pine, slash pine, and Yunnan pine, decocting in water 50 times of the amount of the above materials for 3 hr, concentrating to 6 times, and making into aqueous solution.
And (2) taking the broadleaf holly leaf extract and the glossy privet fruit extract according to the mass ratio of 1.
Experiment 1
In the experiment, the content of the chrysanthemum white rust control agent prepared in the embodiment 1~7 is measured, and the final concentrations of chlorogenic acid, 3,5-O-dicaffeoylquinic acid and luteolin in the control agents are calculated.
The specific content determination method comprises the following steps:
the test drugs are: chlorogenic acid as a control (lot: ST 02150120) purchased from Shanghai Shidande Standard technical service, inc.; luteolin (batch: DST 201102-016), available from Desite organisms; 3,5-O-dicaffeoylquinic acid (batch: DST 210715-036), purchased from Derster organisms. Acetonitrile is chromatographic grade, phosphoric acid, methanol and other reagents are analytical grade, and water is ultrapure water.
Chromatographic conditions are as follows: the column was ZORBAX SB-C18 column (5 μm, 4.6 mm X250 mm, agilent, USA); the mobile phase A is acetonitrile; mobile phase B was 0.1% phosphoric acid solution, gradient elution: 0 to 11min, 10% of A is → 18%;11 to 30min, A is 18% → 20%;30 to 40min, A is 20 percent; volume flow rate: 1.0mL/min; column temperature: 30 ℃; detection wavelength 348nm; sample introduction amount: 20 μ L.
Preparing a test solution: the mixed plant extract required for preparing the chrysanthemum white rust control agent in example 1~7 is precisely weighed as 0.1g, placed in a conical bottle with a plug, dissolved by precisely adding 10mL of ultrapure water, shaken up, filtered by a microporous filter membrane (0.45 mu m), and the filtrate is taken, thus obtaining the test solution.
Drawing a standard curve of chlorogenic acid: preparing chlorogenic acid standard solutions with the concentrations of 7.8125mg/L, 15.625mg/L, 31.25mg/L, 62.5mg/L, 125mg/L and 250mg/L by using 70% methanol solution, and preparing a working curve of the chlorogenic acid standard product to obtain a standard curve equation. Taking the concentration (mg/L) of the reference solution as a horizontal coordinate and the peak area as a vertical coordinate, performing linear regression to obtain a regression equation: y = 32.56x-176.1, R 2 =0.9991, which indicates that the control shows good linear relation in the concentration range of 7.8125 to 250mg/L. Substituting the peak area of the chlorogenic acid in the test sample into the regression equation of the standard curve to obtain the concentration of chlorogenic acid in the test sample solution, wherein the standard curve of chlorogenic acid is shown in FIG. 1.
And (3) drawing a luteolin standard curve: preparing the luteolin standard solutions with the concentrations of 7.8125mg/L, 15.625mg/L, 31.25mg/L, 62.5mg/L, 125mg/L and 250mg/L by using 70% methanol solution, and making a working curve of the luteolin standard product to obtain a standard curve equation. Taking the concentration (mg/L) of the reference solution as a horizontal coordinate and the peak area as a vertical coordinate, performing linear regression to obtain a regression equation: y =49.599x-299.25, and R2=0.999, indicates that the control shows a good linear relationship in the concentration range of 7.8125 to 250mg/L. And (4) substituting the peak area of the luteolin in the test sample into the regression equation of the standard curve to obtain the concentration of the luteolin in the test sample solution. The standard curve of luteolin is shown in figure 2.
Drawing a 3,5-O-dicaffeoylquinic acid standard curve: a70% methanol solution is used for preparing 3,5-O-dicaffeoylquinic acid series standard solutions with the concentrations of 7.8125mg/L, 15.625mg/L, 31.25mg/L, 62.5mg/L, 125mg/L and 250mg/L respectively, a 3,5-O-dicaffeoylquinic acid standard work curve is prepared, and a standard curve equation is obtained. Taking the concentration (mg/L) of the reference solution as a horizontal coordinate and the peak area as a vertical coordinate, performing linear regression to obtain a regression equation: y =38.227x-366.03, and R2=0.9981, indicate that the control shows a good linear relationship in the concentration range of 7.8125 to 250mg/L. And (3) taking the peak area of the 3,5-O-dicaffeoylquinic acid to be tested into the regression equation of the standard curve to obtain the concentration of 3,5-O-dicaffeoylquinic acid in the solution of the test sample. The standard curve of 3,5-O-dicaffeoylquinic acid is shown in FIG. 3.
Content determination of chlorogenic acid, galuteolin and 3, 5-O-dicaffeoylquinic acid: and (4) sampling the test solution for analysis, wherein the chromatographic conditions are as described above. And recording peak area values, substituting the peak area values into the standard curve equation, and respectively calculating the contents of chlorogenic acid, luteoloside and 3,5-O-dicaffeoylquinic acid in the sample. The results are shown in Table 1.
TABLE 1 EXAMPLES 1-8 measurement results of the content of the controlling agent
Experiment 2
Preparation of control solution: precisely weighing appropriate amount of chlorogenic acid, 3,5-O-dicaffeoylquinic acid and luteolin as reference substances, and adding purified water to obtain control solution containing chlorogenic acid 30mg/L, 3,5-O-dicaffeoylquinic acid 30mg/L and luteolin 30mg/L.
In the experiment, 80 chrysanthemum plants with similar white rust are taken as experimental objects, 8 groups of 10 chrysanthemum plants are arranged, the control agent solution of the control agent is taken as a control, the chrysanthemum white rust control agent prepared in the embodiment 1~7 is respectively numbered as 1 # ~7 # Uniformly spraying 2 times/day on the leaves of the chrysanthemum plants, and contrasting and observing the control conditions of the white rust of each group of chrysanthemum plants until the chrysanthemum plants are infectedThe germs on the back of the leaves of the chrysanthemum white rust disease die (the yellow spots and the winter spore piles on the back of the leaves disappear, and the winter spores and the winter spore piles cannot be found under a microscope), when the newly grown leaves are not infected with the white rust disease any more, the inhibition rate reaches 100%, the inhibition time at the moment is recorded, the average value of each group is taken as the observation result, and the observation result is shown in table 5.
TABLE 5 Observation Table for white rust of chrysanthemum
| Numbering | Inhibition rate | Inhibition time (day) |
| 1# | 100% | 18 |
| 2# | 100% | 14 |
| 3# | 100% | 20 |
| 4# | 100% | 18 |
| 5# | 100% | 19 |
| 6# | 100% | 14 |
| 7# | 100% | 17 |
| |
15 |
As can be seen from table 5, the control agent for white rust of chrysanthemum prepared in example 1~7 has a good inhibitory effect on white rust of chrysanthemum; according to the result 4 of experiment 1 # And 6 # The sample extraction method was substantially the same, but the inhibitory effect 6 # The sample is obviously superior to 4 # Sample No. 6 # Superior to the control solution. 7 # And 4 # Comparison, 7 # Chlorogenic acid and 3,5-O-dicaffeoylquinic acid are significantly less than 4 # But inhibiting effect 7 # Sample superiority to 4 # And (3) sampling. 2 # Also better than the reference solution, and is judged from the common characteristics of the three groups: when the solvent is aqueous solution of folium Pini, the effective component capable of killing Puccinia horikoshii exists in the aqueous solution of folium Pini, and has effect of inhibiting white rust of flos Chrysanthemi.
Experiment 3
In order to further enhance the adhesion capability of the chrysanthemum white rust control agent prepared in example 1~7 on chrysanthemum plant leaves and prolong the moisture retention time, the experiment specifically realizes the purpose by adding tween into the control agent.
Preparation of control solution: taking appropriate amount of chlorogenic acid, 3,5-O-dicaffeoylquinic acid, and luteolin as reference substance, and adding purified water to obtain control solution containing chlorogenic acid 30mg/L, 3,5-O-dicaffeoylquinic acid 30mg/L, and luteolin 30mg/L.
Taking 48 chrysanthemum plants with similar white rust as experimental objects, taking the control agent reference solution as a control, dividing 8 groups of the control agent for the chrysanthemum white rust and the control solution prepared in the embodiment 1~7 with the same volume, respectively adding Tween-20 according to the final concentration of 0.2g/L and 0.3g/L, uniformly mixing, setting 3 plants/group according to the concentration, respectively and uniformly spraying to the leaves of each chrysanthemum plant for 2 times/day, comparing and observing the control condition of the white rust of each chrysanthemum plant, and taking the average value of each group as an observation result. The results are shown in Table 6.
TABLE 6 Observation table for controlling white rust of chrysanthemum
| Numbering | Final concentration of tween (mg/L) | Inhibition rate | Inhibition time (day) |
| 1# | 200 | 100% | 11 |
| 300 | 100% | 9 | |
| 2# | 200 | 100% | 10 |
| 300 | 100% | 7 | |
| 3# | 200 | 100% | 12 |
| 300 | 100% | 10 | |
| 4# | 200 | 100% | 11 |
| 300 | 100% | 9 | |
| 5# | 200 | 100% | 11 |
| 300 | 100% | 10 | |
| 6# | 200 | 100% | 10 |
| 300 | 100% | 7 | |
| 7# | 200 | 100% | 11 |
| 300 | 100% | 8 | |
| |
200 | 100% | 12 |
| 300 | 100% | 10 |
The results shown in tables 5 and 6 show that the control agent is uniformly mixed with Tween-20 and then uniformly sprayed on the chrysanthemum plant leaves, so that the effect of controlling the chrysanthemum white rust is remarkably enhanced, the better the acting force of the control agent adhering to the chrysanthemum plant leaves is as the addition of Tween-20 is increased, the longer the contact time of the control agent and the chrysanthemum plant leaves is prolonged, and the control agent is more beneficial to the chrysanthemum plant to absorb the chlorogenic acid, 3,5-O-dicaffeoylquinic acid and luteoloside in the control agent, so that the effects of killing the horizons hirsutum vulgare and inhibiting the chrysanthemum white rust are achieved.
Theoretically, tween-20 is used as one of the tweens, and can be replaced by other tweens, such as Tween-21, tween-40, tween-60, tween-61, tween-80, tween-81 and Tween-85, so as to increase the acting force of the control agent on the leaves of the chrysanthemum plants and prolong the contact time of the control agent and the leaves of the chrysanthemum plants.
Taking 48 chrysanthemum plants with similar white rust as experimental objects, taking the control agent reference solution as a control, dividing 8 groups of the control agent for the chrysanthemum white rust and the control solution prepared in the embodiment 1~7 with the same volume, respectively adding Tween-80 according to the final concentration of 0.2g/L and 0.3g/L, uniformly mixing, setting 3 plants/group according to the concentration, respectively and uniformly spraying to the leaves of each chrysanthemum plant for 2 times/day, comparing and observing the control condition of the white rust of each chrysanthemum plant, and taking the average value of each group as an observation result. The results are shown in Table 7.
TABLE 7 Observation table for controlling white rust of chrysanthemum
| Numbering | Tween Final concentration (mg/L) | Inhibition rate | Inhibition time (day) |
| 1# | 200 | 100% | 12 |
| 300 | 100% | 10 | |
| 2# | 200 | 100% | 11 |
| 300 | 100% | 9 | |
| 3# | 200 | 100% | 16 |
| 300 | 100% | 14 | |
| 4# | 200 | 100% | 16 |
| 300 | 100% | 15 | |
| 5# | 200 | 100% | 16 |
| 300 | 100% | 14 | |
| 6# | 200 | 100% | 12 |
| 300 | 100% | 10 | |
| 7# | 200 | 100% | 16 |
| 300 | 100% | 13 | |
| |
200 | 100% | 14 |
| 300 | 100% | 12 |
From 6 and table 7, the Tween-20 added into the control agent is changed into Tween-80, the Tween-80 is uniformly mixed and then uniformly sprayed on the chrysanthemum plant leaves, so that the effect of controlling the white rust of the chrysanthemum is enhanced, the acting force of the control agent adhered to the chrysanthemum plant leaves is better along with the increase of the addition of the Tween-80, the contact time of the control agent and the chrysanthemum plant leaves is prolonged, the chlorogenic acid, 3,5-O-dicaffeoylquinic acid and the luteoloside in the control agent can be absorbed by the chrysanthemum plants more favorably, and therefore the effects of killing the puccinia horikoshii and inhibiting the white rust of the chrysanthemum are achieved. However, tween-80 was slightly less effective than Tween-20.
Experiment 4
Preparation of control solution: respectively taking appropriate amount of chlorogenic acid, 3,5-O-dicaffeoylquinic acid and luteolin as reference substances, adding purified water, calculating the use amount of Tween-20 according to the final concentration of 0.3g/L, adding Tween-20 to prepare a control agent reference substance solution, wherein the final concentration contains 30mg/L of chlorogenic acid, 30mg/L of 3,5-O-dicaffeoylquinic acid and 30mg/L of luteolin.
In the experiment, chrysanthemum plants with white rust of similar degrees are taken as experimental objects, the control agent reference solution is taken as a control, 3 parts of the control agent for the chrysanthemum white rust prepared in the example 1~7 with the same volume and the control solution are respectively taken as comparison samples, tween-20 is respectively added according to the final concentration of 0.3g/L and is uniformly mixed, the spraying frequency is respectively set to be 1 time/day, 2 times/day and 3 times/day, each spraying frequency is 1 group, 3 chrysanthemum plants are distributed to each group, the influence of the spraying frequency on the control of the chrysanthemum white rust is investigated and analyzed, and the average value of each group is taken as an observation result. The results are shown in Table 8.
TABLE 8 observation table of the influence of spraying frequency on the control of white rust of chrysanthemum
| Number of | Spraying frequency (times/days) | Inhibition rate | Inhibition time (day) |
| 1# | 1 | 100% | 12 |
| 2 | 100% | 9 | |
| 3 | 100% | 9 | |
| 2# | 1 | 100% | 11 |
| 2 | 100% | 7 | |
| 3 | 100% | 7 | |
| 3# | 1 | 100% | 13 |
| 2 | 100% | 10 | |
| 3 | 100% | 10 | |
| 4# | 1 | 100% | 11 |
| 2 | 100% | 9 | |
| 3 | 100% | 9 | |
| 5# | 1 | 100% | 14 |
| 2 | 100% | 10 | |
| 3 | 100% | 10 | |
| 6# | 1 | 100% | 11 |
| 2 | 100% | 7 | |
| 3 | 100% | 7 | |
| 7# | 1 | 100% | 10 |
| 2 | 100% | 8 | |
| 3 | 100% | 8 | |
| Control solution | 1 | 100% | 13 |
| 2 | 100% | 10 | |
| 3 | 100% | 10 |
As can be seen from table 7, the control effect of the chrysanthemum white rust control agent is positively correlated with the spraying frequency when the spraying frequency is 1~2 times/day; when the spraying frequency is 3 times/day, the control effect of the chrysanthemum white rust control agent is the same as or similar to the inhibition effect of the spraying frequency of 2 times/day, and the inhibition time is shorter. Therefore, the spraying frequency of the chrysanthemum white rust control agent is preferably 2~3 times/day.
Experiment 5
In order to analyze the significance of the effects of the chlorogenic acid, 3,5-O-dicaffeoylquinic acid and luteolin on white rust resistance of the chrysanthemum white rust control agent, the experiment further considers the chlorogenic acid, 3,5-O-dicaffeoylquinic acid and a proper amount of a luteolin reference substance, and adds purified water to prepare the following sample solutions:
sample 1: the final concentration contains chlorogenic acid 30mg/L;
sample 2: the final concentration contains 3,5-O-dicaffeoylquinic acid 30mg/L;
sample 3: the final concentration contains luteolin 30mg/L;
sample 4: the final concentration contains chlorogenic acid 30mg/L, 3,5-O-dicaffeoylquinic acid 30mg/L;
sample 5: the final concentration contains chlorogenic acid 30mg/L and luteoloside 30mg/L;
sample 6: the final concentration contains 3,5-O-dicaffeoylquinic acid 30mg/L, luteolin 30mg/L;
sample 7: the final concentration contains chlorogenic acid 30mg/L, 3,5-O-dicaffeoylquinic acid 30mg/L, and luteolin 30mg/L.
Taking 21 chrysanthemum plants with similar white rust as experimental objects, taking 3 chrysanthemum plants as a group, taking the sample 1-7 solution as a control agent, adding Tween-20 according to the final concentration of 0.3g/L, uniformly mixing, uniformly spraying 2 times/day on leaves of the chrysanthemum plants, observing the control relevance of chlorogenic acid, 3,5-O-dicaffeoylquinic acid, luteolin and the chrysanthemum white rust, and taking the average value of each group as an observation result. The results are shown in Table 9.
TABLE 9 Observation table for controlling white rust of chrysanthemum
| Numbering | Composition of ingredients | Inhibition rate | Inhibition time (day) |
| Sample 1 | Chlorogenic acid | 100% | 24 |
| Sample 2 | Chlorogenic acid 3,5-O- |
100% | 22 |
| Sample 3 | |
100% | 23 |
| Sample No. 4 | Chlorogenic acid 3,5-O- |
100% | 18 |
| Sample No. 5 | Chlorogenic acid luteolin | 100% | 20 |
| Sample No. 6 | 3,5-O-dicaffeoylquinic acid luteolin | 100% | 16 |
| Sample 7 | Chlorogenic acid 3,5-O-dicaffeoylquinic acid, and |
100% | 10 |
The results shown in table 9 show that chlorogenic acid, 3,5-O-dicaffeoylquinic acid and luteolin respectively have certain control effects on the white rust of chrysanthemum, and the control effects are remarkable when the chlorogenic acid, the 3,5-O-dicaffeoylquinic acid and the luteolin are used together.
Control solution was prepared under the same conditions as in experiment 5, and the concentration of the sample was changed to a final concentration of 50mg/L chlorogenic acid, 50 mg/L3,5-O-dicaffeoylquinic acid, and 50mg/L luteolin.
Taking 21 chrysanthemum plants with similar white rust as experimental objects, taking 3 chrysanthemum plants as a group, taking the sample 1-7 solution as a control agent, adding Tween-20 according to the final concentration of 0.3g/L, uniformly mixing, uniformly spraying 2 times/day on leaves of the chrysanthemum plants, observing the control relevance of chlorogenic acid, 3,5-O-dicaffeoylquinic acid, luteolin and the chrysanthemum white rust, and taking the average value of each group as an observation result. The results are shown in Table 10.
TABLE 10 Observation table for controlling white rust of chrysanthemum
| Numbering | Composition of ingredients | Inhibition rate | Inhibition time (day) |
| Sample 1 | Chlorogenic acid | 100% | 20 |
| Sample 2 | Chlorogenic acid 3,5-O- |
100% | 18 |
| Sample 3 | |
100% | 21 |
| Sample No. 4 | Chlorogenic acid 3,5-O- |
100% | 14 |
| Sample No. 5 | Chlorogenic acid luteolin | 100% | 16 |
| Sample No. 6 | 3,5-O-dicaffeoylquinic acid luteolin | 100% | 12 |
| Sample 7 | Chlorogenic acid 3,5-O-dicaffeoylquinic acid, and |
100% | 8 |
As can be seen from comparison of the results shown in tables 9 and 10, chlorogenic acid, 3,5-O-dicaffeoylquinic acid and luteolin respectively have certain control effects on the white rust of chrysanthemum, the control effects are remarkable when the chlorogenic acid, the 3,5-O-dicaffeoylquinic acid and the luteolin are used together, but the control effects are more remarkable when the concentrations of the chlorogenic acid, the 3,5-O-dicaffeoylquinic acid and the luteolin are increased.
However, the three components of chlorogenic acid, 3,5-O-dicaffeoylquinic acid and luteolin exist in various natural plants widely, and it is expected that plant extracts obtained by extracting the natural plants rich in chlorogenic acid or 3,5-O-dicaffeoylquinic acid or luteolin are combined to obtain the chrysanthemum white rust disease control agent, wherein the final concentration of the chrysanthemum white rust disease control agent contains 10 to 250mg/L of chlorogenic acid, 10 to 200mg/L of 3,5-O-dicaffeoylquinic acid and 6 to 50mg/L of luteolin, and the chrysanthemum white rust disease control agent is applied to control chrysanthemum white rust disease and has similar or equal control effects.
Therefore, the above-mentioned embodiments are only some preferred embodiments of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered as falling within the technical scope of the present invention, and equivalent substitutions or changes according to the technical solution and the modified concept of the present invention should be covered by the scope of the present invention.
Claims (10)
1. The chrysanthemum white rust control agent is characterized in that: the final concentration of the control agent contains 10 to 250mg/L of chlorogenic acid, 20 to 200mg/L of 3,5-O-dicaffeoylquinic acid and 6 to 50mg/L of luteolin.
2. The chrysanthemum white rust control agent according to claim 1, characterized in that: the chlorogenic acid, 3,5-O-dicaffeoylquinic acid and luteolin are plant extracts.
3. The chrysanthemum white rust disease control agent according to claim 2, wherein the plant extract is composed of jerusalem artichoke extract, artemisia scoparia extract and ixeris denticulata extract in a mass ratio of 1:
(1) Respectively crushing the leaves of the jerusalem artichoke, the stems and leaves of the artemisia scoparia and the whole herb of the ixeris denticulata into powder;
(2) Adding 20-25 times of 50-80% methanol or ethanol solution into the powder obtained in the step (1), performing reflux extraction for 2~3 times, 30-45min each time, filtering, and mixing uniformly to obtain an extracting solution;
(3) And (3) concentrating and drying the extracting solution obtained in the step (2) to obtain the plant extract.
4. The chrysanthemum white rust control agent according to claim 2, characterized in that: the plant extract comprises a broadleaf holly leaf extract and a glossy privet fruit extract according to the mass ratio of 1.
5. The chrysanthemum white rust control agent according to claim 4, characterized in that: the extraction method of the plant extract comprises the following steps:
(1) Grinding the broadleaf holly leaf and the glossy privet fruit into powder respectively;
(2) Adding 20-40 times of 70-80% methanol solution into the powder obtained in the step (1), setting ultrasonic waves at 400W, 30 ℃ and 30min, shaking up for 2 times in the ultrasonic process, standing for 2h, further carrying out ultrasonic treatment for 30min, and filtering to obtain an extracting solution;
(3) And (3) concentrating and drying the extracting solution obtained in the step (2) to obtain the plant extract.
6. The chrysanthemum white rust control agent according to claim 3 or 5, characterized in that: the drying is one of spray drying, freeze drying, constant temperature drying and microwave drying.
7. The chrysanthemum white rust control agent according to claim 1, characterized in that: the control agent comprises a solvent, and the solvent is water or a pine needle water solution.
8. The chrysanthemum white rust control agent according to claim 7, characterized in that: the pine needle water solution is prepared by taking needle leaves of Pinus plants as raw materials, adding water 10-50 times of the needle leaves, decocting for 1-3 h, and concentrating to 6 times of the needle leaves, wherein the Pinus plants are at least one of masson pine, china pine, cedar, korean pine, black pine, chinese pine, slash pine and Yunnan pine.
9. The application method of the chrysanthemum white rust control agent is characterized by comprising the following steps: adding tween into the chrysanthemum white rust control agent of any one of the claims 1~8 at a ratio of 200-1000mg/L, uniformly mixing, and uniformly spraying 2~3 times per day onto the leaves of chrysanthemum plants.
10. The use method of the chrysanthemum white rust control agent according to claim 9, characterized in that: the Tween is at least one of Tween-20, tween-21, tween-40, tween-60, tween-61, tween-80, tween-81 and Tween-85.
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