CN114766487B - Application of butenyl phthalide and butylphthalide in preventing and treating crop southern blight - Google Patents
Application of butenyl phthalide and butylphthalide in preventing and treating crop southern blight Download PDFInfo
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- 229950005197 butylphthalide Drugs 0.000 title claims abstract description 49
- RSKPJCDZAFMWHH-UHFFFAOYSA-N 3-but-1-enyl-3h-2-benzofuran-1-one Chemical compound C1=CC=C2C(C=CCC)OC(=O)C2=C1 RSKPJCDZAFMWHH-UHFFFAOYSA-N 0.000 title claims abstract description 42
- 239000003814 drug Substances 0.000 claims abstract description 29
- 241001530056 Athelia rolfsii Species 0.000 claims abstract description 17
- 239000000463 material Substances 0.000 claims abstract description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 21
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- WMBOCUXXNSOQHM-FLIBITNWSA-N (Z)-3-butylidenephthalide Chemical compound C1=CC=C2C(=C/CCC)/OC(=O)C2=C1 WMBOCUXXNSOQHM-FLIBITNWSA-N 0.000 description 17
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- 241000722363 Piper Species 0.000 description 4
- 235000016761 Piper aduncum Nutrition 0.000 description 4
- 235000017804 Piper guineense Nutrition 0.000 description 4
- 235000008184 Piper nigrum Nutrition 0.000 description 4
- 238000004364 calculation method Methods 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 230000003902 lesion Effects 0.000 description 4
- 239000000375 suspending agent Substances 0.000 description 4
- 230000001018 virulence Effects 0.000 description 4
- 240000008574 Capsicum frutescens Species 0.000 description 3
- 230000004071 biological effect Effects 0.000 description 3
- 239000001390 capsicum minimum Substances 0.000 description 3
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- 239000000126 substance Substances 0.000 description 3
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- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000223221 Fusarium oxysporum Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 241001330975 Magnaporthe oryzae Species 0.000 description 1
- 241001522129 Pinellia Species 0.000 description 1
- 241000813090 Rhizoctonia solani Species 0.000 description 1
- 241000221696 Sclerotinia sclerotiorum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
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- 240000004922 Vigna radiata Species 0.000 description 1
- 235000010721 Vigna radiata var radiata Nutrition 0.000 description 1
- 235000011469 Vigna radiata var sublobata Nutrition 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- KZNIFHPLKGYRTM-UHFFFAOYSA-N apigenin Chemical compound C1=CC(O)=CC=C1C1=CC(=O)C2=C(O)C=C(O)C=C2O1 KZNIFHPLKGYRTM-UHFFFAOYSA-N 0.000 description 1
- 229940117893 apigenin Drugs 0.000 description 1
- XADJWCRESPGUTB-UHFFFAOYSA-N apigenin Natural products C1=CC(O)=CC=C1C1=CC(=O)C2=CC(O)=C(O)C=C2O1 XADJWCRESPGUTB-UHFFFAOYSA-N 0.000 description 1
- 235000008714 apigenin Nutrition 0.000 description 1
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- 230000001225 therapeutic effect Effects 0.000 description 1
- 229910052902 vermiculite Inorganic materials 0.000 description 1
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Classifications
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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
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/06—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom five-membered rings
- A01N43/12—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom five-membered rings condensed with a carbocyclic ring
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Life Sciences & Earth Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses an application of butenyl phthalide and butylphthalide in preventing and treating crop southern blight, and the mixed medicament comprises butenyl phthalide, butylphthalide and auxiliary materials. The invention adopts the application of the butenyl phthalide and the butylphthalide in preventing and treating crop southern blight, and the butenyl phthalide and the butylphthalide aligned sclerotium rolfsii have excellent antibacterial activity.
Description
Technical Field
The invention relates to the technical field of biopesticides, in particular to application of butenyl phthalide and butylphthalide in preventing and treating crop southern blight.
Background
Currently, there is relatively limited research on the biological activity of butylidenephthalide and butylphthalide against phytopathogenic fungi. Butylphthalide, also known as 3-butyl-1 (3H) -isobenzofuranone, apigenin, which is mainly responsible for the flavour and taste of celery. Studies have shown that butylphthalide and butylphthalide have certain inhibitory activities on Rhizoctonia solani, sclerotinia sclerotiorum, alternaria wheat, botrytis cinerea, fusarium oxysporum and Pyricularia oryzae. But the biological activity of the butylidenephthalide and butylidenephthalide aligned sclerotium rolfsii and the control effect on crop southern blight have not been studied.
The sclerotium rolfsii hosts are wide, and can infect more than 500 plants, so that the infected plants turn yellow and wilt, and the rootstock parts turn brown and rot. Especially, the southern blight of peanut, pepper and konjak caused by sclerotium rolfsii has become an important factor for restricting the safe production of the peanut, pepper and konjak, sclerotium produced by the pathogenic bacteria can survive for 5-8 years in soil, and the sclerotium is difficult to control. The agricultural rural area in Henan province in 2020 lists the peanut southern blight as a list of two kinds of plant diseases and insect pests in Henan province. More seriously, the range of sclerotium rolfsii hosts is still expanding. In 2019-2020, the first report of the infection of the chrysanthemum, mung beans, pinellia ternate, mugwort leaves and the like by the sclerotium rolfsii is carried out in China.
At present, the measure for preventing and controlling the crop southern blight caused by the sclerotium rolfsii is to use bactericides, but the bactericides for preventing and controlling the crop southern blight are limited and are mostly chemical bactericides which are registered on the Chinese pesticide information network. The long-term use of the chemical bactericide not only easily causes the drug resistance of the southern blight bacteria, but also causes the problems of pesticide residue, environmental pollution, harm to human health and the like. The search of lead compounds with high biological activity from natural products is one of the main ways of creating new pesticides, so that the search of novel, efficient and safe alternative agents for preventing and treating crop southern blight is urgently needed.
Disclosure of Invention
The invention aims to provide the application of butenyl phthalide and butylphthalide in preventing and treating crop southern blight, and provides a novel, efficient and safe plant source bactericide for reasonable prevention and treatment of the crop southern blight, which aims at the problems that the prevention and treatment agents are limited and most of chemical bactericides exist.
In order to achieve the aim, the invention provides application of butenyl phthalide and butylphthalide in preventing and treating crop southern blight, and the mixed medicament comprises the butenyl phthalide, the butylphthalide and auxiliary materials.
Preferably, the mixed medicament is used for preventing and treating crop southern blight caused by sclerotium rolfsii.
Preferably, the mixed medicament can be powder, wettable powder, soluble powder, granule, aqueous emulsion, microemulsion, water aqua, suspending agent, dispersible oil suspending agent, microcapsule or emulsifiable concentrate, etc.
Preferably, the verification method of butylidenephthalide and butylphthalide for preventing and treating crop southern blight is characterized by comprising the following steps:
s1, dissolving butenyl phthalide in 0.4mL of mixed solution of DMSO and Tween-80, adding sterile water to a volume of 20mL, and then adding the sterile water to sequentially and gradually dilute to different concentrations;
s2, dissolving butylphthalide in 0.4mL of mixed solution of DMSO and Tween-80, adding sterile water to a volume of 20mL, and then adding the sterile water to sequentially and gradually dilute to different concentrations;
s3, determining virulence of the aligned sclerotium rolfsii hyphae growth of the butenyl phthalide and the butylphthalide by adopting a hyphae growth inhibition rate method;
s4, verifying the inhibiting effect of the sclerotium germination of the sclerotium alignment sclerotium of the butenyl phthalide and the butylphthalide.
Preferably, the volume ratio of DMSO to Tween-80 is 1%.
Therefore, the application of the butenyl phthalide and the butylphthalide in preventing and treating crop southern blight has the following beneficial effects compared with the prior art:
1. the butylidenephthalide aligned sclerotium has strong antibacterial activity and also has excellent inhibition effect on sclerotium germination of the butylidenephthalide aligned sclerotium;
2. the butylphthalide aligned micronucleus has excellent antibacterial activity, and the antibacterial activity is stronger along with the increase of the concentration of the butylphthalide;
3. the potted plant control effect test is consistent with the indoor toxicity test result, and shows that the butylphthalide and the butenyl phthalide have good application prospect in preventing and controlling the crop southern blight caused by the sclerotium rolfsii.
The technical scheme of the invention is further described in detail through examples.
Detailed Description
The technical scheme of the invention is further described below by examples.
Unless defined otherwise, technical or scientific terms used herein should be given the ordinary meaning as understood by one of ordinary skill in the art to which this invention belongs. The terms "first," "second," and the like, as used herein, do not denote any order, quantity, or importance, but rather are used to distinguish one element from another. The word "comprising" or "comprises", and the like, means that elements or items preceding the word are included in the element or item listed after the word and equivalents thereof, but does not exclude other elements or items. The terms "connected" or "connected," and the like, are not limited to physical or mechanical connections, but may include electrical connections, whether direct or indirect. "upper", "lower", "left", "right", etc. are used merely to indicate relative positional relationships, which may also be changed when the absolute position of the object to be described is changed.
The application of the butenyl phthalide and the butylphthalide in preventing and treating crop southern blight is disclosed, and the mixed medicament comprises the butenyl phthalide, the butylphthalide and auxiliary materials.
The mixed medicament is used for preventing and treating crop southern blight caused by sclerotium rolfsii.
The mixed medicament can be powder, wettable powder, soluble powder, granules, aqueous emulsion, microemulsion, water aqua, suspending agent, dispersible oil suspending agent, microcapsule or emulsifiable concentrate, etc.
The verification method of butenyl phthalide and butylphthalide for preventing and treating crop southern blight is characterized by comprising the following steps:
s1, dissolving butenyl phthalide in 0.4mL of mixed solution of DMSO and Tween-80, adding sterile water to a volume of 20mL, and then adding the sterile water to sequentially and gradually dilute to different concentrations;
s2, dissolving butylphthalide in 0.4mL of mixed solution of DMSO and Tween-80, adding sterile water to a volume of 20mL, and then adding the sterile water to sequentially and gradually dilute to different concentrations;
s3, determining virulence of the aligned sclerotium rolfsii hyphae growth of the butenyl phthalide and the butylphthalide by adopting a hyphae growth inhibition rate method;
s4, verifying the inhibiting effect of the sclerotium germination of the sclerotium alignment sclerotium of the butenyl phthalide and the butylphthalide.
The volume ratio of DMSO to Tween-80 was 1%.
The verification method for preventing and controlling crop southern blight about butenyl phthalide and butylphthalide comprises the following steps:
1. determining the virulence of the growth of the hyphae of the butylidenephthalide and butylphthalide aligned sclerotium rolfsii by adopting a hyphae growth inhibition rate method:
s1, respectively taking 0.06g of butenyl phthalide (3-n-butenyl phthalide CAS: 551-08-6) and butylphthalide (n-butylphthalide CAS: 6066-49-5) to be dissolved in 0.4mL of mixed solution of DMSO and Tween-80, and adding 19.54mL of sterile water to a volume of 20mL to prepare 3000mg/L mother liquor, wherein the volume ratio of DMSO and Tween-80 is 1%;
s2, carrying out gradient dilution by using sterile water to obtain seven series of liquid medicine with concentration of 1500mg/L, 1000mg/L, 500mg/L, 250mg/L, 125mg/L, 62.5mg/L and 31.25 mg/L;
s3, respectively taking 1mL of liquid medicine with different concentrations, adding the liquid medicine into 9mL of PDA culture medium, and preparing the liquid medicine-containing culture medium with the final concentration of 150mg/L, 100mg/L, 50mg/L, 25mg/L, 12.5mg/L, 6.25mg/L and 3.125mg/L respectively;
s4, taking a non-drug culture medium containing 0.05% DMSO and 0.05% Tween-80 as a control, and repeating each concentration three times;
s5, culturing in the dark in an illumination incubator at 25 ℃ for three days, and measuring the diameter of each treated colony by using a crisscross method;
s6, calculating the inhibition rate of the growth of the butylidenephthalide and butylidenephthalide aligned sclerotium rolfsii mycelia, and obtaining a virulence regression equation, an effective medium concentration (EC 50) and a concentration (EC 90) with the inhibition rate of 90% according to a probability value analysis method;
the calculation method of the hypha growth inhibition rate comprises the following steps:
hypha growth inhibition ratio = [ (control colony diameter-cake diameter) - (treatment colony diameter-cake diameter) ]/(control colony diameter-cake diameter) ×100
Table 1: toxicity of the aligned sclerotium rolfsii mycelia of butenyl phthalide and butylphthalide
| Treatment of | EC 50 (mg/L) | EC 90 (mg/L) | Correlation coefficient r | Toxicity regression equation |
| Butenyl phthalides | 14.0 | 64.4 | 0.9967 | y＝2.7849+1.9334x |
| Butylphthalide | 32.3 | 272.5 | 0.9860 | y＝2.9136+1.3830x |
| Polyoxin | 204.7 | 5763.7 | 0.9006 | y＝5.7643+1.0459x |
As shown in Table 1, the butylidenephthalide and butylidenephthalide aligned micronucleus has good inhibitory activity. With the increase of the concentration of the butenyl phthalide and the butylphthalide, the antibacterial activity is enhanced and is higher than that of the contrast medicament polyoxin.
2. Regarding the inhibition of sclerotium germination of butylidenephthalide and butylidenephthalide:
s1, taking 0.12g of butenyl phthalide (3-n-butenyl phthalide CAS: 551-08-6) and butylphthalide (n-butylphthalide CAS: 6066-49-5), dissolving in 0.4mL of mixed solution of DMSO and Tween-80, adding 19.48mL of sterile water to a volume of 20mL and preparing 6000mg/L mother liquor, wherein the volume ratio of DMSO and Tween-80 is 1%;
s2, diluting the mixture into six series of liquid medicine with concentration of 3000mg/L, 2500mg/L, 2000mg/L, 1500mg/L, 1000mg/L and 500mg/L by using sterile water in a gradient manner;
s3, respectively adding 1mL of liquid medicine with different concentrations into 9mL of PDA culture medium to prepare six kinds of medicine-containing culture mediums with final concentrations of 300mg/L, 250mg/L, 200mg/L, 150mg/L, 100mg/L and 50 mg/L;
s4, taking a non-drug culture medium containing 0.05% DMSO and 0.05% Tween-80 as a control, and repeating each concentration three times;
s5, observing germination conditions of sclerotium after culturing on a drug-containing culture medium for two days, and considering germination when hypha generated by the sclerotium is more than or equal to 2 mm;
the calculation method of the sclerotium germination inhibition rate comprises the following steps:
sclerotium germination inhibition ratio= (control number of sclerotium germination-treated number of sclerotium germination)/(control number of sclerotium germination) ×100.
Table 2: inhibition of sclerotium germination of butylidenephthalide and butylphthalide aligned sclerotium (%)
Table 2 shows that the sclerotium germination of the sclerotium alignment sclerotium of the butenyl phthalide and the butylphthalide also has a good inhibition effect.
Example 1
Potted plant prevention and treatment effect of butenyl phthalide and butylphthalide on peanut southern blight:
s1, selecting peanut seeds with plump grains and uniform sizes as test peanut varieties, placing the peanut seeds in a basin, adding sterile water to submerge one third of the seeds, and placing the seeds in an incubator for seed soaking for half a day;
s2, mixing matrix soil and vermiculite in a volume ratio of 3:1, adding sterile water to adjust the water content of the soil to 70%, placing the soil into a pot, and sowing germinated seed radicle downwards;
s3, artificially inoculating germs: after sowing for one week, selecting healthy peanut plants with consistent growth vigor and size, inoculating 1 bacterial cake on the base of the stem of each plant, and burying with soil;
s4, after hyphae are observed to be generated on the root parts of plant stems and the surrounding soil surfaces, respectively spraying 10mL of each plant of liquid medicine with 100mg/L, 200mg/L of butenyl phthalide, 200mg/L of butylphthalide and 500mg/L of contrast medicament polyoxin liquid medicine;
s5, using sterile water containing 0.1% DMSO and 0.1% Tween-80 as a control, and repeating the steps three times when 15 plants are treated each;
s6, after three days of drug application, identifying the disease conditions according to grading standards, and calculating disease indexes and prevention effects of different treatments;
table 3: potted plant prevention effect of butenyl phthalide and butylphthalide on peanut southern blight
Note that: data in the table are mean ± standard error;
the different letters in the same column indicate that the difference was significant at the P <0.05 level as tested by the least significant difference method.
The disease condition identification criteria are:
level 0: no disease spots;
stage 1: the disease spots are smaller than 1/3 root stems;
3 stages: the disease spots are more than 2/3 of the stems, and a small amount of hyphae are on the ground;
5 stages: the disease spots are more than 2/3 of the stems, and a large number of hyphae are on the ground;
7 stages: the disease spots are more than 2/3 of the stems, a large number of hyphae are on the ground, and wilting of the overground parts occurs;
stage 9: plant withering;
the calculating method of the disease index comprises the following steps:
disease index = Σ (disease strain of each stage×relative grade value)/(total strain number×9) ×100
The calculation method of the control effect comprises the following steps:
control effect = [ (control disease index-treatment disease index)/control disease index ] ×100%.
As shown in Table 3, the potted plant test results are consistent with the indoor toxicity test results, and the control effect of the butenyl phthalide and the butylphthalide is obviously higher than that of the registration agent of 500mg/L polyoxin, so that the butenyl phthalide and the butylphthalide have good effect of preventing and treating peanut southern blight.
Example 2
The control effect of butenyl phthalide and butylphthalide on capsicum and konjak southern blight:
s1, selecting fresh, uniform-size and uniform-quality pepper and konjak fruits, placing the fruits in a sterile plastic box with gauze paved at the bottom and containing sterile water, placing 9 fruits in each box, and inoculating two bacterial cakes to each fruit;
s2, spraying 200mg/L and 100mg/L of butenyl phthalide and butyl phthalide liquid medicine on the fruits of two crops respectively after hyphae or lesions appear around the fruits;
s3, taking 500mg/L polyoxin as a control medicament, taking sterile water containing 0.1% DMSO and 0.1% Tween-80 as a control, wherein each plastic box is treated by one treatment, and each treatment is repeated by 9 times;
s4, measuring the diameter of the disease spots after three days of drug application to calculate the prevention and treatment effect,
the calculation formula of the control effect is as follows:
control effect (%) = (control group lesion diameter-treatment group lesion diameter)/control group lesion diameter×100.
As is clear from Table 4, butenyl phthalide has excellent therapeutic effects on capsicum and konjak southern blight. The control effect of butenyl phthalide on pepper and konjak southern blight is obviously better than that of a control medicament polyoxin at 500mg/L at a dose of 200 mg/L.
Table 4: effect of butylidenephthalide and butylphthalide on preventing and treating capsicum and konjak southern blight
Note that: data in the table are mean ± standard error;
the different letters in the same column indicate that the difference was significant at the P <0.05 level as tested by the least significant difference method.
Therefore, the application of the butenyl phthalide and the butylphthalide with the structure in the control of crop southern blight has excellent antibacterial activity on the sclerotium rolfsii with the butenyl phthalide and the butylphthalide aligned.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention and not for limiting it, and although the present invention has been described in detail with reference to the preferred embodiments, it will be understood by those skilled in the art that: the technical scheme of the invention can be modified or replaced by the same, and the modified technical scheme cannot deviate from the spirit and scope of the technical scheme of the invention.
Claims (2)
1. The application of butenyl phthalide and butylphthalide in preventing and treating crop southern blight is characterized in that: the mixed medicament comprises butenyl phthalide, butylphthalide and auxiliary materials;
the mixed medicament is used for preventing and treating crop southern blight caused by sclerotium rolfsii.
2. The use of butenyl phthalides and butylphthalides according to claim 1 for controlling crop southern blight, characterized in that: the mixed medicament can be powder, granule, water aqua, microemulsion, microcapsule or emulsifiable concentrate.
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