CN116725041A - Plant source biological control agent for controlling cucumber mosaic virus - Google Patents
Plant source biological control agent for controlling cucumber mosaic virus Download PDFInfo
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- CN116725041A CN116725041A CN202310775404.7A CN202310775404A CN116725041A CN 116725041 A CN116725041 A CN 116725041A CN 202310775404 A CN202310775404 A CN 202310775404A CN 116725041 A CN116725041 A CN 116725041A
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- herba
- mosaic virus
- cucumber mosaic
- alternanthera philoxeroides
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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
- 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
-
- 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]
-
- 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
- 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]
- A01N65/22—Lamiaceae or Labiatae [Mint family], e.g. thyme, rosemary, skullcap, selfheal, lavender, perilla, pennyroyal, peppermint or spearmint
-
- 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
- A01P1/00—Disinfectants; Antimicrobial compounds or mixtures thereof
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- 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)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Environmental Sciences (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Agronomy & Crop Science (AREA)
- Biotechnology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Botany (AREA)
- Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Pest Control & Pesticides (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
A plant biological control agent for preventing and treating cucumber mosaic virus disease comprises one or more of Alternaria alternifolia, bidens pilosa, herba Duchesneae Indicae, herba Houttuyniae or herba Menthae. The research of the invention shows that the alternanthera philoxeroides, the bidens pilosa, the herba polygoni multiflori, the herba houttuyniae and the mint have a certain inhibiting effect on the cucumber mosaic virus, wherein the inhibiting effect of the alternanthera philoxeroides on the cucumber mosaic virus is most obvious, and the invention provides more choices for preventing and treating the cucumber mosaic virus.
Description
Technical Field
The invention relates to the technical field of biological control of plant diseases, in particular to a plant source biological control agent for controlling cucumber mosaic virus.
Background
Cucumber mosaic virus (cucumber mosaic virus, CMV) is a typical member of the genus Cucumovirus (Bromoviridae) of the family brome mosaic virus, listed as one of the ten plant viruses in the world. The cucumber mosaic virus has wide hazard range, is one of plant viruses with the largest number of host plants, the largest distribution and the largest economic importance, and can reduce the tomato yield by about 25-50% by statistics of the infection of CMV in China; the yield of the capsicum is reduced by 20 percent. CMV-induced disease poses a serious threat to crops and other commercial crops and causes significant economic losses. Diseases caused by CMV are serious, but are difficult to control, and meanwhile, serious economic losses are caused for crop production in countries around the world.
Plant-derived biocontrol agents are those which extract the active ingredient from artificially cultivated or wild plants, the active ingredient of which is usually some, or even most, of the organic matter in the plant organism; compared with chemical synthesis and other pesticides, the plant source biocontrol agent has the characteristics of environmental friendliness, safety to non-target organisms, difficulty in generating drug resistance, specific action mode, promotion of crop growth, improvement of disease resistance, multiple varieties, multiple development approaches and the like. Therefore, plant-derived biocontrol agents have been widely paid attention to and used in the fields of melons, fruits, vegetables, special crops (tea, mulberry, chinese herbal medicines, flowers, etc.) and organic agriculture. Chinese patent CN105707136a discloses a biological control agent for cucumber mosaic virus diseases, including astragalus root, isatis root, herba artemisiae capillaries, honeysuckle, fructus forsythiae, coptis root, bighead atractylodes rhizome, dandelion and pricklyash peel, plant-derived biological control agents for cucumber mosaic virus diseases are still few at present, so that more plant-derived biological control agents for cucumber mosaic virus are still needed to be provided for selection.
Disclosure of Invention
The invention aims to overcome the defects and the shortcomings in the prior art and provide a new application of any one or more of alternanthera philoxeroides, bidens pilosa, herba houttuyniae and peppermint in resisting cucumber mosaic virus or preparing biocontrol preparations resisting the cucumber mosaic virus, so as to provide more plant-derived biocontrol agents of the cucumber mosaic virus.
The second object of the present invention is to provide a plant-derived biocontrol agent for controlling cucumber mosaic virus diseases.
The above object of the present invention is achieved by the following technical solutions:
alternanthera philoxeroides (Alternanthera philoxeroides (mart.) Griseb.) is a perennial herb of the genus alternanthera of the family amaranthaceae; the stem base part of the plant is creeping, the upper part of the plant is inclined to rise, and the plant is hollow and branched; the leaves are opposite, the leaves are oblong, the front ends of the leaves are sharp or round, the base part of the leaves is gradually narrowed, the upper part of the leaves is provided with attached hair, and the edge of the leaves is provided with eyelashes; the head inflorescence singly grows on the axillary part of the upper part of the stem, is spherical, has a long round shape, is white, and has pink basal part and luster. The alligator alternanthera grassland produces Brazil, after introduction into China, the alligator alternanthera grassland is wild, and is now an invasive plant with great harm; is widely distributed in the south and north slopes of Qinling mountain in China. The alternanthera philoxeroides is warm and warm, and has strong cold resistance; has strong adaptability and can grow on water and land. The propagation mode of the alternanthera philoxeroides is mainly asexual propagation. All the herb of alternanthera philoxeroides is bitter in taste, slightly sweet in flavor and cold in nature; has effects of clearing heat, promoting urination, cooling blood and removing toxic substances.
Bidens pilosa is whole herb of Bidens pilosa L. Distributed in the provinces of east China, middle China, south China and southwest. Has effects of removing toxic substances, relieving swelling, clearing heat, relieving pain, promoting blood circulation, dispelling blood stasis, regulating qi, and resolving food stagnation. Is commonly used for gastroenteritis, heatstroke and abdominal pain, bacillary dysentery, cold and fever, acute laryngitis, stranguria, turbid urine, hemorrhoids, rectocele, stool and urine bleeding, diabetes, snake bite, scapulohumeral periarthritis, traumatic injury, arthritis and the like.
Herba Duchesneae Indicae (Emilia sonchifolia (L.) DC.in light) is an annual or perennial herb of Compositae. The root is vertical, the stem is upright, no hair or short hair is dredged, and the color is grey. The leaf quality is thicker, the top lobe is large, the wide oval triangle is provided with irregular teeth, the side lobe is oblong and provided with wavy teeth, the upper surface is dark green, and the lower surface is always purple; the stem leaves in the middle are free of growth, small and no handle; the upper leaves are few and linear. The head inflorescence sags before flowering and stands upright after flowering; the inflorescence stems are thin, no bract exists, and the total bracts are cylindrical; the total bract is yellow green, has the same length as the floret and has no hair on the back. The pollen is red or purple, and the tube part is slender; the crown hair is rich, white and fine and soft. Wild species are present in south China, southwest and the like. The tea has the advantages of being mild in property, bitter and slightly pungent in taste, cooling blood and detoxifying, taking tender tip tender leaves as a main material, being capable of being fried, being used as soup or being used as chafing dish material, being crisp in texture, being similar to the taste of crowndaisy chrysanthemum, having the effects of clearing heat and detoxifying, promoting blood circulation and dispelling blood stasis and the like, and being capable of treating urinary system infection, sphagitis, sores and the like.
Herba Houttuyniae (Houttuynia cordata Thunb.) is a herb recorded in Chinese pharmacopoeia, and the herb source is dry aerial part of houttuynia cordata Thunb of Saururaceae. The leaves and the flowers are picked up and cut when the leaves and the flowers are exuberant in summer, the impurities are removed, and the leaves and the flowers are dried in the sun. The cordate houttuynia stem is in a flat cylindrical shape, is twisted, and has the length of 20-35 cm and the diameter of 0.2-0.3 cm; the surface is brown yellow, has a plurality of longitudinal edges, has obvious sections, and has residual fibrous roots on the lower section; is crisp and easy to break. She Husheng the blades are curled and contracted, flattened and heart-shaped, and have the length of 3-5 cm and the width of 3-4.5 cm; the tip is tapered and the whole edge is formed; the upper surface is dark yellow-green to dark brown, and the lower surface is gray-green or gray-brown; the petiole is slender, and the base and the supporting leaf are combined to form a sheath shape. The spike grows top and is yellow brown. The grating has fishy smell. Yu xing Cao is pungent in flavor, cold in nature and cool in nature, and enters lung meridian. It has effects in clearing away heat and toxic materials, relieving swelling, treating sore, promoting urination, removing dampness, clearing heat, relieving dysentery, invigorating stomach, and resolving food stagnation, and can be used for treating lung abscess, pyocutaneous disease, hematochezia, spleen and stomach heat caused by excessive heat, toxic heat, damp evil, and disease heat. Modern pharmacological experiments show that it has antibacterial, antiviral, immunity enhancing, and diuretic effects.
Peppermint (Mentha canadensis Linnaeus), also known as guayule, is a perennial herb of the genus Boschniakia of the family Labiatae. The leaves grow, flowers are light purple, lips are shaped, and dark purple brown small fruits are formed after flowers. Warm and moist places with abundant sunlight are usually used in mountain wetlands. The whole plant is fragrant, and is a fragrant crop with special economic value. Peppermint is one of the common traditional Chinese medicines. It is a pungent and cool drug for inducing sweat and relieving fever, and can be used for treating influenza, headache, conjunctival congestion, fever, laryngopharynx, and gum swelling and pain. It can be used for treating neuralgia, skin pruritus, skin rash, and eczema. Peppermint is used for replacing tea in common practice, and is used for clearing heart fire and improving eyesight.
The invention grinds wild plants of alternanthera philoxeroides, bidens pilosa, herba polygoni multiflori, houttuynia cordata and mint into crude juice, grinds the disease leaves of cucumber mosaic virus and the crude juice according to a proportion, and then inoculates healthy pumpkin cotyledons with consistent growth vigor. The test was repeated 3 times and a control was set, and parity leaves were selected 7-12 days after inoculation for symptom observation and statistics. The results show that the alternanthera philoxeroides, the bidens pilosa, the herba polygoni multiflori, the herba houttuyniae and the mint have a certain inhibiting effect on the CMV, wherein the alternanthera philoxeroides has the most obvious inhibiting effect on the CMV. And then the virus concentration in different treated plants is calculated by ELISA method, and the result shows that the treatment of the alligator alternanthera has remarkable CMV inhibition effect, the inhibition rate is 76.20%, the herba Equiseti hiemalis, herba Houttuyniae and herba Menthae all have a certain CMV inhibition effect (10.06-53.72%), and the ELISA result is consistent with the symptom expression result of the treated pumpkin, thus showing that the inhibition effect of the alligator alternanthera on the CMV is remarkable.
Further, carrying out ultrasonic water extraction and alcohol precipitation on the alternanthera philoxeroides to obtain the alternanthera philoxeroides extract. In order to clarify the influence of the alligator alternanthera extract solution on the seeding of the alligator alternanthera with CMV, the extract and CMV disease leaves are ground in proportion and then are inoculated with healthy pumpkin leaves with consistent growth vigor, and the result shows that after the alligator alternanthera extract is inoculated with the pumpkin, the pumpkin leaves have no flower leaf symptoms and the edges of the leaves are slightly yellow; whereas CMV control leaves showed chlorosis and more severe flowers, curled edges and heavier symptoms. The experimental result is consistent with the experimental effect of the alternanthera philoxeroides crude juice, and shows that the alternanthera philoxeroides extract is a component which has an inhibitory effect on CMV in the crude juice. Therefore, the alligator alternanthera, the herba bidentis pilosae, the herba houttuyniae and the mint can be used as a new plant source for preventing and treating cucumber mosaic virus diseases, and provides more choices for the production of CMV prevention and treatment.
Thus, the present invention first protects the following new applications with respect to the above-mentioned wild plants:
application of one or more of Alternanthera philoxeroides, bidens pilosa, herba Duchesneae Indicae, herba Houttuyniae or herba Menthae in resisting cucumber mosaic virus or preparing biological control agent for resisting cucumber mosaic virus is provided.
The application of any one or more of alternanthera philoxeroides, bidens pilosa, herba polygoni multiflori, herba houttuyniae or herba menthae in preventing and treating cucumber mosaic virus or in preparing biological preventing and treating agents for preventing and treating cucumber mosaic virus.
Further, the alternanthera philoxeroides, bidens pilosa, herba polygoni multiflori, herba houttuyniae or herba menthae are crude juice obtained by grinding fresh plants.
Further, the alternanthera philoxeroides is an alternanthera philoxeroides extract and is prepared by a water extraction and alcohol precipitation method.
Further, the preparation method of the alligator alternanthera extract comprises the steps of grinding fresh alligator alternanthera, adding deionized water (the feed-liquid ratio is 1:4-5 g/mL), carrying out ultrasonic intermittent extraction for 2-3 times, each time for 15-25 min, centrifuging the solution, adding 2-4 times of absolute ethyl alcohol in volume into supernatant, and precipitating with 2-6 ℃ ethyl alcohol overnight. And then centrifuging and drying the precipitate to obtain the product.
The invention also provides a plant source biological control agent for controlling cucumber mosaic virus, which contains any one or more of alternanthera philoxeroides, sticktight, herba Duchesneae Indicae, herba houttuyniae or herba Menthae as an active ingredient; the plant source biological control agent can take any one or more of alternanthera philoxeroides, bidens pilosa, herba polygoni multiflori, herba houttuyniae or peppermint as the only active ingredient or can be matched with other known active ingredients to play a role.
Further, the alternanthera philoxeroides, bidens pilosa, herba polygoni multiflori, herba houttuyniae or herba menthae are crude juice obtained by grinding fresh plants.
Further, the alternanthera philoxeroides is an alternanthera philoxeroides extract and is prepared by a water extraction and alcohol precipitation method.
Further, the preparation method of the alligator alternanthera extract comprises the steps of grinding fresh alligator alternanthera, adding deionized water (the feed-liquid ratio is 1:4-5 g/mL), carrying out ultrasonic intermittent extraction for 2-3 times, each time for 15-25 min, centrifuging the solution, adding 2-4 times of absolute ethyl alcohol in volume into supernatant, and precipitating with 2-6 ℃ ethyl alcohol overnight. And then centrifuging and drying the precipitate to obtain the product.
Further, the biocontrol agent also includes adjuvants acceptable to biocontrol agents.
The invention also provides application of any plant source biological control agent in cucumber mosaic virus resistance or in cucumber mosaic virus disease control.
Compared with the prior art, the invention has the following beneficial effects:
the research of the invention shows that the alternanthera philoxeroides, the bidens pilosa, the herba polygoni multiflori, the herba houttuyniae and the mint all have certain inhibition effect on the cucumber mosaic virus, wherein the inhibition effect of the alternanthera philoxeroides on the cucumber mosaic virus is most obvious; therefore, the invention provides the new application of any one or more of alternanthera philoxeroides, bidens pilosa, herba houttuyniae and peppermint in resisting cucumber mosaic virus or preventing and treating cucumber mosaic virus, and provides more choices for preventing and treating cucumber mosaic virus.
Drawings
FIG. 1 shows the symptoms of CMV-vaccinated zucchini after alligator alternanthera treatment. a: blank control; b: treating alternanthera philoxeroides; c: CMV control.
FIG. 2 shows the symptoms of CMV-inoculated zucchini after treatment with Bidens pilosa. a: blank control; b: treating Bidens pilosa; c: CMV control.
FIG. 3 shows the symptoms of CMV-vaccinated zucchini after treatment with Emilia. a: blank control; b: a treatment group of herba Duchesneae Indicae; c: CMV control.
FIG. 4 symptoms of CMV vaccinated zucchini after houttuynia cordata treatment. a: a CMV control; b: treating herba Houttuyniae; c, blank control.
Figure 5 is a symptom of CMV vaccinated zucchini after peppermint treatment. a: blank control; b: mint treatment; c CMV control.
FIG. 6 shows ELISA results of different plants against CMV. And (3) injection: different letters indicate significant differences between the different treatments (P < 0.05).
FIG. 7 shows symptoms of CMV vaccinated zucchini after allium alternifolium extract treatment. a: blank control; b: a treatment group of alternanthera philoxeroides extract; c: CMV control.
Detailed Description
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
Reagents and materials used in the following examples are commercially available unless otherwise specified.
EXAMPLE 1 inhibition effect of crude juice of wild plants on CMV
1. Materials and methods
1. Experimental materials
Pumpkin seeds and various wild plants: alternanthera philoxeroides, bidens pilosa, herba houttuyniae and mint.
2. Experimental method
Grinding the wild plants into crude juice, grinding CMV disease leaves and the crude juice according to a ratio of 1:8, and inoculating healthy pumpkin cotyledons with consistent growth vigor through juice friction. The test was repeated 3 times and a control was set, and parity leaves were selected 7-12 days after inoculation for symptom observation and statistics.
3. ELISA detection application Effect
The virus concentration in the different treated plants was calculated using ELISA. CMV inhibition was calculated as follows: inhibition (%) = (control OD 492 Value-treatment group OD 492 Value)/control OD 492 The value was 100%.
Statistical data analysis is expressed as mean ± Standard Error (SE) of 3 replicates. Data analysis of variance used Duncan's multiple range test, with different letters indicating significant differences at the 0.05 level.
2. Experimental results
1. anti-CMV effect of alternanthera philoxeroides
The CMV resistance effect of the alternanthera philoxeroides is shown in figure 1, leaves of the alternanthera philoxeroides treatment group do not have flowers and leaves and shrink, and the growth condition is good (figure 1 b); CMV control leaves showed chlorosis and more severe flowers, curled edges, and heavier symptoms (fig. 1 c); the control plants grew well (fig. 1 a). The clear effect of the alternanthera philoxeroides on CMV inhibition is shown.
2. anti-CMV effect of Bidens pilosa
The CMV-resisting effect of Bidens pilosa is shown in figure 2, the leaf edges of Bidens pilosa are slightly shrunken, no flower She Zhengzhuang is formed, the leaf edges are slightly yellow, and the leaf is smaller than that of a blank control (figure 2 b); CMV control leaves showed chlorosis and more severe flowers, curled edges, and heavier symptoms (fig. 2 c); the control plants grew well (fig. 2 a). The bidens pilosa has a certain inhibiting effect on CMV and a certain inhibiting effect on the growth of pumpkin.
3. An effect of herba Emiliae on resisting CMV
As shown in figure 3, the effect of the herba Duchesneae Indicae on resisting CMV is shown in the figure, the leaves and flowers are lighter than the control (figure 3 b); CMV control leaves were chlorosis and more severe flowers, leaf edges curled, and symptoms were heavier (fig. 3 c); the control plants grew well (fig. 3 a). It is shown that herba Duchesneae Indicae has certain inhibiting effect on CMV.
4. anti-CMV effect of houttuynia cordata
The effect of houttuynia cordata against CMV is shown in fig. 4, CMV control leaves fade, leaf edges curl, leaves are smaller (fig. 4 a); symptoms such as flowers and leaves, fading and the like appear on the surfaces of the leaves after the cordate houttuynia is treated, but the symptoms are lighter than the CMV control (figure 4 b); the control plants grew well (fig. 4 c). The cordate houttuynia has a certain inhibiting effect on CMV.
5. anti-CMV effect of peppermint
The effect of mint on resisting CMV is shown in figure 5, the surface of the mint treated leaf shows flower leaf and chlorosis symptoms, but the symptoms are not obvious in contrast with CMV, and the leaf is smaller (figure 5 b); CMV control leaves fade and flower and leaf symptoms are heavier (fig. 5 c); the leaves of the placebo group grew well (fig. 5 a). The peppermint has a certain inhibiting effect on CMV and a certain inhibiting effect on the growth of pumpkin.
6. Indirect ELISA method detection results
As shown in FIG. 6, the results of the indirect ELISA method show that the treatment of the alternanthera philoxeroides has remarkable CMV inhibition effect, the inhibition rate is 76.20%, and the herba Equiseti hiemalis, herba Houttuyniae and herba Menthae have a certain CMV inhibition effect (10.06-53.72%). The ELISA result is consistent with the symptom expression result of the treated pumpkin, which shows that the allium macrostemon, the herba polygoni multiflori, the sticktight, the herba houttuyniae and the mint all have a certain inhibiting effect on CMV, wherein the inhibiting effect of the allium macrostemon is most obvious.
Example 2 preparation of Alternanthera philoxeroides extract
Weighing 5g of fresh alligator alternanthera, grinding, adding 20mL of deionized water, and intermittently extracting for 2 times in an ultrasonic instrument for 20min each time. After the above treatment, the solution was centrifuged at 4000r/min for 30min, and 3 volumes of absolute ethanol were added to the supernatant and ethanol was precipitated overnight at 4 ℃. Subsequently, the mixture was centrifuged at 4000r/min for 15min, the precipitate was dried in an oven at 65℃and the dried extract was dissolved in deionized water for further use.
Example 3 inhibition of CMV by Alternanthera philoxeroides extract
In order to clarify the effect of the alligator alternanthera extract solution on the seeding of zucchini with CMV, referring to example 1, step 2, the extract was ground in proportion to CMV disease leaves and then inoculated with healthy zucchini leaves of consistent growth.
As shown in FIG. 7, after the extract of Alternanthera philoxeroides is inoculated into the pumpkin, the leaves of the pumpkin have no flower and leaf symptoms, and the edges of the leaves are slightly yellow (FIG. 7 b); whereas CMV control leaves showed chlorosis and more severe flowers, curled edges, and heavier symptoms (fig. 7 c); the leaves of the placebo group were good (fig. 7 a). The experimental result is consistent with the experimental effect of the alligator alternanthera crude juice, which shows that the alligator alternanthera extract is a component which has an inhibitory effect on CMV in the crude juice.
The results show that the alternanthera philoxeroides, the herba polygoni multiflori, the herba houttuyniae and the mint all have a certain inhibiting effect on the CMV, and compared with the alternanthera philoxeroides, the herba polygoni multiflori, the herba houttuyniae and the mint, the alternanthera philoxeroides has an obvious inhibiting effect on the CMV and the inhibiting effect is the best. Meanwhile, the alternanthera philoxeroides extract is a substance for inhibiting CMV in the crude extract, and has obvious effect of inhibiting CMV. Therefore, the alligator alternanthera, the herba bidentis pilosae, the herba houttuyniae and the mint can be used as a new plant source for preventing and treating cucumber mosaic virus diseases, and provides more choices for the production of CMV prevention and treatment.
Claims (10)
1. Application of one or more of Alternanthera philoxeroides, bidens pilosa, herba Duchesneae Indicae, herba Houttuyniae or herba Menthae in resisting cucumber mosaic virus or preparing biological control agent for resisting cucumber mosaic virus is provided.
2. The application of any one or more of alternanthera philoxeroides, bidens pilosa, herba polygoni multiflori, herba houttuyniae or herba menthae in preventing and treating cucumber mosaic virus or in preparing biological preventing and treating agents for preventing and treating cucumber mosaic virus.
3. Use according to claim 1 or 2, wherein the alternanthera philoxeroides, bidens pilosa, herba Equiseti hiemalis, herba Houttuyniae or herba Menthae is the crude juice of its fresh plant ground.
4. Use according to claim 1 or 2, characterized in that the alternanthera philoxeroides is an aqueous alcohol extract thereof.
5. A plant biological control agent for controlling cucumber mosaic virus disease is characterized by comprising any one or more of alternanthera philoxeroides, bidens pilosa, herba polygoni multiflori, cordate houttuynia or peppermint as an active ingredient.
6. The biocontrol agent of plant origin as claimed in claim 5, wherein the alternanthera philoxeroides, bidens pilosa, herba Duchesneae Indicae, herba Houttuyniae or herba Menthae are crude juices of ground fresh plants thereof.
7. The plant-derived biocontrol agent of claim 5, wherein said allium macrostemon is an aqueous ethanol extract thereof.
8. The plant-derived biocontrol agent of claim 5, further comprising an adjuvant acceptable to the biocontrol agent.
9. The plant biological control agent according to claim 7, wherein the preparation method of the water extracted and alcohol precipitated extract of alternanthera philoxeroides comprises the steps of grinding fresh alternanthera philoxeroides, adding deionized water, intermittently extracting with ultrasound for 2-3 times at a feed liquid ratio of 1:4-5 g/mL for 15-25 min each time, centrifuging the solution, adding 2-4 times volume of absolute ethyl alcohol into the supernatant, precipitating with 2-6 ℃ of ethyl alcohol, centrifuging, and drying the precipitate.
10. Use of a plant-derived biocontrol agent according to any one of claims 5 to 9 for combating cucumber mosaic virus or for controlling cucumber mosaic virus diseases.
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