CN115152798B - Application and composition of Rapex baileyi PR2 extract - Google Patents
Application and composition of Rapex baileyi PR2 extract Download PDFInfo
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 20
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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
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
-
- 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
- A01N43/14—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 six-membered rings
- A01N43/16—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 six-membered rings with oxygen as the ring hetero atom
-
- 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
-
- 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
- A01P21/00—Plant growth regulators
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/645—Fungi ; Processes using fungi
Abstract
The invention relates to the technical field of microbial application, and particularly discloses application of an irpex alba PR2 extract and a composition, particularly relates to application of an ethanol extract of a mycelium of irpex alba PR2 after fermentation in prevention and treatment of plant virus diseases, and further discloses a composition containing the irpex alba PR2 extract and one of amino-oligosaccharin or brown algae oligosaccharide, wherein the composition is mixed and used according to a reasonable proportion, so that the prevention and treatment effect on the plant virus diseases can be remarkably improved, the prevention effect is higher than that of each single agent, and the composition has a remarkable synergistic effect; and after the rakanka pallida PR2 extract is matched with amino-oligosaccharin or alginate-derived oligosaccharide for application, the plant growth promotion effect is also remarkable.
Description
Technical Field
The invention belongs to the technical field of microorganism application, and particularly relates to application and a composition of a Rapex baichii PR2 extract.
Background
In nature, many plants are infected by viruses to cause virus diseases, and the virus diseases are often called plant cancers due to the absolute parasitism of the viruses to hosts; the yield, quality and shelf life of crops can be significantly reduced after the plants are infected with viruses. Due to The rapid mutation rate of Viruses, new virus strains are continuously appeared, about 1300 Viruses are known at present according to data released in 2020 by The International Committee for virus classification (The International Committee on Taxonomy of Viruses, ICTV), and The loss caused by plant Viruses is up to $ 200 billion per year in China due to The virus infection of crops, and is one of The threats of The world population survival.
At present, chemical medicines are mainly adopted for preventing and treating plant virus diseases, and the chemical medicines have certain prevention and control effects on the occurrence and development of the virus diseases, but also bring about the problems of environmental pollution, pesticide residues and the like. The biological antiviral agent is a new biological pesticide, and has the characteristics of no toxicity, no environmental pollution, no residue, no damage to natural enemies, human and animal safety protection and the like; and has the advantages of small dosage, long lasting period, low cost and the like, and is the key for realizing industrialized biological control.
CN106399132B, a Raharomyces baicilis strain and application thereof, provides a Raharomyces baicilis strain (AIrpex lacteus) PR2, the strain is preserved in China general microbiological culture Collection center (CGMCC) for short, and the address is as follows: the collection number of the strain is CGMCC NO.13190, no. 3 of Xilu No.1 of Beijing, chaoyang, beijing, and institute of microbiology of Chinese academy of sciences; the application of the strain extract in promoting the growth of crops or increasing the yield of crops is disclosed, and no report about the application of the irpex cacteus in preventing and treating plant virus diseases exists at present.
Disclosure of Invention
In view of the drawbacks of the prior art, the present invention aims to provide the application and composition of the Rapex baichii PR2 extract.
In order to achieve the technical effects, the invention adopts the following technical scheme:
the invention provides an application of a raking tooth fungus PR2 extract in preventing and treating plant virus diseases; the Rapex baichii PR2 extract is an ethanol extract of Rapex baichii PR2 mycelium.
Preferably, the plant viral disease is tobacco plant PVX viral disease; the pathogeny of the PVX virus disease of the tobacco plant is potato X virus.
Since the PR2 extract induces non-specific resistance, it is theoretically possible to improve the resistance of plants to various viruses including tobacco mosaic virus, potato virus Y, potato virus X, tomato yellow leaf curl virus, tomato de-greening virus, and the like.
The preparation method of the rakanka pallida PR2 extract comprises the steps of fermenting the rakanka pallida PR2, carrying out centrifugal filtration on fermentation liquor to obtain mycelium and extracting the mycelium by adopting ethanol.
Specifically, in the preparation method, the fermentation medium used for fermentation comprises the following raw materials: 1000ml of potato extract, 1.0g of yeast extract, 3.0g of peptone, 15.0g of glucose and 17.0g of agar.
Specifically, the preparation method of the potato extract comprises the following steps: 200g of Pi Maling potatoes are taken out, cut into small pieces, added with 1000ml of water and boiled for 30 minutes, the potato pieces are filtered out, and the filtrate is complemented to 1000ml.
Specifically, in the preparation method, the fermentation conditions are as follows: the inoculation amount is 5 to 15 percent (volume percentage), the fermentation culture temperature is 20 to 30 ℃, and the culture is 4~6 days.
Specifically, in the preparation method, the steps of extracting with ethanol are as follows: drying and crushing the mycelium, adding ethanol, carrying out cold soaking for 20 to 30h, and carrying out ultrasonic extraction for 0.5 to 1.5h; repeatedly extracting 2~4 times, and mixing filtrates.
Specifically, the ethanol extraction is performed by: drying and pulverizing mycelium at 50 deg.C, adding ethanol, cold soaking for 24 hr, and ultrasonic extracting for 2 hr; extracting for 2 times, and mixing filtrates.
The adding amount of the ethanol is 15 to 30 percent of the weight of the mycelium (after drying); the same amount of ethanol was added for each extraction.
Another object of the present invention is to provide a composition for the above use, comprising the above irpex alba PR2 extract in combination with any one of amino-oligosaccharins or fucoidan oligosaccharides.
Preferably, the mass ratio of the rake baileyi PR2 extract to the amino-oligosaccharin in the composition is 1 to 5.
Preferably, the mass ratio of the rake baileyi PR2 extract to the amino-oligosaccharin in the composition is 1.
Preferably, the mass ratio of the Rapex baichisonii PR2 extract to the fucoidan in the composition is 1.
Preferably, the mass ratio of the Rapex baichisonii PR2 extract to the fucoidan in the composition is 1.
Preferably, the concentration of the rakanka baileyi extract in the composition is 7.5mg/ml, and the mass percentage content of the amino-oligosaccharin and the fucoidan oligosaccharide is 90%.
Compared with the prior art, the invention has the following beneficial effects:
the invention provides the application of the Rapex baichii PR2 extract in preventing and treating plant virus diseases, the prevention effect of the Rapex baichii PR2 extract on the plant virus diseases is far higher than that of the existing antiviral preparations (amino-oligosaccharin and chitosan oligosaccharide), a new biological source antiviral preparation is provided, and the preparation is safe, efficient, environment-friendly and wide in application prospect; and the absolute dosage of the extract is small, and the cost is low.
After the rakanka albopictus PR2 extract is applied in a matching way with the amino-oligosaccharin or the fucoidan oligosaccharide, the control effect on plant virus diseases can be obviously improved, the control effect is higher than that of each single agent, and the composition has obvious synergistic effect; and after the rakanka pallida PR2 extract is matched with amino-oligosaccharin or alginate-derived oligosaccharide for application, the plant growth promotion effect is also remarkable.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described with the following embodiments, but is by no means limited thereto. The following is a description of the preferred embodiments of the present invention, and should not be taken as limiting the invention, but rather as embodying the invention in its broadest form and as indicating any variations, equivalents and modifications within the spirit and scope of the present invention.
Preparation of irpex cacteus PR2 extract:
the irpex leucotrichi PR2 extract can be prepared according to a preparation method of a plant resistance inducer disclosed in a patent CN106399132B, and the specific preparation method is as follows:
1. fermentation culture:
inoculating the strain of Rachiomyces baishanensis PR2 stored on the inclined plane of a test tube at 4 ℃ to a PDA culture medium, culturing for a week at 25 ℃, digging agar blocks, inoculating the agar blocks to a 250mL triangular flask filled with 50mL of LPDA culture solution, culturing for 3 days as seeds on a rotary shaking table at 25 ℃ and 180r/min, inoculating 10 percent of the agar blocks to a 500mL triangular flask filled with 150mL of fermentation culture medium, culturing for 5 days under the same condition, and terminating fermentation to obtain a seed solution.
The PDA culture medium formula comprises: 200g of potatoes, 20g of glucose, 20g of agar and 1000ml of water.
The fermentation medium formula comprises: 1.0L of potato extract, 1.0g of yeast extract, 3.0g of peptone, 15.0g of glucose and 17.0g of agar.
Preparing a potato extracting solution: removing Pi Maling potato 200g, cutting into small pieces, adding 1000ml of water, boiling for 30 min, filtering to remove potato pieces, and adding the filtrate to 1000ml.
2. Ethanol extraction:
centrifuging and filtering the fermentation liquor to obtain mycelium, drying and crushing the mycelium at 50 ℃, adding ethanol for cold soaking for 24 hours, and then carrying out ultrasonic extraction for 2 hours; extracting for 2 times, and mixing filtrates. The adding amount of the ethanol is 20% of the weight of the dried mycelium; the same amount of ethanol was added for each extraction.
Example 1 control of PVX Virus disease in indoor tobacco plants
Step one, tobacco culture: taking a seedling raising tray fully paved with nutrient soil, adding water to the bottom of the seedling raising tray for soaking, then, dropping tobacco seeds on the surface of the nutrient soil by using tweezers, dropping four seeds in each hole tray, standing the periphery of the seedling raising tray, covering a preservative film for preserving moisture, placing the seedling raising tray in a 24 ℃ artificial climate chamber for culturing for 16/8h, after the seeds germinate (about fifteen days after dropping), removing the film and continuously culturing to a 3~4 leaf stage, and moving the tobacco seedlings to a small flower pot for continuously culturing to a 4~6 leaf stage for use;
step two, preparing juice with toxin: taking out 0.1g of preserved leaf with virus from a refrigerator at-80 deg.C, adding 2ml of PBS, grinding into homogenate to obtain juice with virus for use;
step three, spraying a medicament:
taking single agents of amino-oligosaccharin, brown alga oligosaccharide and irpex cacteus PR2 extracts as contrast agents, taking the amino-oligosaccharin + irpex cacteus PR2 extracts, the brown alga oligosaccharide + irpex cacteus PR2 extracts as test agents, setting blank contrast, dissolving each treatment agent in 15L of water, quantitatively applying medicines to treat tobacco by using a Bat spray tower, treating six tobacco plants by using each agent, and paying attention to uniform spraying of each leaf; wherein the Ralstonia baichii PR2 extract is a solution with the concentration of 7.5 mg/ml; the amino-oligosaccharin and the alginate-derived oligosaccharide are both raw medicines with the mass percentage of 90 percent.
Step four, virus inoculation: spraying the medicament for two hours, inoculating three true leaves below the tobacco system leaves by a rubbing inoculation method, inoculating 50 mu L of the juice with toxicity prepared in the second step to each true leaf, inoculating by a 600-mesh carborundum rubbing method, and culturing in an artificial climate chamber for 5d after inoculation;
step five, observation and detection: culturing tobacco in artificial climate room for 5 days, observing fluorescence conditions of tobacco system leaf treated by contrast and treatment agent under long-wave ultraviolet lamp as phenotype characteristic (if fluorescence mark appears, it is proved to be infected leaf, picking it, and storing in refrigerator of-80 deg.C for long term); respectively taking 0.1g of samples treated by the control agent and the test agent, adding 1ml of CBS buffer solution for grinding, placing the grinding solution in a centrifuge tube at 5000r/min, centrifuging for 5min, taking supernatant for later use, and measuring the OD value of each treatment under the wavelength of 450nm by using an enzyme-linked immunosorbent assay and an enzyme-linked immunosorbent assay.
Performing statistical analysis on the enzyme-linked result by using SPSS 21.0 software and a Duncan method, wherein the analysis result is shown in tables 1-2; if the significant difference occurs, namely P is less than 0.05, the treatment medicament is regarded as having significant promotion effect on the prevention effect of the tobacco PVX virus disease;
the virus prevention/inhibition rate calculation method comprises the following steps:
control effect/inhibition (%) = [ (control OD) 450 Treatment of OD 450 ) Control OD 450 ]×100%
The control effect was evaluated as the average inhibition rate per treatment, and the test results were as follows:
TABLE 1 preventive Effect of the treatments on PVX Virus disease in tobacco plants
As can be seen from table 1, the control effect of Shi Anji oligosaccharide alone on PVX virus diseases is about 49%, the control effect of PR2 extract alone on PVX virus diseases is about 60%, and after PR2 extract and amino-oligosaccharide are compounded according to the following ratio of 1.
The control effect of 0.3g of single Shi Hezao oligosaccharide on PVX virus diseases is about 46%, the control effect of single PR2 extract on the virus diseases is about 59%, and after the PR2 extract and the alginate oligosaccharide are compounded according to the following ratio of 1.
Example 2
The plant heights of the tobacco plants treated by the optimal treatment and the tobacco plants treated by the control treatment in example 1 were measured and analyzed, and the test results are shown in table 2;
TABLE 2 plant height of the treated plants
As can be seen from Table 2, the plant height of the tobacco treated after the Rapex baileyi PR2 extract is compounded with the amino-oligosaccharin or the brown alginate oligosaccharide is obviously increased compared with that of each single dose, and the difference is obvious, which indicates that the plant growth promotion effect can be realized after the Rapex baileyi PR2 extract is compounded with the amino-oligosaccharin or the brown alginate oligosaccharide.
The above-mentioned embodiments only express the specific embodiments of the present application, and the description thereof is more specific and detailed, but not construed as limiting the scope of the present application. It should be noted that, for those skilled in the art, without departing from the technical idea of the present application, several changes and modifications can be made, which all belong to the protection scope of the present application.
Claims (5)
1. The application of the rakanka albopictus PR2 extract is characterized in that the extract is an ethanol extract of the rakanka albopictus PR2 mycelium and is used for preventing and treating PVX virus diseases of tobacco plants and promoting the growth of the tobacco plants.
2. The application of the rakanka albopictus PR2 extract according to claim 1, wherein the preparation method of the rakanka albopictus PR2 extract comprises the steps of sequentially carrying out culture activation and seed culture on the preserved rakanka albopictus PR2 to obtain a seed solution; inoculating the seed liquid into a fermentation culture medium for fermentation culture to obtain a fermentation liquid; then, centrifugally filtering the fermentation liquor to obtain mycelia; and finally, drying and crushing the mycelium, carrying out cold soaking by using ethanol, and carrying out ultrasonic extraction to obtain an ethanol extract of the mycelium of the Harpagophytum baileyi PR 2.
3. A composition for use according to any one of claims 1~2 wherein the composition is a combination of irpex cacteus PR2 extract and any one of an amino-oligosaccharide or a brown alginate-oligosaccharide; wherein the mass ratio of the Rapex baileyi PR2 extract to the amino-oligosaccharin is 1 to 5; the mass ratio of the rakanka albicans PR2 extract to the brown alga oligosaccharides is 1 to 4.
4. The composition according to claim 3, wherein the mass ratio of the Rapex baichii PR2 extract to the amino-oligosaccharin in the composition is 1.
5. The composition according to claim 3, wherein the mass ratio of the Rapex baichii PR2 extract to the fucoidan oligosaccharide is 1 to 2-1.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH10218718A (en) * | 1997-02-05 | 1998-08-18 | Norin Suisansyo Yasai Chiyagiyou Shikenjo | Antimicrobial composition and control of storage disorder of vegetable and fruits |
| CN106635821A (en) * | 2016-08-13 | 2017-05-10 | 辽宁省农业科学院微生物工程中心 | Mycorrhiza fungi for promoting nutrient absorption of blueberry and preparation method and application of mycorrhiza fungi |
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| CN106399132B (en) * | 2016-12-16 | 2019-09-20 | 山东蓬勃生物科技有限公司 | One plant of Irpex lacteus and its application |
| CN109161488B (en) * | 2018-10-15 | 2021-04-09 | 宜春学院 | White-capsule rake tooth strain for producing cordycepin and culture method thereof |
| CN112899171B (en) * | 2021-04-02 | 2022-06-14 | 河南科技大学 | Rapex leucocytochys LL210, application thereof and biocontrol microbial inoculum |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| JPH10218718A (en) * | 1997-02-05 | 1998-08-18 | Norin Suisansyo Yasai Chiyagiyou Shikenjo | Antimicrobial composition and control of storage disorder of vegetable and fruits |
| CN106635821A (en) * | 2016-08-13 | 2017-05-10 | 辽宁省农业科学院微生物工程中心 | Mycorrhiza fungi for promoting nutrient absorption of blueberry and preparation method and application of mycorrhiza fungi |
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Denomination of invention: Application and Composition of PR2 Extract from Leucocystis serrata Effective date of registration: 20231026 Granted publication date: 20230131 Pledgee: Tai'an Daiyue Rural Commercial Bank Co.,Ltd. Pledgor: SHANDONG PENGBO BIOTECHNOLOGY Co.,Ltd. Registration number: Y2023980062858 |