CN114052048B - Cinnamomum camphora leaf extract and preparation method and application thereof - Google Patents
Cinnamomum camphora leaf extract and preparation method and application thereof Download PDFInfo
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- CN114052048B CN114052048B CN202111238729.9A CN202111238729A CN114052048B CN 114052048 B CN114052048 B CN 114052048B CN 202111238729 A CN202111238729 A CN 202111238729A CN 114052048 B CN114052048 B CN 114052048B
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- leaf extract
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- camphor
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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]
- A01N65/24—Lauraceae [Laurel family], e.g. laurel, avocado, sassafras, cinnamon or camphor
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
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- 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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- General Health & Medical Sciences (AREA)
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- Agronomy & Crop Science (AREA)
- Microbiology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Plant Pathology (AREA)
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- Agricultural Chemicals And Associated Chemicals (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a camphor leaf extract and a preparation method and application thereof. The preparation method is simple and easy to operate, the total yield is between 4.69% and 7.38%, the preparation method is suitable for large-scale production, and the activity of the obtained extract in inhibiting the apple black rot fungi, the watermelon fusarium oxysporum and the fusarium catenulatum is superior to that of the environment-friendly plant source pesticide eugenol which is used in a large amount at present.
Description
Technical Field
The invention relates to the technical field of plant pathogenic bacteria inhibition, in particular to a camphor leaf extract and a preparation method and application thereof.
Background
Pathogenic bacteria have been severely resistant to many chemical agents due to long-term use or abuse of chemical pesticides; in addition, pesticides remained on the surface of agricultural products seriously damage human health, and cause environmental pollution and ecological balance destruction. Many researches have shown that in the long-term struggling process with plant pathogenic fungi, plants can generate secondary metabolites for inhibiting the plant pathogenic fungi, such as eugenol, ethylicin, matrine and the like, have strong inhibition effect on the plant pathogenic fungi and are not easy to generate resistance. Therefore, searching for natural plant source extracts that efficiently inhibit plant pathogenic bacteria has become one of the important ways to replace chemical pesticides and develop environmentally friendly biopesticides.
Apple rot (Valsa canker of apple) caused by apple black rot fungi (Valsa maliMiyabe et Yamada) is an important disease which damages apples in China, and damages the branches and fruits of the apples, so that apple barks rot, the yield and quality of the fruits are seriously reduced, the distribution is wide, the damage is serious, and almost all apple producing areas occur. Gliocladium (Gliocladium catenulatum) is a pathogenic fungus of pea root rot and can attack the plant tissue of clover and leguminous plants.
Cinnamomum camphora Cinnamomum camphora is Lauraceae plant, and is mainly distributed in Fujian, taiwan, jiangxi, guangdong, yunnan, guizhou, sichuan and other areas, and the artificial forest reaches tens of thousands hectares, so that plant resources are rich. However, the resource utilization is only remained in the development of volatile oil (the mass fraction is less than or equal to 5%), a large number of shoots and leaves are extracted and then treated as waste, no other valuable products exist, and the plant utilization rate is low. Through long-term research on development and utilization of camphor plant resources, and through first discovery of research, the camphor leaf extract can well inhibit plant pathogenic bacteria, so that the invention provides the camphor leaf extract, and the preparation method and application thereof.
Disclosure of Invention
The invention aims to provide a preparation method of a camphor leaf extract capable of inhibiting plant pathogenic bacteria and application of the camphor leaf extract in inhibiting plant pathogenic fungi such as apple black rot fungi, watermelon fusarium oxysporum, and gliocladium catenulatum.
In order to achieve the above purpose, the invention provides a preparation method of a camphor leaf extract, which specifically comprises the following steps:
1) Collecting fresh Cinnamomum camphora leaf, oven drying at 50-60deg.C, pulverizing, and sieving with 20-50 mesh sieve to obtain dry powder;
2) 1000g of the powder obtained in the step 1), adding 6000-8000mL of dichloromethane, heating to boiling, extracting under reflux for 50min, filtering, and collecting the extract;
3) Concentrating the extract obtained in the step 2) under reduced pressure to obtain the camphor leaf extract.
The application of Cinnamomum camphora leaf extract in inhibiting plant pathogenic fungi is provided.
The plant pathogenic fungi comprise any one of black rot of apple, fusarium oxysporum, and fusarium catenulatum.
Mixing the camphor leaf extract with water to prepare a liquid medicine with the concentration of 80-260 ppm, and spraying the liquid medicine on leaf surfaces until leaf surfaces drip water; or preparing into 40-130 ppm liquid medicine for root irrigation.
The invention has the remarkable advantages that:
(1) The raw material, namely the camphor wood, used by the invention can be cultivated artificially in a large quantity, the resource is rich, the extraction method is simple and practical to operate, the total yield is between 4.69% and 7.38%, and the large-scale production can be realized.
(2) The extract obtained by the invention has remarkable inhibitory activity on the apple black rot fungi, the watermelon fusarium oxysporum and the chain broom fungus, and the inhibitory activity of the extract is superior to that of the environment-friendly plant-source pesticide eugenol used for preventing and treating plant pathogenic bacteria in a large amount at present under the same concentration.
Detailed Description
A Cinnamomum camphora leaf extract is prepared by the following steps:
1) Collecting fresh Cinnamomum camphora leaf, oven drying at 50-60deg.C, pulverizing, and sieving with 20-50 mesh sieve to obtain dry powder;
2) 1000g of the powder obtained in the step 1), adding 6000-8000mL of methylene dichloride, heating to boiling, extracting under reflux for 50min, filtering, collecting the extracting solution,
3) Concentrating the extract obtained in the step 2) under reduced pressure to obtain the camphor leaf extract.
Research on inhibition of plant pathogenic fungi activity by camphor leaf extract:
the antibacterial activity of the camphor leaf extract is measured by using a mycelium production rate method. The method comprises the steps of preparing a culture plate by using a prepared and sterilized PDA culture medium, and setting a blank control group, a positive control eugenol group and a camphor leaf extract test group; the positive control eugenol group and the camphor leaf extract test group are respectively provided with 3 concentrations, and each concentration is provided with 3 repetitions; inoculating black rot of apple, fusarium oxysporum, and fusarium catenulatum respectively, and sealing the inoculated culture dish with a preservative film to prevent water or impurities in the air generated in the growth process of the strain from affecting the normal growth of the strain. Culturing in intelligent artificial incubator at 28+ -5 deg.c for 3-7 d, and observing every 24 hr. The measurement was performed when the hypha diameter of the control plate strain was as large as about 2/3. During measurement, the colony shape is observed firstly, then the cross measurement method is adopted, the maximum diameter of the irregular colony is measured, and the growth diameters of all groups of colonies are measured and recorded respectively. The bacteriostasis rate was calculated according to the formula (the calculation formula is as follows), and the results are shown in tables 1 to 3.
The antibacterial rate of the eugenol group or the camphor leaf extract test group serving as the positive control agent is= (the colony diameter of the eugenol group or the camphor leaf extract test group serving as the positive control agent-the colony diameter of the blank control group)/(the colony diameter of the blank control group-0.5) ×100%.
Table 1 comparison of the inhibition ratio of eugenol and Cinnamomum camphora leaf extract to apple black rot skin pathogen
Table 2 comparison of bacteriostasis rates of eugenol and Cinnamomum camphora leaf extract against Fusarium oxysporum in watermelon
TABLE 3 comparison of the antibacterial Rate of eugenol and Cinnamomum camphora leaf extract against Scopularillus alternaria
From the results, the inhibition activity of the camphor leaf extract on pathogenic bacteria is obviously better than that of eugenol at the same concentration.
The invention will be better understood from the following examples. However, it will be more readily understood by those skilled in the art that the specific material ratios, process conditions and results thereof described in the examples are illustrative of the present invention and should not be construed as limiting the invention described in detail in the claims.
Example 1
Taking camphor leaves, drying in a baking oven at 55 ℃, then crushing by a crusher, sieving by a 30-mesh sieve, and collecting powder for later use; 1000g of powder is weighed, 8000mL of dichloromethane is added, the mixture is heated to boiling and is extracted under reflux for 50min, and the extracted solution is concentrated under reduced pressure to obtain 73.8g of camphor leaf extract, and the total yield is 7.38%.
The antibacterial activity of the camphor leaf extract is measured by using a mycelium production rate method, and the result shows that: the antibacterial rate of the extract with the concentration of 200 mug/mL to the apple black rot pathogen, the watermelon fusarium oxysporum and the chain spore gliocladium is 91.74+/-3.39%, 96.99+/-3.20% and 97.69+/-3.15% respectively.
Example 2
Taking camphor leaves, drying in a baking oven at 55 ℃, then crushing by a crusher, sieving by a 30-mesh sieve, and collecting powder for later use; 1000g of powder is weighed, 6000mL of dichloromethane is added, the mixture is heated to boiling and is extracted under reflux for 50min, and the extracted solution is concentrated under reduced pressure to obtain 46.9g of camphor leaf extract, and the total yield is 4.69%.
The antibacterial activity of the camphor leaf extract is measured by using a mycelium production rate method, and the result shows that: the antibacterial rate of the extract with the concentration of 200 mug/mL to the apple black rot pathogen, the watermelon fusarium oxysporum and the chain spore gliocladium is 89.79 plus or minus 2.26 percent, 93.99 plus or minus 4.01 percent and 94.02 plus or minus 3.97 percent respectively.
Example 3
Taking camphor leaves, drying in a baking oven at 55 ℃, then crushing by a crusher, sieving by a 30-mesh sieve, and collecting powder for later use; 100g of powder is weighed, 800mL of dichloromethane is added, the mixture is heated to boiling and is extracted under reflux for 50min, and the extracted solution is concentrated under reduced pressure to obtain 6.29g of camphor leaf extract, and the total yield is 6.29%.
The antibacterial activity of the camphor leaf extract is measured by using a mycelium production rate method, and the result shows that: the antibacterial rate of the extract with the concentration of 200 mug/mL to the apple black rot pathogen, the watermelon fusarium oxysporum and the chain spore gliocladium is 90.56+/-4.16%, 92.67+/-2.46% and 96.21 +/-3.05% respectively.
The foregoing description is only of the preferred embodiments of the invention, and all changes and modifications that come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Claims (2)
1. An application of a camphor leaf extract in inhibiting plant pathogenic fungi, which is characterized in that: the plant pathogenic fungi comprise any one of apple black rot fungi, watermelon fusarium oxysporum and gliocladium catenulatum;
the preparation method of the camphor leaf extract specifically comprises the following steps:
1) Collecting fresh Cinnamomum camphora leaf, oven drying at 50-60deg.C, pulverizing, and sieving with 20-50 mesh sieve to obtain dry powder;
2) Adding dichloromethane 6000-8000mL into the powder 1000g obtained in the step 1), heating to boiling, extracting under reflux for 50min, filtering, and collecting the extractive solution;
3) Concentrating the extract obtained in the step 2) under reduced pressure to obtain the camphor leaf extract.
2. Use of the extract of the leaves of Cinnamomum camphora according to claim 1, for inhibiting phytopathogenic fungi, characterized in that: the specific application method comprises the following steps: mixing the camphor leaf extract with water to prepare a liquid medicine with the concentration of 80-260 ppm, and spraying the liquid medicine on leaf surfaces until leaf surfaces drip water; or preparing a liquid medicine with the concentration of 40-130 ppm for root irrigation.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202111238729.9A CN114052048B (en) | 2021-10-25 | 2021-10-25 | Cinnamomum camphora leaf extract and preparation method and application thereof |
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| CN114052048B true CN114052048B (en) | 2023-06-23 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104146055A (en) * | 2014-06-27 | 2014-11-19 | 广西大学 | Plant source fruit and vegetable preservative |
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| KR100611248B1 (en) * | 2004-01-05 | 2006-08-10 | 주식회사 내츄로바이오텍 | Antibacterial and antifungal composition containing plant oil |
| CN103461396B (en) * | 2013-08-22 | 2014-12-10 | 上海交通大学 | Compound botanical fungiticide, and preparation method and application thereof |
| CN110150333B (en) * | 2019-05-16 | 2021-06-01 | 井冈山大学 | Cinnamomum longepaniculatum leaf essential oil bacteriostasis method |
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| CN104146055A (en) * | 2014-06-27 | 2014-11-19 | 广西大学 | Plant source fruit and vegetable preservative |
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