CN114052048A - 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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- CN114052048A CN114052048A CN202111238729.9A CN202111238729A CN114052048A CN 114052048 A CN114052048 A CN 114052048A CN 202111238729 A CN202111238729 A CN 202111238729A CN 114052048 A CN114052048 A CN 114052048A
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- leaf extract
- cinnamomum camphora
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- camphora leaf
- 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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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Mycology (AREA)
- Biotechnology (AREA)
- Agronomy & Crop Science (AREA)
- Microbiology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Plant Pathology (AREA)
- Dentistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a cinnamomum camphora leaf extract, and a preparation method and application thereof. The preparation method is simple and easy to operate, the total yield is 4.69-7.38%, the method is suitable for large-scale production, and the activity of the obtained extract in inhibiting the Humicola reticulata, Fusarium oxysporum and Gliocladium catenulatum is superior to that of the environment-friendly botanical pesticide eugenol which is massively used at present.
Description
Technical Field
The invention relates to the technical field of inhibiting phytopathogens, and in particular relates to a cinnamomum camphora leaf extract and a preparation method and application thereof.
Background
The long-term use or abuse of chemical pesticides has led to the development of serious resistance of pathogenic bacteria to many chemical agents; in addition, the pesticide remained on the surface of agricultural products seriously harms the health of human body, and causes environmental pollution and damage to ecological balance. Many researches have shown that during the long-term fight with plant pathogenic fungi, plants can produce secondary metabolites for inhibiting plant pathogenic fungi, such as eugenol, ethylicin, matrine and the like, which have strong inhibiting effect on the plant pathogenic fungi and are not easy to generate resistance. Therefore, the search for natural plant extracts which can effectively inhibit plant pathogenic bacteria has become one of important ways for replacing chemical pesticides and developing environment-friendly biological pesticides.
Apple rot (Valsa canker of apple) caused by apple pericarp fungus (Valsa maliMiyabe et Yamada) is an important disease harmful to apples in China, and damages branches and fruits of apples, so that apple barks are rotten, the yield and quality of the fruits are seriously reduced, the distribution is wide, the harm is serious, and almost all apple producing areas are generated. Gliocladium catenulatum (Gliocladium catenulatum) is a pathogenic fungus of pea root rot, and can attack plant tissues weakened by clover and beans.
The Cinnamomum camphora is a Cinnamomum plant in Lauraceae, is mainly distributed in areas such as Fujian, Taiwan, Jiangxi, Guangdong, Yunnan, Guizhou, Sichuan and the like in China, reaches ten thousand hectares in artificial forest, and has rich plant resources. However, the resource utilization is only remained in the development of volatile oil (mass fraction is less than or equal to 5%), a large amount of tender branches and leaves are extracted and then are treated as waste, other valuable products are not produced, and the plant utilization rate is low. The invention provides a cinnamomum camphora leaf extract, a preparation method and application thereof, and the method is simple to operate and low in cost, and the obtained extract has obvious activity and extremely good development prospect.
Disclosure of Invention
The invention aims to provide a preparation method of a cinnamomum camphora leaf extract capable of inhibiting plant pathogenic bacteria and application of the cinnamomum camphora leaf extract in inhibiting plant pathogenic fungi such as apple Humicola melanosporum, Fusarium oxysporum f.sp.citrulli, Scopulariopsis alternata and the like.
In order to achieve the purpose, the invention provides a preparation method of a cinnamomum camphora leaf extract, which specifically comprises the following steps:
1) collecting fresh Cinnamomum camphora leaves, oven drying at 50-60 deg.C, pulverizing, and sieving with 20-50 mesh sieve to obtain dried powder;
2) adding 1000g of the powder obtained in the step 1) into 8000mL of dichloromethane 6000-;
3) decompressing and concentrating the extracting solution obtained in the step 2) to obtain the camphor leaf extract.
Application of folium Cinnamomi Camphorae extract in inhibiting plant pathogenic fungi is provided.
The plant pathogenic fungi include any one of Humicola reticulata, Fusarium oxysporum and Gliocladium 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 the leaf surface until the leaf surface drips; or preparing a liquid medicine with the concentration of 40-130 ppm for root irrigation.
The invention has the following remarkable advantages:
(1) the raw material of the cinnamomum camphora can be artificially cultured 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 percent, and the large-scale production can be realized.
(2) The extract obtained by the invention has obvious inhibitory activity on Humicola melanosporum, Fusarium oxysporum f.sp.citrulli and Gliocladium catenulatum, and the inhibitory activity is superior to that of the environmental-friendly botanical pesticide eugenol for preventing and treating the plant pathogenic bacteria in large quantity at the same concentration.
Detailed Description
A Cinnamomum camphora leaf extract is prepared by the following steps:
1) collecting fresh Cinnamomum camphora leaves, oven drying at 50-60 deg.C, pulverizing, and sieving with 20-50 mesh sieve to obtain dried powder;
2) adding 1000g of the powder obtained in the step 1) into 8000mL of dichloromethane 6000-,
3) decompressing and concentrating the extracting solution obtained in the step 2) to obtain the camphor leaf extract.
Research on activity of cinnamomum camphora leaf extract for inhibiting plant pathogenic fungi:
the antibacterial activity of the cinnamomum camphora leaf extract is measured by a hypha production rate method. Specifically, a culture plate is manufactured by using a prepared and sterilized PDA culture medium, and a blank control group, a positive control medicine eugenol group and a camphor leaf extract test group are arranged; the positive control medicine eugenol group and the cinnamomum camphora leaf extract test group are respectively provided with 3 concentrations, and each concentration is provided with 3 repetitions; the method is characterized in that the method comprises the steps of respectively adopting Humicola melanosporum, Fusarium oxysporum and Gliocladium catenulatum to inoculate, and sealing and inverting the inoculated culture dish by using a preservative film so as to prevent water or impurities in the air generated in the growth process of the strain from influencing the normal growth of the strain. Culturing the seeds in an intelligent artificial incubator at the temperature of 28 +/-5 ℃ for 3-7 days, and observing the seeds once every 24 hours. The measurement was carried out when the hypha diameter of the blank plate strain grew to around 2/3. During measurement, the shape of the colonies is observed, then a cross measurement method is adopted, the maximum diameter of the irregular colonies is measured, and the growth diameters of all groups of colonies are measured and recorded respectively. The bacteriostatic rate was calculated according to the formula (the calculation formula is shown below), and the results are shown in tables 1 to 3.
The positive control medicament eugenol group or cinnamomum camphora leaf extract test group has the bacteriostasis rate (the colony diameter of the positive control medicament eugenol group or cinnamomum camphora leaf extract test group-the colony diameter of the blank control group)/(the colony diameter of the blank control group-0.5) multiplied by 100 percent.
TABLE 1 comparison of the bacteriostatic ratio of eugenol and cinnamomum camphora leaf extract to Humicola reticulata
TABLE 2 comparison of the bacteriostatic ratio of eugenol and cinnamomum camphora leaf extract to Fusarium oxysporum f.citrulli
TABLE 3 comparison of the inhibitory rates of eugenol and cinnamomum camphora leaf extract against Gliocladium catenulatum
As can be seen from the above results, the inhibitory activity of the cinnamomum camphora leaf extract on pathogenic bacteria is obviously superior to that of eugenol at the same concentration.
The invention will be better understood from the following examples. However, it will be readily understood by those skilled in the art that the specific material ratios, process conditions and results thereof described in the examples are merely illustrative of the present invention and should not, nor should they be construed as limiting the invention as detailed in the claims.
Example 1
Taking camphor leaves, drying the camphor leaves in a drying oven at 55 ℃, then crushing the camphor leaves by using a crusher, sieving the camphor leaves by using a 30-mesh sieve, and collecting powder for later use; weighing 1000g of powder, adding 8000mL of dichloromethane, heating to boil, extracting under reflux for 50min, and concentrating the extract under reduced pressure to obtain 73.8g of Cinnamomum camphora leaf extract with total yield of 7.38%.
The antibacterial activity of the cinnamomum camphora leaf extract is measured by a hypha production rate method, and the result shows that: the bacteriostasis rates of the extract with the concentration of 200 mu g/mL on the Humicola reticulata, the Fusarium oxysporum pratense and the Gliocladium catenulatum are respectively 91.74 +/-3.39%, 96.99 +/-3.20% and 97.69 +/-3.15%.
Example 2
Taking camphor leaves, drying the camphor leaves in a drying oven at 55 ℃, then crushing the camphor leaves by using a crusher, sieving the camphor leaves by using a 30-mesh sieve, and collecting powder for later use; weighing 1000g of powder, adding 6000mL of dichloromethane, heating to boil, extracting under reflux for 50min, and concentrating the extract under reduced pressure to obtain 46.9g of camphor leaf extract, wherein the total yield is 4.69%.
The antibacterial activity of the cinnamomum camphora leaf extract is measured by a hypha production rate method, and the result shows that: the bacteriostasis rates of the extract with the concentration of 200 mu g/mL on the Humicola reticulata, the Fusarium oxysporum pratense and the Gliocladium catenulatum are respectively 89.79 +/-2.26%, 93.99 +/-4.01% and 94.02 +/-3.97%.
Example 3
Taking camphor leaves, drying the camphor leaves in a drying oven at 55 ℃, then crushing the camphor leaves by using a crusher, sieving the camphor leaves by using a 30-mesh sieve, and collecting powder for later use; weighing 100g of powder, adding 800mL of dichloromethane, heating to boil, extracting under reflux for 50min, and concentrating the extract under reduced pressure to obtain 6.29g of camphor leaf extract, wherein the total yield is 6.29%.
The antibacterial activity of the cinnamomum camphora leaf extract is measured by a hypha production rate method, and the result shows that: the bacteriostasis rates of the extract with the concentration of 200 mu g/mL on the Humicola lanuginosa, the Fusarium oxysporum f.citrulli and the Gliocladium catenulatum are respectively 90.56 +/-4.16%, 92.67 +/-2.46% and 96.21 +/-3.05%.
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
Claims (4)
1. A preparation method of a cinnamomum camphora leaf extract is characterized by comprising the following steps: the method specifically comprises the following steps:
1) collecting fresh Cinnamomum camphora leaves, oven drying at 50-60 deg.C, pulverizing, and sieving with 20-50 mesh sieve to obtain dried powder;
2) adding 1000g of the powder obtained in the step 1) into 8000mL of dichloromethane 6000-;
3) decompressing and concentrating the extracting solution obtained in the step 2) to obtain the camphor leaf extract.
2. The use of the cinnamomum camphora leaf extract obtained by the preparation method according to claim 1 for inhibiting plant pathogenic fungi.
3. The use of the cinnamomum camphora leaf extract according to claim 2, wherein: the plant pathogenic fungi include any one of Humicola reticulata, Fusarium oxysporum and Gliocladium catenulatum.
4. The use of the cinnamomum camphora leaf extract according to claim 3, wherein: 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 the leaf surface until the leaf surface drips; 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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| CN202111238729.9A CN114052048B (en) | 2021-10-25 | 2021-10-25 | Cinnamomum camphora leaf extract and preparation method and application thereof |
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| CN114052048A true CN114052048A (en) | 2022-02-18 |
| CN114052048B CN114052048B (en) | 2023-06-23 |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20050071981A (en) * | 2004-01-05 | 2005-07-08 | 주식회사 내츄로바이오텍 | Antibacterial and antifungal composition containing plant oil |
| CN103461396A (en) * | 2013-08-22 | 2013-12-25 | 上海交通大学 | Compound botanical fungiticide, and preparation method and application thereof |
| CN104146055A (en) * | 2014-06-27 | 2014-11-19 | 广西大学 | Plant source fruit and vegetable preservative |
| CN110150333A (en) * | 2019-05-16 | 2019-08-23 | 井冈山大学 | A kind of oil camphor tree leaf essential oil bacteriostasis method |
-
2021
- 2021-10-25 CN CN202111238729.9A patent/CN114052048B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20050071981A (en) * | 2004-01-05 | 2005-07-08 | 주식회사 내츄로바이오텍 | Antibacterial and antifungal composition containing plant oil |
| CN103461396A (en) * | 2013-08-22 | 2013-12-25 | 上海交通大学 | Compound botanical fungiticide, and preparation method and application thereof |
| CN104146055A (en) * | 2014-06-27 | 2014-11-19 | 广西大学 | Plant source fruit and vegetable preservative |
| CN110150333A (en) * | 2019-05-16 | 2019-08-23 | 井冈山大学 | A kind of oil camphor tree leaf essential oil bacteriostasis method |
Non-Patent Citations (3)
| Title |
|---|
| WEIDONG HUANG ET AL.: "Inhibition of Fusarium oxysporum by AgNPs biosynthesised using Cinnamomum camphora fruit extract", 《IET NANOBIOTECHNOLOGY》 * |
| 解福双等: "樟树枝叶提取物的不同组分的抗病虫害活性研究", 《农业开发与装备》 * |
| 顾丽莎等: "香樟树叶水提物对三七根腐病菌的抑制", 《云南师范大学学报（自然科学版）》 * |
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