CN108142450B - Herba schizonepetae extract and application thereof as plant antibacterial - Google Patents
Herba schizonepetae extract and application thereof as plant antibacterial Download PDFInfo
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- CN108142450B CN108142450B CN201711324937.4A CN201711324937A CN108142450B CN 108142450 B CN108142450 B CN 108142450B CN 201711324937 A CN201711324937 A CN 201711324937A CN 108142450 B CN108142450 B CN 108142450B
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- extract
- alcohol extract
- botrytis cinerea
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- 239000000284 extract Substances 0.000 title claims abstract description 44
- 230000000844 anti-bacterial effect Effects 0.000 title abstract description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 36
- 241000951473 Schizonepeta Species 0.000 claims abstract description 17
- 241000196324 Embryophyta Species 0.000 claims abstract description 16
- 238000001035 drying Methods 0.000 claims abstract description 5
- 238000005303 weighing Methods 0.000 claims abstract description 5
- 238000000605 extraction Methods 0.000 claims abstract description 4
- 239000000706 filtrate Substances 0.000 claims abstract description 4
- 239000000843 powder Substances 0.000 claims abstract description 4
- 238000002791 soaking Methods 0.000 claims abstract description 4
- 238000002137 ultrasound extraction Methods 0.000 claims abstract description 4
- 241000123650 Botrytis cinerea Species 0.000 claims description 17
- 230000005764 inhibitory process Effects 0.000 claims description 17
- 241000222199 Colletotrichum Species 0.000 claims description 10
- 241000228245 Aspergillus niger Species 0.000 claims description 6
- 244000052616 bacterial pathogen Species 0.000 claims description 6
- 239000003814 drug Substances 0.000 claims description 6
- 125000003158 alcohol group Chemical group 0.000 claims description 2
- 241000193738 Bacillus anthracis Species 0.000 claims 1
- 241000894006 Bacteria Species 0.000 claims 1
- 230000001775 anti-pathogenic effect Effects 0.000 claims 1
- 238000003828 vacuum filtration Methods 0.000 claims 1
- 240000000560 Citrus x paradisi Species 0.000 abstract description 4
- 238000002156 mixing Methods 0.000 abstract description 4
- 238000001914 filtration Methods 0.000 abstract description 3
- 235000013305 food Nutrition 0.000 abstract description 3
- 241001465754 Metazoa Species 0.000 abstract description 2
- 239000006286 aqueous extract Substances 0.000 abstract description 2
- 230000000853 biopesticidal effect Effects 0.000 abstract description 2
- 231100000956 nontoxicity Toxicity 0.000 abstract description 2
- 238000001228 spectrum Methods 0.000 abstract description 2
- 241000219095 Vitis Species 0.000 description 12
- 235000009754 Vitis X bourquina Nutrition 0.000 description 12
- 235000012333 Vitis X labruscana Nutrition 0.000 description 12
- 235000014787 Vitis vinifera Nutrition 0.000 description 12
- TWFZGCMQGLPBSX-UHFFFAOYSA-N Carbendazim Natural products C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 description 11
- 239000006013 carbendazim Substances 0.000 description 11
- JNPZQRQPIHJYNM-UHFFFAOYSA-N carbendazim Chemical compound C1=C[CH]C2=NC(NC(=O)OC)=NC2=C1 JNPZQRQPIHJYNM-UHFFFAOYSA-N 0.000 description 11
- 238000000034 method Methods 0.000 description 9
- 201000010099 disease Diseases 0.000 description 7
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 7
- 230000002401 inhibitory effect Effects 0.000 description 7
- 239000000575 pesticide Substances 0.000 description 7
- 239000001963 growth medium Substances 0.000 description 5
- 241001529387 Colletotrichum gloeosporioides Species 0.000 description 4
- 230000002265 prevention Effects 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 241000233866 Fungi Species 0.000 description 3
- 241000238631 Hexapoda Species 0.000 description 3
- 241000607479 Yersinia pestis Species 0.000 description 3
- 230000003385 bacteriostatic effect Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 241000207923 Lamiaceae Species 0.000 description 2
- 241001529733 Nepeta Species 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 244000052769 pathogen Species 0.000 description 2
- 230000001717 pathogenic effect Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000125121 Aspergillus carbonarius Species 0.000 description 1
- 241001465180 Botrytis Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 208000012895 Gastric disease Diseases 0.000 description 1
- 231100000678 Mycotoxin Toxicity 0.000 description 1
- 240000009215 Nepeta cataria Species 0.000 description 1
- 235000010679 Nepeta cataria Nutrition 0.000 description 1
- 241000219094 Vitaceae Species 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 244000053095 fungal pathogen Species 0.000 description 1
- 235000021021 grapes Nutrition 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 150000002611 lead compounds Chemical class 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000005065 mining Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002636 mycotoxin Substances 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 238000010298 pulverizing process Methods 0.000 description 1
- 238000007711 solidification Methods 0.000 description 1
- 230000008023 solidification Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 208000018556 stomach disease Diseases 0.000 description 1
Classifications
-
- 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]
- A01N65/22—Lamiaceae or Labiatae [Mint family], e.g. thyme, rosemary, skullcap, selfheal, lavender, perilla, pennyroyal, peppermint or spearmint
Landscapes
- 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)
- Medicines Containing Plant Substances (AREA)
- Food Preservation Except Freezing, Refrigeration, And Drying (AREA)
Abstract
The invention relates to a schizonepeta extract, which is an aqueous extract or an alcohol extract, wherein the extraction method of the alcohol extract comprises the following steps: weighing 800g of a crushed sample, soaking plant dry powder in 5 times of ethanol, and standing at room temperature for 12 h; ultrasonic extraction is carried out for 2 h; vacuum filtering, continuously extracting for 3 times, mixing filtrates, concentrating at 45 deg.C with rotary evaporator, drying, and storing at 4 deg.C; also includes the application of the schizonepeta extract as a plant antibacterial. The schizonepeta extract used by the invention is a botanical biopesticide, has the characteristics of high antibacterial activity, wide antibacterial spectrum, difficult generation of resistance and no toxicity to human and animals, and can improve the food safety of grape fruits.
Description
Technical Field
The invention relates to the field of biological pesticides, in particular to a botanical biological pesticide, and more particularly relates to a schizonepeta extract and application thereof as a plant antibacterial.
Background
The grape diseases and insect pests are the same as other horticultural crops, the serious damage of the diseases and insect pests is the limiting factor of the healthy development of the industry, the common grape diseases with serious damage such as grape gray mold, white rot, anthracnose and the like seriously affect the quality and the yield of the grape, and are the difficulties which must be solved in the grape production in China and the worldwide grape production. In agricultural production, a common prevention and control method is a chemical prevention and control method using chemical pesticides, but chemical prevention and control has the risks of causing drug resistance, residual pollution, rampant and the like of grape pathogenic bacteria. The alternative matching of various methods for comprehensive prevention and control is a development direction for preventing and controlling plant diseases and insect pests and preventing mycotoxin pollution and a way for avoiding risks. Therefore, the search for other ways and methods for non-chemical pesticides is an important task for the development of industry and an important subject of scientific research. The method for mining plant sources from plants, including developing plant source pesticides, searching lead compounds from plant source compounds and even directly utilizing plant leaching liquor, is an important content in the development direction.
Herba Schizonepetae (Nepeta cataria L.) is a plant of Nepeta of Labiatae (Labiatae) genus Nepeta, and is produced in Xinjiang, Gansu, Shanxi, Henan, Shanxi, Shandong, Hubei, Guizhou, Sichuan and Yunnan provinces; most of them live at the house or in the bush, and the altitude is generally not more than 2500 m. It is used for preventing and treating gastropathy, anemia and other diseases in various parts of Europe. The roots of schizonepeta contain ingredients that strongly stimulate the nervous system. The whole herb is used for preventing and treating cold, which is sometimes cultivated and used in China. But reports and researches on applying the compound as a biopesticide to control grape diseases are not found.
Disclosure of Invention
The invention mainly solves the technical problem of providing the schizonepeta extract and the application thereof as a plant antibacterial, and can improve the food safety of grapes.
In order to solve the technical problems, the invention adopts a technical scheme that:
the schizonepeta extract is an alcohol extract, and the extraction method of the alcohol extract comprises the following steps: weighing 800g of a crushed sample, soaking plant dry powder in 5 times of ethanol, and standing at room temperature for 12 h; ultrasonic extraction is carried out for 2 h; vacuum filtering, continuously extracting for 3 times, mixing filtrates, concentrating at 45 deg.C with rotary evaporator, drying, and storing at 4 deg.C.
The schizonepeta extract has obvious bacteriostatic activity on pathogenic bacteria of following diseases, mainly including botrytis cinerea, botrytis cinerea and aspergillus niger.
Further, the colletotrichum gloeosporioides comprises a highly-resistant colletotrichum gloeosporioides strain to carbendazim, and the botrytis cinerea comprises a highly-resistant botrytis cinerea strain to carbendazim.
Further, the concentration of the alcohol extract is 0.3125mg/mL-5 mg/mL.
Furthermore, when the concentration of the alcohol extract is 5mg/mL, the inhibition rates of the alcohol extract on white rot pathogen, colletotrichum, botrytis cinerea and aspergillus niger chartarum are 94.57%, 70.23%, 71.15% and 20.06%, respectively, the inhibition rates on sensitive colletotrichum strain FJZZ-62, coll-9 and coll-20 are 62.96%, 68.27% and 56.25%, the inhibition rates on high-resistance colletotrichum strain FJND-9, FJND-40 and GZSD-89 are 64.10%, 68.73% and 72.94%, respectively, the inhibition rates on sensitive grape botrytis cinerea strains GXGL-7, GXNN-6 and GXNN-7 are 60.50%, 62.77% and 63.18%, and the inhibition rates on high-resistance grape botrytis cinerea strains HBLF-10, GXZY-4 and GXZY-7 are 57.47%, 66.83% and 64.90%, respectively.
The invention has the beneficial effects that: the invention relates to a sterilizing medicine with schizonepeta extract as an active ingredient, which is a botanical biological pesticide, belongs to one of biological medicines, and meets the requirements of modern society on new pesticides. Secondly, the schizonepeta extract has the characteristics of high antibacterial activity, wide antibacterial spectrum, difficult generation of resistance and no toxicity to human and animals. In addition, the grape fruit preservative has an inhibiting effect on common pathogenic bacteria of grape fruits, and can improve the food safety of the grape fruits.
Detailed Description
The following detailed description of the preferred embodiments is provided to enable those skilled in the art to more readily understand the advantages and features of the present invention, and to clearly and unequivocally define the scope of the invention.
Example 1:
firstly, preparation of alcohol extract
The schizonepeta of the invention is purchased from the Anhui Bozhou medicinal material market. Drying the materials in the sun, pulverizing, and storing in a ventilated and dry place. The extraction method of the schizonepeta alcohol extract comprises the following steps: weighing 800g of a crushed sample, soaking the plant dry powder in 5 times of ethanol, and standing at room temperature for 12 h; ultrasonic extraction is carried out for 2 h; vacuum filtering, continuously extracting for 3 times, mixing filtrates, concentrating at 45 deg.C with rotary evaporator, drying, and storing at 4 deg.C.
Secondly, bacteriostasis of alcohol extract
The bacteriostatic activity of aqueous extract and alcohol extract of herba Schizonepetae on Botrytis cinerea, Botrytis viticola, Colletotrichum viticola and Aspergillus carbonarius is determined by hypha growth rate method. The method comprises the following specific steps:
recovering the frozen strains at room temperature, picking a small amount of colonies by using an inoculating needle to a PDA-containing culture medium, and performing inverted culture in an incubator at the temperature of 25 ℃ and the humidity of 73%;
then weighing a certain amount of extract, and preparing the alcohol extract into solutions of 50mg/mL, 25mg/mL, 12.5mg/mL, 6.25mg/mL and 3.125mg/mL respectively by using sterile water. Sterile water was used as a blank control and 5. mu.g/mL carbendazim was used as a positive control. Mixing the prepared liquid medicine with PDA culture medium at a ratio of 1:9 in a sterile clean workbench to obtain culture medium with medicine, pouring the culture medium with medicine at concentrations of 5mg/mL, 2.5mg/mL, 1.25mg/mL, 0.625mg/mL and 0.3125mg/mL into a culture dish with diameter of 90mm, and standing for solidification.
Activating strain, uniformly perforating at the position where hyphae grow vigorously by using a sterile perforator (phi is 5mm), preparing fungus cakes, inoculating the fungus cakes onto a culture medium with the strain, inoculating one fungus cake to the center of each culture dish, carrying out inverted culture in an incubator at the temperature of 25-28 ℃ and the humidity of 73% for 2-7d, measuring the diameter of the colony by using a cross method, calculating the inhibition rate, and setting 3 times for each treatment.
The results show that: the alcohol extract shows different degrees of bacteriostatic effects on 4 grape pathogenic fungi, and when the concentration of the alcohol extract is 5mg/mL, the inhibition rates on white rot pathogen, colletotrichum gloeosporioides, botrytis cinerea and aspergillus niger chartarkii are 94.57%, 70.23%, 71.15% and 20.06% respectively. The inhibitor has the best inhibiting effect on white rot germs and the worst inhibiting effect on carbon aspergillus niger germs.
Third, the inhibiting effect of the alcohol extract on multi-bacterium sensitive colletotrichum gloeosporioides strains
When the concentration of the schizonepeta alcohol extract is 5mg/mL, the inhibition rates on sensitive colletotrichum strain FJZZ-62, coll-9 and coll-20 are respectively 62.96%, 68.27% and 56.25%.
Fourthly, the inhibiting effect of the alcohol extract on carbendazim strain with high resistance to colletotrichum
The inhibition rates of the carbendazim with the concentration of 100 mu g/mL on the high-antibacterial strains FJND-9, FJND-40 and GZSD-89 are respectively 9.80%, 21.50% and 10.01%. The schizonepeta alcohol extract has a strong inhibiting effect on carbendazim strains with high resistance to colletotrichum. When the concentration of the alcohol extract is 5mg/mL, the inhibition rates of the alcohol extract on high-resistance colletotrichum anthracnose bacterial strains FJND-9, FJND-40 and GZSD-89 are 64.10%, 68.73% and 72.94% respectively, and the inhibition rates are obviously superior to 100 mu g/mL carbendazim. The effect of 0.625mg/mL schizonepeta alcohol extract is equivalent to 100 mug/mL carbendazim.
Fifthly, inhibiting effect of alcohol extract on multiple sensitive botrytis cinerea strains
When the concentration of the alcohol extract is 5mg/mL, the inhibition rates of the sensitive grape botrytis cinerea strains GXGL-7, GXNN-6 and GXNN-7 are 60.50%, 62.77% and 63.18% respectively.
Sixthly, the inhibition effect of the alcohol extract on the highly anti-botrytis cinerea bacterial strain of carbendazim
The inhibition rate of 100 mu g/mL carbendazim on the strains HBLF-10, GXZY-4 and GXZY-7 is 53.45%, 55.11% and 14.76%, respectively. When the concentration of the alcohol extract is 5mg/mL, the inhibition rates of the alcohol extract on the high-resistance botrytis cinerea strains HBLF-10, GXZY-4 and GXZY-7 are 57.47%, 66.83% and 64.90% respectively, and the inhibition effect is more than 100 mu g/mL carbendazim.
The above description is only an embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by the present specification, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (2)
1. The application of the schizonepeta extract as a plant anti-pathogenic bacteria medicament is characterized in that: the schizonepeta extract is an alcohol extract, and the extraction method of the alcohol extract comprises the following steps: weighing 800g of a crushed sample, soaking the plant dry powder in 5 times of ethanol, and standing at room temperature for 12 h; ultrasonic extraction is carried out for 2 h; carrying out vacuum filtration under reduced pressure, continuously extracting for 3 times, combining filtrates, concentrating at 45 ℃ by using a rotary evaporator, drying, and storing at 4 ℃ for later use, wherein pathogenic bacteria are botrytis cinerea, botrytis cinerea and aspergillus niger chartarum, and the concentration of the alcohol extract is 0.3125mg/mL-5 mg/mL.
2. The use according to claim 1, wherein the inhibition rates of the alcohol extract at a concentration of 5mg/mL are 94.57%, 70.23%, 71.15% and 20.06% for white rot, colletotrichum, botrytis cinerea and aspergillus niger cinerea respectively, 62.96%, 68.27% and 56.25% for sensitive colletotrichum strains FJZZ-62, coll-9 and coll-20 respectively, 64.10%, 68.73% and 72.94% for highly anthrax resistant strains FJND-9, jnd-40 and GZSD-89 respectively, 60.50%, 62.77% and 63.18% for sensitive botrytis cinerea strains GXGL-7, GXNN-6 and GXNN-7 respectively, and 57.47%, 66.83% and 64.90% for highly resistant botrytis cinerea strains gxzlf-10, gxzlf-4 and GXZY-7 respectively.
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| CN201711324937.4A CN108142450B (en) | 2017-12-13 | 2017-12-13 | Herba schizonepetae extract and application thereof as plant antibacterial |
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| CN201711324937.4A CN108142450B (en) | 2017-12-13 | 2017-12-13 | Herba schizonepetae extract and application thereof as plant antibacterial |
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| CN108142450B true CN108142450B (en) | 2020-09-25 |
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| CN201711324937.4A Expired - Fee Related CN108142450B (en) | 2017-12-13 | 2017-12-13 | Herba schizonepetae extract and application thereof as plant antibacterial |
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| CN108850002A (en) * | 2018-06-28 | 2018-11-23 | 兰州世创生物科技有限公司 | It is a kind of for preventing and treating the botanical pesticide preparation of crops gray mold |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009235049A (en) * | 2008-03-25 | 2009-10-15 | Yanagi Tomotama | Composition for controlling plant pest |
| CN103734204A (en) * | 2013-12-27 | 2014-04-23 | 成都新朝阳作物科学有限公司 | Pesticide composition containing schizonepeta and tobacco extracts as well as preparation method and application of pesticide composition |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2009235049A (en) * | 2008-03-25 | 2009-10-15 | Yanagi Tomotama | Composition for controlling plant pest |
| CN103734204A (en) * | 2013-12-27 | 2014-04-23 | 成都新朝阳作物科学有限公司 | Pesticide composition containing schizonepeta and tobacco extracts as well as preparation method and application of pesticide composition |
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