CN108184920B - Solanum nigrum extract and application thereof as plant antibacterial - Google Patents
Solanum nigrum extract and application thereof as plant antibacterial Download PDFInfo
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- CN108184920B CN108184920B CN201711324940.6A CN201711324940A CN108184920B CN 108184920 B CN108184920 B CN 108184920B CN 201711324940 A CN201711324940 A CN 201711324940A CN 108184920 B CN108184920 B CN 108184920B
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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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- 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]
- A01N65/38—Solanaceae [Potato family], e.g. nightshade, tomato, tobacco or chilli pepper
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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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- Agronomy & Crop Science (AREA)
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- Natural Medicines & Medicinal Plants (AREA)
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- Medicines Containing Plant Substances (AREA)
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Abstract
The black nightshade extract is an aqueous extract or an alcohol extract, wherein the alcohol extract extraction method 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 12h; ultrasonic extraction is carried out for 2h; 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 black nightshade extract as the plant antibacterial. The solanum nigrum extract used in the invention is a botanical biological pesticide, 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 black nightshade 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.
Solanum nigrum L is Solanum genus (Solanum) of Solanaceae, and is distributed almost everywhere in China. It is fond of living near the edges of the fields, wastelands and villages. Is widely distributed in the temperate zone of Europe, asia and America to tropical regions. The whole plant can be used as a medicine for dissipating blood stasis and detumescence, and clearing away heat and toxic material. 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 a black nightshade extract and application thereof as a plant antibacterial, which can improve the safety of grape food.
In order to solve the technical problems, the invention adopts a technical scheme that:
the black nightshade extract is an aqueous extract or an alcohol extract, and the preparation method of the aqueous extract comprises the following steps: weighing 100g of crushed and dried solanum nigrum, adding 1000mL of distilled water, and soaking for 30min. Boiling with strong fire, decocting with slow fire, and extracting for 40min. Filtering the liquid medicine with 4 layers of gauze while the liquid medicine is hot; extracting for one time repeatedly, mixing filtrates for 2 times, concentrating with slow fire to 1000mg/mL, and storing at 4 deg.C; the method for extracting 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 12h; ultrasonic extraction is carried out for 2h; 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 black nightshade extract has obvious bacteriostatic activity to 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 water extract is 6.25mg/mL-100mg/mL, and the concentration of the alcohol extract is 0.3125mg/mL-5mg/mL.
Further, when the concentration of the aqueous extract is 100mg/mL, the bacteriostatic rates on colletotrichum and white rot are respectively 78.77% and 64.56%; when the concentration of the alcohol extract is 5mg/mL, the inhibition rates of the alcohol extract on colletotrichum, white rot and botrytis are respectively 80.15%, 87.54% and 18.69%, the inhibition rates on multi-bacterium sensitive colletotrichum strains FJZZ-62, coll-9 and coll-20 are respectively 69.44%, 75.92% and 77.08%, the inhibition rates on high-resistance colletotrichum strains FJND-9, FJND-40 and GZSD-89 are respectively 60.83%, 84.37% and 44.12%, the inhibition rates on high-resistance botrytis strain GXGL-7, GXNN-6 and GXNN-7 are respectively 30.38%, 34.70% and 27.36%, and the inhibition rates on high-resistance botrytis strain GXGL-10, GXZLF-4 and GXZY-7 are respectively 18.10%, 29.17% and 29.80%.
The invention has the beneficial effects that: the invention relates to a sterilization medicine taking a black nightshade 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 black nightshade 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 is given with reference to preferred embodiments so that advantages and features of the present invention will be readily understood by those skilled in the art, and the scope of the present invention will be clearly and unequivocally defined.
Example 1:
the black nightshade of the present invention is purchased from Anhui Bozhou medicinal material market. Drying the materials in the sun, pulverizing, and storing in a ventilated and dry place.
Botrytis cinerea (Botrytis cinerea), botrytis viticola (Coniothyrium diplodiella), botrytis cinerea (Glomerella cingulata) and Aspergillus niger (Aspergillus carbonarius) are all provided by the plant protection institute of the Chinese academy of agricultural sciences.
The preparation of the black nightshade water extract comprises weighing 100g of black nightshade crushed dry product, adding 1000mL of distilled water, and soaking for 30min. Boiling with strong fire, decocting with slow fire, and extracting for 40min. The liquid medicine is filtered with 4 layers of gauze while it is hot. Extracting once again, mixing 2 times of filtrates, and concentrating with slow fire to 1000mg/mL. Storing at 4 deg.C for use.
Preparing the alcohol extract, weighing 800g of a crushed sample, soaking plant dry powder in 5 times of ethanol, and standing at room temperature for 12h; ultrasonic extraction is carried out for 2h; vacuum filtering, extracting continuously for 3 times, mixing filtrates, concentrating at 45 deg.C with rotary evaporator, drying, and storing at 4 deg.C.
The bacteriostatic activity of black nightshade water extract and alcohol extract on Botrytis cinerea, botrytis viticola, gluconobacter littoralis, glucocephalum cingula 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 number of colonies with an inoculating needle to the central area of a culture dish containing a PDA culture medium, sealing, and inversely culturing in an incubator at the temperature of 25 ℃ and the humidity of 73% without illumination.
The aqueous extract stock solutions were prepared as solutions of 1000mg/mL, 500mg/mL, 250mg/mL, 125mg/mL, 62.5mg/mL, respectively, with sterile water. Sterile water was used as a blank control and 5. Mu.g/mL carbendazim was used as a positive control. In a sterile clean workbench, the prepared liquid medicine is respectively mixed with PDA culture medium according to the proportion of 1:9, the concentration of the culture medium with the medicine is 100mg/mL, 50mg/mL, 25mg/mL, 12.5mg/mL and 6.25mg/mL, and the mixture is poured into a culture dish with the diameter of 90mm and placed for solidification.
Weighing a certain amount of the 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. In a sterile clean workbench, the prepared liquid medicine is respectively mixed with PDA culture medium according to the proportion of 1:9, the concentrations of the culture medium with the medicine are 5mg/mL, 2.5mg/mL, 1.25mg/mL, 0.625mg/mL and 0.3125mg/mL, and the mixture is poured into a culture dish with the diameter of 90mm and placed for solidification.
Activating strain, uniformly perforating at the position where hyphae grow vigorously by using a sterile perforator (phi =5 mm) to obtain bacterial cakes, inoculating the bacterial cakes onto a culture medium with the drug, inoculating one bacterial cake to the center of each culture dish, performing inverted culture in an incubator at the temperature of 25-28 ℃ and the humidity of 73% for 2-7 days, measuring the diameter of the bacterial colony by using a cross method, and calculating the inhibition rate. Each treatment was set to 3 replicates.
The result shows that the bacteriostatic effect of the water extract on colletotrichum is better than that of white rot pathogen, and the water extract has no inhibiting effect on carbon aspergillus niger and botrytis cinerea. When the concentration of the water extract is 100mg/mL, the bacteriostasis rates of the anthracnose pathogen and the white rot pathogen are respectively 78.77 percent and 64.56 percent.
The alcohol extract shows different effects on 4 grape pathogenic fungi. When the concentration of the alcohol extract is 5mg/mL, the inhibition rates of the alcohol extract on colletotrichum, white rot and gray mold are respectively 80.15%, 87.54% and 18.69%. Has no inhibiting effect on carbon black aspergillosis.
When the concentration of the alcohol extract is 5mg/mL, the inhibition rates of the multi-strain sensitive anthrax bacteria strains FJZZ-62, coll-9 and coll-20 are 69.44%, 75.92% and 77.08%, respectively.
The inhibition rate of carbendazim with the concentration of 100 mu g/mL on the strains FJND-9, FJND-40 and GZSD-89 is 9.80%, 21.50% and 10.01% respectively. The black nightshade alcohol extract has stronger inhibiting effect on carbendazim which has high resistance to anthracnose pathogen strains. When the concentration of the alcohol extract is 5mg/mL, the inhibition rates of the alcohol extract on high-resistance colletotrichum bacteria strains FJND-9, FJND-40 and GZSD-89 are respectively 60.83%, 84.37% and 44.12%, the inhibition effect on FJND-40 is the best, and the alcohol extract is obviously superior to 100 mu g/mL carbendazim. The inhibitory effect of 0.625mg/mL alcohol extract is equivalent to 100 μ g/mL carbendazim.
When the concentration of the alcohol extract is 5mg/mL, the inhibition rates of the botrytis cinerea strains GXGL-7, GXNN-6 and GXNN-7 on the sensitive multiple strains are respectively 30.38%, 34.70% and 27.36%.
The inhibition rate of carbendazim with the concentration of 100 mu g/mL on the strains HBLF-10, GXZY-4 and GXZY-7 is 53.45 percent, 55.11 percent and 14.76 percent respectively. When the concentration of the alcohol extract is 5mg/mL, the inhibition rates of the carbendazim on the botrytis cinerea strains HBLF-10, GXZY-4 and GXZY-7 are respectively 18.10%, 29.17% and 29.80%, and the effect is not as good as that of the 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 black nightshade extract as the plant antibacterial is characterized in that: the black nightshade extract is an aqueous extract, and the preparation method of the aqueous extract comprises the following steps: weighing 100g of pulverized dry Solanum nigrum, adding 1000mL of distilled water, soaking for 30min, boiling with strong fire, decocting with slow fire for 40min, and filtering the medicinal liquid with 4 layers of gauze while it is hot; extracting for one time, mixing the filtrates for 2 times, concentrating with slow fire to 1000mg/mL, and storing at 4 deg.C; the concentration of the water extract is 100mg/mL, the black nightshade water extract has an inhibition effect on the white rot pathogen of grapes, and when the concentration of the water extract is 100mg/mL, the inhibition rate on the white rot pathogen is 64.56%.
2. The application of the black nightshade extract as the plant antibacterial is characterized in that: the black nightshade 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 12h; ultrasonic extraction is carried out for 2h; carrying out vacuum filtration under reduced pressure, continuously extracting for 3 times, combining filtrates, concentrating by using a rotary evaporator at 45 ℃, drying, storing at 4 ℃ for later use, wherein the concentration of the alcohol extract is 5mg/mL, the black nightshade alcohol extract has remarkable bacteriostatic activity on white rot of grapes, and high-resistance anthracnose pathogen strains FJND-9, FJND-40 and GZSD-89, when the concentration of the alcohol extract is 5mg/mL, the inhibition rate on white rot is 87.54%, and the inhibition rates on high-resistance anthracnose pathogen strains FJND-9, FJND-40 and GZSD-89 are respectively 60.83%, 84.37% and 44.12%.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711324940.6A CN108184920B (en) | 2017-12-13 | 2017-12-13 | Solanum nigrum extract and application thereof as plant antibacterial |
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| CN201711324940.6A CN108184920B (en) | 2017-12-13 | 2017-12-13 | Solanum nigrum extract and application thereof as plant antibacterial |
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| CN108184920B true CN108184920B (en) | 2023-01-06 |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106577844A (en) * | 2016-11-15 | 2017-04-26 | 中国农业科学院麻类研究所 | Preparation for prevention and cure of pepper phytophthora blight, and production and application thereof |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106577844A (en) * | 2016-11-15 | 2017-04-26 | 中国农业科学院麻类研究所 | Preparation for prevention and cure of pepper phytophthora blight, and production and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| "56种植物抑菌活性筛选试验";冯俊涛 等;《西北农林科技大学学报(自然科学版)》;20010425;第29卷(第2期);摘要，第2.1小节 * |
| "龙葵水提物对2种常见致病菌的抑制作用研究";赵锦慧 等;《周口师范学院学报》;20130315;第30卷(第2期);第1.2.2小节 * |
| "龙葵等19种野生植物提取物的离体抑菌活性研究";姬妍茹 等;《中国农学通报》;20140905;第30卷(第25期);第1.3.1小节，第3小节 * |
| 姬妍茹 等."龙葵等19种野生植物提取物的离体抑菌活性研究".《中国农学通报》.2014,第30卷(第25期),第1.3.1小节. * |
| 赵锦慧 等."龙葵水提物对2种常见致病菌的抑制作用研究".《周口师范学院学报》.2013,第30卷(第2期),第1.2.2小节. * |
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