CN118696960A - Use of paulownia flower extract or composition containing the same in bacteriostasis - Google Patents
Use of paulownia flower extract or composition containing the same in bacteriostasis Download PDFInfo
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- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention belongs to the technical field of prevention and control of plant diseases and air harmful bacteria, in particular to application of paulownia flower extract or a composition containing the paulownia flower extract in bacteriostasis, which is proved by bacteriostasis tests, the essential oil extracted from paulownia flower has obvious antibacterial activity and has obvious inhibition effect on the anthracnose of white wax tree, fusarium solani, aromatic fusarium, fusarium graminearum, cladosporium and aspergillus flavus.
Description
Technical Field
The invention belongs to the technical field of prevention and control of plant diseases and air harmful bacteria, and particularly relates to application of a paulownia flower extract or a composition containing the paulownia flower extract in bacteriostasis.
Background
Paulownia flowers are flowers of paulownia (Paulowniafortunei (Seem.) Hemsl.) and paulownia (P.tomotosa (Thunb.) Steud.) belonging to the Scrophulariaceae, are known as "spring users", are one of traditional Chinese medicinal materials in China, and have extremely high medicinal value and ornamental value. The outer surface of the paulownia flower corolla is light purple, the inner surface is white, and purple stripes are formed. The flower and fruit period is 4-9 months. The paulownia flower medicinal materials are crushed by shrinkage, and only the corolla and the calyx are rare; weak smell and light taste, dry body, complete flowers and no impurity. Paulownia flower has bitter taste and cold property, and has the effects of clearing lung-heat, relieving sore throat, removing toxin and detumescence. It is used for treating cough due to lung heat, acute tonsillitis, bacillary dysentery, acute enteritis, acute conjunctivitis, parotitis, furuncle, and tinea. Modern pharmacological researches have shown that paulownia flower also has antibacterial and antiviral, antitussive, expectorant, antiasthmatic, anticancer, blood pressure lowering and insecticidal effects.
Researches show that the paulownia flower extract has antibacterial effects on staphylococcus aureus, tetranecticoccus, escherichia coli and other bacteria, aspergillus niger, penicillium citrinum, rhizopus nigricans and other fungi. Wei Xiying and the like, which are subjected to in-vitro bacteriostasis tests on different extraction parts of paulownia flower extract by adopting a filter paper sheet method, show that the different extraction parts have different degrees of inhibition on staphylococcus aureus, escherichia coli and bacillus subtilis, wherein the inhibition on staphylococcus aureus is strongest, and the inhibition on aspergillus niger, beer yeast and penicillium chrysogenum is not obvious. Studies in Qianjin and the like show that the paulownia flower fat-soluble component has a strong inhibition effect on staphylococcus aureus, and the water-soluble component has a strong inhibition effect on escherichia coli.
The invention aims to extract plant essential oil from paulownia flower petals by collecting the paulownia flower petals and observe the broader antibacterial effect.
Disclosure of Invention
Based on the technical purposes, the invention provides an application of paulownia flower extract or a composition containing the paulownia flower extract in bacteriostasis. Antibacterial tests prove that the paulownia flower essential oil extract has obvious antibacterial effects on the anthracnose of the ash tree, fusarium, aromatic fusarium, fusarium graminearum and aspergillus flavus.
The technical scheme provided by the invention is as follows:
in a first aspect, the invention provides an application of paulownia flower extract or a composition containing the same in bacteriostasis, wherein the paulownia flower extract is extracted according to the following steps: extracting paulownia flower by a steam distillation method, and collecting essential oil, namely the paulownia flower extract;
The paulownia flower extract is used for inhibiting white wax tree anthracnose, eggplant disease fusarium, aromatic fusarium, fusarium layering fusarium, cladosporium, aspergillus flavus, blue fungus or mucor.
Preferably, the ratio of paulownia flower to water is 1g: 20-30 mL.
Preferably, the paulownia flower extract or the composition containing the same is used for preparing pesticides for preventing and treating plant diseases.
Further preferably, the paulownia flower extract or a composition containing the same is used for controlling plant anthracnose or plant root rot.
Preferably, the paulownia flower extract or the composition containing the same is used for preparing a preservative, and the preservative is used for preventing the spoilage of materials such as grains, foods, medicinal materials or feeds.
Preferably, the paulownia flower extract or a composition containing the same is used for preparing an air freshener for killing air harmful fungi.
The anthracnose of the white wax tree mainly causes plant anthracnose, and fusarium solani, fusarium aromaticum and fusarium layering mainly cause plant root rot. The cladosporium, aspergillus flavus, blue-like fungus and mucor are common air fungi, and mainly cause putrefaction of materials such as grains, foods, medicinal materials, feeds and the like. The invention verifies that the paulownia flower extract has inhibition effect on pathogenic bacteria or air fungi, so the paulownia flower extract can be used for preparing products for inhibiting the pathogenic bacteria, and the product forms can be classified into pesticides or preservatives, antistaling agents, air freshening agents and the like according to different strains.
Preferably, the composition is a 100-2000 fold dilution of paulownia flower extract.
Preferably, the composition comprises an acceptable adjuvant or carrier. The excipients are specifically selected according to the dosage form from which the composition is ultimately prepared. Exemplary agropharmaceutically preparable dosage forms are emulsions, microemulsions, suspensions, granules, powders, and the like. Exemplary, additional adjuvants are diluents, suspending agents, osmotic adjusting agents, colorants, pH adjusting agents, stabilizers, preservatives, and the like.
Compared with the prior art, the invention has the beneficial effects that:
The invention provides an application of paulownia flower extract in bacteriostasis. Antibacterial tests prove that the essential oil extracted from paulownia flowers has remarkable antibacterial activity, has obvious inhibition effect on anthracnose of white wax trees, fusarium solani, aromatic fusarium, fusarium layering, cladosporium pullulans and aspergillus flavus, and has small inhibition effect on mucor and blue-like bacteria.
According to the principle of the DPPH free radical scavenging method, positive control data are used as control standards, so that DPPH free radical scavenging data of the water extract remained after extracting and collecting essential oil of paulownia flowers are obtained, and the water extract has a certain DPPH free radical scavenging effect although the DPPH free radical scavenging data are extremely lower than DPPH free radical scavenging data of the positive control group, so that the paulownia flowers are verified to have oxidation resistance.
Drawings
FIG. 1 is a colony diameter of Fusarium solani (Fusarium solani) under the action of paulownia flower essential oil;
FIG. 2 shows colony diameters of Fusarium (Fusariumproliferatum) under the action of paulownia flower essential oil;
FIG. 3 is the colony diameter of Fusarium aromaticum (Fusarium redolens) under the action of paulownia flower essential oil;
FIG. 4 is colony diameter of the ash tree anthrax (Colletotrichum spaethianum) under the action of paulownia flower essential oil;
FIG. 5 is a colony diameter of Acremonium pullulans (Cladosporium cladosporioides) under the action of paulownia flower essential oil;
FIG. 6 is a colony diameter of Aspergillus flavus (Aspergillusflavus) under the action of paulownia flower essential oil;
FIG. 7 is a colony diameter of blue fungus (Talaromycespurpureogenus) under the action of paulownia flower essential oil;
FIG. 8 is a colony diameter of Mucor sp under the influence of paulownia flower essential oil;
FIG. 9 is the effect of paulownia flower aqueous extract on DPPH radical scavenging rate.
Detailed Description
The invention will be further illustrated with reference to specific examples, which include but are not limited to.
1. Experimental materials
Petals of Taraxacum taishanense are collected from a teacher apartment of a college of Taraxacum taishanense the Eastern Mountain in Mount Tai of Taian City of Shandong in 2024, and are paved flat and dried in the shade for later use.
Plant pathogenic bacteria: fusarium solani (Fusarium solani), fusarium aromaticum (Fusarium redolens), fusarium layering (Fusariumproliferatum), and fraxinus chinensis anthrax (Colletotrichum spaethianum); air fungus: mucor sp., acremonium pullulans Cladosporium cladosporioides, aspergillus flavus Aspergillusflavus and Penicillium Talaromycespurpureogenus. All are stored in the Taishan academy of Chinese medicine quality detection laboratory.
2. Experimental reagent and instrument
The reagents used in the experiments are shown in Table 1.
TABLE 1 test agents
The apparatus used for the experiment is shown in table 2.
Table 2 test instrument
3. Experimental method
3.1 Activation of pathogenic bacteria
Under the aseptic operation of an ultra-clean workbench, inoculating the target strain to a culture dish, and inversely culturing for 5-7 d at 25 ℃ for later use.
3.2 Preparation of Paulownia flower essential oil and Water extract
Collecting petals of paulownia flower, separating sepals, preparing two volumetric flasks of 500ml and 1000ml respectively, and mixing the petals with distilled water 1:15 proportion, extracting essential oil by steam distillation, adjusting the temperature to 260 ℃, cutting off the power supply of the electric heating sleeve after the liquid in the round bottom flask boils for 2 hours, cleaning and repeating the steam distillation step. Purifying the collected essential oil again, putting 400ml of the collected liquid containing the paulownia flower essential oil into a 500ml volumetric flask for purification each time, and concentrating the finally collected essential oil into a small bottle for marking for later use.
Pouring the solution in the round bottom flask after the extraction into a beaker to obtain an aqueous extract to be detected, cooling the aqueous extract, filling the aqueous extract into a sealing bag for sealing, and marking for later use.
3.3 Preparation of essential oil-containing Medium
In an ultra-clean workbench, absorbing 0.5ml of paulownia flower essential oil by a liquid-transferring gun, adding tween 20 with mass fraction of 0.5% for emulsification, and diluting to concentration gradients of two mother solutions of 100x and 200 x.
Cooling sterilized PDA culture medium to about 50deg.C, adding diluted gradient essential oil mother liquor, and mixing thoroughly to dilute the essential oil concentration to 1000x and 2000x. Pouring the diluted culture medium into a flat plate, and cooling for standby.
3.4 Inoculation and measurement of colonies
Under the aseptic operation of the ultra-clean workbench, the target strain is uniformly perforated along the edge of the flora by a puncher, and the bacterial slices are picked by an inoculating needle and inoculated to the center of the culture dish. Adding 2 drops of 0.5% Tween reagent for emulsifying the strains with more spores, sucking 50 mu L (or 1-2 drops) of bacterial liquid from the dissolved tube, dripping the bacterial liquid into the center of the solid designated culture medium, and inoculating the bacterial liquid into the center of the culture medium. Each gradient was repeated 3 times.
Placing the culture dish with the inoculated bacteria in an incubator to be inversely cultured for 5-7 d, observing the colony on the culture dish, and measuring the diameter of the colony by a ruler. And filling the measured diameter data into a table, and calculating statistical data such as average diameter, standard deviation and the like.
Antibacterial ratio (%) = [ control colony average diameter-6 mm) - (treated colony average diameter-6 mm) ]/(control colony average diameter-6 mm) ×100%
3.5 Measurement of antioxidant Activity
DPPH solution is prepared by using 95% ethanol until the final concentration of DPPH is 0.1mmol/L, and the solution is preserved in dark place. Preparation of VC solution: 2.2g of ascorbic acid powder is weighed into a beaker, added with a proper amount of distilled water for dissolution, and poured into a 250ml volumetric flask for constant volume.
The method comprises the steps of taking an aqueous extract of petals of paulownia flower as a liquid to be detected, referring to the documents Lv Tingting, tao Juan and Xie Ji, and the like, performing enzymolysis-ultrasonic assisted double water phase method to extract total flavonoids of the paulownia flower, optimizing the process, researching in vitro antioxidant capacity [ J/OL ]. Feed industry, and measuring absorbance at 517nm by a method of 1-14[2024-05-26 ]; distilled water is used for replacing the liquid to be tested, as a blank control; VC was used as a positive control instead of the test solution. A 1 is water extract of paulownia flower petal and DPPH ethanol (1:1 mixture), and the absorbance at 517nm is measured in dark place for 30 min. A 2 is Paulownia flower petal water extract and ethanol (1:1 mixture), and is protected from light for 30min, and absorbance at 517nm is measured. A 0 is ethanol+DPPH ethanol (1:1 mixture), and the absorbance at 517m is measured. Calculation of DPPH radical clearance (%):
DPPH radical scavenging rate (D) = (a 0-(A1-A2))/A0 x 100%
Wherein: a 0 -absorbance of 1ml ethanol+ mlDPPH solution;
a 1 -absorbance of 1ml Paulownia flower petal aqueous extract + mlDPPH;
A 2 -absorbance of 1ml Paulownia flower petal aqueous extract+1 ml ethanol.
4. Data analysis
EC50 (EFFECTIVE CONCENTRATION) 50): i.e. half the inhibitory concentration, also called half maximal effect concentration, EC50 is often used as a drug safety indicator. It means that the concentration of the drug has a certain obvious effect on 50% of individuals of the tested strain. The term "test" refers to the concentration of essential oil that causes 50% of individuals of the test species to be inhibited.
Data conditioning and analysis were performed using Microsoft Excel 2019, SPSS 21.0.
5. Results and analysis
5.1 Effect of paulownia flower essential oil on plant pathogen
After 4d inoculation, the average diameter of the bacterial colonies of fusarium solani in PDA culture medium reaches 4.87cm, and the bacterial colony diameters in 1000 times and 2000 times of paulownia flower essential oil dilutions are slightly smaller, namely 4.45cm and 4.6cm respectively. As can be seen from FIG. 1, the colony growth of Fusarium solani in PDA medium was significantly higher than in 2000 times of paulownia flower essential oil dilution, and the colony growth in PDA medium was significantly higher than in 1000 times of paulownia flower essential oil dilution. Therefore, the essential oil extracted from the paulownia flowers has obvious inhibition effect on fusarium solani.
After 4d inoculation, the average diameter of the colonies of Fusarium in PDA culture medium reached 4.27cm, and the diameters of the colonies in 1000 times and 2000 times of paulownia flower essential oil dilutions were slightly smaller than 2.15cm and 2.4cm respectively. As can be seen from FIG. 2, the colony growth of Fusarium on PDA medium was significantly higher than on 1000-fold and 2000-fold dilutions of paulownia flower essential oil. From the above, the paulownia flower essential oil has obvious inhibition effect on fusarium layering.
After 4d inoculation, the average diameter of the colonies of Fusarium aromaticum in PDA medium reached 4.0cm, and the diameters of the colonies in 1000 times and 2000 times of paulownia flower essential oil dilutions were slightly smaller, respectively 3.07cm and 3.33cm. As can be seen from FIG. 3, the colony growth of Fusarium aromaticum in PDA medium was significantly higher than in 1000-fold and 2000-fold dilutions of paulownia flower essential oil. From the results, the paulownia flower essential oil has obvious inhibition effect on fusarium aromaticum. According to the data analysis of the three fusarium species, the plant essential oil extracted from the paulownia flowers has obvious inhibition effect on the fusarium species.
After 4d inoculation, the average diameter of bacterial colonies of the white wax tree anthrax in the PDA culture medium reaches 4.47cm, the bacterial colony diameter in the 2000 times of paulownia flower essential oil diluent is slightly smaller, and the bacterial colony diameter in the 1000 times of paulownia flower essential oil diluent is respectively 2.27cm and 3.8cm. As can be seen from FIG. 4, the colony growth of the anthrax of Fraxinus mandshurica in PDA medium is significantly higher than that in 2000 times of paulownia flower essential oil dilution, and at the same time, there is no significant difference from that in 1000 times of paulownia flower essential oil dilution. From this, it is clear that the paulownia flower essential oil does not show a significant inhibition effect on the colletotrichum gloeosporioides.
5.2 Effect of paulownia flower essential oil on air fungi
After 4d inoculation, the average colony diameter of the dendritic branch spore bacteria in the PDA culture medium reaches 2.47cm, and the colony diameters in the 1000-time and 2000-time paulownia flower essential oil dilutions are slightly smaller than 2.05cm and 2.13cm respectively. As can be seen from FIG. 5, the colony growth of the dendritic fungus in PDA medium was significantly higher than in 1000-fold and 2000-fold paulownia flower essential oil dilutions, and the colony growth in 1000-fold paulownia flower essential oil dilutions was not significantly different from the colony growth in 2000-fold paulownia flower essential oil dilutions. From the results, the paulownia flower essential oil has obvious inhibition effect on the amycolatopsis.
After 4d inoculation, the average diameter of the colonies of the Aspergillus flavus in the PDA medium reached 4.8cm, and the diameters of the colonies in the 1000-fold and 2000-fold dilutions of paulownia flower essential oil were slightly smaller, respectively 3.67cm and 4.0cm. As can be seen from FIG. 6, the colony growth of Aspergillus in PDA medium was significantly higher than in 1000-fold and 2000-fold dilutions of paulownia flower essential oil. From the results, the paulownia flower essential oil has obvious inhibition effect on aspergillus flavus.
After 4d inoculation, the average diameter of the bacterial colonies of the blue fungus in the PDA culture medium reaches 2.47cm, and the diameters of the bacterial colonies in the 1000-time and 2000-time paulownia flower essential oil dilutions are similar and are respectively 2.35cm and 2.55cm. As can be seen from FIG. 7, there was no significant difference between the colony growth of the blue fungus in PDA medium and the colony growth in 1000-fold and 2000-fold dilutions of paulownia flower essential oil. And, the colony diameter of blue fungus in 2000 times paulownia flower essential oil dilution was slightly higher than that in PDA medium. From this, it is clear that the paulownia flower essential oil does not exhibit a significant inhibitory effect on the cyanobacteria.
After 4d inoculation, the average diameter of the bacterial colony of the mucor in the PDA culture medium is 7.8cm, and the diameter of the bacterial colony in the 1000 times of paulownia flower essential oil diluent is slightly larger than that of the bacterial colony in the 2000 times of paulownia flower essential oil diluent, and the diameters of the bacterial colony in the PDA culture medium are respectively 8.0cm and 7.8cm. As can be seen from FIG. 8, the colony growth of Mucor in PDA medium was not significantly different from that in 1000 times of paulownia flower essential oil dilution, and the colony growth in 1000 times of paulownia flower essential oil dilution was higher than that in PDA medium. From this, it is clear that the paulownia flower essential oil does not show a significant inhibition effect on Mucor.
5.3 Inhibiting effect of paulownia flower essential oil on pathogenic bacteria
According to the obtained EC 50 value, the arrangement sequence of the inhibition effect of the paulownia flower essential oil on five pathogenic bacteria from excellent to poor is as follows: fusarium roseum > aspergillus flavus > fusarium bud branch spore bacteria > fusarium solani, and the corresponding EC 50 values are 0.000853413mL/L (dilution 1172 times), 0.001750027mL/L (dilution 571 times), 0.001784391mL/L (dilution 560 times), 0.002140137mL/L (dilution 467 times) and 0.005072909mL/L (dilution 197 times) respectively, namely, the inhibition effect on fusarium roseum is relatively best, and the inhibition effect on fusarium solani is relatively worst.
TABLE 3 EC 50 values of plant pathogens under paulownia flower essential oils
5.4 Effect of Paulownia flower aqueous extract on DPPH radical scavenger
As can be seen from fig. 9, the DPPH radical scavenging rate of the positive control group was significantly higher than that of the paulownia flower aqueous extract experimental group, but the paulownia flower aqueous extract experimental group still showed scavenging effect on DPPH radicals. From this, it is clear that paulownia flower has oxidation resistance.
At present, the research on the antibacterial effect of paulownia flowers mainly focuses on various paulownia flower extracts. In the observation and understanding process of the paulownia flower, the petals of the paulownia flower can be determined to have a certain fragrance, the relevant plant essential oil extraction literature is consulted, the plant essential oil is tried to be extracted from the petals of the paulownia flower in a milder mode, the antibacterial effect of the essential oil is observed, and a thought is provided for the subsequent research direction of the paulownia flower. Meanwhile, in order to ensure that the collected paulownia flowers are not excessively different from the previous experimental materials, the DPPH free radical elimination method is utilized to test the water extract of the paulownia flowers, and the oxidation resistance of the paulownia flowers is determined again.
The study utilizes a steam distillation method to extract and collect paulownia flower essential oil, and 4 pathogenic bacteria and 4 air fungi are respectively inoculated to observe the antibacterial effect of the paulownia flower. Experimental data show that the essential oil extracted from paulownia flowers has remarkable antibacterial activity, has remarkable inhibition effect on anthracnose of white wax tree, fusarium solani, aromatic fusarium, fusarium graminearum, cladosporium pullulans and aspergillus flavus, and has a certain inhibition effect on mucor and blue-like bacteria but has an insignificant effect. The characteristic makes the paulownia flower essential oil a novel medicine resource. Meanwhile, according to the principle of a DPPH free radical scavenging method, positive control data are used as control standards, so that DPPH free radical scavenging data of the paulownia flower water extract are obtained to be extremely lower than DPPH free radical scavenging data of a positive control group, and the paulownia flower has oxidation resistance.
The above embodiment is only one of the preferred embodiments of the present invention, and should not be used to limit the scope of the present invention, but all the insubstantial modifications or color changes made in the main design concept and spirit of the present invention are still consistent with the present invention, and all the technical problems to be solved are included in the scope of the present invention.
Claims (7)
1. Use of paulownia flower extract or a composition containing the same in bacteriostasis, characterized in that the paulownia flower extract is extracted according to the following steps: extracting paulownia flower by a steam distillation method, and collecting essential oil, namely the paulownia flower extract;
The paulownia flower extract is used for inhibiting white wax tree anthracnose, eggplant disease fusarium, aromatic fusarium, fusarium layering fusarium, cladosporium, aspergillus flavus, blue fungus or mucor.
2. Use according to claim 1, characterized in that the ratio of paulownia flower to water is 1g: 20-30 mL.
3. The use according to claim 1, characterized in that the paulownia flower extract or the composition containing the same is used for preparing a pesticide for controlling plant diseases.
4. The use according to claim 3, characterized in that the paulownia flower extract or the composition containing it is used for controlling plant anthracnose or plant root rot.
5. Use according to claim 1, characterized in that the paulownia flower extract or the composition containing the same is used for the preparation of a preservative for preventing spoilage of grains, foods, medicinal materials or feeds or an air freshener for killing air harmful fungi.
6. The use according to claim 1, wherein the composition is a 100-2000 fold dilution of paulownia flower extract.
7. The use according to claim 1, wherein the composition comprises acceptable excipients.
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