阅读说明：本技术 一种用于杨梅的精油防腐驱虫剂 (Essential oil antiseptic insect repellent for waxberries ) 是由 宋达峰 李自成 何帆 于 2020-02-25 设计创作，主要内容包括：本发明公开了一种用于杨梅的精油防腐驱虫剂,其特征在于,按重量百分比计,所述精油防腐驱虫剂包括以下成分：1％-30％植物精油、0.1％-10％吐温80、其余为去离子水。所述植物精油优选为为山苍籽精油、桔子精油和龙脑樟精油,其质量配比为3∶2∶1。实验证明,采用这三种植物精油按照适合的比例混合,可产生协同作用,从而更好地实现功能。本发明精油防腐驱虫剂中的精油为植物精油,属于食品添加剂,对杨梅和人体安全,对环境污染小,另外,本发明中植物精油展着力比较好,可以使药液均匀粘附在杨梅表面,使保鲜效果均匀。(The invention discloses an essential oil antiseptic insect repellent for waxberries, which is characterized by comprising the following components in percentage by weight: 1 to 30 percent of vegetable essential oil, 0.1 to 10 percent of Tween 80 and the balance of deionized water. The plant essential oil is preferably litsea cubeba essential oil, orange essential oil and borneol camphor essential oil, and the mass ratio of the plant essential oil to the borneol essential oil is 3:2: 1. Experiments prove that the three plant essential oils are mixed according to a proper proportion to generate a synergistic effect, so that the functions are better realized. The essential oil in the essential oil anti-corrosion insect repellent belongs to food additives, is safe to waxberries and human bodies, and has small environmental pollution.)

1. An essential oil antiseptic insect repellent for waxberries, which is characterized by comprising the following components in percentage by weight: 1 to 30 percent of vegetable essential oil, 0.1 to 10 percent of Tween 80 and the balance of deionized water.

2. The essential oil antiseptic insect repellent for red bayberries according to claim 1, wherein the plant essential oil is one or more of litsea cubeba essential oil, orange essential oil and borneol camphor essential oil.

3. The essential oil antiseptic insect repellent for waxberries according to claim 2, wherein the plant essential oil is litsea cubeba essential oil, orange essential oil and borneol camphor essential oil in a mass ratio of 3:2: 1.

4. The essential oil antiseptic insect repellent for red bayberries according to claim 3, wherein the plant essential oil content is 10%.

5. An insect-repelling, antiseptic and fresh-keeping method for waxberries, which is characterized in that the fresh waxberries are fumigated by the essential oil antiseptic insect repellent according to any one of claims 1 to 4 for 2 to 6 hours.

6. An insect-repelling, antiseptic and fresh-keeping method for waxberries is characterized in that 15 days, 10 days and 5 days before the waxberries are picked, the essential oil antiseptic insect-repelling agent according to any one of claims 1 to 4 is uniformly sprayed on waxberry trees.

Technical Field

The invention relates to the technical field of preservation, and particularly relates to an essential oil antiseptic insect repellent for waxberries.

Background

The waxberry is one of the special fruits in China, and has the reputation of being questioned for one thousand of gold. The waxberry has bright color, more juice, proper sweetness and sourness and high nutritive value, and the ripe waxberry is just a slack season of the fruit, thereby providing edible fresh fruit for the market and being deeply popular with consumers. However, the red bayberries are easy to rot and not resistant to storage, and are very easy to attract insects (fruit flies), so that the corrosion prevention and insect repelling of the red bayberries are very important links in the production, processing and sale processes of the red bayberries. The existing anticorrosion and insect-repellent technology mainly comprises a physical method and a chemical method, the chemical method comprises medicament spraying, soaking treatment and the like, the physical preservation cost is high, and the taste of the waxberries is adversely affected, so that chemical reagents are frequently used in the existing anticorrosion and insect-repellent technology to carry out early-stage treatment on the waxberries, but the components of certain chemical reagents are harmful to the bodies, the drug resistance is easy to generate, and certain pollution is caused to the environment.

The compound essential oil preservative and insect repellent is prepared by mixing several essential oils according to different components, and the various raw materials have strong antibacterial activity and antioxidant activity under the combined action, so that the spoilage of the waxberries by microorganisms is inhibited, and the preservative and fresh-keeping effects of the fruits are achieved. Meanwhile, the essential oil has the functions of killing and expelling insects, and can better reduce the quantity of fruit flies and other insects in the waxberry. At present, no compound essential oil is used as a fresh-keeping insect repellent for the waxberries in the existing research.

Therefore, in order to solve the defects of the prior art, the invention provides the preservative and insect repellent which has good preservative and fresh-keeping effects and strong insect repelling effect, and is safe and environment-friendly for waxberries and human bodies.

Disclosure of Invention

The invention aims to provide an essential oil antiseptic insect repellent for waxberries, which can effectively reduce the spoilage and insect production rate of the waxberries and improve the quality of the waxberries.

In order to achieve the purpose, the invention provides the following technical scheme:

an essential oil antiseptic insect repellent for waxberries, which is characterized by comprising the following components in percentage by weight: 1 to 30 percent of vegetable essential oil, 0.1 to 10 percent of Tween 80 and the balance of deionized water.

Further, the plant essential oil is one or a combination of more of litsea cubeba essential oil, orange essential oil and borneol camphor essential oil.

Further, the plant essential oil is litsea cubeba essential oil, orange essential oil and borneol camphor essential oil, and the mass ratio of the plant essential oil to the borneol essential oil is 3:2: 1. Experiments prove that the three plant essential oils are mixed according to a proper proportion to generate a synergistic effect (the synergistic effect refers to the effect of various common efforts which can be found in the decision of resource allocation, namely the effect of '1 +1> 2'), so that the functions are better realized. The litsea cubeba essential oil and the orange essential oil are combined to improve the antibacterial and insect-resistant performance, and when the litsea cubeba essential oil is the orange essential oil: when the borneol camphor essential oil is equal to 3:2:1, the synergistic effect is the best.

The invention also provides a pest expelling, corrosion preventing and fresh keeping method for the waxberries, which is characterized in that the essential oil corrosion preventing and pest expelling agent is adopted to fumigate the waxberries, and the fumigating time is 2-6 hours.

Another method for expelling parasites, preserving and preserving waxberries comprises uniformly spraying the essential oil-containing antiseptic and insect-repellent agent on waxberries 15 days, 10 days and 5 days before the waxberries are picked.

The plant essential oil is extracted by an extraction method or a distillation method commonly used in the field.

The invention has the following advantages and beneficial effects:

1. the essential oil in the essential oil antiseptic insect repellent belongs to food additives, is safe to waxberries and human bodies, and has little pollution to the environment.

2. The essential oil anti-corrosion insect repellent can quickly sterilize the surfaces of the waxberries, reduce the residue of active microorganisms and is not easy to decay.

3. The essential oil anti-corrosion insect repellent can greatly reduce the quantity of fruit fly larvae in the waxberries and improve the quality of the waxberries.

4. The compound essential oil has good spreading force, and can make the liquid medicine uniformly adhere to the surface of the waxberry, so that the preservation effect is uniform.

5. The plant essential oil selected by the invention has low cost, large production capacity in China and easy acquisition.

Drawings

Fig. 1 is a structural schematic diagram of a Y-shaped olfactometer.

Fig. 2 is a schematic representation of the effect of essential oil # 3 on survival (%) e.coli (a) and salmonella (b).

Fig. 3 is a graph showing the extracellular relative conductivity (%) of e.coli (a) and salmonella (b) after treatment with # 3 essential oil.

Fig. 4 is a schematic of the nucleic acid leakage (OD260) of e.coli (a) and salmonella (b) after treatment with # 3 essential oil.

Coli and Salmonella variations of 3# essential oil treatment.

Fig. 6 shows the decrease in cell wall damage (AKP activity) after survival after treatment of e.coli with different concentrations of essential oil # 3: a is a control group under an optical microscope with 400 times; b, C, D are e.coli after treatment with different concentrations of essential oil under a 400-fold fluorescence microscope (control, MIC, MBC), respectively.

Detailed Description

The invention discloses an essential oil antiseptic insect repellent for waxberries, which can be realized by appropriately improving parameters by referring to the content in the text. It is expressly intended that all such similar substitutes and modifications which would be obvious to one skilled in the art are deemed to be included in the invention. While the invention has been described in terms of preferred embodiments, it will be apparent to those skilled in the art that the invention can be practiced and applied by modifying or appropriately combining the methods and applications described herein without departing from the spirit and scope of the invention.

In order to better illustrate the invention, reference will now be made to specific examples.

1. Zone of inhibition test

And (3) measuring the antibacterial activity of the essential oil by adopting an Oxford cup diffusion method, pouring a solid culture medium and semi-solid bacterial liquid into the culture dish in sequence (the solid culture medium and the semi-solid bacterial liquid are poured into the culture dish after the solidification of the solid culture medium and the semi-solid bacterial liquid), adding 50 mu L pure essential oil into the Oxford cup, taking sterile water as a blank control, and measuring the size of the antibacterial zone (minus the diameter of the Oxford cup) after culturing for 24 hours at 37.

2. Determination of Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC)

Measuring MIC and MBC of different combination essential oil by a 96-well plate method, diluting with ethanol by 10 times to obtain essential oil with the concentration range of 0.01-1%, using absolute ethanol as a control group, culturing overnight at 37 ℃, using an enzyme-labeling instrument to measure the optical density value (OD) at 600nm, comparing with the control group, dipping the essential oil with the lowest concentration of OD value smaller than the control hole as MIC, using an inoculating loop to dip the mixture of the essential oil with the MIC concentration and a bacterial liquid by times, inoculating on L B solid culture medium, culturing overnight at 37 ℃, observing, and using the lowest concentration without bacterial colony as MBC.

3. Drawing growth curve

Adding MIC, 3 × MIC essential oil and overnight culture solution into 96-well plate, culturing at 37 deg.C overnight, and recording OD at 0h, 4h, 8h, 12h, 16h, 20h, and 24h₆₀₀On the abscissa of time, OD₆₀₀Growth curves were plotted for the ordinate.

4. Determination of the relative conductivity (REC)

The overnight cultured bacterial liquid is centrifuged for 8min at 4 ℃ and 6000G, then the isotonic bacteria is obtained by washing with 5% glucose solution, then the bacterial liquid is treated with essential oil with MIC and MBC concentration, the conductivity is measured every 2h at 37 ℃ and is recorded as L2, the conductivity of 0h is L1, L0 is the conductivity measured after the bacterial liquid in the glucose solution is boiled for 5min, and REC (%) is calculated as (L2-L1)/L0 × 100 according to the following formula.

5. Determination of nucleic acid leakage

Centrifuging overnight culture solution at 4 deg.C and 6000G for 8min, washing with phosphate buffer solution for 3 times, resuspending, adding compound essential oil with MIC and MBC concentrations, culturing at 37 deg.C for 24 hr, collecting supernatant every 4 hr, and measuring OD with enzyme-labeling instrument₂₆₀。

6. Determination of alkaline phosphatase (AKP) leakage

Adding 120 μ L R1 solution, 30 μ L R2 solution (R1, R2 solution is kit: Nanjing institute of bioengineering), 3 μ L washed bacterial suspension (method is the same as 1.6), adding MIC and MBC concentration compound essential oil, and immediately measuring OD with microplate reader₄₀₅And the absorbance was measured again after 2 min.

7. Propidium Iodide (PI) assay

The determination was carried out by the PI staining kit method (BBI L ife Sciences, consisting of components A and B). The buffer solution of component B was diluted 10-fold with sterile water, the treated bacterial solution (method 1.6) was stained, incubated for 5-30min at rest, the stained bacterial suspension was prepared into a slide, and the staining was observed by a fluorescence microscope.

8. Experiment for oviposition of fruit fly imago

Using 1% Tween 80 to respectively dilute 9 essential oil combinations to 50 times of liquid (the concentration is 20m L/L), collecting 200 female fruit flies of 10 days old, 100 female fruit flies of a control group and an experimental group respectively, 5 female fruit flies of each tube with each concentration, using a fruit fly corn culture medium for the experiment, adding a brilliant blue solution with the concentration of 200 mu L and the concentration of 10% on the surface of food, using 100 mu L as the control group and using 100 mu L essential oil diluent as the experimental group, placing the fruit flies on the food after the treatment, placing the food in the culture medium, clearing the female fruit flies after 22 hours, counting the number of fruit eggs under a microscope, and using each essential oil combination as an experiment.

9. Selection and determination of fruit fly 'Y' -shaped tube behavior

A Y-shaped olfactometer is adopted to test the behavioral reaction of fruit flies to essential oil combination (litsea cubeba essential oil: orange essential oil: borneol camphor essential oil: 3:2:1), the instrument consists of an oxygen increasing instrument, a connector, an air humidifier, an activated carbon air filtering device and a Y-shaped tube, all the components are connected by a silicone tube, the length of 2 side arms of the olfactometer is 15 cm. straight tube length 10cm, the included angle of two arms is 70 degrees (as shown in figure 1), when in test, one arm of the olfactometer is used for processing, a test reagent is 3# essential oil combination, 5, 10 and 15 mu L3 # essential oil combination is respectively diluted to different concentrations by 1m L acetone, and then a trace sample injector is used for absorbing equivalent amount of essential oil and uniformly dripping the essential oil on 2cm²The method comprises the steps of dropping equal amount of acetone on filter paper to serve as a control, placing the filter paper in air, after the acetone volatilizes, respectively placing treated and control filter paper into corresponding smell test arms, wherein the air flow in the test arms is 200m L/min, introducing 20 female adults from the tail end of a straight pipe of a Y-type olfactometer, observing and recording behavior reactions of the adults, observing for 10min each time, determining that the adults staying in a base arm do not react in the test process, determining that the adults entering a treatment arm do not react selectively, respectively determining the reactions of the female insects to 3# essential oil combinations with different concentrations by using the method, and treating for 10 times at each concentration.

And the directions of the two arms of the Y-shaped pipe are changed after each test. To eliminate the effect of geometric position on the behavior of fruit flies. After each treatment, the entire olfactometer was immediately cleaned with 95% ethanol and dried in an oven at 100 ℃ for further use. All tests were conducted in a dedicated olfactory testing chamber with clean air and relatively closed.

10. Preservation experiment for waxberry

(1) Selecting a plurality of parts of waxberries with uniform size and no mechanical injury or pest damage, wherein each part has the mass of 1000g, and carrying out the following treatment. The 5cm square filter paper was immersed with 10% (volume fraction) of each of the 9 essential oil composition solutions and water was used as a control. The method comprises the following steps of attaching 4 pieces of filter paper to the bottom and the peripheral box walls of a fumigation box at certain intervals, putting the waxberries in a porous placing box, and then putting the waxberries in a corresponding fumigation box with a support to prevent the surfaces of the waxberries from directly contacting with a liquid bacteriostatic agent. After sealing treatment, marking, fumigating in a 4 ℃ constant temperature climate box for 4h, subpackaging and marking the preservation boxes for each group of waxberries, and then placing in the 4 ℃ constant temperature climate box. And respectively measuring the hardness, the fruit yield and the fruit fly maggot number of the waxberry fruits of each treatment group at 5d and 10 d.

(2) Selecting several similar waxberry trees which have basically the same growth condition and are in proper growth environment. The control group was not treated at all and was allowed to grow normally; the experimental group utilized 1% 3# essential oil solution diluted to be uniformly sprayed on the waxberry trees for 3 times respectively 15 days, 10 days and 5 days before picking. After picking, the fruits are placed in a constant temperature climate box at 4 ℃, and the rotting level and the number of the fruit flies and maggots are measured every 5 days.