An attractant for oil-tea camellia pollinating insects, use method and application thereof

Abstract

The present invention discloses an attractant for oil-tea camellia pollinating insects, a method of use and an application thereof, wherein the attractant comprises a chlorogenic acid solution with a concentration of 0.1-1 mg / g, wherein the solvent is water. The method of use of the attractant for oil-tea camellia pollinating insects of the present invention comprises the steps of: loading the prepared attractant into a sprayer, and spraying the oil-tea camellia flowers that are just opening or in full bloom. The present invention also provides the application of the attractant for oil-tea camellia pollinating insects in oil-tea pollination. The attractant of the present invention significantly enhances the visual flower-visiting signal of oil-tea camellia, increases the number of flower-visiting insects of pollinating insects, and enhances the pollination of oil-tea camellia. The attractant has simple ingredients and low cost. The method of use of the attractant of the present invention is easy to operate, and can treat oil-tea camellia plants on a large scale. The number of visits to a single flower of oil-tea camellia can be increased by up to 4 times, and the number of fruits can be increased by 1.7 to 5 times.

Description

Technical Field

[0001] The invention belongs to the biological field, and specifically relates to an attractant for oil-tea pollinating insects, a using method and an application thereof. Background Art

[0002] Camellia oleifera Camellia oleifera Camellia oleifera belongs to the Camellia genus of the family Camellia oleifera. It is a shrub or tree. It is the main edible oil tree species in southern my country and one of the four major woody edible oil tree species in the world. Camellia oleifera seeds are rich in unsaturated fatty acids and vitamin E and can be used for oil extraction. Camellia oleifera seed oil has the effects of preventing atherosclerosis and anti-tumor. It is an excellent edible oil with both dietary and therapeutic effects.

[0003] Camellia oleifera is self-incompatible and highly dependent on pollinating insects. The flowering period of Camellia oleifera is from late autumn to winter, and there are few visiting insects, which makes it difficult to meet the pollination needs of Camellia oleifera, resulting in the phenomenon of "flowers but no fruits", "one fruit per thousand flowers", and a low fruit setting rate. In order to increase the fruit setting rate of Camellia oleifera, fruit farmers usually use artificial pollination. However, artificial pollination is labor-intensive and time-consuming, especially for species such as Camellia oleifera with high planting density, large number of flowers, and high flower density. Artificial pollination is more difficult, resulting in high pollination time and labor costs.

[0004] Therefore, there is an urgent need to develop an attractant for oil-tea camellia pollinating insects, a method of use and its application, so as to increase the number of pollinating insects visiting flowers of oil-tea camellia, promote outcrossing of oil-tea camellia to enable it to be fully pollinated, and thus further increase the yield of oil-tea camellia. Summary of the invention

[0005] The technical problem to be solved by the present invention is to provide an attractant for oil-tea camellia pollinating insects, a method of use and application thereof, so as to increase the number of pollinating insects visiting flowers of oil-tea camellia, promote outcrossing of oil-tea camellia to achieve full pollination, and thus further increase the yield of oil-tea camellia.

[0006] In order to achieve the above object, the present invention adopts the following technical scheme:

[0007] An attractant for oil-tea pollinating insects comprises a chlorogenic acid solution with a concentration of 0.1-1 mg / g, wherein the solvent is water.

[0008] Based on the above scheme, in another improved scheme, a chlorogenic acid solution with a concentration of 0.4-1 mg / g is included, wherein the solvent is water.

[0009] On the basis of the above scheme, in another improved scheme, a mixed solution of a chlorogenic acid solution with a concentration of 0.4-1 mg / g and a sucrose solution with a concentration of 5%-7% is included, wherein the solvent is water.

[0010] The invention also provides a method for using an attractant for oil-tea pollinating insects, comprising the following steps: loading the prepared attractant into a sprayer, and spraying the attractant on the oil-tea flowers that are just opening or in full bloom.

[0011] Based on the above scheme, in another improved scheme, the newly opened camellia oleifera flower refers to a state in which the stigma is exposed, and the fully bloomed camellia oleifera flower refers to a state in which the stigma is exposed and the anthers are cracked.

[0012] Based on the above scheme, in another improved scheme, spraying is carried out in the morning or at noon on non-rainy days.

[0013] Based on the above scheme, in another improved scheme, 2-3 days after the first spraying of the attractant, the oil tea flowers are supplementally sprayed with the attractant.

[0014] The invention also provides application of the attractant for oil-tea camellia pollinating insects in oil-tea camellia pollination.

[0015] The beneficial effects of the present invention include:

[0016] After the attractant for oil-tea camellia pollinating insects is sprayed on oil-tea camellia flowers, sucrose and chlorogenic acid are attached to petals, stamens or stigmas. Under sunlight, chlorogenic acid emits blue fluorescence that can be captured by insects, and sucrose releases a sweet smell, which can enhance the stimulation of oil-tea camellia flowers to insects, thereby attracting more pollinating insects to visit the flowers, thereby making the pollination of oil-tea camellia more complete and increasing the yield of oil-tea camellia.

[0017] The attractant of the present invention significantly enhances the visual flower-visiting signal of camellia oleifera and increases the number of flower-visiting pollinating insects, thereby further enhancing the pollination of camellia oleifera. The method for using the attractant of camellia oleifera pollinating insects provided by the present invention is simple to prepare, easy to operate, and has a low application cost, and has a significant effect on attracting camellia oleifera pollinating insects.

[0018] The attractant of the present invention has simple ingredients, low cost, simple operation, can be used to treat tea plants on a large scale, and saves labor costs; the method is fast in effect, has a long stable time, does not cause damage to the plants, and does not require direct human contact with the plants, which can avoid damage to the plants during manual operation. After spraying tea flowers with the attractant for pollinating insects of the present invention, the number of visits to a single tea flower can be increased by up to 4 times, and the number of fruits can be increased by 1.7 to 5 times. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 The fluorescence performance of the oil-tea camellia petals under a fluorescence microscope in Experiment 2 of the present invention, wherein A is the morphology of the outer side of the oil-tea camellia petals, B is the morphology of the outer side of the oil-tea camellia petals under ultraviolet light, C is the morphology of the inner side of the oil-tea camellia petals, and D is the morphology of the inner side of the oil-tea camellia petals under ultraviolet light;

[0020] Figure 2 The excitation spectrum and emission spectrum of the oil-tea camellia petals in Experiment 2 of the present invention, wherein A is the excitation spectrum of the outer side of the oil-tea camellia petals, B is the emission spectrum of the outer side of the oil-tea camellia petals, C is the excitation spectrum of the inner side of the oil-tea camellia petals, and D is the emission spectrum of the inner side of the oil-tea camellia petals;

[0021] Figure 3 is a molecular structure diagram of chlorogenic acid in the present invention;

[0022] Figure 4 These are photos of the 0.3 mg / g chlorogenic acid solution and undissolved chlorogenic acid particles in experiment three emitting blue fluorescence under ultraviolet light. DETAILED DESCRIPTION

[0023] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0024] In addition, the technical solutions between the various embodiments of the present invention can be combined with each other, but it must be based on the fact that ordinary technicians in the field can implement it. When the combination of technical solutions is contradictory or cannot be implemented, it should be deemed that such combination of technical solutions does not exist and is not within the scope of protection required by the present invention.

[0025] The contents not described in detail in this specification belong to the prior art known to professional and technical personnel in this field.

[0026] The main experimental process is recorded below in the order in which the research was carried out. The following experimental content has included multiple specific embodiments of the present invention.

[0027] Test site conditions:

[0028] The Camellia oleifera planting base of Central South University of Forestry and Technology in Yuelu District, Changsha City, Hunan Province, has longitude and latitude of 28°13′48″ N, 112°55′53″ E. It has a subtropical monsoon climate with mild climate, abundant precipitation, simultaneous rainfall and heat, and four distinct seasons. The average annual temperature is 16.9-17.5℃, the average annual precipitation is 1360-1400 mm, the altitude is 60-80m, the vegetation is mainly subtropical evergreen broad-leaved forest, and the soil is mainly red soil.

[0029] Test methods

[0030] Experiment 1: Comparison of the ability of oil-tea camellia flower colors and odors to attract pollinating insects

[0031] This experiment was designed to study whether the oil-tea tree attracts pollinating insects mainly based on vision or smell. During the flowering period of oil-tea, two oil-tea trees with similar growth status, age and location were selected. Three branches were selected in the same direction of the two oil-tea trees, and one oil-tea flower with the same opening status was selected on each branch. A transparent plastic bag was put on the flowers of one oil-tea tree, and a black plastic bag with small holes was put on the flowers of the other oil-tea tree. Then the number of flower-visiting insects within 10 minutes after bagging was counted and the flower-visiting insects were classified.

[0032] As shown in the statistical results in Table 1 below, only one fly visited the flowers on the branches covered with black plastic bags with small holes within 10 minutes, while five flies, six aphid flies, six bees and one wasp visited the flowers on the branches covered with transparent plastic bags within 10 minutes. The results of this experiment show that Camellia oleifera attracts pollinators by transmitting visual signals through petals that are stronger than olfactory signals.

[0033] Table 1 Comparison of the ability of oil-tea camellia flower colors and scents to attract pollinating insects

[0034]

[0035] Experiment 2: The mechanism by which Camellia oleifera petals send visual signals to pollinators

[0036] In November 2020 (Camellia oleifera flowering period), fresh Camellia oleifera petals were collected, and the fluorescence performance of the petals under ultraviolet light was observed under a fluorescence microscope and photographed and recorded. See attached for photos. Figure 1 After the observation, the collected fresh petals were cut into two parts in the middle and placed in a 60°C oven for 3 days. After drying, they were ground into powder and their fluorescence spectra under ultraviolet light were measured. The results are shown in the attached Figure 2 .

[0037] See attached Figure 1 and attached Figure 2 The test results show that the best excitation wavelength for the inner side of the oil-tea camellia petals is 400nm, and the best excitation wavelength for the outer side is 450nm. Under the corresponding excitation wavelength conditions, the oil-tea camellia petals will release blue fluorescence. Since insects have visual synapses that can sense the ultraviolet band (≤400nm), the blue fluorescence released by the inner side of the oil-tea camellia petals under ultraviolet light can be recognized by insects, and this blue fluorescence can attract pollinating insects. This is also the first research discovery in this field that oil-tea camellia flowers rely on blue fluorescence to attract pollinating insects.

[0038] Effects of spraying chlorogenic acid solution on camellia oleifera flowers on pollinating insects

[0039] Chlorogenic acid is an indicator component of many drugs. It exists in higher dicotyledons and ferns. Its content is relatively high in plants such as chrysanthemum, honeysuckle, and capillaries. The molecular structure is shown in the attached Figure 3 The content of chlorogenic acid in honeysuckle can be as high as 4%, which is one of the main antibacterial and antiviral effective pharmacological ingredients of honeysuckle. Chlorogenic acid is highly stable, has a maximum absorption peak at 380nm, and can emit blue fluorescence after absorbing ultraviolet light.

[0040] The purchased analytically pure chlorogenic acid solid was dissolved in water to form a 0.3 mg / g chlorogenic acid solution, and then placed under ultraviolet light, observed under a fluorescence microscope and photographed. See the attached photo. Figure 4 , it can be seen that the chlorogenic acid solution and undissolved chlorogenic acid particles emit blue fluorescence under ultraviolet light.

[0041] During the flowering period of Camellia oleifera in November 2020, a 0.3 mg / g chlorogenic acid aqueous solution was prepared and loaded into a sprayer. On a sunny day between 11:00 and 13:00 noon, three trees were selected from the experimental site, and three branches with newly opened or blooming Camellia oleifera flowers were selected from the target tree. One was left untreated, one was sprayed evenly with pure water, and one was sprayed with 0.3 mg / g chlorogenic acid solution. Immediately after spraying, the number of insect visits to the three branches within 30 minutes was recorded (each branch was guaranteed to have 3 flowers).

[0042] As shown in the statistical results in Table 2 below, the number of insects visiting flowers on branches sprayed with chlorogenic acid is significantly higher than that on branches in natural state and branches sprayed with pure water. Based on the principle that chlorogenic acid emits blue fluorescence under ultraviolet light, by spraying 0.3mg / g chlorogenic acid solution, the number of visits of four types of pollinating insects to oil-tea can be increased by more than 1.3 times.

[0043] Table 2 The attraction of blue fluorescence to pollinating insects

[0044]

[0045] Experiment 4 Effects of different concentrations of chlorogenic acid solution and sucrose solution on oil-tea camellia pollinating insects

[0046] In order to find the best concentration of chlorogenic acid solution to attract pollinating insects, different concentrations of chlorogenic acid solution were designed to spray on oil-tea camellia flowers. In order to study the effect of sucrose solution on the pollinating insects of oil-tea camellia, different concentrations of sucrose were designed to spray on oil-tea camellia flowers.

[0047] During the camellia flowering period in December 2020, 16 trees were selected from the experimental site at 11:00-13:00 noon, and two adjacent branches with newly opened or blooming camellia flowers were selected from each target tree. One was left untreated and the other was evenly treated with the solution. The number of insect visits to a single flower of the two branches within 10 minutes of spraying was recorded (the total number of branch insect visits divided by the number of flowers). The treatment solutions were 0.01 mg / g, 0.05 mg / g, 0.1 mg / g, 0.2 mg / g, 0.4 mg / g, 0.8 mg / g, 1 mg / g, 2 mg / g of chlorogenic acid solution and 1%, 2%, 3%, 4%, 5%, 6%, 7% sucrose solution.

[0048] Referring to the statistical results in Table 3 below, when the concentration of the chlorogenic acid solution is lower than 0.1 mg / g and equal to 2.0 mg / g, there is no attraction effect or a weak effect on pollinating insects after spraying; when the concentration of the chlorogenic acid solution is 0.1 mg / g-1 mg / g, the visit of pollinating insects increases significantly after spraying; the effect of 0.4 mg / g-1 mg / g chlorogenic acid solution is significant.

[0049] Spraying 1% sucrose solution has no effect on attracting pollinating insects, while 2%-7% sucrose solution has a good effect on attracting pollinating insects, among which 5%-7% sucrose solution has a significant effect.

[0050] Table 3 Effects of different concentrations of chlorogenic acid and sucrose on the number of visits to a single flower by pollinators in 30 minutes

[0051]

[0052] Experiment 5: Effects of different concentrations of attractants on pollinating insects of Camellia oleifera

[0053] This experiment was designed to study the attraction of the mixture of chlorogenic acid solution and sucrose solution to oil-tea camellia pollinating insects. According to the experimental results, three chlorogenic acid solution concentrations and sucrose solution concentrations with the best effects were screened out, and they were mixed in a ratio of 1:1 to prepare 9 different solutions. The experiment was carried out again on untreated branches according to the above method, and the number of insect visits to a single flower on the branch within 30 minutes was recorded. Finally, a combination with good effects was screened out based on the results.

[0054] Referring to the statistical results in Table 4 below, compared with the plants not sprayed, the number of insect visits to the plants sprayed with attractants can be increased by more than 1.5 times within 30 minutes after spraying, and can be increased up to 4 times.

[0055] Table 4 Effects of spraying different combinations of chlorogenic acid and sucrose mixed solutions on oil-tea camellia pollinating insects

[0056]

[0057] Experiment 6 Effects of different concentrations of attractants on fruit setting rate of Camellia oleifera

[0058] This experiment was designed to study the effects of different concentrations of attractants on the fruit setting rate of Camellia oleifera. In December 2020 (Camellia oleifera flowering period), another 4 target trees were selected from 11:00 to 13:00 on a sunny day at noon. 50 flowers were selected from the north, south, northwest and north of each target tree. Five target flowers were sprayed with each of the nine mixed agents selected in Experiment 5. The remaining five flowers were not treated as a natural control. The ovary development of the target flowers was checked in March 2021 to determine the number of fruits for each treatment.

[0059] As shown in the statistical results in Table 5 below, the number of fruits of plants sprayed with attractants can be increased by more than 1.7 times, and up to nearly 5 times, compared with plants not sprayed. The best ratio is 5%-6% sucrose + 0.4-1 mg / g chlorogenic acid solution, and the best ratio is 6% sucrose + 0.4-1 mg / g chlorogenic acid.

[0060] Table 5 The number of fruit set in target flowers in different directions of Camellia oleifera under different treatments

[0061]

[0062] The attractant of the present invention mainly utilizes the blue fluorescence released by chlorogenic acid under ultraviolet light to attract pollinators, so the reagent can only play a better role near noon. At 10:00-11:00 in the morning on a sunny day during the flowering period, select the newly opened (stigma exposed) or blooming Camellia oleifera flowers (stigma exposed and anthers cracked), spray them with insect attractants, and spray them evenly without any overlap or leakage. Since the chlorogenic acid structure will degrade over time at room temperature, it is necessary to perform additional spraying 2-3 days after the first spraying to ensure that the attractant can play a stable role during the flowering period. In addition, if it rains, rainwater will wash away the chlorogenic acid and sucrose sprayed on the flowers, and additional spraying must be performed on the second day after the rain or after it is relatively dry. There are differences in the flowering time of different Camellia oleifera plants, different branches and different flowers, so the attractant can be sprayed according to the degree of flowering, and the flowers in the open state can be sprayed as much as possible.

[0063] The innovations of the present invention mainly include the following points:

[0064] 1. In the field of camellia research, the present invention has for the first time found that camellia uses visual signals to attract pollinating insects, and clarified that the mechanism of visual signals used by camellia to attract pollinating insects is that the petals emit blue fluorescence under ultraviolet light. The present invention sprays chlorogenic acid solution on camellia flowers, significantly enhances the blue fluorescence of camellia petals, and significantly enhances the visual flower-visiting signals of camellia flowers, thereby increasing the number of pollinating insects that visit the flowers, thereby further enhancing the pollination and fruit setting rate of camellia.

[0065] 2. The present invention adopts the method of dissolving chlorogenic acid in water to obtain a chlorogenic acid aqueous solution, which has a simple preparation method and is non-toxic and harmless to the environment. Increasing the mixing of a certain concentration of sucrose further increases the number of insects visiting the oil-tea camellia flowers without causing negative effects on the oil-tea camellia plants, pollinating insects and the environment, and can significantly increase the pollination effect of the oil-tea camellia without harming the plants and the ecological environment.

[0066] 3. The present invention uses strict experimental design to test that the attractant has a good attracting effect on four major pollinating insects of oil-tea, namely flies, aphid flies, bees and wasps. The attractant can attract multiple pollinating insects at the same time, and the diversification of pollination groups can achieve better pollination. The attractant of the present invention utilizes the mechanism of oil-tea flowers themselves to attract flower-visiting insects, and has significant effects and is green and environmentally friendly.

Claims

1. Use of an attractant for oil-tea camellia pollinating insects in promoting oil-tea camellia pollination, characterized in that: The attractant is a chlorogenic acid aqueous solution with a concentration of 0.3 mg / g, and the oil-tea pollinating insects are aphid flies that pollinate oil-tea. The prepared attractant is loaded into a sprayer and sprayed on the oil-tea flowers that are just opening or in full bloom. The just opening oil-tea flowers refer to the flowers with exposed stigmas, and the flowers in full bloom refer to the flowers with exposed stigmas and cracked anthers. The attractant increases the number of visits of oil-tea pollinating insects by enhancing the visual flower-visiting signals of oil-tea.

2. The use of the attractant for oil-tea camellia pollinating insects as claimed in claim 1 in promoting oil-tea camellia pollination, characterized in that: The attractant is sprayed on the camellia flowers in the morning or at noon on a non-rainy day.

3. The use of the attractant for oil-tea camellia pollinating insects as claimed in claim 2 in promoting oil-tea camellia pollination, characterized in that: 2-3 days after the first spraying of the attractant, the oil-tea camellia flowers are sprayed with the attractant.

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