Field pollination method for megachile megachile

Abstract

The invention relates to the technical field of application of agricultural pollination insects, in particular to a field pollination method for megachile megachile. The invention provides a field pollination method for megachile megachile. The method comprises the following steps: a preparation stage: preparing incubation, arranging a honeycomb and preparing a nest plate; in the pollination stage, megachile clover is released, nesting conditions are observed, and disease and pest control is carried out; and a recovery treatment stage. The method can effectively improve the fruiting performance of alfalfa.

Description

Technical Field

[0001] The invention relates to the technical field of agricultural pollination insect application, and in particular to a method for field pollination of alfalfa leafcutter bees. Background Art

[0002] Alpinia phalaenopsis is a perennial leguminous plant, a rare shrub. Pinnate compound leaves, alternate; 3 leaflets, usually serrated on the margins. Racemes are axillary, sometimes capitate or solitary, with small flowers; bracts are small or absent; calyx is bell-shaped or tubular; corolla is yellow, purple, violet, brown, etc. Pods are spirally twisted, kidney-shaped, sickle-shaped or nearly straight, longer than calyx, with dorsal sutures often ridged or thorny; with 1 to many seeds.

[0003] As an important forage and green manure crop, the seed quality and yield of clover are directly related to the benefits of agricultural production. In modern agricultural production, pollination is a key link in improving crop yield and quality. Traditional pollination methods mainly rely on wind pollination and insects, but these methods are limited by environmental factors and are not efficient. In order to improve pollination efficiency, people began to study and use specific insects for field pollination.

[0004] The clover leafcutter bee (Megachile rotundata) is an effective pollinating insect, but its application technology in the field is not yet mature, resulting in low pollination efficiency and unstable pollination effect in the field. Therefore, optimizing its field pollination technology is still an urgent problem to be solved. Summary of the invention

[0005] In view of this, the purpose of the present invention is to provide a method for field pollination of alfalfa leafcutter bees, so as to solve the problems of low field pollination efficiency and unstable pollination effect of alfalfa leafcutter bees in the prior art, thereby improving the fruit setting performance of alfalfa, and further improving the yield and seed quality of alfalfa.

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

[0007] In a first aspect, the present invention provides a method for field pollination of alfalfa leafcutter bees, the method comprising the following steps:

[0008] Step S1. Preparation stage:

[0009] S11. Hatching preparation: 25 days before the alfalfa seed field blooms, the bee cocoons are placed in an artificial climate chamber at 30°C and 50% relative humidity for hatching;

[0010] S12. Set up beehives: beehives shall be set up in the alfalfa seed field. The beehive doors shall be opened and face the southeast direction. The distance between two adjacent beehives shall be 160 meters.

[0011] It should be noted that when setting up beehives in the alfalfa seed field, the beehive doors should be opened and facing the southeast so that the alfalfa leafcutter bees can better receive sunlight and fly. The distance between two adjacent beehives is 160 meters to ensure the effective collection range of the alfalfa leafcutter bees.

[0012] S13. Prepare nest boards: Set up several nest boards in the hive, and the nesting surfaces on the nest boards are painted with different patterns and logos;

[0013] Step S2. Pollination stage:

[0014] S21. Release alfalfa leafcutter bees: spray alfalfa with attractants during the initial flowering period of alfalfa; feed the alfalfa leafcutter bees with attractants when the maturity rate reaches more than 50%; then use hatching trays to transport the alfalfa leafcutter bees to the field for release; release should be done on a sunny day between 10:00 and 15:00;

[0015] S22. Observe the nesting situation: After release, regularly observe the nesting situation of the alfalfa leafcutter bees; by observing the activities of the alfalfa leafcutter bees and the changes on the nest board, the adaptation situation and pollination efficiency of the alfalfa leafcutter bees can be learned;

[0016] S23. Pest and disease control: Pay attention to the prevention and control of field pests and diseases; when using pesticides, choose pesticides that are safer for alfalfa leafcutter bees and stay at least 100 meters away from alfalfa leafcutter bee hives;

[0017] Step S3. Recycling and processing stage:

[0018] S31. Recycling of cocoons:

[0019] After the pollination season is over, the cocoons of the alfalfa leafcutter bees are collected; when the humidity of the nest board drops below 30%, the alfalfa leafcutter bee cocooning machine is used to remove the cocoons indoors.

[0020] It should be noted that after the pollination season ends, the cocoons of alfalfa leafcutter bees should be collected in time. This helps to protect alfalfa leafcutter bee resources and prepare for the next year's pollination work.

[0021] It should be noted that after the alfalfa flowering period ends, stop spraying attractants, stop feeding attractants, and evacuate the pollinating bee colonies from the alfalfa fields.

[0022] Preferably, in terms of mass fractions, the attractant consists of 30-35 parts of alfalfa pollen, 2-4 parts of alfalfa flower exosomes, 5-7 parts of alfalfa exosomes, 15-20 parts of white sugar and 50-60 parts of water.

[0023] Preferably, the attractant consists of 15-20 parts of jasmonic acid, 2-4 parts of alfalfa flower exosomes and 40-50 parts of water in parts by mass.

[0024] More preferably, in terms of mass percentage, the attractant consists of 32 parts of alfalfa pollen, 3 parts of alfalfa flower exosomes, 6 parts of alfalfa exosomes, 18 parts of white sugar and 52 parts of water.

[0025] More preferably, the attractant consists of 16 parts of jasmonic acid, 3 parts of alfalfa flower exosomes and 43 parts of water in terms of mass.

[0026] Preferably, the release density in step S21 is controlled at 3000 to 10000 heads per mu.

[0027] Preferably, the nesting surface on the nest board in step S13 is set to bluish black.

[0028] It should be noted that setting the nesting surface on the nest board to blue-black helps attract alfalfa leafcutter bees and improve their nesting efficiency.

[0029] In the present invention, the preparation method of alfalfa flower exosomes is as follows:

[0030] ① Add 300 mL of sterilized PBS to 260 g of fresh alfalfa flowers, crush them with a crusher, centrifuge at 5°C, 3000×g for 40 min, remove the larger residues, and take the yellow transparent solution in the middle layer;

[0031] ② Centrifuge the yellow transparent solution obtained in step ① at 5°C, 10,000×g for 60 min, and take the supernatant;

[0032] ③ The supernatant obtained in step ② was centrifuged at 5°C, 120,000×g for 60 min, and then the precipitate was resuspended in 3 mL of sterile PBS and centrifuged at 5°C, 150,000×g for 65 min, and the precipitate was resuspended in 3 mL of sterile PBS again to obtain alfalfa flower exosomes.

[0033] In the present invention, the preparation method of alfalfa exosomes is as follows:

[0034] ① Add 350 mL of sterilized PBS to 300 g of fresh alfalfa, crush it with a crusher, centrifuge it at 5°C, 3000×g for 45 min, remove the larger residues, and take the green transparent solution in the middle layer;

[0035] ② Centrifuge the green transparent solution obtained in step ① at 5°C, 10,000×g for 65 min, and take the supernatant;

[0036] ③ The supernatant obtained in step ② was centrifuged at 5°C, 120,000×g for 65 min, and then the precipitate was resuspended in 3 mL of sterile PBS and centrifuged at 5°C, 150,000×g for 70 min, and the precipitate was resuspended in 3 mL of sterile PBS again to obtain alfalfa exosomes.

[0037] In a second aspect, an alfalfa leafcutter bee is provided for performing field pollination according to the method described in the present invention.

[0038] In a third aspect, the invention provides application of the method described in the invention in pollination by alfalfa leafcutter bees.

[0039] In a fourth aspect, the invention provides application of the method described in the present invention in field pollination of alfalfa leafcutter bees.

[0040] Compared with the prior art, the beneficial effects of the present invention are:

[0041] The present invention has been concluded through a large number of experimental screenings that by feeding alfalfa leafcutter bees with an attractant composed of alfalfa pollen, alfalfa flower exosomes, alfalfa exosomes, white sugar and water, this attractant can help improve the fruit setting rate of the first clover. During the experimental screening process, the inventor conducted a large number of experimental screenings. The inventor tried to replace alfalfa pollen, alfalfa flower exosomes, and alfalfa exosomes with other substances for experiments, and found that the replaced technical solution was not effective in improving the fruit setting rate of the first clover; the inventor tried to replace alfalfa flower exosomes with alfalfa flowers or alfalfa flower extracts, and found that the replaced technical solution was also generally effective in improving the fruit setting rate of the first clover; the inventor tried to replace alfalfa exosomes with alfalfa or alfalfa extracts, and found that the replaced technical solution was still not as effective as the present invention in improving the fruit setting rate of the first clover. The inventors speculate that alfalfa flower exosomes and alfalfa exosomes, as microvesicles secreted by cells, may contain bioactive substances that are beneficial to alfalfa leafcutter bees, such as growth factors, signal molecules, etc. These substances play an important role in intercellular communication, signal transmission, etc. These substances may promote the growth and reproduction of alfalfa leafcutter bees, thereby improving their pollination efficiency, and then significantly improving the fruit setting rate of alfalfa. In addition, the inventors speculate that alfalfa flower exosomes and certain substances in alfalfa exosomes may also have the effect of regulating the behavior of alfalfa leafcutter bees. They may prompt alfalfa leafcutter bees to visit alfalfa flowers more frequently for nectar collection and pollination activities. This change in behavior helps to increase the frequency of pollination and improve the quality of pollination, thereby improving the fruit setting rate of alfalfa. In contrast, alfalfa flower extracts may simply contain certain components in alfalfa flowers, and not necessarily these microvesicles with biological activity.

[0042] The present invention has been screened through a large number of experiments. By spraying an attractant composed of jasmonic acid, alfalfa flower exosomes and water on alfalfa during the initial flowering period of alfalfa, the pollination efficiency can be enhanced. The inventors speculate that the active ingredients in jasmonic acid and alfalfa flower exosomes may have the effect of attracting pollinating insects (alfalfa leaf-cutting bees). These ingredients can guide pollinating insects to visit alfalfa flowers more frequently through their unique smell or chemical substances, thereby increasing the chance of pollination. Furthermore, by spraying the attractant of the present invention, pollinating insects may stay on the flowers for a longer time, or be more focused on pollination activities, thereby improving the quality of pollination. On the other hand, the growth of alfalfa can be promoted. The inventors speculate that alfalfa flower exosomes contain rich nutrients, such as protein, amino acids, vitamins, etc. After these ingredients are absorbed by alfalfa leaves, they can provide necessary nutritional support for their growth and promote the healthy growth of alfalfa plants. Furthermore, jasmonic acid and certain ingredients in alfalfa flower exosomes may have the effect of regulating plant physiological functions, such as promoting root development and improving photosynthesis efficiency. The improvement of these physiological functions helps alfalfa to better utilize nutrients and light energy, thereby increasing its fruiting rate. On the other hand, stress resistance can be improved. The inventors speculate that jasmonic acid and certain components in alfalfa flower exosomes may have antibacterial and antiviral biological activities, which can enhance the disease resistance of alfalfa plants; when the disease occurs, these components can reduce the damage of the disease to alfalfa and maintain its normal growth and fruiting ability. Furthermore, by spraying the attractant of the present invention, it may be possible to improve the adaptability of alfalfa to adversities such as drought and salinity to a certain extent; this helps alfalfa maintain stable growth and fruiting performance under adverse environmental conditions.

[0043] In addition, the present invention utilizes natural biological resources for pollination, reduces dependence on chemical pesticides, and is also beneficial to protecting the ecological environment and promoting sustainable agricultural development. DETAILED DESCRIPTION

[0044] The technical solution of the present invention is clearly and completely described below in conjunction with the embodiments. 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.

[0045] Example 1

[0046] This embodiment provides a method for field pollination of alfalfa leafcutter bees, the method comprising the following steps:

[0047] Step S1. Preparation stage:

[0048] S11. Hatching preparation: 25 days before the alfalfa seed field blooms, the bee cocoons are placed in an artificial climate chamber at 30°C and 50% relative humidity for hatching;

[0049] S12. Set up beehives: beehives should be set up in the alfalfa seed field. The beehive doors should be opened and face the southeast direction. The distance between two adjacent beehives is 160 meters.

[0050] S13. Prepare nest boards: Set up several nest boards in the hive, and the nesting surfaces on the nest boards are painted with different patterns and logos;

[0051] Step S2. Pollination stage:

[0052] S21. Release alfalfa leafcutter bees: spray alfalfa with attractants during the initial flowering period of alfalfa; feed the alfalfa leafcutter bees with attractants when the maturity rate reaches more than 50%; then use hatching trays to transport the alfalfa leafcutter bees to the field for release; release should be done on a sunny day between 10:00 and 15:00;

[0053] S22. Observe the nesting situation: After release, regularly observe the nesting situation of the alfalfa leafcutter bees; by observing the activities of the alfalfa leafcutter bees and the changes on the nest board, the adaptation situation and pollination efficiency of the alfalfa leafcutter bees can be learned;

[0054] S23. Pest and disease control: Pay attention to the prevention and control of field pests and diseases; when using pesticides, choose pesticides that are safer for alfalfa leafcutter bees and try to stay more than 100 meters away from the alfalfa leafcutter bee hive;

[0055] Step S3. Recycling and processing stage:

[0056] S31. Recycling of cocoons:

[0057] After the pollination season is over, the cocoons of the alfalfa leafcutter bees are collected; when the humidity of the nest board drops below 30%, the alfalfa leafcutter bee cocooning machine is used to remove the cocoons indoors;

[0058] According to the weight percentage, the attractant is composed of 30 parts of alfalfa pollen, 2 parts of alfalfa flower exosomes, 5 parts of alfalfa exosomes, 15 parts of white sugar and 50 parts of water;

[0059] In terms of weight, the attractant is composed of 15 parts of jasmonic acid, 2 parts of alfalfa flower exosomes and 40 parts of water;

[0060] The release density in step S21 is controlled at 3000 to 10000 heads / mu;

[0061] The nesting surface on the nest board in step S13 is set to be bluish black.

[0062] Example 2

[0063] This embodiment provides a method for field pollination of alfalfa leafcutter bees, the method comprising the following steps:

[0064] Step S1. Preparation stage:

[0065] S11. Hatching preparation: 25 days before the alfalfa seed field blooms, the bee cocoons are placed in an artificial climate chamber at 30°C and 50% relative humidity for hatching;

[0066] S12. Set up beehives: beehives should be set up in the alfalfa seed field. The beehive doors should be opened and face the southeast direction. The distance between two adjacent beehives is 160 meters.

[0067] S13. Prepare nest boards: Set up several nest boards in the hive, and the nesting surfaces on the nest boards are painted with different patterns and logos;

[0068] Step S2. Pollination stage:

[0069] S21. Release alfalfa leafcutter bees: spray alfalfa with attractants during the initial flowering period of alfalfa; feed the alfalfa leafcutter bees with attractants when the maturity rate reaches more than 50%; then use hatching trays to transport the alfalfa leafcutter bees to the field for release; release should be done on a sunny day between 10:00 and 15:00;

[0070] S22. Observe the nesting situation: After release, regularly observe the nesting situation of the alfalfa leafcutter bees; by observing the activities of the alfalfa leafcutter bees and the changes on the nest board, the adaptation situation and pollination efficiency of the alfalfa leafcutter bees can be learned;

[0071] S23. Pest and disease control: Pay attention to the prevention and control of field pests and diseases; when using pesticides, choose pesticides that are safer for alfalfa leafcutter bees and try to stay more than 100 meters away from the alfalfa leafcutter bee hive;

[0072] Step S3. Recycling and processing stage:

[0073] S31. Recycling of cocoons:

[0074] After the pollination season is over, the cocoons of the alfalfa leafcutter bees are collected; when the humidity of the nest board drops below 30%, the alfalfa leafcutter bee cocooning machine is used to remove the cocoons indoors;

[0075] According to the mass percentage, the attractant is composed of 35 parts of alfalfa pollen, 4 parts of alfalfa flower exosomes, 7 parts of alfalfa exosomes, 20 parts of white sugar and 60 parts of water;

[0076] In terms of weight, the attractant is composed of 20 parts of jasmonic acid, 4 parts of alfalfa flower exosomes and 50 parts of water;

[0077] The release density in step S21 is controlled at 3000 to 10000 heads / mu;

[0078] The nesting surface on the nest board in step S13 is set to be bluish black.

[0079] Example 3

[0080] This embodiment provides a method for field pollination of alfalfa leafcutter bees, the method comprising the following steps:

[0081] Step S1. Preparation stage:

[0082] S11. Hatching preparation: 25 days before the alfalfa seed field blooms, the bee cocoons are placed in an artificial climate chamber at 30°C and 50% relative humidity for hatching;

[0083] S12. Set up beehives: beehives should be set up in the alfalfa seed field. The beehive doors should be opened and face the southeast direction. The distance between two adjacent beehives is 160 meters.

[0084] S13. Prepare nest boards: Set up several nest boards in the hive, and the nesting surfaces on the nest boards are painted with different patterns and logos;

[0085] Step S2. Pollination stage:

[0086] S21. Release alfalfa leafcutter bees: spray alfalfa with attractants during the initial flowering period of alfalfa; feed the alfalfa leafcutter bees with attractants when the maturity rate reaches more than 50%; then use hatching trays to transport the alfalfa leafcutter bees to the field for release; release should be done on a sunny day between 10:00 and 15:00;

[0087] S22. Observe the nesting situation: After release, regularly observe the nesting situation of the alfalfa leafcutter bees; by observing the activities of the alfalfa leafcutter bees and the changes on the nest board, the adaptation situation and pollination efficiency of the alfalfa leafcutter bees can be learned;

[0088] S23. Pest and disease control: Pay attention to the prevention and control of field pests and diseases; when using pesticides, choose pesticides that are safer for alfalfa leafcutter bees and try to stay more than 100 meters away from the alfalfa leafcutter bee hive;

[0089] Step S3. Recycling and processing stage:

[0090] S31. Recycling of cocoons:

[0091] After the pollination season is over, the cocoons of the alfalfa leafcutter bees are collected; when the humidity of the nest board drops below 30%, the alfalfa leafcutter bee cocooning machine is used to remove the cocoons indoors;

[0092] In terms of weight, the attractant consists of 32 parts of alfalfa pollen, 3 parts of alfalfa flower exosomes, 6 parts of alfalfa exosomes, 18 parts of white sugar and 52 parts of water.

[0093] In terms of weight, the attractant is composed of 16 parts of jasmonic acid, 3 parts of alfalfa flower exosomes and 43 parts of water;

[0094] The release density in step S21 is controlled at 3000 to 10000 heads / mu;

[0095] The nesting surface on the nest board in step S13 is set to be bluish black.

[0096] Comparative Example 1

[0097] Compared with Example 3, no alfalfa flower exosomes were added to the attractant, and the rest was the same as Example 3.

[0098] Comparative Example 2

[0099] Compared with Example 3, alfalfa exosomes were not added to the attractant, and the rest was the same as Example 3.

[0100] Comparative Example 3

[0101] Compared with Example 3, the alfalfa flower exosomes in the attractant are replaced with alfalfa flower extract, and the rest is the same as Example 3; wherein, the alfalfa flower is soaked in ethanol and stirred to dissolve the target component in the ethanol to obtain the alfalfa flower extract.

[0102] Effect verification

[0103] Test method: The test set up six treatments: Example 1, Example 2, Example 3, Comparative Example 1, Comparative Example 2, and Comparative Example 3, each treatment had one cell, and each cell was 8m 2 Five cameras were set up in each plot to record the flowering and pollination process of alfalfa.

[0104] Measurement items and methods: Before the test, 50 inflorescences were randomly selected in each plot for marking, and the number of florets in each inflorescence was recorded. After pollination, the number of pods in a single inflorescence was counted and the pod-setting rate was calculated. After the seeds matured, the number of seeds and thousand-grain weight in a single pod were measured. Pod-setting rate = number of pods / number of florets × 100%.

[0105] The experimental results are shown in Table 1.

[0106] Table 1 Fruiting situation of each treatment group

[0107] Treatment Group Pod setting rate / % Number of seeds / pod Thousand grain weight / g Example 1 73.31 5.15 2.31 Example 2 71.26 5.02 2.29 Example 3 82.15 5.97 2.86 Comparative Example 1 41.30 2.40 0.82 Comparative Example 2 40.69 2.53 0.79 Comparative Example 3 36.87 2.31 0.66

[0108] The pollination effect was evaluated from the pod setting rate, the number of grains per pod and the 1000-grain weight of alfalfa. The pod setting rate, the number of grains and the 1000-grain weight of alfalfa can be significantly improved by the treatment methods in Examples 1-3, while Comparative Examples 1-3 cannot improve the pollination effect due to the lack of effective methods or unfavorable conditions. Moreover, the pod setting rate, the number of grains and the 1000-grain weight of alfalfa obtained by the treatment method in Example 3 are all higher than those in Examples 1 and 2, and are all significantly higher than those in each comparative example. These experimental data show that the pod setting performance of alfalfa can be effectively improved by the specific treatment method of the present invention.

[0109] The embodiments described above are only some embodiments of the present invention, not all embodiments. The detailed description of the embodiments of the present invention is not intended to limit the scope of the invention claimed for protection, but only represents selected embodiments of the present invention. All other embodiments obtained without creative work and related deductions and substitutions made by ordinary technicians in the field under the conditions of the concept of the present invention belong to the scope of protection of the present invention.

Claims

1. A method for field pollination by alfalfa leafcutter bees, characterized in that: The method comprises the following steps: Step S1. Preparation stage: S11. Hatching preparation: 25 days before the alfalfa seed field blooms, the bee cocoons are placed in an artificial climate chamber at 30°C and 50% relative humidity for hatching; S12. Set up beehives: beehives shall be set up in the alfalfa seed field. The beehive doors shall be opened and face the southeast direction. The distance between two adjacent beehives shall be 160 meters. S13. Prepare nest boards: Set up several nest boards in the hive, and the nesting surfaces on the nest boards are painted with different patterns and logos; Step S2. Pollination stage: S21. Release alfalfa leafcutter bees: spray alfalfa with attractants during the initial flowering period of alfalfa; feed the alfalfa leafcutter bees with attractants when the maturity rate reaches more than 50%; then use hatching trays to transport the alfalfa leafcutter bees to the field for release; release should be done on a sunny day between 10:00 and 15:00; S22. Observe the nesting situation: After release, regularly observe the nesting situation of the alfalfa leafcutter bees; by observing the activities of the alfalfa leafcutter bees and the changes on the nest board, the adaptation situation and pollination efficiency of the alfalfa leafcutter bees can be learned; S23. Pest and disease control: Pay attention to the prevention and control of field pests and diseases; when using pesticides, choose pesticides that are safer for alfalfa leafcutter bees and stay at least 100 meters away from alfalfa leafcutter bee hives; Step S3. Recycling and processing stage: S31. Recycling of cocoons: After the pollination season is over, the cocoons of the alfalfa leafcutter bees are collected; when the humidity of the nest board drops below 30%, the alfalfa leafcutter bee cocooning machine is used to remove the cocoons indoors.

2. The method according to claim 1, characterized in that: In terms of weight, the attractant consists of 30-35 parts of alfalfa pollen, 2-4 parts of alfalfa flower exosomes, 5-7 parts of alfalfa exosomes, 15-20 parts of white sugar and 50-60 parts of water.

3. The method according to claim 1, characterized in that In terms of weight, the attractant consists of 15-20 parts of jasmonic acid, 2-4 parts of alfalfa flower exosomes and 40-50 parts of water.

4. The method according to claim 2, characterized in that: In terms of weight, the attractant consists of 32 parts of alfalfa pollen, 3 parts of alfalfa flower exosomes, 6 parts of alfalfa exosomes, 18 parts of white sugar and 52 parts of water.

5. The method according to claim 3, characterized in that: In terms of weight, the attractant consists of 16 parts of jasmonic acid, 3 parts of alfalfa flower exosomes and 43 parts of water.

6. The method according to claim 1, characterized in that The release density in step S21 is controlled at 3,000 to 10,000 heads per mu.

7. The method according to claim 1, characterized in that The nesting surface on the nest board in step S13 is set to be bluish black.

8. An alfalfa leafcutter bee, characterized in that Field pollination is carried out according to the method according to any one of claims 1 to 7.

9. Use of the method according to claim 1 in pollination of alfalfa leafcutter bees.

10. Use of the method according to claim 1 in field pollination of alfalfa leafcutter bees.