Method for accelerating directional breeding of bumblebees

Abstract

The invention provides a method for accelerating directional breeding of bumblebees, which specifically comprises the following steps: selecting worker bees with excellent characters as female parent worker bees for directional breeding, laying eggs and hatching the worker bees into male bees in a dark environment with the temperature of 27-29 DEG C and the humidity of 50%-60%, obtaining sperms of the male bees after sexual maturity, and then putting the sperms into a bumblebee breeding box to breed the bumblebees. And inseminating female parent worker bees by using the obtained sperms, establishing a queen bee breeding group by using the inseminated female parent worker bees and more than 30 worker bees which just emerge and come out of the house to produce offspring queen bees, and mating the offspring queen bees with the male bees to obtain an offspring queen bee colony. According to the method, the queen bee colony can be stably and rapidly obtained only through a single female parent worker bee, all genes come from the female parent worker bee, rapid propagation of bumblebees and breeding of excellent characters are facilitated, and the method has important application prospects in natural resource protection and agricultural production.

Description

Technical Field

[0001] The present invention relates to the technical field of bumblebee breeding, and in particular to a method for accelerating the directional breeding of bumblebees. Background Art

[0002] Bumblebee pollination has been proven to be an essential supporting technology for the safe and efficient production of greenhouse fruits and vegetables. It not only replaces manual labor and saves labor, but also reduces the use of hormones and pesticides, thereby improving product quality and safety, increasing farmers' incomes, and positively impacting the ecological environment. In the 1990s, developed countries in Europe and the United States achieved major breakthroughs in bumblebee successive propagation technology, enabling multiple generations of propagation per year and factory-scale production. This led to the emergence of a number of specialized breeding companies, supplying pollinating bumblebees to the global market.

[0003] However, currently, up to 90% of the bumblebees used for pollination in the greenhouse fruit and vegetable industry rely on imports, with both import quantity and price strictly restricted. In contrast, domestic bumblebee breeding companies are generally small in scale and relatively backward in technology, resulting in low colony breeding efficiency, poor colony quality and stability, and short pollination periods. Furthermore, key bumblebee breeding technologies are strictly kept secret abroad, which has become a key factor hindering the overall improvement of the bumblebee pollination industry and the sustainable and healthy development of agriculture.

[0004] The root of these problems lies in the complexity and long-term nature of bumblebee breeding. As social insects, bumblebees exhibit unique caste differentiation. Worker bees play a crucial "worker" role, never mating and unable to produce female offspring. Their key performance, especially the crucial pollination ability, is primarily embodied in worker bees, making direct breeding from queen bees difficult. Therefore, bumblebee breeding presents a greater challenge. Recently, researchers have reported that worker bees possess reproductive characteristics similar to those of queen bees (Zhuang et al., 2023), providing new insights into bumblebee breeding. Summary of the Invention

[0005] The present invention provides a method for accelerating the directional breeding of bumblebees. The method can use a single worker bee with excellent traits as a mother parent to quickly and stably breed a daughter queen bee colony with the properties of the worker bee.

[0006] In a first aspect, the present invention provides a method for accelerating the directional breeding of bumblebees, comprising the following steps: (1) Selecting the female parent: Select worker bees with target traits as the female parent for targeted breeding; (2) Producing male bees: causing the female worker bees to produce offspring male bees to obtain male bee colonies of male bees; (3) Breeding daughter queen bees: using sperm from any male bee in the male bee colony of the father to inseminate the female worker bees, breeding daughter queen bees, and then forming a queen breeding colony; (4) After the offspring queen bee reaches sexual maturity, it mates and lays eggs with any male bee in the male bee colony of the father obtained in step (2) to obtain a directed breeding bee colony.

[0007] Preferably, the target traits in step (1) of the above method include pollination performance, stress resistance, nectar collection ability, and lifespan.

[0008] Preferably, the method for obtaining the male bee colony of the father parent in step (2) of the above method is: The female worker bees are placed in a dark environment with a constant temperature and humidity of 27-29°C and 50%-60% humidity in a rearing room, and are fed with mixed pollen and diluted honey as food sources; and / or Each mother worker bee colony is accompanied by 5-10 newly emerged worker bees from other colonies.

[0009] Further preferably, the 5-10 newly emerged worker bees from other bee colonies should be replaced every 5-7 days, preferably every 6 days.

[0010] The primary task of these companion bees is to assist with nesting and other tasks. To prevent competition among worker bees for egg-laying, companion bees are replaced every six days. To accurately record the age of male bees, once a male bee leaves the hive, it should be immediately removed and housed individually, with the date of its departure clearly marked on the rearing box.

[0011] Preferably, the method for breeding daughter queen bees in step (3) of the above method comprises: collecting sperm from any male bees in the male bee colony of the father parent during the period of 6-13 days after leaving the hive, and inseminating the female worker bees; and / or The female worker bees are placed in a dark environment with a constant temperature and humidity of 27-29°C and 50%-60% humidity in a rearing room, and are fed with mixed pollen and diluted honey as food sources; and / or Select more than 30 newly emerged worker bees from other bee colonies to accompany the mother worker bees. There is no need to replace the worker bees during this stage until the daughter queen bee is successfully bred.

[0012] The present invention has found that only the first batch of larvae cultivated by the fertilized mother worker bee after forming a queen breeding colony with more than 30 worker bees will develop into queen bees. Otherwise, they may be ordinary worker bees. The daughter queen bee produced can mate with its brother male bees to obtain a daughter queen bee colony. The genetic material of this bee colony all comes from the mother worker bee, which is very suitable for the selection and breeding of excellent traits of the bee colony.

[0013] Preferably, step (4) of the above method further includes a method for raising the queen bee progeny after mating, wherein the queen bee is treated with carbon dioxide to break its diapause state and wait for it to wake up for subsequent feeding and management. This process requires anesthesia for two consecutive days, with anesthesia lasting 10 minutes each day, during which carbon dioxide must be supplied continuously. For example, the VE-AIIOQB-H1.0-A insect insemination system of Shanghai Suosheng Biological releases about 300 bubbles of carbon dioxide per minute.

[0014] Further preferably, the queen bee after waking up should be accompanied by two worker bees that have just emerged from the cell to induce the daughter queen bee to lay eggs. Before the first batch of worker bees leave the cell, the two accompanying worker bees of the queen bee need to be removed to avoid confusion between them and the daughter worker bees of the queen bee.

[0015] Preferably, the above method further comprises step (5), specifically, eliminating those poorly developed directed breeding colonies, retaining colonies in good development and meeting the standards, and at the same time, selecting worker bees with target traits as the maternal worker bees for breeding the next generation, and obtaining new varieties or new strains of bumblebees with stable traits and meeting the breeding goals through continuous backcrossing, father-daughter crosses, and inbreeding between generations for multiple generations.

[0016] Preferably, the bumblebee is a ground bumblebee, a Lanzhou bumblebee, a dense forest bumblebee, or a red light bumblebee.

[0017] In a second aspect, the present invention provides the use of the above method and the bee colony cultivated by the above method in the directed breeding of bumblebees.

[0018] The beneficial effects of the present invention are: Based on the reproductive characteristics of worker bees and their unique role as the embodiment of bee species traits, the present invention cleverly utilizes worker bees as maternal parents and their offspring male bees as paternal parents to implement efficient backcrossing, father-daughter crosses, and inbreeding directional breeding strategies between generations. This method aims to accelerate the cultivation of new bee colonies with the characteristics of maternal worker bees. In traditional breeding, queen bees often find it difficult to directly display key traits such as pollination performance. However, the present invention uses worker bees as maternal parents for directional breeding, successfully overcoming this problem. It cleverly utilizes the characteristics of newly emerged worker bees that do not compete for egg laying and egg consumption, and using 30 newly emerged worker bees, the first batch of larvae can be quickly and effectively cultivated into queen bees. Compared with traditional methods, this method is more accurate and efficient, greatly shortens the breeding cycle, and accelerates the overall process of bumblebee directional breeding. The implementation of the present invention not only points the way for technological innovation and industrial upgrading in the field of bumblebee breeding, but also is expected to cultivate more new bumblebee strains or varieties with excellent traits and strong adaptability in the future, thereby effectively promoting the vigorous development of bumblebee-related industries. DETAILED DESCRIPTION

[0019] To make the objectives, technical solutions, and advantages of the present invention more clear, the technical solutions of the present invention are described clearly and completely below. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. All other embodiments derived by persons of ordinary skill in the art based on the embodiments of the present invention without inventive effort are within the scope of protection of the present invention.

[0020] Example 1 Taking the targeted breeding of bumblebees as an example, the specific steps include: S1 selects worker bees, and chooses worker bees with strong pollination performance and stress resistance in the tomato greenhouse as female parents, and the male offspring bees they produce as male parents; S2 forms a male-producing group, and groups the worker bees selected according to the above S1 standards and places them in a breeding room. The environmental conditions in the breeding room are set as follows: a dark environment, maintaining a constant temperature and humidity, specifically, the temperature is controlled within the range of 28℃±1℃, and the humidity is maintained between 50% and 60%. In addition, these worker bees are provided with mixed pollen and diluted honey as a food source. In addition, 5 newly emerged worker bees are selected from other bee colonies in each group of worker bees to accompany these selected worker bees. It should be noted that the main task of these 5 accompanying worker bees is to assist in feeding and other nest work. In order to prevent competition for egg laying among worker bees, the accompanying worker bees need to be replaced every 6 days. In order to accurately record the age of the male bees, once the male bees leave the room, they should be immediately caught and raised separately, and the date of leaving the room should be clearly marked on the breeding box; S3: Sperm collection: Male bees can be dissected to collect sperm between 6 and 13 days after leaving the hive; S4 artificial insemination: After collecting sperm, immediately use the insect artificial insemination instrument to artificially inseminate the female parent. After the insemination is completed, wait for the female parent worker bees to wake up; S5: After the insemination worker bees wake up, newly emerged worker bees are selected from other bee colonies. Each insemination worker bee needs to select 30 newly emerged worker bees to form a queen rearing colony. During this stage, the worker bees do not need to be replaced until the daughter queen is successfully bred. The rearing conditions follow the standards described in step 2 above. The S6 daughter queen bee is mated when the daughter queen bee reaches sexual maturity and is selected when the queen bee is 5 to 7 days old. The male bees required for mating come from the male bees previously produced by its mother, that is, these male bees are brothers of the daughter queen bee in the same group; S7 daughter queen bee colony rearing, in the process of rearing daughter queen bees, the queen bee needs to use carbon dioxide to break the queen bee's diapause state after mating. It should be noted that this process requires anesthesia for two consecutive days, and anesthesia lasts for 10 minutes each day. During this period, carbon dioxide must be supplied uninterruptedly, releasing about 300 bubbles of carbon dioxide per minute. Each queen bee that wakes up should be accompanied by two worker bees that have just emerged from the cell to induce it to lay eggs. At the same time, it should be noted that before the first batch of worker bees leave the cell, the two accompanying worker bees of the queen bee need to be removed to avoid confusion between them and the queen bee's daughter worker bees; S8 selects daughter queen bee colonies, eliminates those with poor development, and retains only those with good development.

[0021] Repeated experiments have shown that the breeding method of Example 1 is used to selectively breed bumblebees to obtain bee colonies with excellent traits (strong pollination performance and stress resistance).

[0022] Example 2 The difference from Example 1 is that the 30 newly emerged Bombus terrestris worker bees in S5 are replaced by 35 newly emerged Bombus terrestris worker bees.

[0023] Example 3 The difference from Example 1 is that the 30 newly emerged Bombus terrestris worker bees in S5 are replaced by 40 newly emerged Bombus terrestris worker bees.

[0024] Comparative Example 1 The difference from Example 1 is that the 30 newly emerged Bombus terrestris worker bees in S5 are replaced by 1 newly emerged Bombus terrestris worker bee.

[0025] Comparative Example 2 The difference from Example 1 is that the 30 newly emerged Bombus terrestris worker bees in S5 are replaced by 10 newly emerged Bombus terrestris worker bees.

[0026] Comparative Example 3 The difference from Example 1 is that the 30 newly emerged Bombus terrestris worker bees in S5 are replaced by 15 newly emerged Bombus terrestris worker bees.

[0027] Comparative Example 4 The difference from Example 1 is that the 30 newly emerged Bombus terrestris worker bees in S5 are replaced by 20 newly emerged Bombus terrestris worker bees.

[0028] Comparative Example 5 The difference from Example 1 is that the 30 newly emerged Bombus terrestris worker bees in S5 are replaced by 25 newly emerged Bombus terrestris worker bees.

[0029] Test Example 1 This test example observed the first batch of larvae laid by the female worker bees after fertilization in Examples 1, 2, 3 and Comparative Examples 1-5.

[0030] Because the inseminated female worker bees are post-ovipositors, they typically lay eggs on the same day or the next day. Therefore, queen rearing colonies were formed according to the specific requirements of each group, with each group containing 20 biological replicates. The development of the first larvae laid by the inseminated worker bees into queens was observed, and the results are shown in Table 1.

[0031] Table 1 The development of queen bees from the first batch of larvae

[0032] As can be seen from the above table, after the inseminated worker bees laid eggs, in comparative example 1, one worker bee was used to feed the larvae, and the first batch did not develop into a queen bee; in comparative example 2, 10 worker bees were used to feed the larvae, and the first batch did not develop into a queen bee; in comparative example 3, 15 worker bees were used to feed the larvae, and the first batch did not develop into a queen bee; in comparative example 4, 20 worker bees were used to feed the larvae, and the first batch of larvae in some groups of bee colonies were able to develop into queen bees; in comparative example 5, 25 worker bees were used to feed the larvae, and the first batch of larvae in some groups of bee colonies were able to develop into queen bees; and in embodiment 1, when the number of worker bees reached 30 for feeding, all the first batch of larvae successfully developed into queen bees; in embodiment 2, 35 worker bees were used to feed the larvae, and all the first batch of larvae developed into queen bees; in embodiment 3, 40 worker bees were used to feed the larvae, and all the first batch of larvae also developed into queen bees. This indicates that after the mother worker bee fertilizes a queen, at least 30 additional workers are needed to ensure that all the larvae produced in the first batch develop into queens. This demonstrates that this method is feasible and effective for accelerating the directed breeding process of bumblebees.

[0033] Test Example 2 Queen bees bred using the methods described in Examples 1-3 were mated with their male siblings at 7 days old. Observation of their mating patterns revealed that the daughter queens produced by these worker bees were able to mate normally with the male bees. This demonstrates that this approach is feasible and effective for accelerating the directed breeding process of bumblebees.

[0034] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the various embodiments of the present invention.

Claims

1. A method for accelerating the directional breeding of bumblebees, characterized in that: The following steps are involved: (1) Selecting the maternal parent: Selecting worker bees with target traits as maternal worker bees for targeted breeding; (2) Producing male bees: causing the female worker bees to produce male bees of the next generation to obtain male bee colonies of male bees; (3) Breeding queen bees of the next generation: using sperm from any male bees in the male bee colony of the father to inseminate the worker bees of the mother parent to breed queen bees of the next generation, and then forming a queen breeding colony; (4) After the offspring queen bee reaches sexual maturity, it mates with any male bee in the male bee colony obtained in step (2) to lay eggs to obtain a directed breeding bee colony.

2. The method according to claim 1, characterized in that: The target traits in step (1) include pollination performance, stress resistance, nectar collection ability, and lifespan.

3. The method according to claim 1, characterized in that The method for obtaining the male bee colony of the father parent in step (2) is: The female worker bees are placed in a breeding room with a constant temperature and humidity of 27°C-29°C and a humidity of 50%-60% in a dark environment, and mixed pollen and diluted honey are used as food sources; and / or In each mother worker bee colony, there are 5-10 newly emerged worker bees from other colonies accompanying the mother worker bees.

4. The method according to claim 3, characterized in that The 5-10 worker bees that have just emerged from other bee colonies should be replaced every 5-7 days.

5. The method according to claim 1, characterized in that The method for breeding queen bees of the next generation in step (3) comprises: collecting sperm from any male bees in the male bee colony of the father during the period of 6-13 days after leaving the hive, and inseminating the female worker bees; and / or The female worker bees are placed in a breeding room with a constant temperature and humidity of 27°C-29°C and a humidity of 50%-60% in a dark environment, and mixed pollen and diluted honey are used as food sources; and / or Select more than 30 newly emerged worker bees from other bee colonies to accompany the mother worker bee. There is no need to replace the worker bees during this stage until the daughter queen bee is successfully bred.

6. The method according to claim 1, characterized in that Step (4) also includes a method for raising the offspring queen bee after mating, which is to treat the queen bee after mating with carbon dioxide to break its dormant state and wait for it to wake up so as to carry out subsequent feeding and management.

7. The method according to claim 6, characterized in that After waking up, the queen bee should be accompanied by two worker bees that have just emerged from the cell to induce the daughter queen bee to lay eggs.

8. The method according to claim 1, characterized in that: The method also includes step (5), wherein those colonies with poor development are eliminated, and colonies with good development and meeting the standards are retained. At the same time, worker bees with target traits are selected from the colonies to serve as the mother worker bees for breeding the next generation. By implementing backcrossing, father-daughter crosses, and inbreeding between generations for multiple consecutive generations, a new variety or strain of bumblebees with stable traits that meet the breeding goals is obtained.

9. The method according to any one of claims 1 to 8, characterized in that: The bumblebees are ground bumblebees, Lanzhou bumblebees, dense forest bumblebees, and red-light bumblebees.

10. Use of the method according to any one of claims 1 to 8 and the bee colony bred by the method in the directed breeding of bumblebees.

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