A method for accelerating directional breeding of bumblebees

Abstract

The present invention provides a method for accelerating the directional breeding of bumblebees, specifically, selecting worker bees with excellent traits as maternal worker bees for directional breeding, laying eggs and hatching them into male bees in a dark environment with a temperature range of 27°C-29°C and a humidity maintained between 50% and 60%, obtaining sperm from the male bees after sexual maturity, using the obtained sperm to fertilize the maternal worker bees, and forming a breeding group with more than 30 worker bees that have just emerged from the room to produce daughter queen bees, so that the daughter queen bees mate with the aforementioned male bees to obtain daughter queen bee colonies. This method can stably and quickly obtain a queen bee colony only through a single maternal worker bee, and all genes come from the maternal worker bee, which is conducive to the rapid reproduction of bumblebees and the breeding of excellent traits, and has important application prospects in natural resource protection and agricultural production.

Description

Technical Field

[0001] The 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 important supporting technology for the safe and efficient production of greenhouse fruits and vegetables. It can not only replace manual labor and save labor, but also reduce the use of hormones and pesticides, thereby improving product quality and safety, increasing farmers' income, and having a positive impact on the ecological environment. In the 1990s, developed countries in Europe and the United States made major breakthroughs in bumblebee succession breeding technology, achieving multi-generation breeding and factory production in one year, and a number of professional breeding companies emerged to supply pollination bumblebees to the global market.

[0003] However, up to 90% of the bumblebees used for pollination in the current greenhouse fruit and vegetable industry rely on foreign imports, and the import quantity and price are 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 stability, and short pollination time. What is more serious is that foreign countries strictly keep the key technologies of bumblebee breeding confidential, which has become a key factor restricting the comprehensive 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. Bumblebees are social insects with unique caste differentiation characteristics, among which worker bees play a vital role as "workers". They do not mate for life and cannot produce female offspring. Their key performance, especially the crucial pollination performance, is mainly reflected in worker bees, which is difficult to breed directly from queen bees. Therefore, the breeding of bumblebees faces greater challenges. Recently, the inventors reported that worker bees have reproductive characteristics similar to queen bees (Zhuang et al., 2023), which provides new ideas for bumblebee breeding. Summary of the invention

[0005] The 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:

[0007] (1) Selecting the maternal parent: Selecting worker bees with target traits as maternal worker bees for targeted breeding;

[0008] (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;

[0009] (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;

[0010] (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.

[0011] Preferably, the target traits in step (1) of the above method include pollination performance, stress resistance, nectar collection ability, and lifespan.

[0012] Preferably, the method for obtaining the male bee colony of the father parent in step (2) of the above method is:

[0013] 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

[0014] In each mother worker bee colony, there are 5-10 newly emerged worker bees from other colonies accompanying the mother worker bees.

[0015] Further preferably, the 5-10 worker bees that have just emerged from other bee colonies should be replaced every 5-7 days, preferably every 6 days.

[0016] The main task of these accompanying worker bees is to assist in feeding and other work in the nest. In order to prevent competition among worker bees for laying eggs, the accompanying worker bees need to be replaced every 6 days. In order to accurately record the age of male bees, once the male bees leave the hive, they should be caught and raised separately, and the date of leaving the hive should be clearly marked on the feeding box.

[0017] Preferably, the method for breeding offspring queen bees in step (3) of the above method 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

[0018] 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

[0019] 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.

[0020] The present invention finds that only the first batch of larvae cultivated by the fertilized mother worker bee after forming a queen breeding group with more than 30 worker bees will develop into queen bees, otherwise they may be ordinary worker bees, and the produced daughter queen bees can mate with their brother male bees to obtain a daughter queen bee colony. The genetic material of the colony all comes from the mother worker bee, which is very suitable for the selection and breeding of excellent traits of the colony.

[0021] Preferably, the above method step (4) also includes a method for raising the queen bee offspring 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 for subsequent feeding and management. This process requires anesthesia for two consecutive days, and the anesthesia lasts for 10 minutes each day. During this period, carbon dioxide needs to be supplied uninterruptedly. Taking the VE-AIIOQB-H1.0-A insect insemination system of Shanghai Suosheng Biology as an example, about 300 bubbles of carbon dioxide are released per minute.

[0022] Further preferably, the queen bee after waking up should be accompanied by two worker bees that have just emerged from the cell, so as 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.

[0023] Preferably, the method further comprises step (5), specifically, removing those poorly developed directed breeding colonies, retaining colonies with good development and meeting the standards, and selecting worker bees with target traits as the maternal worker bees for breeding the next generation, and obtaining a new variety or strain of bumblebees with stable traits and meeting the breeding objectives by implementing backcrossing, father-daughter crosses, and inbreeding between generations for multiple consecutive generations.

[0024] Preferably, the bumblebee is a ground bumblebee, a Lanzhou bumblebee, a dense forest bumblebee, or a red-light bumblebee.

[0025] In a second aspect, the present invention provides the use of the above method and the bee colony bred by the above method in the directed breeding of bumblebees.

[0026] The beneficial effects of the present invention are:

[0027] The present invention is based on the reproductive characteristics of worker bees and their unique role as the embodiment of bee species traits, cleverly using worker bees as maternal parents and their offspring male bees as paternal parents, and implementing efficient backcrossing, father-daughter crosses, and inbreeding directional breeding strategies between generations. This method aims to accelerate the cultivation of new bee colonies with maternal worker bee characteristics. In traditional breeding, queen bees are often difficult to directly show key traits such as pollination performance, and the present invention uses worker bees as maternal parents for directional breeding, successfully overcoming this problem. The characteristics of worker bees that have just emerged from the room will not compete for laying eggs and eating eggs are cleverly utilized, and 30 newly emerged worker bees can quickly and effectively cultivate the first batch of larvae 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 out the direction for technological innovation and industrial upgrading in the field of bumblebee breeding, but also is expected to cultivate more new strains or new varieties of bumblebees with excellent traits and strong adaptability in the future, thereby effectively promoting the vigorous development of bumblebee-related industries. DETAILED DESCRIPTION

[0028] In order to make the purpose, technical solution and advantages of the present invention clearer, the technical solution of the present invention will be described clearly and completely below. Obviously, the described embodiments are 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.

[0029] Example 1

[0030] Taking the targeted breeding of bumblebees as an example, the specific steps include:

[0031] S1 selects worker bees, and selects worker bees with strong pollination performance and stress resistance in the tomato greenhouse as female parents, and the male bees produced by them as male parents;

[0032] S2 forms a male production group. The worker bees selected according to the above S1 standards are grouped and placed in a breeding room. The environmental conditions in the breeding room are set as follows: a dark environment, maintaining a constant temperature and humidity state, 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 worker bees that have just emerged from the cell 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 work in the nest. 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 male bees, once the male bees leave the cell, they should be caught immediately for separate feeding, and the date of leaving the cell should be clearly marked on the breeding box;

[0033] S3: Sperm collection: Male bees can be dissected to collect sperm between 6 and 13 days after leaving the hive;

[0034] S4 artificial insemination: after collecting sperm, immediately use the insect artificial insemination instrument to artificially inseminate the female parent, and wait for the female parent worker bees to wake up after the insemination is completed;

[0035] S5: Formation of a queen rearing colony. After the insemination worker bees wake up, select the newly emerged worker bees from other bee colonies. Each insemination worker bee needs to select 30 newly emerged worker bees to form a colony. There is no need to replace the worker bees at this stage until the daughter queen bee is successfully bred. The rearing conditions follow the standards described in step 2 above.

[0036] The S6 queen bee daughters will mate when they reach sexual maturity, and the mating will be carried out when the queen bee reaches 5 to 7 days old. The male bees needed for mating come from the male bees previously laid by their mother, that is, these male bees are brothers of the same group of the queen bee daughters;

[0037] S7 colony rearing of daughter queens. In the process of rearing daughter queens, the queen bee needs to use carbon dioxide to break the queen bee's dormant state after mating. It should be noted that this process requires anesthesia for two consecutive days, and the anesthesia lasts for 10 minutes each day. During this period, carbon dioxide needs to 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 daughter worker bees of the queen bee;

[0038] S8 selects daughter queen colonies, removes those that are poorly developed, and retains only those that are in good development condition.

[0039] 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).

[0040] Example 2

[0041] The difference from the first embodiment is that the 30 newly emerged Bombus terrestris worker bees in S5 are replaced by 35 newly emerged Bombus terrestris worker bees.

[0042] Example 3

[0043] The difference from the first embodiment is that the 30 newly emerged Bombus terrestris worker bees in S5 are replaced by 40 newly emerged Bombus terrestris worker bees.

[0044] Comparative Example 1

[0045] The difference from the first embodiment is that the 30 newly emerged Bombus terrestris worker bees in S5 are replaced by 1 newly emerged Bombus terrestris worker bee.

[0046] Comparative Example 2

[0047] The difference from the first embodiment is that the 30 newly emerged Bombus terrestris worker bees in S5 are replaced by 10 newly emerged Bombus terrestris worker bees.

[0048] Comparative Example 3

[0049] The difference from the first embodiment is that the 30 newly emerged Bombus terrestris worker bees in S5 are replaced by 15 newly emerged Bombus terrestris worker bees.

[0050] Comparative Example 4

[0051] The difference from the first embodiment is that the 30 newly emerged Bombus terrestris worker bees in S5 are replaced by 20 newly emerged Bombus terrestris worker bees.

[0052] Comparative Example 5

[0053] The difference from the first embodiment is that the 30 newly emerged Bombus terrestris worker bees in S5 are replaced by 25 newly emerged Bombus terrestris worker bees.

[0054] Test Example 1

[0055] 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.

[0056] Since the female worker bees that inseminate are post-ovipositors, they will generally lay eggs on the same day or the next day, so the queen rearing groups were directly formed according to the specific requirements of each group, and each group contained 20 biological replicates. The first batch of larvae laid by the inseminated worker bees were observed to develop into queens, and the results are shown in Table 1.

[0057] Table 1 The first batch of larvae developed into queen bees

[0058]

[0059] As can be seen from the above table, after the fertilized worker bees laid eggs, in Comparative Example 1, 1 worker bee was used to feed the larvae, and the first batch of larvae did not develop into queen bees; in Comparative Example 2, 10 worker bees were used to feed the larvae, and the first batch of larvae did not develop into queen bees; in Comparative Example 3, 15 worker bees were used to feed the larvae, and the first batch of larvae did not develop into queen bees; 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 could 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 could develop into queen bees; and in Example 1, when the number of worker bees reached 30 for feeding, all the first batch of larvae successfully developed into queen bees; in Example 2, 35 worker bees were used to feed the larvae, and the first batch of larvae all developed into queen bees; in Example 3, 40 worker bees were used to feed the larvae, and the first batch of larvae also all developed into queen bees. This means that in the formation of a queen breeding colony after the female worker bee fertilizes, at least 30 other worker bees are needed to ensure that all the first batch of larvae produced develop into queen bees. This proves that this plan is feasible and effective in accelerating the directional breeding process of bumblebees.

[0060] Test Example 2

[0061] The queen bee bred by the method described in Examples 1-3 was mated with its brother male bees when it reached 7 days old. The mating was observed and it was found that the daughter queen bees produced by the worker bees were able to mate normally with the male bees. This proves that the scheme is feasible and effective for accelerating the directional breeding process of bumblebees.

[0062] 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 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 of the father obtained in step (2) to lay eggs to obtain a directed breeding bee colony; When breeding the queen bee offspring in step (3), more than 30 newly emerged worker bees are selected from other bee colonies to accompany the mother worker bees; The directional breeding is to use a single worker bee with excellent traits as the mother parent to quickly and stably breed a colony of queen bees with the properties of the worker bee.

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 in directed breeding of bumblebees.

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