Sectional artificial breeding method for gleditsia sinensis bruchid based on host preference

Abstract

The invention belongs to the technical field of coleoptera bean weevil subfamily insect breeding, and particularly relates to a gleditsia sinensis bean weevil sectional type artificial breeding method based on host preference, and a breeding device comprises an egg period insect breeding box, a double-layer insect breeding box connecting table, an adult egg laying box, a hollow attractant storage box, an opening gauze box cover and a top gauze box cover; mature and undamaged wild gleditsia sinensis seeds are selected and put into a larva feeding box, gleditsia sinensis bruchid eggs are inoculated, larvae and pupae develop in the gleditsia sinensis seeds, eclosion adults are transferred into an oviposition box, gleditsia sinensis seeds are put into the oviposition box, the adults naturally mate and oviposition, egg particles are collected and transferred into a new larva feeding box, and after the eggs are hatched, the gleditsia sinensis bruchid eggs are put into the new larva feeding box. The gleditsia sinensis seeds are replaced with gleditsia sinensis seeds for larva stage breeding according to the step 4, and circulation breeding is achieved. The method is easy and convenient to operate and low in cost, the survival rate of the larvae and the egg laying amount of the imagoes can be remarkably increased, large-scale continuous breeding of the gleditsia sinensis bruchid is achieved, and a stable and high-quality insect source is provided for scientific research tests.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of feeding of liquid Coleoptera Bruchidae insects, and particularly relates to a segmented artificial feeding method for Megabruchidius dorsalis based on host preference. BACKGROUND

[0002] Megabruchidius dorsalis belongs to Coleoptera, Chrysomelidae, and Bruchinae, is a main pest of Gleditsia fruit, and lays eggs on the fruit in the forest to cause damage. The main damage stage is completely hidden, has strong damage to the seeds of Gleditsia, completely loses the activity of the seeds, and has a very short external intervention window because the life history is completed in the seeds, so that field prevention and control cannot be implemented. Megabruchidius dorsalis has strong reproductive capacity, can have 3-4 generations in a year in a storage environment, and causes great loss. The hidden property and high reproductive capacity directly lead to great difficulty in prevention and control.

[0003] Megabruchidius dorsalis mainly feeds on the seeds of Gleditsia plants, is important experimental material in the fields of research on plant and insect coevolution and insect food preference, and the like. At present, indoor breeding research on Megabruchidius dorsalis is relatively scarce, and the existing breeding methods mostly use single type of seeds as feed without considering the preference difference of Megabruchidius dorsalis to host seeds in different growth and development stages (larval stage and adult stage), so that the breeding efficiency is low, the population stability is poor, and the demand of large-scale experiments cannot be met. For example, if single Gleditsia seed is used all the time, the survival rate of larvae is low; if wild Gleditsia seed is used all the time, the egg laying amount of adults is insufficient, and the population cannot be efficiently expanded. Therefore, in view of the food preference and egg laying preference of Megabruchidius dorsalis in different development stages, designing a segmented indoor breeding method becomes the key to solve the current breeding problem of Megabruchidius dorsalis. SUMMARY

[0004] The application aims to solve the problem of lack of high-quality insect sources in scientific experiments. The method relies on the host plant preference of Megabruchidius dorsalis for directional breeding, is simple and low in cost, can realize large-scale and long-term continuous breeding of Megabruchidius dorsalis, provides stable and sufficient insect source guarantee for comprehensive prevention and control related research, and provides a segmented artificial feeding method for Megabruchidius dorsalis based on host preference, including the following steps: Step 1. Rearing device manufacturing: make a rearing device including an egg stage rearing box, a double-layer rearing box connecting table, an adult oviposition box, a hollow attractant storage box, an open mesh box cover, and a top mesh box cover, the rearing device body is a plastic rectangular container, the lower layer is an egg stage rearing box, the upper layer is an open mesh box cover, the middle layer is a double-layer rearing box connecting table, the double-layer rearing box connecting table supports an adult oviposition box, the adult oviposition box has a hollow attractant storage box, and the adult oviposition box has an open mesh box cover; Step 2. Egg stage: select mature and undamaged Gleditsia japonica seeds, place them in the larval rearing box, and the seeds are laid with a thickness of 2-3 cm, and the Callosobruchus chinensis eggs are introduced; Step 3. Larval stage rearing: larvae and pupae develop inside the Gleditsia japonica seeds; Step 4. Adult stage rearing and oviposition: transfer the emerged adults to the oviposition box, place Gleditsia japonica seeds in the oviposition box, and lay the seeds with a thickness of 2-3 cm, allowing the adults to naturally mate and lay eggs; Step 5. Next generation larval cultivation: collect the egg masses and transfer them to a new larval rearing box, replace the Gleditsia japonica seeds after the eggs hatch, and follow Step 4 for larval stage rearing to achieve cyclic reproduction.

[0005] Further, the size of the open mesh box cover 6 is 21 cm long, 13 cm wide, and 5 cm high.

[0006] Further, the size of the open mesh box cover 6 is 21 cm long, 13 cm wide, and 5 cm high.

[0007] Further, the size of the adult oviposition box 3 is 16 cm long, 8 cm wide, and 5 cm high.

[0008] Further, the larval stage rearing conditions are 28±1 ℃, relative humidity 65±5 %, and photoperiod L:D=10:14 h.

[0009] Further, the Gleditsia japonica and Gleditsia japonica seeds used in Steps 3 and 4 for rearing larvae and adults for oviposition are stored in a refrigerator at -20 ℃ for 48 h before use.

[0010] Further, the rearing device is made of transparent plastic, and the open mesh box cover 5 and the top mesh box cover 6 have air vents with a pore size of 0.5 cm, and the air vents are covered with insect-proof mesh.

[0011] Further, in Step 4, the adults are transferred to the oviposition box using attractants after emergence.

[0012] Compared with the existing technology, the segmented artificial rearing method of Callosobruchus chinensis based on host preference and the method have the following advantages: 1、The application is based on the host preference characteristics of Callosobruchus chinensis at different growth stages, and adopts a segmented feeding strategy accordingly. The number of holes on the seed coat of wild soybean is the largest, which is significantly higher than that of Japanese soybean and soybean, being 3.7 times and 2.3 times that of the latter, respectively. The egg stage, intracapsular stage and pre-adult stage of Callosobruchus chinensis on wild soybean are 6.83 d, 25.38 d and 32.19 d, respectively, which are lower than those on Japanese soybean and soybean. The emergence rate is 0.42, which is higher than that on Japanese soybean and soybean. The egg stage and larval stage select wild soybean seeds as feed, which meets the growth and development needs of larvae, can significantly improve the survival rate and development rate of larvae, and reduce the mortality rate of larvae.

[0013] 2、The application is based on the host preference characteristics of Callosobruchus chinensis at different growth stages, and adopts a segmented feeding strategy accordingly. The number of holes on the seed coat of wild soybean is the largest, which is significantly higher than that of Japanese soybean and soybean, being 3.7 times and 2.3 times that of the latter, respectively. The egg stage, intracapsular stage and pre-adult stage of Callosobruchus chinensis on wild soybean are 6.83 d, 25.38 d and 32.19 d, respectively, which are lower than those on Japanese soybean and soybean. The emergence rate is 0.42, which is higher than that on Japanese soybean and soybean. The egg stage and larval stage select wild soybean seeds as feed, which meets the growth and development needs of larvae, can significantly improve the survival rate and development rate of larvae, and reduce the mortality rate of larvae.

[0014] 3、The application is based on the host preference characteristics of Callosobruchus chinensis at different growth stages, and adopts a segmented feeding strategy accordingly. The number of holes on the seed coat of wild soybean is the largest, which is significantly higher than that of Japanese soybean and soybean, being 3.7 times and 2.3 times that of the latter, respectively. The egg stage, intracapsular stage and pre-adult stage of Callosobruchus chinensis on wild soybean are 6.83 d, 25.38 d and 32.19 d, respectively, which are lower than those on Japanese soybean and soybean. The emergence rate is 0.42, which is higher than that on Japanese soybean and soybean. The egg stage and larval stage select wild soybean seeds as feed, which meets the growth and development needs of larvae, can significantly improve the survival rate and development rate of larvae, and reduce the mortality rate of larvae. BRIEF DESCRIPTION OF DRAWINGS

[0015] Figure 1 is a front view of the feeding device in the segmented artificial feeding method of Callosobruchus chinensis based on host preference provided by the application; Figure 2 is a top view of the feeding device in the segmented artificial feeding method of Callosobruchus chinensis based on host preference provided by the application; Figure 3 is the preference selection of Callosobruchus chinensis larvae on different host foods in the segmented artificial feeding method of Callosobruchus chinensis based on host preference provided by the application; Figure 4 is the preference selection of Callosobruchus chinensis adults on different hosts in the segmented artificial feeding method of Callosobruchus chinensis based on host preference provided by the application. DETAILED DESCRIPTION

[0016] The following examples are for illustrative purposes only and are not intended to limit the scope of the application.

[0017] The application provides a segmented artificial feeding method for Callosobruchus chinensis based on host preference, which comprises the following steps. Step 1. Feeding device manufacturing: referring to Figures 1-2 , the feeding device is manufactured, and the feeding device comprises an egg stage insect rearing box 1, a double-layer insect rearing box connecting table 2, an adult egg laying box 3, a hollow attractant storage box 4, an open mesh box cover 5 and a top mesh box cover 6. The main body of the feeding device is a plastic cuboid container. The lower layer is the egg stage insect rearing box 1 with a length of 21 cm, a width of 13 cm and a height of 5 cm. The upper layer is the open mesh box cover 6 with a length of 21 cm, a width of 13 cm and a height of 7 cm. The middle layer is the double-layer insect rearing box connecting table 2 supporting the adult egg laying box 3 with a length of 16 cm, a width of 8 cm and a height of 5 cm. The adult egg laying box 3 has the hollow attractant storage box 4. The adult egg laying box 3 has the open mesh box cover 5. The feeding device is made of transparent plastic. The open mesh box cover 5 and the top mesh box cover 6 are provided with air holes with a diameter of 0.5 cm, and the air holes are covered with insect-proof nets.

[0018] The egg stage insect rearing box 1 is used for feeding Callosobruchus chinensis in the egg stage, the larva stage and the pupa stage. The double-layer insect rearing box connecting table 2 is detachable and used for supporting the adult egg laying box 3. The adult egg laying box 3 provides food types different from those in the egg stage and is used for Callosobruchus chinensis in the adult stage to lay eggs. The hollow attractant storage box 4 is used for attracting Callosobruchus chinensis adults after emergence into the egg laying box. The open mesh box cover 5 is provided with a through hole in the middle part, so that Callosobruchus chinensis adults can easily enter the adult egg laying box. The top mesh box cover 6 prevents Callosobruchus chinensis from flying away, and the mesh can provide sufficient air. All components can be disassembled and cleaned, repeated use is supported, and the long-term feeding cost is reduced. The whole device is placed in an artificial climate box, and the standard feeding conditions are set as follows: temperature 28±1 ℃, relative humidity 65±5%, light cycle L:D = 10:14 h, and the summer and autumn season environment of the natural habitat is simulated. Step 2. Egg stage: mature wild soybean seeds without damage and stored in a refrigerator at-20 ℃ for 48 h are selected and placed in the larva rearing box. The seeds are laid with a thickness of 2-3 cm to avoid over-thick stacking and cause hypoxia or uneven temperature and humidity. Then, the Callosobruchus chinensis eggs collected in the laboratory or produced by the previous generation are evenly connected to the surface of the seeds. Step 3. Larva stage feeding: the larvae and pupae develop in the wild soybean seeds, and the whole larva stage and subsequent pupa stage are completed in the seeds without additional intervention. The formation of insect holes on the seed surface is observed regularly, which is an intuitive index of larva survival and development progress. Step 4. Adult feeding and oviposition: After the adult emergence, most individuals will climb and enter the upper oviposition box through the through-hole driven by the volatile odor of the attractant. The mature Gleditsia japonica seeds that have been refrigerated at -20℃ for 48 hours are placed in the oviposition box. The seeds are laid in a thickness of 2-3 cm. The stronger oviposition attraction of Gleditsia japonica to the adults is used to promote their concentrated mating and oviposition, thereby significantly improving the egg density and collection efficiency. Step 5. Next-generation larval cultivation: Fresh egg masses attached to the surface of the Gleditsia japonica seeds are carefully collected using a soft hairbrush or tweezers and transferred to a new larval rearing box. After the eggs are hatched, the wild Gleditsia japonica seeds are replaced and the larval rearing is performed according to step 4 to realize continuous and stable expansion of the population.

[0019] Table 1 G. heterophylla G. japonica G. sinensis Egg stage (d) 6.83±0.09 7.26±0.08 9.35±0.05 Inside pod stage (d) 25.38±0.12 25.62±0.17 27.81±0.20 Preadult duration (d) 32.19±0.17 32.85±0.21 37.06±0.20 Fledge rate (%) 42.00±4.00 35.00±2.00 35.00±2.00 Note: Table 1 is the influence of different hosts on the growth and development of Callosobruchus chinensis according to the embodiments of the present application. Figures 3-4 In the table, Gh, Gj and Gs represent wild Gleditsia japonica seeds, Japanese Gleditsia japonica seeds and Gleditsia japonica seeds, respectively, and different lowercase letters represent significant differences (P<0.05).

[0020] The above is only a preferred embodiment of the present application and is not intended to limit the present application. Any modification, equivalent replacement, improvement, etc. made within the spirit and principles of the present application shall be included in the protection scope of the present application.

Claims

1. A segmented artificial rearing method of Callosobruchus chinensis based on host preference, characterized by, The method comprises the following steps: Step 1. Rearing device manufacturing: a rearing device is manufactured, which comprises an egg stage insect rearing box (1), a double-layer insect rearing box connecting table (2), an adult egg laying box (3), a hollow attractant storage box (4), an open mesh box cover (5) and a top mesh box cover (6). The rearing device body is a plastic cuboid container, the lower layer is the egg stage insect rearing box (1), the upper layer is the open mesh box cover (6), and the middle layer is the double-layer insect rearing box connecting table (2). The double-layer insect rearing box connecting table (2) supports the adult egg laying box (3), the adult egg laying box (3) has the hollow attractant storage box (4) therein, and the adult egg laying box (3) has the open mesh box cover (5) thereon; Step 2. Egg stage: mature and undamaged wild Gleditsia japonica seeds are selected and placed in the larva rearing box, and the seeds are laid with a thickness of 2-3 cm, and the Callosobruchus chinensis eggs are introduced; Step 3. Larva stage rearing: the larvae and pupae develop in the wild Gleditsia japonica seeds; Step 4. Adult stage rearing and egg laying: the hatched adults are transferred to the egg laying box, Gleditsia japonica seeds are placed in the egg laying box, the seeds are laid with a thickness of 2-3 cm, and the adults are allowed to naturally mate and lay eggs; Step 5. Next generation larva cultivation: the egg grains are collected and transferred to a new larva rearing box, and after the eggs are hatched, wild Gleditsia japonica seeds are replaced for larva stage rearing according to step 4 to realize cyclic propagation.

2. The segmented rearing method of Callosobruchus chinensis based on host preference according to claim 1, characterized in that, The size of the open mesh box cover (6) is 21 cm in length, 13 cm in width and 5 cm in height.

3. The segmented rearing method of Callosobruchus chinensis based on host preference according to claim 1, characterized in that, The size of the open mesh box cover (6) is 21 cm in length, 13 cm in width and 5 cm in height.

4. The segmented rearing method of Callosobruchus chinensis based on host preference according to claim 1, characterized in that, The size of the adult egg laying box (3) is 16 cm in length, 8 cm in width and 5 cm in height.

5. The segmented rearing method of Callosobruchus chinensis based on host preference according to claim 1, characterized in that, The larva stage rearing conditions are 28±1 ℃, relative humidity 65±5 %, and light cycle L:D=10:14 h.

6. The segmented rearing method of Callosobruchus chinensis based on host preference according to claim 1, wherein, The wild Gleditsia japonica and Gleditsia japonica seeds provided for rearing larvae and adults to lay eggs in steps 3 and 4 are used after being refrigerated at-20 ℃ for 48 h.

7. The segmented rearing method of Callosobruchus chinensis based on host preference according to claim 1, characterized in that, The rearing device is made of transparent plastic material, and the open mesh box cover (5) and the top mesh box cover (6) are provided with air vents with a pore size of 0.5 cm, and the air vents are covered with an insect-proof net.

8. The segmented rearing method of Callosobruchus chinensis based on host preference according to claim 1, wherein, In step 4, the adults are transferred to the egg laying box using the attractant after hatching.

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