CN113519468B - Method for feeding phyllotreta striolata in indoor generation - Google Patents
Method for feeding phyllotreta striolata in indoor generation Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
Abstract
The invention discloses an indoor generation feeding method of phyllotreta striolata, which is used for continuously feeding four insect states of one generation of phyllotreta striolata at 26+/-1 ℃ and relative humidity of 60% -80%. Wrapping wet cotton in the rape root, wrapping black cotton cloth, placing into a feeding bottle, and inoculating male and female flea beetles; after the adults spawn, collecting egg grains on black cotton cloth under a stereoscopic vision, placing the egg grains on black cotton cloth slices with agar as a substrate in a 24-hole culture plate hole, and placing radish discs as larva foodstuff; eggs are hatched on black cotton cloth and then become adults after larva, pre-pupa and pupa stages, the adults are collected by a self-made insect fluke device and are differentiated into male and female, and then the eggs enter the next generation of breeding. The invention has simple operation, high breeding efficiency and low cost, can keep the breeding and population reproduction of continuous generation in the phyllotreta striolata chamber, is convenient for breeding, observing and taking the test insects in each growth period, thereby meeting the requirements of population genetic research, further meeting the continuous supply of the test insects in each development period required by scientific research and providing an important basis for scientific research of the phyllotreta striolata and improvement of field control strategies.
Description
Technical Field
The invention relates to a method for feeding phyllotreta striolata in an indoor generation.
Background
Yellow flea beetles Phyllotreta striolata (Fabricius), belonging to the Coleoptera (Coleoptera) phyllotoferae (Chrysomelidae) flea beetle (altecina), are important crucifer vegetable pests that are widely distributed in asia, europe, and america. Yellow flea beetles are serious in the south of China, and the level of harm to cruciferous vegetables in Guangdong and Fujian in individual years has been approaching or even exceeding that of plutella xylostella. Yellow flea beetles are different in generation 2-8 in one year in China, have peaks in spring and autumn, and can reproduce in Guangdong warm and humid areas all the year round.
The phyllotreta oligovora is most favored by cruciferous crops such as rape, cabbage, radish, cabbage, mustard and the like. The adults start to move at 14 ℃, can jump and fly quickly at about 18 ℃, eat a lot of the adults at 20-34 ℃, reach the movement peak at the average daily air temperature of 18-24 ℃, lay eggs in soil seams around plant roots, the hatching temperature of eggs is 26 ℃, and the optimal relative humidity is 100%. The yellow flea beetle grows in four stages of egg, larva, pupa and adult, the larva is in three stages, before entering the pupa stage, the larva has a period of no food, no movement and shortened thickening of the organism, and the period from egg to pupa is soil. The yellow flea beetles are harmful by feeding adults and larvae on the ground and the underground parts respectively and overlapping generations, the adults mainly feed on growing points and leaves of plants to form a large number of holes, and the seedlings are destroyed in a large area when serious; the larvae eat the roots of plants to cause rotten roots, so that the plants wilt and die, and the soft rot of cruciferous vegetables can be transmitted, the yield and quality of the vegetables are seriously affected, and the income of farmers is reduced. At present, phyllotreta striolata mainly depends on pesticide control, and the drug resistance of the phyllotreta striolata is rapidly increased due to unreasonable application of chemical pesticides, so that control is difficult.
Because the space distribution of the phyllotreta striolata in different development periods has specificity, the phyllotreta striolata is harmful on the adult ground and spawns on the shallow soil layer root system, and the larva is harmful underground and pupates in the soil, the indoor generation feeding of the phyllotreta striolata is very difficult, and the development of the biology, ecology, toxicology research and field control technology of the phyllotreta striolata is limited.
At present, the existing feeding method has certain defects. In the method for raising the experimental population of phyllotreta striolata (ZL 200910037640.9), the adult is raised by an insect raising box, the larva is raised by an insect raising box or a culture dish, a pupation place is the culture dish added with wet sand, the method can obtain test insects in different insect states, but the method needs to transfer adults or egg black cloth for many times, the loss is easy to cause, and the larva raising box (culture dish) paved with the egg black cloth is directly put into radish skin, the mould is easy to cause in the box, the larva is easy to damage by replacing the radish skin or transferring the larva, then the mature larva is required to be picked up and pupated in the screened wet sand, the workload is large, and the pupation condition is difficult to observe. Chinese patent 'an artificial feeding method of phyllotreta striolata adults' (ZL 201410164446.8) takes cabbage leaves as food materials and transparent mineral water bottles as feeding bottles, so that adult living bodies screened by the agents can be obtained, but larvae in all ages and other insect states cannot be obtained. In addition, there are many methods of breeding in the prior art, and there are some disadvantages. In the aspect of adult feeding, the simple vegetable She Weiyang is easy to lose water and needs to be replaced for many times; when laying cloth in the spawning box (bottle) to collect eggs, the eggs are easy to die due to water loss of the dry eggs on the cloth; the method of paving fine sand under the cover bottle is easy to cause adults to be stained on the sand, which is unfavorable for activities and mating. In the aspect of egg storage, the method of directly paving black cloth or filter paper in a culture dish is difficult to control humidity, the black cloth is easy to dry or mold, the development of eggs is not facilitated, and the incubation of the eggs cannot be directly observed; the method for collecting the eggs from the soil is low in efficiency, and large loss is easy to cause in the process of washing eggs by using a buoyancy method. In the aspect of larva feeding, a method of directly placing radish slices or seedlings with roots on black cloth in a spawning box (bottle) is easy to cause radish spoilage or seedling wilting, needs to be replaced for a plurality of times, has large workload, is extremely easy to cause damage to the bodies of the insects in the replacement process, and is not beneficial to larva development; the method of putting the larvae into the finger-shaped pipe after wrapping the plant roots is that the roots are easy to rot, the larvae are similar in color to the roots and are good in concealment, the larvae are difficult to observe and take, and the foodstuff is complicated to replace and easy to damage; the method for feeding the larvae into the white porcelain trays with vegetable seedlings is difficult to find and pick after the larvae enter soil, is difficult to observe the development age, has low taking efficiency and large loss, and is easy to influence the larvae in non-target age; the method for feeding the larvae into the radish slices or the radish concave holes in the culture dish has the defects of difficult control of humidity, large workload of replacing foodstuff and easy damage to the larvae. In the aspect of preservation of pupa, the method for transferring mature larva or pupa into fine sand has large workload, is easy to damage during transfer, pupa in sand is not easy to find, the eclosion time is difficult to grasp, and the method for transferring larva or pupa into plant root for wrapping needs to spray water for multiple times for moisturizing, so that the eclosion condition of the pupa can not be observed.
The feeding of the indoor generation of the phyllotreta striolata has important significance for the research of the growth and development rule of the phyllotreta striolata, can lay a foundation for the research of biology, physiology, biochemistry and molecular toxicology, and provides a basic research material for the development of the phyllotreta striolata field control technology and the improvement of the control strategy.
Disclosure of Invention
Aiming at the problems in the field, the invention provides an indoor generation feeding method for phyllotreta striolata, which can realize generation continuous feeding, ensure indoor continuation of population, provide four insect states for phyllotreta striolata in each growth period and facilitate the progress of a plurality of important scientific researches. Meanwhile, the breeding method is simple and efficient, and can realize indoor subculture of the population and continuous supply of test insects in each growth period by adopting a simple laboratory device.
In order to achieve the purpose, the technical scheme of the method for indoor generation feeding of phyllotreta striolata comprises the following steps:
(1) Breeding and egg collection of phyllotreta striolata adults
(1) Digging out the whole rape with roots, cleaning and wiping the leaves and the roots;
(2) wrapping the root with wet cotton, wrapping with black cotton strips until no cotton is exposed, and vertically placing the plant into an insect-raising bottle;
(3) the same number of yellow stripe flea beetles female and male adults are inoculated into the insect-raising bottle, and the insect-raising bottle is tightly covered;
(4) preparing a sterile 24-well culture plate, and adding 1mL of agar solution containing preservative with a certain proportion into the well by using a pipetting gun;
(5) cooling agar, and adding black cotton cloth wafer into the hole to make it adhere to agar;
(6) when receiving eggs, firstly, temporarily collecting the adults by using a fluke device, then taking down black cloth wrapped by plant roots, placing under a stereoscopic vision mirror, adjusting the multiple of the stereoscopic vision mirror, picking egg grains on the inner surface and the edge of the stereoscopic vision mirror by using a fine hair pen while observing, placing eggs on a black cloth wafer in a culture plate hole at a position close to the hole wall, and covering a transparent culture plate cover;
(2) Egg hatching and raising of larvae of different ages
A. On day 3 after egg collection, round radish slices were placed in the center of the wells of the oval-containing culture plates;
B. after the eggs are hatched, the larvae of all ages are actively fed before the initial hatching and the pre-pupation;
(3) Pupa management and adult collection
a. The larvae are pre-pupated and pupated on black cotton cloth pieces beside the radish pieces in the holes, and the radish pieces in the holes can be taken out after pupation;
b. after the pupae emerged as adults, the adults were collected with a fluke trap and sex was differentiated under a body view mirror.
In the steps (1) - (3), the adults are fed at 26+/-1 ℃ and with humidity of 60% -80%, and the illumination is 12L:12D in a climatic chamber; the culture plates with eggs, larvae and pupae are placed in a light-tight storage box and also placed in a climate chamber under the above conditions.
In the step (1), the rape is in 8-9 leaf stage, the leaves are hypertrophic, and the drought resistance of plants is high.
In the step (1), the root of the defatted cotton with the thickness of 0.5cm is wetted and wrapped by sterile water, and the cotton is wrapped by black cotton strips layer by layer, is tight and loose in the inside and the outside, so that a spawning space is provided for adults.
In the step (1), the insect-raising bottle is a transparent plastic bottle with the diameter of 22cm and the height of 23cm, and is sterilized by 0.1% calcium hypochlorite before use, 3-5 holes with the diameter of 2cm are formed in the bottle cover, and 400 meshes of gauze are adhered below the holes; 30 yellow stripe flea beetles are respectively connected into the insect-raising bottle.
In the step (1), the egg collection period can be determined according to the requirement, and the single egg collection period is not more than 3 days.
In the step (1), after ovum collection, new wet black cloth is taken to wrap the root of the plant for continuous feeding, and the plant replacement period is 3 days.
In the step (1), the inner diameter of a single hole of a 24-hole culture plate is 0.8cm and the height is 2cm.
In the step (1), the diameter of the black cotton cloth wafer is 0.8cm.
In the step (1), the number of eggs placed in each hole of the 24-hole culture plate is determined according to the requirement, and 1-5 eggs can be placed.
In the step (1), the pipette is a 5mL micro-pipette, and the tip of the used pipette needs to be cut off by 1cm.
In the step (1), the preservative in the agar and the content of the preservative are respectively 80 ten thousand units of penicillin 0.019 percent and potassium sorbate 0.19 percent.
In the step (2), the diameter of the round radish slices placed in the culture plate holes is 0.5cm, and the thickness is 3.5mm.
In the step (2), if the larvae do not need to be hatched, circular radish slices can be put in the holes of the culture plates while eggs are put in.
In the steps (2) - (3), larvae, pre-pupae, pupae and adults in each age can be observed through the plate cover without opening the culture plate.
In the step (3), whether the radish slices are taken out after pupation or not can be determined according to the fresh state.
The invention relates to an indoor generation feeding method of phyllotreta striolata, wherein the manufacturing method of the trematode device comprises the following steps:
1) Taking a transparent 50mL centrifuge tube, and punching two holes with the diameter of 0.4cm on the middle position of a tube cover by using a hot-melting device;
2) One hole on the tube cover is inserted with a rubber tube with the inner diameter of 0.4cm and the length of 30cm, one end of the tube extends out of the inner side of the cover for 0.3cm and is sealed by a 400-mesh gauze, and the other end is inserted with a pipette tip (with the specification of 1 mL) with the tip of 1cm cut off as a suction nozzle;
3) A rubber tube with an inner diameter of 0.4cm and a length of 15cm is inserted into the other hole of the tube cover, one end of the tube extends out of the inner side of the cover by 0.3cm, and the other end of the tube is inserted into a pipette tip (with a specification of 0.2 mL) with a tip end of 0.4cm cut off to serve as an insect inlet.
The beneficial effects of the invention are as follows:
the invention realizes the feeding of four insect states and the reproduction of population in the indoor generation of the phyllotreta striolata, and has the following beneficial effects. (1) In the invention, adult rape plants are fed into a light-transmitting and breathable insect-breeding bottle, and lay eggs on black collecting cloth at the root, which is similar to the field conditions; the cotton wrapped by the plant root has strong moisture retention, ensures moisture supply, maintains freshness and reduces replacement times; meanwhile, the black cotton cloth for collecting eggs at the root is tight and loose, the mode of wrapping the eggs layer by layer can provide an effective spawning space for adults, and the wet black cloth avoids the water loss of the eggs, improves the survival rate, and has strong contrast ratio to the color of the eggs. (2) In the invention, the larvae are fed into the 24-hole culture plate taking agar as a substrate, fresh radish slices are taken as foodstuff, and the agar substrate (containing preservative) can provide proper environmental humidity for the fresh radish slices and the growth of eggs, larvae and pupae on one hand, and on the other hand, the growth of harmful microorganisms in the environment is effectively avoided, the radish spoilage is prevented, and the insect test is prevented from being infected by pathogenic bacteria. (3) In the invention, the self-made insect collector is used for collecting the emerging adults, the collecting pipe of the insect collector can be replaced at any time, classification of different populations is facilitated, and meanwhile, the transparent pipe wall is convenient for distinguishing the gender of the adults under the body view mirror.
Compared with other feeding methods, the invention has the following advantages. (1) The feeding method is suitable for the growth requirement of phyllotreta striolata, and the adopted artificial climate has good indoor air circulation and is convenient for temperature and humidity control; adults and larvae are fed separately, so that the arrangement is clear, and the deterioration of feeding environment and the vertical transmission of germs are avoided; the breeding method in each growth period is simple, the breeding efficiency is high, and the test insects are convenient to take; the device for raising is prepared in laboratory and has low cost. (2) In the invention, the states of adults, the development of larvae, the pre-pupation, the pupation and the eclosion can be directly observed through the insect-raising bottle or the culture plate cover, and the black cotton cloth disc in the hole provides a good background for the observation and positioning of the bodies of the insects. (3) When the adults are bred, the black cotton cloth at the root of the plant is wrapped with a wet cotton layer, so that the maintenance time of the plant is prolonged, proper humidity is provided for egg adhesion, and meanwhile, the state of the plant in the insect breeding bottle is similar to that in the field, so that the adults can conveniently spawn at the root; when eggs are collected, the number of the placed holes of the culture plate can be flexibly adjusted according to the needs, and the placement time of the radish slices can be determined according to the requirements on the initially hatched larvae; during the raising process, larva foodstuff is only required to be added once until pupation occurs, so that damage caused by foodstuff replacement and larva transfer is avoided. (4) The agar substrate has good moisturizing and antiseptic properties, the larva breeding does not need moisturizing treatment, and the food spoilage does not need to be worried about. (5) The pupa is developed at the same place as the larva, the humidity is proper, the pupa is prevented from being transferred, and the workload is reduced. (6) The collecting tube of the insect suction device is transparent, so that sex of the adults can be distinguished under the body view mirror conveniently.
In summary, the invention has the following characteristics and meanings: the yellow flea beetle indoor generation feeding method is convenient and quick in flow, successfully realizes generation feeding and breeding of indoor population, provides good growth conditions for each growth period of test insects, and can observe the insect states of each growth period at any time; the feeding device used in the invention has low manufacturing difficulty and strong practicability; the indoor breeding method can maintain continuous multi-generation breeding of population, breaks through the current situation of difficult indoor subculture breeding of phyllotreta striolata, solves the technical problem of damage to adults caused by foodstuff replacement or conversion of breeding devices in other breeding modes, furthest avoids the loss and damage of test insects, realizes long-term supply of indoor test insects, and provides a basis for research of biology, ecology and toxicology of phyllotreta striolata and development of field control technology.
Drawings
Figure 1 is a schematic cross-sectional view of a larval raising apparatus of the invention,
wherein 1-round radish slices, 2-black cotton cloth discs and 3-agar substrates;
FIG. 2 is a schematic diagram of the structure of the fluke device,
wherein, the device comprises a 4-tube cover, a 5-centrifuge tube, a 6-suction nozzle, a 7-insect inlet and an 8-gauze;
figure 3 is a flow chart of a feeding process,
figure 4 yellow flea beetle indoor generation feeding picture,
wherein A is an adult feeding device, B is eggs on black cloths in culture plate holes, C is a feeding larva, D is a pre-pupa, E is a pupa, and F is an adult.
Detailed Description
The present invention will be described in further detail with reference to examples.
Preparing materials:
1. the method for breeding the yellow flea beetles in the indoor generation comprises the following steps of:
(1) Agar strips (agar powder) were added to sterile water with an agar content of 2.5%.
(2) Heating with microwave oven or electromagnetic oven, and completely thawing agar.
(3) When the temperature of the agar solution is below 60 ℃, 80 ten thousand units of penicillin and potassium sorbate are added to make the final contents of the penicillin and the potassium sorbate be 0.019 percent and 0.19 percent respectively.
(4) A5 mL sterile pipette tip was taken and the tip was trimmed about 1cm.
(5) A pipette is used to prepare a short 5mL tip, and more than 1mL of agar solution is sucked and added into the holes of a 24-hole culture plate to enable the liquid level to be high.
(6) The 24-hole culture plate added with the agar is placed at a ventilation position, and the agar is stored in a refrigerator at 4 ℃ after the agar is solidified and the pore wall is free of water vapor.
2. The invention relates to an indoor generation feeding method for phyllotreta striolata, which comprises the following steps of:
(1) Cleaning fresh radix Raphani, and drying.
(2) The radish was cut into 3.5mm pieces by a slicer, and the pieces were made into round pieces by a punch with a diameter of 0.5 cm.
(3) The round radish slices are sucked by water absorbing paper for standby, and are prepared at present.
3. The invention relates to an indoor generation feeding method of phyllotreta striolata, wherein the larva feeding device is shown in figure 1:
black cotton cloth disks 2 are placed on the agar substrate 3 of each culture hole, eggs are placed at positions close to the hole wall, and then round radish slices 1 are placed.
4. The invention relates to an indoor generation feeding method of phyllotreta striolata, wherein the manufacturing method of the trematode device comprises the following steps:
(1) Taking a transparent 50mL centrifuge tube 5, and punching two holes with the diameter of 0.4cm in the middle of the tube cover 4 by using a hot-melting device;
(2) One hole of the tube cover 4 is inserted with a rubber tube with the inner diameter of 0.4cm and the length of 30cm, one end of the tube extends out of the inner side of the cover by 0.3cm and is sealed by a 400-mesh gauze 8, and the other end of the tube is inserted with a pipette tip (1 mL) with the tip of 1cm cut off as a suction nozzle 6;
(3) A rubber tube with an inner diameter of 0.4cm and a length of 15cm was inserted into the other hole of the tube cap 4, one end of the tube was extended from the inner side of the cap by 0.3cm, and the other end of the tube was inserted with a pipette tip (0.2 mL) with a shearing tip of 0.4cm as the inlet 7. The fluke device is shown in figure 2. Example (feeding conditions: adult: 26.+ -. 1 ℃, humidity 60% -80%, light 12L:12D; eggs, larvae and pupae: 26.+ -. 1 ℃, darkness) the feeding process of the present invention is shown in FIG. 3.
(1) Rape cultivation and plant acquisition: selecting rape varieties with thick leaves and strong drought resistance, and cultivating the rape varieties in a flowerpot for a glass greenhouse, wherein the attention is paid to soil sterilization, thinning, water and fertilizer and pest management; when the plant reaches the 8-9 leaf stage, carefully digging out the plant with the root, cleaning and wiping the leaf and the root, and taking the plant at present.
(2) Moisture retention of plants: wrapping plant roots with wet absorbent cotton for moisturizing, wrapping the outer layer with wet black cotton strips layer by layer, and making the outer layer loose and inner tight. The wrapped plants were placed in spawning bottles and allowed to stand. See fig. 4 a.
(3) Raising adults: the same number of yellow leaf-striped flea beetles are inoculated into the insect-raising bottle. See fig. 4 a.
(4) Collection of eggs: the egg laying time of the adult in one day is not fixed, and eggs can be collected once every day or every other day, and the egg laying time is not more than 3 days. When the eggs are collected, whether the rape is replaced or not is determined according to the feeding condition of the rape, if the rape is not required to be replaced, new wet black cloth can be taken to wrap the roots of the plants for continuous feeding, and the replacement period is generally 3 days. When eggs are collected, adults in the insect-raising bottle are collected and temporarily placed by a self-made insect-sucking device, black cloth strips at the roots of plants are taken down and are spread on an observation table of a stereoscopic vision mirror, the multiple of the stereoscopic vision mirror is adjusted, 1-5 eggs are picked by a fine hair pen while observation, and the eggs are placed at positions, close to the hole wall, on black cotton cloth discs in holes of a 24-hole culture plate, as shown in a B in fig. 4.
(5) Feeding larvae: on the 3 rd day after egg collection, a round radish slice is placed in the middle position on the black cotton cloth wafer in the culture plate hole, and the larvae automatically feed after hatching, as shown in the figure 4C; the larvae before pre-pupation were pre-pupated on black cotton discs in the culture wells for about every 3 days for 2-3 days, as shown in fig. 4D.
(6) Storing and feathering the pupae: the pre-pupal stage larvae were pupated on black cotton discs beside the radish slices in the culture holes, as shown in E in FIG. 4. Taking out the radish slices in the holes, and allowing pupa to emerge after 5-6 days.
(7) Collection of adults: collecting newly emerged adults in the culture plate holes by self-made fluke devices, distinguishing sexes under the body vision mirror, and collecting male and female adults respectively for later use. Adults are shown in fig. 4 at F.
(8) Data collection and statistics: observing the eggs every day after collecting eggs, counting the calendar of eggs, larvae, pre-pupae and pupae, and counting the hatching rate of eggs, the pre-pupae rate of larvae, the pupation rate and the emergence rate of pupae.
(9) Example analysis:
TABLE 1 yellow leaf curl flea first Nanjing population development calendar period for each insect state
Data in the table are mean ± standard error.
The breeding method can clearly observe the development conditions of eggs, larvae, pre-pupal larvae and pupae and count. The results show that under the feeding method of the invention, the total calendar period of the phyllotreta striolata from eggs to adults is about 24 days, wherein the developmental calendar period of the eggs is about 5 days, the calendar period of the larvae is about 11 days, the pre-pupation period is about 2 days and the pupation period is about 6 days. Under the feeding method, the total development period of the phyllotreta striolata from eggs to adults is shortened by about 1 week compared with the field records, which is favorable for shortening the scientific research period, improving the research efficiency of the biology, ecology and toxicology of the phyllotreta striolata and accelerating the research and development of the field control technology and improving the pace.
TABLE 2 development rate of yellow stripe flea beetle Nanjing population in each insect state
Data in the table are mean ± standard error.
The results show that under the feeding condition of the invention, the egg hatching rate of the phyllotreta striolata is about 60%, the pre-pupation rate of the larvae is about 70%, the pupation rate of the pre-pupation larvae is about 80%, and the emergence rate of the pupas is about 65%. As a result of feeding the total number of male and female adults to about 40, it was estimated that the development rate at each stage was further improved by increasing the number of adults. The development rate of each stage of the breeding method is 55-80%, and the requirements of generation breeding, scientific research and insect test supply and research of population genetics of indoor population are completely met.
The results show that under the application of the invention, the phyllotreta striolata can develop from eggs to adults within 24 days, and the development calendar period and the generation time of each growth period are shortened compared with the fields. Meanwhile, the invention breaks through the current situation that the phyllotreta striolata is difficult to breed in a room in a subculture way, realizes the continuous breeding of the generation and the population in the room, ensures the population maintenance and the insect test supply in the scientific research work, can provide important materials for the research of the phyllotreta striolata field control technology, and lays an important foundation for the deeper biological research of the phyllotreta striolata.
Claims (4)
1. A method for feeding phyllotreta striolata in indoor generation comprises the following steps:
1. breeding and egg collection of phyllotreta striolata adults
(1) Digging out the whole rape with roots, cleaning and wiping the roots and leaves, wherein the rape in the first step is 8-9 leaves, the leaves are hypertrophic, and the drought resistance of plants is strong;
(2) wrapping the root with wet cotton, wrapping with black cotton strips until no cotton is exposed, and vertically placing the plant into an insect-raising bottle;
(3) the method comprises the steps of (1) inoculating the same number of yellow trepang flea beetles, namely, female and male adults, and tightly covering the yellow trepang flea beetles, wherein 30 yellow trepang flea beetles are inoculated into the insect-raising bottle;
(4) sterile 24-well plates were prepared and 1mL of preservative-containing agar solution was added to the wells using a pipette; the content of the preservative in the agar is respectively 80 ten thousand units of penicillin 0.019 and potassium sorbate 0.19 percent; adding preservative when the temperature of the agar solution is below 60 ℃;
(5) cooling agar, and adding black cotton cloth wafer into the hole to make it adhere to agar;
(6) when receiving eggs, firstly, temporarily collecting the adults by using a fluke device, then taking down black cloth wrapped by plant roots, placing under a stereoscopic vision mirror, adjusting the multiple of the stereoscopic vision mirror, picking egg grains on the inner surface and the edge of the stereoscopic vision mirror by using a fine hair pen while observing, placing eggs on a black cloth wafer in a culture plate hole at a position close to the hole wall, and covering a transparent culture plate cover;
2. egg hatching and raising of larvae of different ages
A. On day 3 after egg collection, round radish slices were placed in the center of the wells of the oval-containing culture plates;
B. after the eggs are hatched, the larvae of all ages are actively fed before the initial hatching and the pre-pupation;
3. pupa management and adult collection
a. The larvae are subjected to pupation and pupation on black cotton cloth pieces beside the radish pieces in the holes, and the radish pieces are taken out after pupation;
b. after the pupae eclosion into adults, collecting the adults by a fluke device and distinguishing gender under a body view mirror;
in the first to third steps, the adults are fed at 26+/-1 ℃ and with humidity of 60% -80%, and the illumination is 12L:12D in a climatic chamber; placing the culture plate with eggs, larvae and pupae in an opaque storage box, and also placing the culture plate in a condition of 26+/-1 ℃ and humidity of 60% -80%, and irradiating with 12L:12D in a climatic chamber;
in the first step, defatted cotton with the thickness of 0.5cm is used for wetting and wrapping rape roots by sterile water, and cotton is wrapped layer by black cotton strips, so that a spawning space is provided for adults;
the egg collection in the first step is carried out for a single egg collection period of not more than 3 days, new wet black cloth is taken to wrap the root of the plant for continuous feeding after egg collection, and the plant replacement period is 3 days;
the manufacturing method of the insect suction device comprises the following steps:
1) Taking a transparent 50mL centrifuge tube, and punching two holes with the diameter of 0.4cm on the middle position of a tube cover by using a hot-melting device;
2) One hole on the tube cover is inserted with a rubber tube with the inner diameter of 0.4cm and the length of 30cm, one end of the tube extends out of the inner side of the cover by 0.3cm and is sealed by a gauze with 400 meshes, and the other end of the tube is inserted with a 1mL pipette tip with the tip cut off by 1cm to be used as a suction nozzle;
3) A rubber tube with an inner diameter of 0.4cm and a length of 15cm is inserted into the other hole of the tube cover, one end of the tube extends out of the inner side of the cover by 0.3cm, and the other end of the tube is inserted into a 0.2mL pipette tip with a tip end cut off by 0.4cm to serve as an insect inlet.
2. The method for feeding yellow flea beetles in the indoor generation of the yellow flea beetles according to claim 1, wherein the insect-breeding bottle in the first step is a transparent plastic bottle with the diameter of 22cm and the height of 23cm, the bottle is sterilized with 0.1% calcium hypochlorite before use, 3-5 holes with the diameter of 2cm are formed in the bottle cover, and 400 meshes of gauze are adhered below the holes.
3. The method for feeding the yellow flea beetles in the indoor generation of the yellow flea beetles according to claim 1, wherein the single hole inner diameter of the 24-hole culture plate is 0.8cm and the height is 2cm; the diameter of the black cotton cloth wafer is 0.8cm; 1-5 eggs are placed in each hole of the culture plate; round radish slices placed in the culture plates have a diameter of 0.5cm and a thickness of 3.5mm.
4. The method for feeding yellow flea beetle in the indoor generation of claim 1, wherein the pipette in the first step is a 5mL micropipette, and the tip of the pipette is cut off by 1cm.
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