CN116406650A - A method for artificially multiplying a large number of Drosophila spp. - Google Patents

Abstract

The invention relates to the field of biological pest control, in particular to a method for artificially and rapidly propagating drosophila melanogaster horner bees in large quantity, which comprises the following steps: 1) Preparing a feeding tank for Drosophila bifidus and Drosophila mallet; 2) Collecting the fruit fly pupae; 3) Drosophila melanogaster horn fine bee parasitic double thorn drosophila melanogaster pupa; 4) Eclosion and feeding of the drosophila bicolor pupae parasitized by the drosophila bicolor hammer; 5) Raising adult drosophila melanogaster; 6) And (3) repeating the steps 1) to 5), so that a large number of drosophila hammer angle bees can be obtained in a short period. The method provided by the invention is time-saving, labor-saving, space-saving, simple in required device, capable of breeding a large amount of drosophila melanogaster hornet in a short time and meeting the application requirements of biological characteristic research tests or field control of the drosophila melanogaster.

Description

A method for artificially multiplying a large number of Drosophila spp.

technical field

The invention belongs to the field of biological control of pests, and in particular relates to a method for artificially and rapidly multiplying a large number of Drosophila chinensis.

Background technique

Drosophila suzukii Matsumura , also known as Suzuki's Drosophila, Cherry Drosophila, Blueberry Drosophila, etc., belongs to Diptera, Cyclorrhapha, Drosophilidae, Drosophila, Drosophila , Fruit flies of the subgenus Subgenus Sophophora . Native to Southeast Asia, it is one of the few important pests in the genus Drosophila that can harm fresh and healthy fruits. In recent years, the insect has spread around the world and seriously harmed more than 240 economic crops such as blueberries, strawberries, cherries, grapes, and red bayberry. Cause huge loss to the fruit industry of the whole world.

At present, the control of Drosophila spotted wing is still dominated by chemical control, but the disadvantages of chemical control are also emerging year by year. Therefore, in recent years, efforts have been made to carry out research work on the biological control of Drosophila spotted wing at home and abroad, in order to form a sustainable comprehensive control method that combines biological control with other pollution-free control methods. Drosophila Trichopriadrosophilae Perkins parasitizes the pupae of Drosophila spotted wing, which is widely distributed in Asia, Europe, and North America. It has broad application prospects and needs to be artificially propagated in large quantities for production.

Spot wing Drosophila endangers healthy fresh fruit, and it is difficult to artificially raise a large number of them. Using fresh fruit to raise spotted wing Drosophila and then using spotted wing Drosophila to artificially multiply Drosophila japonica has high cost and low efficiency, which cannot meet the needs of production. The invention adopts Drosophila bispinus as an alternative host, and provides a method for rapidly multiplying the Drosophila japonica. When using Drosophila bispinus to breed Drosophila spp., compared with the existing patent of using D. The advantages of the host are: 1. Drosophila bispinus has strong dryness, and when pupating, it will drill into the corn flour to pupate, and the pupae will not stick to each other, so it is easy to collect; while the secretion of Drosophila melanogaster when pupating The pupae stick together, making it difficult to separate and collect. The quality of the pupae will be affected after separation by instruments; Therefore, compared with Drosophila melanogaster, the number of parasitic wasps obtained with the same pupa weight and quantity is more; 3. Studies have shown that Drosophila spp. The lifespan of bees is longer than that of male and female bees with Drosophila melanogaster as host; 4. In host selection, the results show that Drosophila melanogaster is more preferred to parasitize the original host (ie, Drosophila melanogaster pupae), while Drosophila spp. bred with Drosophila bispinus had no obvious host preference. Therefore, in the field application, Drosophila spp. with double-spined Drosophila as a host has a better control effect on Drosophila spotted wing.

Contents of the invention

The object of the present invention is to provide a method for artificially multiplying Drosophila spp. in large numbers and rapidly.

To achieve the above object, the present invention adopts the following technical solutions:

The invention discloses a method for artificially multiplying Drosophila spp. in large quantities rapidly, which uses Drosophila bispinus as a host to multiply the Drosophila spp.

Above-mentioned a kind of method of artificially a large number of rapid multiplication Drosophila japonica comprises the following steps:

1) The preparation of the breeding tanks of Drosophila bispinus and Drosophila japonica;

2) Collection of Drosophila pupae;

3) Drosophila japonica parasitizes the pupa of Drosophila bispinus;

4) Eclosion and rearing of Drosophila bispinatus pupae parasitized by Drosophila spp.;

5) Rearing of adults of Drosophila japonica;

6) Repeat steps 1) to 5) to obtain a large number of Drosophila spp. in a short period of time.

In the above step 1), the adults of Drosophila bispinus and the adults of Drosophila spp. are raised in the same kind of transparent plastic tanks, which need to be cleaned and dried before raising the insects. When using, the breeding tanks need to be placed and fixed on the table Go up and seal the rearing jar with 140 mesh nylon net and rubber band.

The above step 2) is as follows: place the pupae of Drosophila bispinus in a rearing tank for feeding, and provide artificial feed and clear water for feeding in the rearing tank; after Drosophila biscarina begins to emerge for 5 days, use a plastic box with artificial feed in the Place in the rearing tank for 24 hours to collect enough eggs; place the plastic box with mature larvae and pupae of Drosophila bispinus in a multi-functional fruit fly adult cage for raising, and provide clear water in the multi-functional fruit fly adult cage and yeast powder to provide nutrition for the larvae, and use the desiccation ability of the mature larvae to collect and isolate the same-day-old pupae of Drosophila bispinus through cornstarch for parasitism.

The above step 3) is specifically as follows: on the next day after the emergence of Drosophila spp. 3 boxes of fruit fly pupae, each containing 12,500 grains of double-spined fruit fly pupae, provided for 7,500 fruit flies to parasitize for 24 hours; every 24 hours to parasitize a batch, each batch of fruit flies can be parasitized continuously for 25- 30 boxes of Drosophila pupae were used for mass multiplication.

The above-mentioned step 4) is as follows: transfer each batch of Drosophila bispinus pupae parasitized by Drosophila spp. The parasitized Drosophila double-spined was eclosion and starved to death, the nylon net was removed, and the plastic box containing the Drosophila double-spined pupae parasitized by Drosophila spp. .

The above step 5) is as follows: after the emergence of Drosophila spp., the breeding tank of Drosophila spp. was provided with 10wt% honey water, and replaced with fresh 10wt% honey water every 5 days.

Significant advantage of the present invention is:

Drosophila melanogaster is used as a host in the prior art, but the present invention uses Drosophila bispinus as a host to multiply Drosophila spp., which has the advantages of:

1) Drosophila bispinus has a strong tendency to dryness. When pupating, it will drill into corn flour to pupate, and the pupae do not adhere to each other, so they are easy to collect. However, the secretion of Drosophila melanogaster when pupating will cause the pupae to stick together , not easy to separate and collect, and the quality of pupae will be affected after separation by instruments;

2) The weight and size of D. melanogaster pupae are smaller than that of Drosophila melanogaster, but it does not affect the quality of Drosophila melanogaster. Therefore, compared with Drosophila melanogaster, the same pupae weight and quantity are used The number of parasitic wasps is more; 3) The life span of the male and female bees of the Drosophila spp. which is bred with Drosophila double-stinger as the host is longer than that of the male and female bees which use Drosophila melanogaster as the host;

4) D. melanogaster D. melanogaster breeding D. melanogaster D. melanogaster prefers to parasitize the original host (that is, D. melanogaster pupae), while the Drosophila D. melanogaster bred with double stinger has no obvious Host preference, therefore, the control effect of Drosophila spp. with Drosophila bispinus as host in field application is better.

Description of drawings

Figure 1: Rearing tanks for Drosophila biscarina and Drosophila spp.

Figure 2: Schematic diagram of the same-day-old pupae of Drosophila bispinus obtained by collecting and isolating cornstarch using the desiccation ability of mature larvae.

Detailed ways

The present invention will be further described in detail below in conjunction with specific examples.

In the present invention, the formula of the artificial feed is: cornstarch 50g/L, white granulated sugar 50g/L, yeast 25g/L, agar powder 5g/L.

Example 1

Step 1: Preparation of Drosophila biscarina and Drosophila spp.

Adults of Drosophila bispinus and adult Drosophila spp. were reared in the same transparent plastic tank (Figure 1), with a capacity of 20 L, a height of 41 cm, a diameter of the bottom of the tank of 28 cm, and a diameter of the tank mouth of 13 cm. Before raising insects, the insect raising tank needs to be cleaned and dried. When in use, the rearing tank needs to be placed on the table so that the mouth of the tank is perpendicular to the table. Two rubber tubes are used to stick the body of the tank to the table where the insect tank touches the table so that it is fixed on the table to prevent the insect tank from rolling when in use. The mouth is covered with a 140-mesh nylon mesh and fixed with a rubber band.

Step 2: Collection of Drosophila pupae

Put 25ml of Drosophila bispinus pupae (about 500 Drosophila bispinus pupae per ml) in a prepared rearing tank for development. The rearing temperature is 25±1°C, relative humidity is 60±10%, light The cycle is 14L:10D, and a plastic box with artificial feed and a conical bottle with clear water are placed in the breeding tank for feeding. The plastic box is 10 cm long, 10 cm wide, and 4 cm high, with an unsealed upper cover. There is a long strip of absorbent cotton extending from the mouth of the bottle to the bottom of the bottle. After Drosophila bispinus began to emerge for 5 days, a plastic box (length 10 cm, width 10 cm, height 4 cm, with an open lid) containing artificial feed was placed in the breeding tank for 24 hours to collect enough eggs. The plastic box containing the artificial feed that collected the mature larvae and pupae of Drosophila bispinus was transferred and placed in a multifunctional adult fruit fly breeding cage (CN201220177838.4) for development. The breeding temperature was 25±1°C and the relative humidity was 60±10%, the photoperiod is 14L:10D, a Erlenmeyer flask with clear water and a petri dish with yeast powder are placed in the multifunctional fruit fly adult cage to provide nutrition for the larvae, and the larvae with a diameter of 18cm Qualitative filter paper was folded into a triangular funnel shape, placed in an empty plastic box (length 10cm, width 10cm, height 4cm, and the upper cover was not sealed), and a layer of dry cornstarch was spread inside the filter paper funnel, and then the mature Plastic boxes containing artificial feed for larvae and pupae and empty plastic boxes with filter paper funnels were stacked in a staggered direction at 45 degrees (Figure 2). The plastic boxes containing artificial feed for mature larvae and pupae of D. On the top, the empty plastic box with a filter paper funnel is placed on the bottom. The hatched larvae will mature after 2-3 days; Put it into the cornstarch laid flat in the filter paper funnel and wait for pupation; after pupation, pour the cornflour in the filter paper funnel into a 40-mesh sieve with an aperture of 0.45mm, shake it gently from side to side, and sieve off the excess cornstarch. The obtained D. bispinus pupae were poured into an empty plastic box (17cm x 10cm x 1cm) and spread flat, and the pupae were screened every 24 hours for subsequent parasitism.

Step 3: Drosophila spp. parasitizing Drosophila pupae

Drosophila bispinus pupae were provided for parasitism on the day after the emergence of Drosophila spp. Place 25ml of 1-day-old Drosophila bispinus pupae (about 500 Drosophila bispinus pupae per milliliter) in an empty plastic box (17cm x 10cm x 1cm, with the upper cover not sealed), and spread them on each fruit fly. Place 3 boxes of double-spined Drosophila pupae in the rearing tank of the fly hammer horn, and provide the parasitism of the fruit fly hammer horn for 24 hours. The mouth of the rearing tank is covered with a 140-purpose nylon net and fixed with a rubber band ring; each fruit fly hammer horn There are 7,500 Drosophila japonicas in the bee breeding tank. One batch can be parasitized every 24 hours, and each batch of Drosophila spp. can continuously parasitize 25-30 boxes of Drosophila bispinus pupae for large-scale propagation.

Step 4: Eclosion and rearing of Drosophila bispinus pupae parasitized by Drosophila spp.

Transfer each batch of Drosophila double-spined pupae parasitized by Drosophila chinensis to an empty transparent plastic box (16cm x 16cm x 20cm), seal it with 140-mesh nylon mesh and tighten it with a rubber band, and wait for 6 days before being parasitized The double-spined Drosophila erupted and starved to death, removed the nylon net, and placed the pupae box in the Drosophila spp. It is 14L:10D, waiting for the emergence of parasitoids; after 14-15 days of feeding, the male parasitoids begin to emerge, the males of parasitoids begin to emerge, the females begin to emerge the next day, and the males of parasitoids begin to emerge.

Step 5: Rearing of adults of Drosophila japonica

After the emergence of Drosophila spp., a 50ml conical flask with a capacity of 50ml was placed in the breeding tank of Drosophila spp., and the configured 10wt% honey water was added to the conical flask, and absorbent cotton was inserted at the bottom. Extend to the mouth of the bottle to feed on the Drosophila spp., and replace it with fresh honey water every 5 days.

Step 6:

By repeating the above steps 1 to 5, a large number of Drosophila chinensis can be obtained in a short period of time for field application.

The relevant parasitic indicators are shown in the table below:

Note: The data in the table are mean ± standard error

As can be seen from the above table, using the pupae of Drosophila bispinus as a host to breed Drosophila chinensis, the parasitic rate can reach 82.64%±0.88, and the eclosion rate can reach 76.21%±1.06. 37,875-39,375 pupae of Drosophila bispinus can be placed each time, and 23,282.76-25,724.39 adults of Drosophila chinensis can be obtained for each batch of parasitism, including 13,038.37-15,949.12 female wasps.

This set of parasitic wasp breeding technology can quickly and easily multiply a large number of Drosophila spp., the process is easy to master and carry out, and the double-spined Drosophila is easy to raise, the egg production is large, the pupa has strong drying tendency, and it is easy to collect and store , suitable for rapid and large-scale breeding of bees, meeting the application requirements of biological characteristics research experiments and field control of spotted winged fruit flies.

Finally, it should be declared that the above descriptions are only preferred embodiments of the present invention, and all equivalent changes and modifications made according to the scope of the patent application of the present invention shall fall within the scope of the present invention.

Claims (7)

1. A method for manually and rapidly propagating a large number of drosophila melanogaster horners is characterized by comprising the following steps: the Drosophila bifidus is used as a host to propagate the Drosophila bifidus hammer horned bees.

2. The method for artificially and rapidly propagating a large number of drosophila melanogaster horners according to claim 1, wherein the method comprises the following steps: the method comprises the following steps:

1) Preparing a feeding tank for Drosophila bifidus and Drosophila mallet;

2) Collecting the fruit fly pupae;

3) Drosophila melanogaster horn fine bee parasitic double thorn drosophila melanogaster pupa;

4) Eclosion and feeding of the drosophila bicolor pupae parasitized by the drosophila bicolor hammer;

5) Raising adult drosophila melanogaster;

6) And (3) repeating the steps 1) to 5), so that a large number of drosophila hammer angle bees can be obtained in a short period.

3. The method for artificially and rapidly propagating a large number of drosophila melanogaster horners according to claim 2, which is characterized in that: in the step 1), the adult drosophila spinosa and the adult drosophila melanogaster are raised by using the same transparent plastic tank, the raising tank is required to be cleaned and dried before raising the insect, the raising tank is required to be horizontally placed and fixed on a table surface when in use, and the raising tank is required to be sealed by using a 140-mesh nylon net and rubber band.

4. The method for artificially and rapidly propagating a large number of drosophila melanogaster horners according to claim 2, which is characterized in that: the step 2) is specifically as follows: placing the Drosophila bifidus pupae in a feeding tank for feeding, and providing artificial feed and clear water in the feeding tank for feeding; after the drosophila bifidus begins to emerge for 5 days, placing the drosophila bifidus in a feeding tank for 24 hours by using a plastic box filled with artificial feed to collect enough eggs; placing the plastic box with the mature larvae and pupae of the Drosophila bifidus in a multifunctional adult fruit fly feeding cage for feeding, providing clear water and yeast powder in the multifunctional adult fruit fly feeding cage for providing nutrition for the larvae, and collecting and separating the mature larvae through corn starch by utilizing the drier property of the mature larvae to obtain the pupae of the Drosophila bifidus at the same day age for parasitism.

5. The method for artificially and rapidly propagating a large number of drosophila melanogaster horners according to claim 2, which is characterized in that: the step 3) is specifically as follows: providing double thorn drosophila pupae for parasitism at the beginning of the following day of the emergence of the drosophila hammer horned fine bees, spreading the double thorn drosophila pupae in an empty plastic box, selecting a double thorn drosophila pupae 3 box with the age of 1 day each time, wherein each box contains 12500 grains of double thorn drosophila pupae, and providing 7500 drosophila hammer horned fine bees for parasitism for 24 hours; every 24 hours, a batch of drosophila melanogaster with the hammer feet can be continuously parasitic to 25-30 boxes of drosophila melanogaster pupas for mass propagation.

6. The method for artificially and rapidly propagating a large number of drosophila melanogaster horners according to claim 2, which is characterized in that: the step 4) is specifically as follows: transferring the two thorn fruit fly pupas of each batch of parasitic fruit fly with the hammer horn of fruit fly into an empty plastic box, sealing the plastic box by using a 140-mesh nylon net and rubber bands, allowing the two thorn fruit fly to eclosion and starve after 6 days, removing the nylon net, placing the plastic box filled with the two thorn fruit fly pupas of parasitic fruit fly with the hammer horn of fruit fly into a raising tank for development, and waiting for the eclosion of parasitic fruit fly.

7. The method for artificially and rapidly propagating a large number of drosophila melanogaster horners according to claim 2, which is characterized in that: the step 5) is specifically as follows: after emerging from the drosophila melanogaster, 10wt% of honey water is provided in the raising tank of the drosophila melanogaster adult bees, and fresh 10wt% of honey water is replaced every 5 days.

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