Method for mass rearing of cyrtopimpla spp.

Abstract

The present application relates to a kind of methods for mass propagation of tea corner miridae in laboratory, comprising the following technical steps: collecting tea garden tea corner miridae adult, cultivating fall kui seedling, putting seedling and leading insect, tea corner miridae habitat and feeding, mating and oviposition and nymph hatching and feeding.The present application uses fall kui seedling as the main body of feeding raw material, and the leaf of raw material is relatively thick, strong in harm resistance, not easy to dry, which helps to improve the hatching rate of tea corner miridae eggs, reduce the frequency of raw material supply and replacement, at the same time, the present application saves time and effort, can be realized in laboratory low cost and sustainability fast, efficient, mass propagation of tea corner miridae test insect source.

Description

Technical Field

[0001] This invention relates to the field of artificial insect rearing, specifically to a method for large-scale laboratory breeding of the tea-horned mirid bug. Background Technology

[0002] The tea horned bug (Helopeltis theivora waterhouse), also known as the tea thorn bug, cocoa hammer bug, and cashew horned bug, is a major pest of various economic crops such as tea, cocoa, and cashew in tropical regions. It is characterized by its voracious appetite and destructive power. This insect has a wide host range, but tea trees are its preferred feeding and oviposition host. Compared to other plants, the tea horned bug has the highest reproductive rate, growth rate, and survival rate on tea trees. The tea horned bug infests more than 80% of tea gardens in India, making it the number one pest in India's three major tea-producing regions. In Hainan tea-producing areas of my country, the tea horned bug also seriously damages the growth and development of tea trees, especially during the summer and autumn tea harvesting period from August to November.

[0003] Indoor research on the life cycle and biological characteristics of the tea mirid bug is a prerequisite for studying its control techniques. In the integrated management of the tea mirid bug, insecticide screening, the development of new biological pesticides, and parasitic wasp breeding all require large-scale breeding of standard test insects with consistent nutritional status. However, current research on the tea mirid bug is limited both domestically and internationally, with the most significant technical obstacle being the difficulty in breeding tea mirid bugs in the laboratory. On one hand, both adults and nymphs of the tea mirid bug feed by piercing and sucking sap from the tender parts of tea tree shoots. Due to their large feeding volume and destructive power, the damaged shoots typically turn completely brown and necrotic within 2-3 days, eventually drying out. On the other hand, female tea mirid bugs mainly lay eggs inside the epidermis of tender tea shoots, with an incubation period of 7-10 days. Most eggs laid inside the tender shoots fail to hatch normally, and the few that do hatch are hampered by insufficient sap supply from the tender shoots, resulting in a low survival rate for the nymphs. Therefore, obtaining a sustainable source of insects through economical, simple, and easy-to-operate methods is crucial for scientific research.

[0004] Currently, the primary method for indoor rearing of tea mirid bugs is still using tea shoots. However, this method requires consuming a large amount of tea shoots to produce a relatively small number of bugs, resulting in low cost-effectiveness and falling far short of the needs of scientific research experiments. A more suitable approach is to find alternative host plants. These plants should be easier to obtain and more resistant to mirid bug infestations, allowing the tea mirid bug to successfully complete its life cycle activities, including habitat, feeding, mating, and egg-laying.

[0005] In the existing technology, Chinese patent CN108967364B provides a technique for raising tea mirid bugs using the tender shoots of Forsythia suspensa. This method achieves an average egg hatching rate of 50%, and is simple to operate, low in cost, and can continuously raise more than eight generations. However, tea mirid bugs are mainly distributed in tropical regions, while Forsythia suspensa is less common in tropical areas and difficult to cultivate. Furthermore, when using this method to raise tea mirid bugs, the tender shoots of Forsythia suspensa need to be replaced every three days, which is still relatively time-consuming and labor-intensive.

[0006] Basella alba L. is an annual, climbing herbaceous plant belonging to the genus Basella of the family Basellaceae. Its stems are hairless and entirely fleshy; its deciduous leaves are ovate or nearly round; its flowers are spike-like with very small bracts. Basella alba prefers warm temperatures and tolerates high humidity. It is not particular about soil conditions, as long as it is loose and fertile, with sandy loam rich in organic matter being most suitable.

[0007] Based on the characteristics of Malabar spinach, a new method for large-scale laboratory breeding of tea horned mirid bugs was developed using Malabar spinach seedlings as the main feed material. This method has important application value for obtaining a continuous source of insects to solve related scientific problems. Summary of the Invention

[0008] To address the above problems, the present invention provides the following technical solution:

[0009] This invention provides an application of the plant Malabar spinach for the large-scale propagation of the tea horned mirid bug in the laboratory. The plant is used as the main feed for the tea horned mirid bug, and propagation is achieved through both sowing and cutting. Figure 1 This method allows for the rapid acquisition of tender shoots for the tea horned mirid bug to feed on and lay eggs, and successfully enables the reincarnation of the tea horned mirid bug, solving the technical challenge of large-scale indoor propagation of the plant.

[0010] This invention also provides a method for large-scale laboratory breeding of the tea-horned mirid bug, comprising the following steps:

[0011] (1) Collecting adult tea mirid bugs in tea gardens: Collect 30 adult tea mirid bugs and place them in fruit fly tubes. Place one adult bug in each fruit fly tube. Put 1-2 tender tea shoot leaves in the fruit fly tube in advance and plug the opening of the fruit fly tube with breathable cotton plug.

[0012] (2) Cultivating Malabar spinach seedlings: After soaking and germinating 50g of Malabar spinach seeds, sow the seeds evenly in the planting area, cover them with soil to a depth of 1.5-2cm, and cover them with plastic film to maintain soil temperature and humidity; cultivate the Malabar spinach seedlings to the 3-4 leaf stage for feeding on the tea horned mirid bug. Among them, the 3-leaf stage Malabar spinach seedlings can be used to feed the tea horned mirid bugs, and the 4-leaf stage seedlings are most suitable for female insects to lay eggs.

[0013] (3) Introducing insects by releasing seedlings: Place 2-3 Malabar spinach seedlings with 3-4 leaves in a wooden insect raising cage and keep them moist with purified water or floral foam. Figure 2 ), put the 30 tea-horned mirid bugs collected in step (1) into the insect rearing cage, put the petri dish containing 10ml of 10% (w / v) honey water into the insect rearing cage, and the ambient temperature of the insect rearing cage is 26±2℃ and the air humidity is 70±3%.

[0014] (4) Habitat and feeding of the tea mirid bug: Observe the feeding of adult bugs, the growth of Malabar spinach seedlings and the damage to the leaves every day. Replace the Malabar spinach seedlings every 3-5 days. Spray 20ml of purified water on the Malabar spinach seedlings every day so that there are some water droplets attached to the seedlings for the tea mirid bugs to drink. Add 3-5ml of 10% (w / v) honey water every day for the tea mirid bugs to supplement their nutrition. When the adult tea mirid bugs feed too much and cause the Malabar spinach buds to dry out prematurely, it is necessary to replenish the new Malabar spinach seedlings in time for the tea mirid bugs to feed on.

[0015] (5) Mating and oviposition: Adult tea mirid bugs reach sexual maturity 3-5 days after emergence and then begin mating. Females lay eggs 3-5 days after mating. Most eggs are scattered in the epidermis of tender stems, and a small number are laid on petioles and veins. Two white bristles, one long and one short, are attached to the tip of each egg. The oviposition period lasts 30-40 days, with an average of 6-7 eggs laid per female per day.

[0016] (6) Nymphal hatching and feeding: The egg stage of the tea horned mirid bug is 5-8 days and the nymphal stage is 10-15 days. After the first instar nymphs hatch, a small amount of Malabar spinach buds are added as food to facilitate the piercing and sucking of the young nymphs. After the nymphs molt twice and grow to the third instar, Malabar spinach seedlings with 3-4 leaves are added as food. After the nymphs molt 4 times, they become adults.

[0017] Furthermore, in step (1), among the 30 adult tea-horned mirid bugs, the ratio of female to male adults is 2:1.

[0018] Furthermore, in step (2) during the cultivation of Malabar spinach seedlings, the number of Malabar spinach seedlings cultivated to the 3-4 leaf stage is 30.

[0019] Furthermore, in step (3), the insect cage has dimensions of 40cm×40cm×40cm.

[0020] The beneficial effects of this invention are:

[0021] This invention utilizes Malabar spinach seedlings as the main feed material for the breeding of tea horned mirid bugs, and is used to breed and raise tea horned mirid bug nymphs and adults in large quantities in the laboratory to meet the requirements of indoor experimental standard test insects.

[0022] (1) The Malabar spinach leaves provided by this invention are thicker, with well-developed leaf mesophyll cells and strong resistance to pests. Compared with the tea tree shoots currently used as feed, they are less prone to drying out and more durable.

[0023] (2) This invention improves the hatching rate of tea horn mirid bug eggs. In the current process of using tea shoots for breeding, the hatching rate of tea horn mirid bug eggs is often low because the tender shoots are very easy to dry out too early. However, the tender stems of Malabar spinach are relatively thick, so the hatching rate of eggs is higher, reaching more than 90%. At the same time, it is easier to obtain insect sources, which helps to carry out experiments with a large number of insects at any time.

[0024] (3) This invention reduces the supply and replacement frequency of the main raw material, Malabar spinach leaves. When using tea shoots for feeding, fresh tea shoots need to be replaced or added almost every day, while when using Malabar spinach seedlings for feeding, they only need to be replenished every 3-5 days. Attached Figure Description

[0025] Figure 1 Seedlings (A) propagated from Malabar spinach seeds and new shoots (B) propagated from stem cuttings.

[0026] Figure 2 The images show Malabar spinach seedlings and shoots grown directly in water (A) and Malabar spinach seedlings grown using floral foam to retain water (B).

[0027] Figure 3 Adult tea mirid bugs collected from a tea garden.

[0028] Figure 4 The images show adult tea mirid bugs feeding on Malabar spinach leaves (A) and feeding damage (B).

[0029] Figure 5 The images show male and female tea-horned mirid bugs mating.

[0030] Figure 6 These are eggs laid by the female insect on the tender stems of Malabar spinach.

[0031] Figure 7 These are young nymphs of the tea-horned mirid bug.

[0032] Figure 8 The image shows older nymphs feeding on Malabar spinach seedlings. Detailed Implementation

[0033] The specific embodiments of the present invention are described below to enable those skilled in the art to understand the present invention. However, it should be understood that the present invention is not limited to the scope of the specific embodiments. For those skilled in the art, various changes are obvious as long as they are within the spirit and scope of the present invention as defined and determined by the appended claims. All inventions utilizing the concept of the present invention are protected.

[0034] Example 1: Mass breeding of the tea-horned mirid bug in the laboratory

[0035] (1) Collection of adult tea mirid bugs from tea gardens: In January 2024, a number of adult tea mirid bugs were collected from the tea garden of Dongchang Tea Factory in Haikou and brought back to the laboratory. Twenty female bugs and ten male bugs with good activity were selected. The tested adult bugs were temporarily stored in fruit fly tubes, with one bug in each tube. One to two tender tea shoot leaves were placed in the tube beforehand, and the tube opening was plugged with breathable cotton. Figure 3 ).

[0036] (2) Cultivation of Malabar spinach seedlings: The main method of propagation is by seed. In early January 2024, 50g of Malabar spinach seeds were soaked and germinated. The seeds were then evenly sown in the planting area. Before sowing, the land was prepared and fertilized. After sowing, the soil was covered with 1.5-2cm of fine soil and covered with plastic film to maintain the temperature and humidity of the soil. Two weeks later, when the Malabar spinach seedlings grew to the 3-4 leaf stage, the whole plant was harvested for insect rearing. Finally, a total of 30 Malabar spinach seedlings were obtained for the test.

[0037] (3) Release seedlings to attract insects: Take 2-3 Malabar spinach seedlings with 3-4 leaves and place them in a wooden insect raising cage (40cm×40cm×40cm). Use flower mud to keep them moist and cultivate them. Put 30 tea horned mirid bugs collected in step (1) into the insect raising cage. Put a petri dish containing 10ml of 10% (w / v) honey water into the insect raising cage. The ambient temperature of the insect raising cage is 26±2℃ and the air humidity is 70±3%.

[0038] (4) Habitat and feeding of the tea mirid bug: Adult tea mirid bugs mainly pierce and suck the sap from the tender parts of Malabar spinach leaves. The feeding behavior of adults, the growth status of Malabar spinach seedlings, and the damage to leaves are observed daily. Figure 4 Replace the Malabar spinach seedlings every 3-5 days. Spray 20ml of purified water on the seedlings daily to ensure some water droplets adhere to them, providing sustenance for the tea horned mirid bugs. Add 3-5ml of 10% (w / v) honey water daily to supplement the tea horned mirid bugs' nutrition. If the adult tea horned mirid bugs consume too much food, causing the Malabar spinach buds to dry out prematurely, promptly replenish the seedlings with new Malabar spinach seedlings for the bugs to feed on.

[0039] (5) Mating and oviposition: Adult tea mirid bugs reach sexual maturity 3-5 days after emergence, and then begin mating. Male and female bugs mate in a "I" shape. Figure 5 Each mating session lasts for more than 2 hours; the female lays eggs 3-5 days after mating, with most eggs scattered inside the epidermis of tender stems, and a small number laid on petioles and veins. Each egg has two white bristles, one long and one short, clearly visible to the naked eye at the tip. Figure 6 The egg-laying period can last for more than a month, with each female laying an average of 6-7 eggs per day.

[0040] (6) Nymphal hatching and molting: Under laboratory conditions, the egg stage of the tea mirid bug is 5-8 days, and the nymphal stage is 10-15 days; the nymphs have a total of 5 instars and need to undergo 4 molts. Newly hatched nymphs are orange-red, and as they grow older, their body color gradually turns light yellow to light green; after the first instar nymphs hatch, a small amount of Malabar spinach shoots are added as food to facilitate the piercing and sucking of food by the young nymphs. Figure 7 After the nymphs have molted twice and reached the 3rd instar, supplement their diet with Malabar spinach seedlings at the 3-4 leaf stage. Figure 8 The nymphs molt four times before becoming adults.

[0041] The test results are as follows:

[0042] After steps (3) and (4), all adult tea horn mirid bugs survived, and no dead bugs were found in the breeding cages;

[0043] After step (5), a total of 4020 eggs of the tea horn mirid bug were collected;

[0044] After step (6), a total of 3,698 first-instar nymphs were hatched, with a hatching rate of 92.0%. After the nymphal stage, a total of 3,550 adults were obtained. Both nymphs and adults can be used directly as standard test insects for indoor experiments.

[0045] Specific embodiments have been used to illustrate the principles and implementation methods of this invention. The descriptions of the embodiments above are only for the purpose of helping to understand the method and core ideas of this invention. At the same time, for those skilled in the art, there will be changes in the specific implementation methods and application scope based on the ideas of this invention. Therefore, the content of this specification should not be construed as a limitation of this invention.

Claims

1. A method of mass rearing of Dendrothrips tea in the laboratory, characterized by, The method comprises the following steps: (1) collecting tea angle stink bugs: 30 adult tea angle stink bugs are collected and placed in a fruit fly tube, with 1 adult tea angle stink bug in each fruit fly tube, 1-2 pieces of tea shoot tender leaves are placed in the fruit fly tube in advance, and the fruit fly tube is plugged with a breathable cotton plug; (2) cultivating amaranth seedlings: 50 g of amaranth seeds are soaked and germinated, and then evenly sown in the planting site, with a soil covering depth of 1.5-2 cm, and a plastic film is covered to maintain the soil temperature and humidity; the amaranth seedlings are cultivated to the 3-4 leaf stage for the tea angle stink bugs to feed on; (3) placing seedlings to attract insects: 2-3 amaranth seedlings at the 3-4 leaf stage are placed in a wooden insect rearing cage, watered with flower mud, and the 30 adult tea angle stink bugs collected in step (1) are placed in the insect rearing cage, and a culture dish containing 10 ml of 10% (w / v) honey water is placed in the insect rearing cage; the air temperature in the environment of the insect rearing cage is 26±2°C, and the air humidity is 70±3%; (4) tea angle stink bug habitat and feeding: the feeding behavior of adult tea angle stink bugs and the growth status of amaranth seedlings are observed every day, the amaranth seedlings are replaced every 3-5 days, and 20 ml of pure water is sprayed on the amaranth seedlings every day to form water droplets on the amaranth seedlings for the tea angle stink bugs to drink; 3-5 ml of 10% (w / v) honey water is added every day as a nutritional supplement for the tea angle stink bugs; when the adult tea angle stink bugs have a large appetite and cause the amaranth tender buds to dry out prematurely, new amaranth seedlings are added in time for the tea angle stink bugs to feed on; (5) mating and oviposition: adult tea angle stink bugs mature 3-5 days after eclosion, then start mating, and female insects lay eggs 3-5 days after mating; most of the eggs are scattered in the epidermis of the tender stems, a small part is laid in the petiole and leaf vein, and the top of the egg is attached to a long and short white bristle; the oviposition period is 30-40 days, and the average number of eggs laid by each female insect per day is 6-7; (6) nymph hatching and rearing: the egg stage of the tea angle stink bug is 5-8 days, and the nymph stage is 10-15 days; a small amount of amaranth tender buds is added as food for the 1st instar nymphs to facilitate the low-age nymphs to feed by piercing and sucking; 3-4 leaf stage amaranth seedlings are added as food for the nymphs after they have molted twice to reach the 3rd instar; the nymphs become adult after 4 molts.

2. A method of mass rearing of Cyrtopimpla spp. according to claim 1, characterized in that, In step (1), the number ratio of female adult tea angle stink bugs to male adult tea angle stink bugs is 2:

1.

3. A method of mass rearing of Cyrtopimpla spp. in the laboratory as claimed in claim 1, wherein, In step (2), the number of amaranth seedlings cultivated to the 3-4 leaf stage is 30.

4. The method for mass rearing of D. zicuhy in the laboratory as claimed in claim 1 wherein, In step (3), the size of the insect rearing cage is 40 cm×40 cm×40 cm.

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