CN107410213A - A kind of method for propagating Monochamus alternatus artificially - Google Patents

Abstract

The invention discloses a method for artificially breeding Monochamus alternatus, which is used for raising and breeding Monochamus alternatus under laboratory conditions. The method includes: (1) artificial breeding of sub-mature Monochamus alternatus individuals; (2) equipment of an artificial delivery room for Monochamus alternatus; (3) artificial pairing breeding and induced breeding of Monochamus alternatus sexually mature individuals; 4) Preparation of artificial feed for Monochamus alternatus larvae at various ages; (5) Separation and acquisition of monochamus monochamus larvae; (6) Subsequent cultivation of Monochamus alternatus larvae transferred to artificial feed; (7) Monochamus alternata larvae Cow pupal stage and eclosion management. The method for artificially cultivating Monochamus alternatus adopted in the present invention constructs a set of systems suitable for artificially cultivating Monochamus alternatus at each growth stage under laboratory conditions. The survival rate of each growth stage of cattle, eliminate the influence of seasons on the growth and reproduction of Monochamus alternatus, shorten the growth cycle of Monochamus alternatus, and form a stable source of experimental insects.

Description

A kind of method of artificially breeding Monochamus alternatus

technical field

The invention belongs to the field of experimental animal breeding, and in particular relates to a method for artificially breeding Monochamus alternatus.

Background technique

Monochamus alternatus is a major pest of pine trees, widely distributed in my country and even in the world. The larvae eat the trunk and the adults eat the bark, leading to weakening of the tree. In severe cases, the whole pine tree may die and break. Simultaneously, the monochamus alternata is also an effective carrier of the pine xylophilus nematode, the direct culprit of the dead disease of pine trees. The pine wood nematode migrates between the pine forests with the monochamus alternatus by parasitizing in the trachea of the monochamus alternata. The plant is burnt and withered. Monochamus monochamus and pine wood nematode disease cause huge direct economic losses to the forestry industry every year, so more effective control of monochamus alternatus and pine wood nematode disease is imminent.

The methods currently used to control Monochamus alternatus mainly include trapping, destruction of pest trees, pesticide spraying, and biological control, among which biological control includes natural enemy animal control and fungal control, Beauveria bassiana, Bacillus thuringiensis, etc. Monochamus monochamus has a certain poisonous effect, which is effective against both adults and larvae of Monochamus alternatus, and has lower adverse effects on the environment than other control methods. In the process of finding efficient control methods, it is undoubtedly necessary to use a large number of monochamus alternatus in various stages for experiments. The traditional methods of collecting monochamus alternatus adults in the field and isolating monochamus alternatus larvae from infected trees are time-consuming and laborious , and due to seasonal factors, it is difficult to obtain Monochamus alternatus in various stages at any time of the year. If artificial cultivation of Monochamus alternatus can be realized under laboratory conditions, it will bring great help to the research work of Monochamus alternatus control.

Aiming at the biological characteristics of Monochamus alternatus, a method of artificially breeding Monochamus alternatus is used to raise and reproduce each stage of Monochamus alternatus under laboratory conditions, so as to reduce the consumption of various aspects and solve the influence of seasonal factors To solve the experimental problems, establish a stable laboratory population of Monochamus alternatus and provide a strong guarantee for the research work on the prevention and control of Monochamus alternatus and pine wood nematode disease.

Contents of the invention

The purpose of the present invention is to provide a method for artificially cultivating Monochamus alternatus, get rid of the influence of seasonal factors on Monochamus alternatus related experiments, reduce the consumption of all aspects of collecting Monochamus alternata in the field, and build a stable Monochamus alternatus artificial population.

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

A method for artificially cultivating Monochamus alternatus, said method comprising the following steps:

(1) Artificial breeding of sub-mature Monochamus alternatus;

(2) The equipment of the artificial delivery room of Monochamus alternatus;

(3) Artificial mating and induced breeding of sexually mature individuals of Monochamus alternatus;

(4) Preparation of artificial feed for larvae of different ages of Monochamus alternatus;

(5) Separation and acquisition of newborn larvae of Monochamus alternatus;

(6) Subsequent cultivation of monochamus monochamus larvae transferred to artificial feed;

(7) Management of the pupal stage and eclosion of Monochamus alternatus.

Specific steps are as follows:

1. Hang the monochamus alternatus trap in the pine forest in the area where the monochamus monochamus occurs; or separate the mature monochamus alternatus larvae from the infected wood, take a 12 ml EP test tube and fill it with natural Moisten wood pulp paper towels with water, put the mature monochamus alternatus larvae in this tube, and place the test tube containing the mature monochamus alternatus larvae in a constant temperature box with a temperature of 25~28°C and a humidity of 65~80%. Cultured in medium darkness to promote pupation and eventual eclosion of mature larvae.

2. Prepare a gauze cage with a length and width of 50-60 cm and a height of 80-120 cm as the long-term beetle delivery room. Put a 500ML triangular bottle in the center of the cage, fill it with clean water, and insert fresh pine branches into the triangular bottle for hydroponics as Nutritious branches; place a fresh healthy masson pine band bark wood section with a diameter of 5-10 cm, a length of 15-20 cm and both ends sealed in the four corners of the cage as the spawning wood. Place the above-mentioned Monochamus alternatus farrowing room in a greenhouse with a temperature of 25-28°C, a humidity of 65-80%, and a photoperiod of light:dark=12:12h.

3. Put the adults of Monochamus alternatus into a group of two females and one male, and put them into the prepared delivery room, and let them mate and lay eggs randomly. Five to six groups of Monochamus alternata adults were placed in each delivery room. Replace the vegetative shoots and spawning wood every two days, and mark the time on the spawning wood.

4. After the mature larvae of Monochamus alternata isolated from the phytophthora erupted, take a small plastic goldfish tank as a breeding box, put paper towels on the bottom as a moisturizing material, and add water to moisten it; intercept a section of about 10cm nutrient tree branch and put it in The box serves as feed for newly eclovened subadult adults. Put the eclosioned subadult adults into the above-mentioned breeding boxes, one in each box, and place them in the same environment as the delivery room to raise them for seven to ten days until they are sexually mature. Cage feeding and breeding.

5. Place the replaced oviposition wood in the delivery room in a greenhouse under the same conditions for further cultivation for 12 to 15 days, then carefully peel off the bark and xylem with a wood carving knife, and take out the primary monochamus larvae resident in the phloem.

6. The bark and xylem of the oviposition wood of the isolated Monochamus alternata larvae are crushed to below 40 mesh respectively, and prepared into xylem powder and bark powder for subsequent use; allocation is configured into 1-2 instar Monochamus alternatus Feed for young larvae and feed for older larvae of Monochamus alternatus after the 3rd instar;

The 1-2 instar young larvae feed of Monochamus alternatus comprises the following raw materials in parts by weight: 45-55 parts of xylem powder, 90-110 parts of bark powder, corn flour, soybean powder, wheat bran, edible fungus chaff 60~70 parts of one or several kinds of benzoic acid, sodium benzoate, sorbic acid, potassium sorbate 2~3 parts, one or several kinds of dry yeast, yeast powder, and yeast extract species 20 ~ 30 parts, 25 ~ 35 parts of agar strips, 8 ~ 15 parts of shrimp shell powder, 35 ~ 45 parts of sucrose, 280 ~ 320 parts of single distilled water; after the described 3 ages, the old larvae feed of Monochamus alternatus comprises the following Raw materials in parts by weight: 140-160 parts of xylem powder, 60-70 parts of one or more of corn flour, soybean flour, wheat bran, and edible fungus bran, benzoic acid, sodium benzoate, sorbic acid, and potassium sorbate 2~3 parts of one or several kinds, 20~30 parts of one or several kinds of dry yeast, yeast powder, and yeast extract, 25~35 parts of agar strips, 8~15 parts of shrimp shell powder, 35~45 parts of sucrose , 280～320 parts of single distilled water.

7. Put the above-mentioned ingredients except xylem powder and phloem powder in an aluminum pot and heat until the agar strip dissolves, and mix the ingredients into a uniform paste; take 45~55 parts of xylem powder, 90~110 parts of bark powder and The above-mentioned ingredients are mixed with a paste, cooled, filled into a 12MLEP test tube to a third of the volume from the mouth of the tube, and sealed to make artificial feed for young larvae of Monochamus alternatus. Take 140~160 parts of xylem powder and mix it with the above-mentioned ingredients, cool it and fill it into a 50ML centrifuge tube to a third of the volume from the mouth of the tube, seal the cap, and make the 3-year-old Alpinia monochamus. Artificial feed for middle and senior larvae. The above two artificial feeds are put into a sterilizing pot, sterilized at 121 degrees Celsius for 30 minutes, and then taken out and cooled for later use.

8. Use sterilized tweezers to pierce a small hole in the center of the subpackaged artificial feed for young larvae, put the separated newborn Monochamus alternata larva head down into the small hole, cover the centrifuge tube cover, and place it in the same They were cultured in an artificial climate box with the same environmental conditions in the delivery room. Regularly observe the feeding situation of Monochamus alternata larvae, and carefully take out Monochamus alternata larvae that have grown to about the early stage of the third instar two weeks later, complete the rearing of the young stage, and then use the same method above to remove the monochamus alternatus larvae The larvae of monochamus alternatus are transferred to the artificial diet of middle and old larvae for subsequent cultivation, and the feeding and growth of monochamus alternatus are regularly observed, until the pupal chamber is flattened out of the artificial diet of monochamus alternatus, and finally a new generation emerges adults of

9. Put the newly emerged adults into the goldfish tank feeding box and raise them in a single cage for 7-10 days, then transfer them to the delivery room to mate and lay eggs, and complete a round of feeding. By batch production of monochamus alternatus adults, the growth stages of monochamus monochamus are staggered, so as to achieve the monochamus alternatus maintaining each insect state and growth stage at the same time. The above process is continuously circulated to realize the laboratory artificial breeding of Monochamus alternatus.

The advantages of the present invention are:

1, by adopting a kind of method of artificially raising Monochamus alternatus described in the present invention, with existing patent (CN 10228853B; CN 101983580 A; CN 102715377 A) and reported article "Indoor Monochamus alternatus artificial breeding Compared with the method of artificially raising Monochamus alternatus described in "Observation of its biological characteristics", the utilization rate of pine wood has been improved, the use of cheap raw materials such as fungus chaff and wheat bran has saved the cost of feeding and manpower and material resources, A proper amount of chitin-supplementing shrimp shell ingredients are added to the larval diet of Monochamus alternata, and other components are also more scientific. The feed for adults is also more in line with the feeding habits of Monochamus alternatus, which is more conducive to Cattle growth. Under laboratory conditions, the subculture of Monochamus alternatus has been realized, a stable experimental population of Monochamus alternatus has been established, the growth cycle of Monochamus alternatus has been shortened, and it is not affected by seasonal factors. The age of Monochamus alternatus was used for experimental research. It greatly reduces the consumption of manpower, material and financial resources caused by the traditional field collection and isolation of Monochamus alternatus, accelerates the process of research on the prevention and control of Monochamus alternatus and pine wood nematode, and makes great contributions to the forestry industry.

2. By adopting the method of the present invention, the purification of Monochamus alternata strains can be carried out. Compared with the monochamus alternatus collected in the wild, the monochamus alternatus raised by this method can reduce the influence of the pathogenic microorganisms carried by the monochamus alternatus itself on the experiment as much as possible, and is a better experimental material.

Description of drawings

Figure 1 Schematic diagram of single-cage rearing (left picture) and delivery room (right picture).

Fig. 2 Schematic diagram of artificial feed for larvae (left image) and middle-aged and older larvae (right image) growing in the feed.

Figure 3. Monochamus alternatus mating (left) and a female monochamus laying eggs (right).

detailed description

Example 1

During September to October, go to the occurrence area of Monochamus alternatus to collect branches and trunks eaten by Monochamus alternatus, separate Monochamus alternatus from them, and place them in EP test tubes with moist paper towels, and set the ambient temperature to 27 ℃, humidity 70%, cultured in a dark box until eclosion into adults. Place the newly emerged adults in a feeding box lined with moist paper towels, and feed them with fresh branches. After feeding for ten days, put them into the prepared delivery room in groups of two females and one male, and put five groups of pine moths in each delivery room. Adult beetle. Change the vegetative branches and the wood that lay eggs in the delivery room every two days, put the time mark on the wood that lays eggs, place and continue to cultivate for 15 days under the same conditions as the delivery room, peel off the bark of the wood that lays eggs with a carving knife, Take out the young monochamus alternatus larvae born in the phloem, transfer them to the prepared monochamus alternatus juvenile larvae feed and cultivate them for two weeks, and then transfer them to the prepared monochamus alternatus middle-aged and senior larvae feed to cultivate until eclosion. Enter the next feeding cycle. By staggering the breeding time of the monochamus alternatus adults, the growth stages of the offspring monochamus alternatus are staggered, so that the monochamus alternatus of each insect state and each growth stage can be kept at the same time. The eclosion rate of Monochamus alternatus cultivated in this embodiment reached 83.5%.

The 1-2 instar Monochamus alternatus young larva feed comprises the following raw materials in parts by weight: 50 parts of xylem powder, 100 parts of bark powder, 65 parts of corn flour, 2 parts of benzoic acid, 25 parts of dry yeast, 30 parts of agar bar, shrimp 10 parts of shell powder, 40 parts of sucrose, 300 parts of single distilled water;

The 3-year-old long-term larva feed comprises the following raw materials in parts by weight: 150 parts of xylem powder, 65 parts of soybean meal, 3 parts of sodium benzoate, 25 parts of yeast powder, 30 parts of agar strips, 10 parts of shrimp shell powder, 40 parts of sucrose Part, 300 parts of single distilled water.

Example 2

1. In the pine forest where the monochamus alternatus occurs, hang the monochamus alternatus trap to trap the wild monochamus alternatus live.

2. Prepare a gauze cage with a length and width of 60 cm and a height of 120 cm as the delivery room of longhorn beetles. Put a 500ML triangular bottle in the center of the cage, fill it with clean water, and insert fresh pine branches into the triangular bottle for hydroponics as nutrient branches; Four jiaos in the cage respectively place a diameter that is 10CM, length is 20CM and the fresh healthy masson pine band bark wood section of two ends sealing as laying eggs wood. The above-mentioned Monochamus alternatus farrowing room was placed in a greenhouse with a temperature of 25°C, a humidity of 65%, and a photoperiod of light:dark=12:12h.

3. Put the adults of Monochamus alternatus into a group of two females and one male, and put them into the prepared delivery room, and let them mate and lay eggs randomly. Six groups of Monochamus alternata adults were placed in each delivery room. Replace the vegetative shoots and spawning wood every two days, and mark the time on the spawning wood.

4. After the mature larvae of Monochamus alternata isolated from the phytophthora erupted, take a small plastic goldfish tank as a breeding box, put a paper towel on the bottom as a moisturizing material, and add water to moisten it; cut a 10CM nutrient branch and put it in the box as feed for newly emerged subadult adults. Put the eclosioned subadult adults into the above-mentioned breeding boxes, one in each box, and place them in the same environment as the delivery room to raise them for seven to ten days until they are sexually mature. Cage feeding and breeding.

5. Place the replaced oviposition wood in the delivery room in a greenhouse under the same conditions to continue culturing for 15 days, then carefully peel off the bark and xylem with a wood carving knife, and take out the first-born Monochamus alternatus larvae resident in the phloem.

6. The bark and xylem of the oviposition wood of the isolated Monochamus alternata larvae are crushed to below 40 mesh respectively, and prepared into xylem powder and bark powder for subsequent use; allocation is configured into 1-2 instar Monochamus alternatus Feed for young larvae and feed for older larvae of Monochamus alternatus after the 3rd instar;

Described 1～2 instar Monochamus alternatus young larvae feed comprises following weight portion raw material: 45 parts of xylem powders, 90 parts of bark powders, 60 parts of soybean powders, 2 parts of sorbic acid, 20 parts of yeast powders, 25 parts of agar strips 8 parts, 8 parts of shrimp shell powder, 35 parts of sucrose, 280 parts of single distilled water; Described after the 3rd age, the larvae feed of Monochamus alternatus comprises the following raw materials in parts by weight: 140 parts of xylem powder, 60 parts of wheat bran, sorbic acid 3 parts of potassium, 20 parts of yeast powder, 25 parts of agar strips, 8 parts of shrimp shell powder, 35 parts of sucrose, 280 parts of single distilled water.

7. Put the ingredients other than the xylem powder and phloem powder in an aluminum pot and heat until the agar strip dissolves, mix the ingredients into a uniform paste, add an appropriate amount of sulfuric acid to adjust to pH=5; take the xylem powder, bark powder and The above-mentioned ingredients are mixed with a paste, cooled, filled into a 12MLEP test tube to a third of the volume from the mouth of the tube, and sealed to make artificial feed for young larvae of Monochamus alternatus. Take the xylem powder and mix it with the above ingredients, cool it and fill it into a 50ML centrifuge tube to a third of the volume from the mouth of the tube, seal the cap, and make the middle-aged and senior-aged larvae of Monochamus alternatus after it has transferred to the third instar Artificial feed. The above two artificial feeds are put into a sterilizing pot, sterilized at 121 degrees Celsius for 30 minutes, and then taken out and cooled for later use.

8. Use sterilized tweezers to pierce a small hole in the center of the subpackaged artificial feed for young larvae, put the separated newborn Monochamus alternata larva head down into the small hole, cover the centrifuge tube cover, and place it in the same They were cultured in an artificial climate box with the same environmental conditions in the delivery room. Regularly observe the feeding situation of Monochamus alternata larvae, and carefully take out Monochamus alternata larvae that have grown to about the early stage of the third instar two weeks later, complete the rearing of the young stage, and then use the same method above to remove the monochamus alternatus larvae The larvae of monochamus alternatus are transferred to the artificial diet of middle and old larvae for subsequent cultivation, and the feeding and growth of monochamus alternatus are regularly observed, until the pupal chamber is flattened out of the artificial diet of monochamus alternatus, and finally a new generation emerges adults of

9. Put the newly emerged adults into a single cage in a goldfish tank feeding box and raise them for 10 days, then transfer them to the delivery room for mating and laying eggs, and complete a round of raising. By batch production of monochamus alternatus adults, the growth stages of monochamus monochamus are staggered, so as to achieve the monochamus alternatus maintaining each insect state and growth stage at the same time. The above process is continuously circulated to realize the laboratory artificial breeding of Monochamus alternatus. The eclosion rate of Monochamus alternatus cultivated in this embodiment reaches 86.0%.

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 (8)

1. a method for artificially breeding Monochamus alternatus, is characterized in that, described method comprises as follows: (1) Artificial breeding of sub-mature Monochamus alternatus; (2) The equipment of the artificial delivery room of Monochamus alternatus; (3) Artificial mating and induced breeding of sexually mature individuals of Monochamus alternatus; (4) Preparation of artificial feed for larvae of different ages of Monochamus alternatus; (5) Separation and acquisition of newborn larvae of Monochamus alternatus; (6) Subsequent cultivation of monochamus monochamus larvae transferred to artificial feed; (7) Management of the pupal stage and eclosion of Monochamus alternatus. 2. A method for artificially breeding Monochamus alternatus according to claim 1, characterized in that: step (1) is: hanging Monochamus alternatus traps in the pine forest in the area where Monochamus alternata occurs to trap wild Monochamus alternatus Live cattle; or separate mature Monochamus alternatus larvae from infected wood, take a 12ml EP test tube, fill it with natural wood pulp paper towels, add water to moisten, put the mature Monochamus alternatus larvae in this tube, and The test tubes containing the mature larvae of Monochamus alternata were placed in an incubator with a temperature of 25-28°C and a humidity of 65-80% for dark cultivation, to promote the mature larvae to pupate and finally emerge into adults. 3. A kind of method for artificially cultivating Monochamus alternatus according to claim 1, characterized in that: the equipment of the artificial delivery room of Monochamus alternatus: the preparation length and width are respectively 50～60CM, and the height is 80～120CM. The gauze cage is used as the delivery room of the beetle, and a 500ML triangular bottle is placed in the center of the cage, filled with clean water, and fresh pine branches are inserted into the triangular bottle for hydroponics as nutrient branches; a diameter of 5~10CM, The length is 15 ~ 20CM and the fresh healthy masson pine band bark wood section of both ends sealing is used as spawning wood; The above-mentioned pine nut farrowing room is placed in a temperature of 25 ~ 28 ℃, a humidity of 65 ~ 80%, and a photoperiod of Lighting: dark = 12:12h in the greenhouse. 4. a kind of method for artificially cultivating Monochamus alternatus according to claim 1 is characterized in that: the artificial mating of Monochamus alternatus sexually mature individuals and the induced breeding are: the Monochamus alternata adults are divided into two females and one male Make it into a group, put it into the prepared delivery room, let it randomly mate and lay eggs; put five to six groups of Monochamus alternatus adults in each delivery room; replace the vegetative branches and the egg-laying wood every two days, and Mark the time on the egg wood. 5. a kind of method for artificially cultivating Monochamus alternatus according to claim 1, is characterized in that: the separation of Monochamus alternatus primary larva is obtained for placing the egg-laying wood replaced in the delivery room in the greenhouse of the same condition After continuing to cultivate for 12 to 15 days, carefully peel off the bark and xylem with a wood carving knife, and take out the first-born Monochamus alternata larvae resident in the phloem. 6. a kind of method for artificially cultivating Monochamus alternatus according to claim 1 is characterized in that: the follow-up cultivation of Monochamus alternatus larvae into artificial feed is: separating the egg-laying wood of Monochamus alternatus larvae The bark and xylem were crushed to less than 40 meshes respectively, and prepared into xylem powder and bark powder for subsequent use; distributed and configured as feed for young larvae of Monochamus alternatus 1-2 ages and advanced larvae of Monochamus alternatus after 3 ages Feed; Prick a small hole with sterilized tweezers in the feed center of the centrifuge tube of the subpackaged young larvae feed, put the isolated monochamus alternatus larva head down into the small hole, cover the centrifuge tube cover, Place them in an artificial climate box with the same environmental conditions as the delivery room; regularly observe the feeding situation of Monochamus alternatus larvae, and carefully take out Monochamus alternatus larvae that have grown to the early 3rd instar two weeks later to complete the rearing of the young stage , then use the above-mentioned same method to transfer the taken-out monochamus alternatus larvae into the feed of middle and old larvae for follow-up cultivation, regularly observe the feeding situation and growth of monochamus alternatus until it is added to the artificial diet of monochamus alternatus Level off the pupal chamber to pupate, and finally emerge a new generation of adults. 7. a kind of method for artificially cultivating Monochamus alternatus according to claim 6, is characterized in that: described 1～2 instar Monochamus alternatus low-age larvae feed comprises the following raw materials in parts by weight: xylem powder 45～55 90-110 parts of bark powder, 60-70 parts of one or more of corn flour, soybean flour, wheat bran, and edible mushroom bran, one of benzoic acid, sodium benzoate, sorbic acid, and potassium sorbate 2-3 parts of one or more species, 20-30 parts of one or more species of dry yeast, yeast powder, and yeast extract, 25-35 parts of agar strips, 8-15 parts of shrimp shell powder, 35-45 parts of sucrose, 280～320 parts of simple distilled water; The described 3 ages later the long-age larvae feed of Monochamus alternatus comprises the following parts by weight of raw materials: 140～160 parts of xylem powder, corn flour, soybean powder, wheat bran, edible fungus fungus chaff 60-70 parts of one or more, 2-3 parts of one or more of benzoic acid, sodium benzoate, sorbic acid, and potassium sorbate, 20 parts of one or more of dry yeast, yeast powder, and yeast extract ~30 parts, 25~35 parts of agar strips, 8~15 parts of shrimp shell powder, 35~45 parts of sucrose, 280~320 parts of single distilled water. 8. a kind of method for artificially cultivating Monochamus alternatus according to claim 1, is characterized in that: Monochamus alternatus pupal stage and eclosion management are: the adult of newly eclosion is put into the rearing box and single-cage raises 7～ After 10 days, transfer to the delivery room for mating and laying eggs, and complete a round of feeding; through the batch production of Monochamus alternatus adults, the growth stages of Monochamus alternatus are staggered, so as to achieve the simultaneous preservation of Monochamus alternatus in various stages and growth stages. Beetle; the above process is continuously circulated to realize the artificial breeding of Songmo longifer in the laboratory.