CN108575918B - Large-scale propagation method for Chouioia cunea Yang - Google Patents
Large-scale propagation method for Chouioia cunea Yang Download PDFInfo
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- CN108575918B CN108575918B CN201810540470.5A CN201810540470A CN108575918B CN 108575918 B CN108575918 B CN 108575918B CN 201810540470 A CN201810540470 A CN 201810540470A CN 108575918 B CN108575918 B CN 108575918B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
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Abstract
An application of cultured Chouioia cunea Yang in preventing and treating the Chouia cunea Yang, and provides a large-scale propagation method of the Chouia cunea Yang, which relates to the technical field of forestry protection; aims at effectively and biologically preventing and controlling the fruit tip striped rice borer, other needle leaf tree species and branch tip pests, and the expanding propagation method comprises the following steps: step one, collecting and storing bee seeds; step two, breeding the bee seeds; the host is soaked in the hypochlorous acid aqueous solution, and the parasitic capacity of the red pine tip borer wasp is improved by gradient temperature reduction and preservation.
Description
Technical Field
The invention relates to the technical field of forestry protection, in particular to prevention and treatment of fruit tip striped rice borer and a large-scale propagation method for preventing and treating red pine tip striped rice borer wasp.
Background
Korean pine (Pinus koraiensis) is an evergreen arbor of Pinus of Pinaceae, is not only a precious tree species, but also a dominant species of natural forests in lesser Khingan mountains and Changbai mountains in northeast China, and is one of main afforestation and greening tree species in northeast. Meanwhile, the Korean pine seeds are rich in grease and protein, are deeply favored by domestic and foreign consumers due to extremely high nutritional and health-care values, and are important economic resources in northeast forest regions. In recent years, the increasing price of pine nuts makes economic forest of red pine become one of the important ways for people in forest areas to become rich, and is one of the important pulling forces for rapid development of forestry industry. However, in recent years, the area of the ball pests such as the pink borer (diryctria pryeri), the yellow borer (d. abietella), the diamond-back moth (eupithelia abiitaria) and the like in the dew river, the yellow mud river, the second white river and the like in the Changbai mountain area is getting bigger and bigger, the damage to the red pine is becoming serious day by day, the growth vigor of the trees is severely inhibited, the yield and the quality of the red pine seeds are greatly reduced, the ball to be matured can be seriously damaged to have no economic value in serious situations, the natural renewal of the red pine forest and the value of the red pine forest are greatly damaged, the huge economic loss is caused, and the local ecological environment is more seriously damaged. Among them, the damage of the pinkeye is the most serious.
The Pinctada martensii Karsch is also called Pinctada martensii Karsch, Pinctada sinensii Karsch, Pinctada martensii Karsch, and is an arthropod of Lepidoptera, Bocartialidae, Pinctada martensii Karsch. China occurs in northeast, northwest, Anhui, Jiangsu, Zhejiang, Sichuan and Taiwan, and is mainly harmful to Chinese pine, black pine, red pine, Chinese red pine, China mountain pine, larch, spruce and the like, and the normal growth and seed yield of trees are seriously influenced.
There are few examples of applying natural enemy insects to control forestry pests in the field of forest protection. The reason is mainly as follows: firstly, effective natural enemies are difficult to discover; secondly, natural enemy insects are not easy to propagate on a large scale; and thirdly, the natural enemy insects can not achieve the effect of continuous control after being released in the field. At present, only trichogramma, scleroderma guani, trichogramma wasp and ladybug are used for large-scale propagation and release utilization of natural enemy insects in China. At present, no biological control means capable of effectively controlling the fruit tip striped rice borer exists.
The Korean pine forest belongs to an ecosystem seriously interfered, and the ecological balance is artificially broken (felling by human beings, collecting Korean pine seeds, cultivating economic crops under the forest, releasing various pesticides, and the like). When the pesticide is used by human beings to control pests, the drug resistance of the pests is gradually improved, but a large number of natural enemies of the pests are killed due to smaller individuals and lower drug resistance, so that the pests are lack of control of the natural enemies, and a large number of outbreaks of the pests are easily caused. Therefore, in order to restore ecological balance, reduce the harm to the environment and improve food safety, the adoption of biological control means becomes the most ideal solution. Biological control, however, requires submerged control in order to achieve the desired results, which requires the provision of sufficient natural enemies of the pests to kill them.
Disclosure of Invention
The invention aims to effectively and biologically control the fruit tip borer, other needle leaf tree species and branch tip pests, the invention cultivates the red pine tip borer wasp to control the fruit tip borer and protect the application of the red pine forest tree, and also provides a large-scale propagation method of the red pine tip borer wasp.
The invention discloses a red pine tip spot borer wasp (Baryscapus dioycriae) which is an inventor for 11 years, and is finally discovered by indoor culture after continuously collecting red pine cone pests, wherein the red pine tip spot borer wasp can take the red pine tip spot borer as a host although the host is the red pine tip spot borer and has a good control effect on the red pine tip spot borer and other various needle leaf tree species and branch tip pests.
In order to better control the fruit tip borer, a large amount of red pine tip borer wasps needs to be cultivated, the biological distribution of the red pine tip borer wasps is less, the red pine tip borer wasps is concentrated in a needle forest area, and needle trees are high and have great collection difficulty, so the invention also provides a cultivation method of the red pine tip borer wasps, which comprises the following steps: the method comprises the following steps:
step one, bee species collection and low-temperature preservation: collecting pupa of Pinus koraiensis Chun in forest at pupa stage, placing into finger-shaped glass tube, culturing the finger-shaped tube containing pupa in room or incubator at 25-28 deg.C under 16L/8D, identifying bee species after pupa apis cerana Humifera emergence, and storing at low temperature;
step two, propagation and preservation of bee species: taking out the red pine tip borer wasps stored at low temperature by adopting a finger tube bee breeding method or an open bee breeding method, inoculating 2-30 heads of the red pine tip borer wasps into each host pupa, and then putting the host pupas inoculated with the red pine tip borer wasps into a thermostat or a greenhouse for culturing and storing;
wherein, before the second step of inoculating the bees, the host pupae is firstly put into a hypochlorous acid aqueous solution with the concentration of 0 to 25 percent for soaking, then taken out and washed cleanly by distilled water, and naturally dried at room temperature.
Because the Chouioia cunea Yang is small and is difficult to distinguish from various other vespids, the accurate identification method is as follows:
the red pinworm nibea webworm brood is mainly characterized as follows: the length of a female body is 1.8-2.2 mm; the whole body is dark blue-green and has metallic luster; the face, the back plate of the middle breast, the scutellum of the middle breast and the scutellum of the middle breast have copper luster; the antennary depressions are dark yellow brown; the antenna sections are yellow brown, and the handle sections have green metallic luster; compound eye ochre red, single eye yellowish; each podocarpus section has the same body color; leg sections are brown and slightly have blue-green metal luster; the tibioes and tarsal nodes are yellow, but the base 2/3 of the hind podophyllum is slightly brownish, and the distal nodes of each tarsal node are brownish; transparent wings and light brown wing veins; the anterior wing is brownish. The back of the head is slightly dumbbell-shaped, the head width is 2.1 times of the length, the width is basically equal to the chest width (42:41), and is 1.1 times of the abdomen width (varieties: the newly eclosized adult specimen adult head is narrower than the abdomen and is 0.9 times of the abdomen); the upper cheek length is 0.3 times of the compound eye length; POL is 2.5 times OOL; the back head is provided with a complete shallow and thin middle longitudinal groove without a back head ridge; the setae in the monocular region are sparse, and the periphery of the monocular region is provided with an obvious shallow eye surrounding ditch; the head top is provided with dense seta; the reticular pattern on the top of the head and the face is obvious. The front width of the head is 1.2 times of the height; the face width is 1.3 times of the compound eye height; the lower edge of the antennal fossa is positioned on the connecting line of the lower edge of the compound eye; the space between the antenna nests is the same as the transverse diameter of the antenna nests; the antennary fossa protrudes in a triangular shape up to 1/3 the height of the antennary fossa. The antennal depression is deep and reaches the front edge of the middle monocular; the hollow side area is bulged and has dense bristles.
The compound eye has dense short fiber hairs, and the compound eye is narrow and obvious around the eye groove; the palate sulcus is straight and obvious, and the length of the palate sulcus is 0.57 times of the height of the compound eye; the face area is provided with sparse setae, and the middle part of the face area is slightly raised; the distance from the lower edge of the antennal fossa to the base end of the lip is 0.54 times of the distance from the lower edge of the antennal fossa to the front edge of the middle monocular; the basal labial part is shallow and double-leaf. The tentacle handle section is slightly shorter, and the end part does not reach the middle monocular (relative length: handle section length 23, handle section base to middle monocular leading edge distance 26); the back of the handle section has carved patterns and dense felt hair, the ventral surface is smooth, and the length is 3.3 times of the width; the sum of the length of the stalk and the whiplash is shorter than the head width and is 0.88 times of the head width; the length of the stalk section is 2 times of the width, and the stalk section, the rope section and the rod section have dense felt hair; 1 section of annular section, 3 sections of cable section and 3 sections of rod section; each section is provided with 1 row of strip-shaped sensors; the cable joint is gradually widened towards the rod joint until the rod joint 1 is widest; the relative length ratio of the sections 1, 2 and 3 of the rope section is 7:5:5, and the width ratio of each section is 6:6.5: 7.0; the relative length of the bar sections 1, 2 and 3 is 6:4:4, and the width ratio is 8:7: 6; the end of the end bar section is provided with a remarkable end needle. Carrying on the back and observing the chest: the back surface is raised and protruded, and the length of the back surface is 1.2 times of the width.
The anterior dorsal back plate is narrow and steeply declines from the rear edge all the way forward without ridges, and is provided with reticular sculpture and sparse setae. The mesothorax shield is provided with compact concave ridge-shaped carving patterns which are in a fine longitudinal stripe shape; the shield longitudinal groove is deep and wide. The middle area of the middle thoracic shield has a shallow and obvious middle longitudinal sulcus, and both sides of the middle longitudinal sulcus are respectively provided with about 15 setae; the width of the mesothorax scutellum is 1.7 times of the length, the width of the front edge of the middle area is 1.2 times of the length, and the width of the front edge is 1.9 times of the width of the rear edge; the side area is provided with convex ridge net-shaped carved patterns and sparse seta; the protrusion of the triangular area is obvious and reaches 0.38 times of the length of the middle thoracic shield; the surface of the triangular plate is provided with convex ridge net-shaped carving patterns. The width of the middle chest scutellum is larger than the length, and the width is 1.3 times of the length; two of the inferior-medial sulci are obvious, and the other two are obvious lateral sulci; the small shield is provided with a concave ridge net-shaped sculpture which is thinner and denser than that of the middle-chest shield; the medial posterior part of the lateral area of the inferior sulcus of the scutellum has 3 bristles. The back board of the chest is crescent, the surface is weak and smooth.
The middle length of the posterior sternum scutellum is 0.18 times of the length of the middle sternum scutellum. The thoracic and abdominal joints are inclined backwards downwards, and the width is 9.1 times of the length; the middle part is 0.22 times as long as the scutellum of the middle chest; has a middle longitudinal ridge and no side fold ridge; the middle area is provided with concave ridge net-shaped carving patterns; the valve is large, the front edge is almost close to the front edge of the juxtaco-ventral ganglion and grows, and the distance from the front edge to the rear edge is 1.36 times of the length of the valve; the side calluses are provided with about 5 setae. The front wing extends backwards slightly beyond the abdomen and is 2.13 times as long as the width; the relative length ratio of the inferior anterior marginal pulse, the marginal pulse and the nevus pulse is 96:102: 38; 3-4 setae are arranged on the inferior anterior border; the front edge of the basal chamber has complete setae, and the end part of the lower edge of the basal chamber has 2-3 setae; the setae on the elbow pulse is complete; cilia are distributed on the wing surface longer and more prominently. The lateral posterior crus is longer, its length is 1.33 times of the tarsal and 1.14 times of the leg. Abdomen: long oval shape, pointed end; the length is 1.85 times of the width, the width is 1.14 times of the chest, and the head width is 1.11 times of the head width; the back edge of each section of the abdomen backboard is in a shallow arc shape and protrudes backwards, and the surface of the abdomen backboard is provided with obvious convex ridge net-shaped carving lines; the middle of each section of the back plate is provided with no seta, and the side surface is provided with sparse seta; the relative length of the sections 1-7 is 40:20:25:30:28:12:14, and the ratio of the rear edge width of the sections 1-6 is 76:88:92:72:44: 18; the tail section back plate is triangular, and the length of the tail section back plate is basically equal to the width of the base; 4 bristles are arranged on the tail hair. The lower genital plate ends are located 0.56 times the length of the ventral surface.
Male bodies are 1.3-1.6mm long, similar to females. The relative width of head, chest and abdomen is 25:25:22, and the relative length is 12:36: 36. The tentacle handle is light brown, and the rest sections are light earthy yellow; the relative length ratio of the stalk section, the cord section 1, 2, 3 and 4 sections and the rod section 1, 2 and 3 sections is 20:5:5:6:6:5:4:6:5, and the relative width ratio of the sections is 8:4:5:6:6.5:6.5: 6.5: 5.5: 4.5; the back of the stem node is provided with sparse seta, the ventral surface is smooth, and the lower edge of the length of the base node 3/4 is provided with callus which is dark brown; the ring-shaped bristles on each section of the cord sections 1-4 are also yellow and do not grow, and only extend forwards to the base of the ring-shaped bristles on the next section; the end of the bar section has a distinct end needle, the length of which is 1/3 of the last bar section.
In the second step of the invention, the host pupa is one or more of the fruit tip spotted borer, the tussah, the corn borer, the elm moth, the willow moth, the wax moth, the rice borer or the silkworm.
The biological distribution of the red pine tip borer wasps is less, the red pine tip borer wasps are concentrated in a needle forest area, and the needle trees are higher, so that the collection difficulty is extremely high (unfortunate occurs in the collection process, and personal injury and death are caused). The Chouioia cunea Yang is only one grain of dust, and only the damaged cones are collected back to the laboratory for dissection, the pupae of the Chouioia cunea Yang is taken out and put into a glass tube for culture, and the Chouioia cunea Yang is identified, collected and stored after the Chouioia cunea Yang eclosises.
In the northeast region, the conifer carpesiella hubner is in the stage from egg to pupa at the beginning of 6 months every year, and conifer forests without medicament control are selected for collection, and because the wild population quantity of the Chouioia cunea Yang is small, only 20 pupas can emerge from 100 host pupas when the Chouia cunea Yang is collected. The natural population quantity of the Chouioia cunea Yang is too low, and the population quantity must be expanded for submerged prevention and treatment, so that the propagation is expanded.
Although the Chouioia cunea Yang is a parasitic wasp of the obligate parasitic needle leaf tree fruit borer, the Chouioia cunea Yang can still continue to develop after being inoculated into a substitute host; and the alternative hosts can be selected from the fruit tip spotted borer, the tussah, the corn borer, the elm moth, the willow moth, the wax moth, the rice borer, the cotton bollworm, the silkworm or the pine moth, and are not limited to the conifer coccinella furnacalis, so that the difficulty of cultivation and propagation is greatly reduced.
The culture method of the red pine shoot borer wasp is easy to operate, efficient and systematic. The technical problem of breeding a large number of Chouioia cunea Yang is solved by screening excellent breeding bees, and the method has great practical significance for the biological control technology of the fruit tip Sichuan moth. The effective effects obtained by the invention are also embodied in that:
(1) the inventor finds that the Chouioia cunea Yang wasp can not pierce the epidermis to lay eggs and cause parasitic failure for the host with thicker and harder epidermis, and hypochlorous acid has softening effect on the epidermis of pupae and improves parasitic rate of the parasitic wasp by soaking in hypochlorous acid aqueous solution; for small pupae, the epidermis is softened after hypochlorous acid treatment, and the parasitic rate of parasitic wasps can be greatly improved.
(2) Aiming at the parasitic capacity of the red pine tip spaghetti walker after preservation on the fruit tip spaghetti walker, in order to preserve brood bees and prolong the shelf life of the red pine tip spaghetti walker and accumulate the number of individuals, the inventor needs to preserve the wasps at low temperature before releasing the wasps. Placing the alternate host pupae after inoculating the bees for 36 hours into a container with the temperature of 25-28 ℃ and the illumination of 16L:8D, culturing in a thermostat with the humidity of 45% -60% or a greenhouse for 7-10 days, transferring into a greenhouse with the temperature of 15 ℃, and irradiating with 12L: 12D, culturing in an incubator or a room with humidity of 45-60% for 3 days, transferring to a room or an incubator with temperature of 4 ℃ and full darkness illumination, and storing. By the gradient cooling preservation, on one hand, the parasitic capacity of the red tip pinworm nibalism on the fruit tip pinworm can be maintained, and the parasitic rate is hardly reduced; on the other hand, the shelf life of the Chouioia cunea Yang can be prolonged, and the Chouioia cunea Yang can be stored for 180 days at most.
The Chouioia cunea Yang bred by the method is put in a forest area with needle-leaved trees, so that the population quantity of the needle-leaved trees Cnaphalocrocis medinalis is effectively controlled.
Drawings
FIG. 1 is a graph showing the relationship between the development duration and temperature of Chouioia cunea Yang;
FIG. 2 is a graph showing the egg laying parasitism of Chouioia cunea Yang (Megacephalus daniell) under a microscope;
FIG. 3 is a diagram of the parasitic host situation of Chouioia cunea Yang;
FIG. 4 is a diagram of the bee-exiting situation after the wasp is received;
FIG. 5 is a diagram of a field releasing process of Chouioia cunea;
Detailed Description
The alternative host pupae described in the specification refers to the host pupae inoculated with the Chouioia cunea Yang.
Rejuvenation described in the specification means that after the reproductive capacity of the wild parasitic wasp is reduced after a plurality of generations (more than 10 generations) of the wild parasitic wasp is continuously propagated, other alternative host pupas propagated indoors are placed in the wild, and the wild parasitic wasp is taken back indoors after being parasitic, so that a wild parasitic wasp population is obtained and is mated with indoor wasp species, and the rejuvenation effect is achieved. And rejuvenating the bee species after artificial propagation for 6-10 generations to avoid the bee species from degeneration, thereby directly influencing the quality and the control effect of the propagated red pine tip pinworm Chouioia cunea.
The first embodiment is as follows: the method for breeding the red pine tip webworm nibea dorsalis comprises the following steps:
step one, bee species collection and preservation: collecting and identifying bee species at pupal stage of the fruit tip borer, and storing at low temperature;
step two, bee species reproduction: taking out the red pine tip borer wasps stored at low temperature by adopting a finger tube bee breeding method or an open bee breeding method, inoculating 2-30 heads of the red pine tip borer wasps into each host pupa, and then putting the host pupas parasitized by the red pine tip borer wasps into a thermostat or a greenhouse for culturing.
Wherein, before the second step of inoculating the bees, the host pupae is firstly put into hypochlorous acid aqueous solution with the concentration less than or equal to 25 percent for soaking, then taken out and washed clean by distilled water, and dried.
The curve of the development duration of Chouioia cunea Yang as a function of temperature is shown in FIG. 1.
Hypochlorous acid has a softening effect on the epidermis of the pupae, particularly for the relatively large individual pupae such as tussah pupae, parasitic wasps cannot pierce the epidermis to lay eggs, so that parasitic failure is caused, and for the relatively small individual pupae, the epidermis is softened after hypochlorous acid treatment, so that the parasitic rate of the parasitic wasps can be greatly improved.
The bee breeding method of the embodiment can effectively realize the propagation of the red pine tip striped rice borer wasps, and the parasitic rate reaches 100%.
The second embodiment is as follows: the present embodiment differs from the first embodiment in that: in the second step, the effective accumulated temperature of the bred Chouioia cunea Yang is 1-3 days of female bees, and the humidity is 50-75%. Other steps and parameters are the same as those in the first embodiment.
Table 1 shows the development starting point temperature and effective accumulated temperature of the Chouioia cunea Yang (using the fruit tip spotted borer as the host).
TABLE 1
The third concrete implementation mode: the present embodiment is different from the first or second embodiment in that: and in the second step, the young wasps of the fruit tip borer are female bees which emerge on the same day or 1-2 days after emergence. Other steps and parameters are the same as in one or both embodiments.
The fourth concrete implementation mode: the present embodiment is different from the first to third embodiments in that: in the second step, the host pupa is one or more of the fruit tip striped rice borer, the tussah, the corn borer, the wax moth, the rice borer or the silkworm. Other steps and parameters are the same as those in one of the first to third embodiments.
The fifth concrete implementation mode: the present embodiment is different from one of the first to fourth embodiments in that: in the first step, the preservation temperature of the Chouioia cunea Yang is-10 ℃. The other steps and parameters are the same as those in one of the first to fourth embodiments.
The sixth specific implementation mode: the present embodiment is different from one of the first to fifth embodiments in that: also comprises the rejuvenation of bee species, which comprises the following specific operations:
after the Chouioia cunea Yang is bred indoors for 10 generations, putting the alternative host pupae into a glass bottle, hanging the glass bottle in a forest, and recovering for feeding after several days; or covering the plants which substitute the hosts and are favored with gauze, putting a plurality of head substitute host larvae into the plants, taking back the larvae to be bred after the larvae are pupated and the larvae of the red pine tip borer in nature are parasitized in the pupae, and then rejuvenating once every 10 generations of breeding. Other steps and parameters are the same as those in one of the first to fifth embodiments.
The seventh embodiment: the present embodiment differs from one of the first to sixth embodiments in that: and 2-9 pinus koraiensis nakai and Chouioia cunea Yang are inoculated into each host pupa. Other steps and parameters are the same as those in one of the first to sixth embodiments.
The influence of different bee-inoculating ratios of the present embodiment on the net growth rate of the population of the pinus koraiensis wald's larva is shown in table 2.
TABLE 2
The specific implementation mode is eight: the present embodiment is different from the first to seventh embodiments in that: in the second step, the temperature of the hypochlorous acid aqueous solution is 25 +/-5 ℃; soaking host pupa in hypochlorous acid water solution for 60 + -10 s; and after inoculating the bees for 36-48 hours, putting the host pupae into a container with the temperature of 25-28 ℃ and the illumination of 16L: culturing in a thermostat or a greenhouse with the humidity of 45-60% for 7-10 days at 8D, transferring to a container with the temperature of 15 ℃ and the illumination of 12L: culturing in an incubator or room with humidity of 45% for 3 days at 12D, and transferring to a room or incubator at 4 deg.C and dark light. Other steps and parameters are the same as those in one of the first to seventh embodiments.
Example 1
The pupa of the fruit tip borer parasitized by the red pine tip borer Chouioia cunea Yang is characterized by dark gray color and no luster; the abdomen can not swing; the pupa shell is semi-transparent, thin and fragile. Selecting parasitized pupae from the collected fruit tip sipper nubilalis pupae according to the characteristics for breeding adult wasps of the red pine tip sipper nubilalis.
If the young fruit tip spotted borer pupae containing the young adult larvae of the red pine tip spotted borer are collected in winter and stored at the temperature of minus 5 ℃. Before bee breeding, the pupae are taken out and placed indoors at the room temperature of 24-28 ℃ until the pupae feather.
Example 2
The cultivation of the Chouioia cunea Yang is carried out according to the following steps:
step one, initial bee species collection: in the pupa stage of the Pinus koraiensis Chu, the Pinus koraiensis Chu pupas are collected in a forest, taken back to the indoor, put into a finger-shaped glass tube with the diameter of 1cm-2cm and the height of 5cm-7cm, put into filter paper which is wetted by distilled water in a square of 1cm, and then the mouth of the tube is plugged by a cotton ball, and 1 pupa is produced in each tube. Culturing the finger-shaped tube containing pupa in room or incubator at 25-28 deg.C under 16L/8D, identifying bee species after pupa apis cerana and apis cerana are emerged, and propagating;
step two, softening the alternative host pupae: before inoculation, the host pupae to be replaced are put into hypochlorous acid aqueous solution with different concentrations and at the temperature of 25 ℃, taken out after 60 seconds, washed cleanly by distilled water, put into a blast drying oven at the temperature of 25 ℃ and dried for 24 hours for later use, and the host pupae to be replaced in the following steps are all pupae treated in the next step.
Step three, subculturing and propagating the bee species: adopting a finger tube bee breeding method or an open bee breeding method, when the finger tube bee breeding method is used, putting the finger tube containing the bee seeds in the step one into a cuboid closed glass container with the height, the length and the width of 20cm, 35cm and 20cm respectively, and left and right symmetrical round holes with the diameter of 5cm right in front, putting 10 left and right wasps, namely 1000 plus 2000, in the initial bee receiving process, holding the finger tube containing the alternative host pupa in the step one by one hand and stretching the finger tube into the glass container from one round hole, sweeping the wasps crawling on the four walls of the glass container into the finger tube after taking a fine hairbrush from the other round hole, according to the size of the alternative host (when the fruit tip borer is used as the host, each tube is connected with 2-4, the number of other alternative host bees can be determined according to the volume of the fruit tip borer), inserting 2-30 red pine borers into each tube, plugging the opening of the small wasps, after inoculating the bees for 36 hours, putting the finger-shaped tube into a tube with the temperature of 25-28 ℃ and the illumination of 16L: and 8D, culturing in an incubator or a greenhouse with the humidity of 45-60% until bees emerge. When the open bee breeding method is used for breeding bees, the surface of the A4 paper is uniformly brushed with a layer of 10% starch paste with the width of 4cm and the vertical interval of 1cm, and then the alternative host pupae are uniformly adhered to the surface. The A4 paper stuck with the alternative host pupa is flatly placed in a wooden box with the length and the width slightly larger than the A4 paper and the bottom surface of the wooden box is provided with 100 meshes of gauze or wire gauze. Then the wooden box is moved into a room with the periphery closed and light-tight and the red bulb as the light source, the fingerling bees in the finger-shaped tube are evenly swept into the box by the fine hairbrush, and then the upper cover is covered for parasitism. According to the size of the alternative host (when the fruit tip borer is used as the host, 1000 plus 1500 heads are connected into each box, the number of the other alternative hosts can be determined according to the volume multiple of the fruit tip borer), 1000-100000 heads of the red pine tip borer wasps are connected into each tube, the temperature is 25-28 ℃ after the wasps are connected for 36 hours, and the illumination is 16L: and 8D, culturing in an incubator or a greenhouse with the humidity of 45-60% until bees emerge.
Step four, preserving bee species: in order to preserve the brood bees and prolong the shelf life of the red pine tip spotted borer wasps and accumulate the number of individuals, low-temperature preservation is needed before the wasps are released. Placing the alternate host pupae after inoculating the bees for 36 hours into a container with the temperature of 25-28 ℃ and the illumination of 16L:8D, culturing in a thermostat with the humidity of 45% -60% or a greenhouse for 7-10 days, transferring into a greenhouse with the temperature of 15 ℃, and irradiating with 12L: 12D, culturing in an incubator or a room with humidity of 45-60% for 3 days, transferring to a room or an incubator with temperature of 4 ℃ and full darkness illumination, and storing. Aiming at the parasitic capacity of the red pine tip spaghetti walker after preservation on the fruit tip spaghetti walker, in order to preserve brood bees and prolong the shelf life of the red pine tip spaghetti walker and accumulate the number of individuals, the inventor needs to preserve the wasps at low temperature before releasing the wasps. On one hand, the parasitic capacity of the red tip pinworm nibalisma on the fruit tip pinworm can be maintained, and the parasitic rate is hardly reduced; on the other hand, the shelf life of the Chouioia cunea Yang can be prolonged, and the Chouioia cunea Yang can be stored for 180 days at most.
Step five, diapause relief of bee species: and determining diapause relieving time according to the required quantity and the required time of the control bees. Firstly, transferring the brood bees stored at low temperature in the fourth step to a temperature of 15 ℃, and irradiating the brood bees with 16L:8D, culturing in a room or an incubator with humidity of 45% -60% for 3 days, then transferring to a room or an incubator with temperature of 25 ℃ again, and illuminating for 16L:8D, culturing in a room or an incubator with the humidity of 45-60% until emergence, and then beginning propagation or releasing in the field after 2 weeks of culture.
TABLE 3 the test results of the Chouioia cunea Yang test for the host reproduction of the Pinus koraiensis by using different concentrations of hypochlorous acid
Example 3
The present embodiment is different from embodiment 2 in that step two is not included.
Example 4
The pupae of the fruit tip borer can not be artificially fed, the field collection cost is high, the quantity is unstable, and the requirements of production control can not be met due to the influence of seasons.
In this example, bee breeding tests were carried out using pupae of insects such as tussah (Antheraea pernyi Guerin-Meneville), corn borer (Pyrausta nubilalis, Hubern), greater wax moth (Galleria mellonella L.), rice borer (Chilotraa aurilia), cotton bollworm (Helicoverpa armigera (Hubner)), silkworm (Bombyxmori L.), pine moth (Dendrolimus) as alternative hosts. And (3) respectively using the pupae replacing hosts to carry out bee inoculation and bee breeding, arranging one pupae in each test tube or finger-shaped tube, inoculating the bee species (the number of the bee species can be determined according to the size of the pupae and is 2-30), plugging the tube opening with absorbent cotton, and placing at room temperature for breeding.
And (3) during artificial bee breeding, using female bees emerging on the same day to receive bees, or using female bees 1-2 days after emergence to receive bees. And after 24-48 hours, finishing the oviposition process of the Chouioia cunea Yang, putting the host parasitized by the Chouia cunea Yang into a thermostat or into a bee breeding room with controllable temperature for culturing, wherein the temperature is controlled at 24-28 ℃.
When in bee breeding, the alternative host cocoons are taken out from the refrigerating chamber, and robust pupas are selected for bee breeding according to indexes such as appearance, cocoon weight, skull and buttock plate color of the pupas and the like. Pupa cocoons were peeled off and then cultivated by the methods of example 2 and example 3, respectively. And (3) putting 3-4 pupas into the finger-shaped tube during bee breeding, then inoculating the wasps, and plugging the cotton plug. 20-30 small bees are put into a large test tube for pupae of tussah, silkworm and the like, and the test tube is sealed by non-woven fabrics. After about 18-30 days, the bees emerge, and the number and the parasitic rate are counted respectively, and the results are shown in tables 4 and 5.
TABLE 4 propagation of Chouioia cunea Yang test results using different hosts of example 2
Except for bollworm and pine moth, other hosts can breed normal bees. It shows that 8 insect pupae except the fruit tip borer can be used as the alternative hosts of artificial bee breeding. The best effect is achieved by breeding bees of tussah, silkworm, corn borer, greater wax borer and rice borer, wherein the tussah, silkworm, corn borer and greater wax borer are easy to obtain materials and low in price, and the bred small bees are large in quantity, strong and robust, and very economical.
The parasitic pupae can still normally emerge after being stored for about 90 days at the temperature of 4-10 ℃, so the normal need of bee breeding in the emergence stage of the fruit tip borer in one year can be ensured.
TABLE 5 propagation of Pinus koraiensis Pink-Protoyod Chouioia test results using different hosts of example 3
Comparing the experimental data in tables 4 and 5, it can be seen that the parasitic wasp parasitizing rate can be greatly improved by processing the host with the hypochlorous acid concentration, especially for the substituted hosts of tussah and silkworm, the tussah is increased from 0% to 86%, and the silkworm is increased from 14% to 70%, so that the breeding effect of the wasp is greatly improved, and therefore, the technical effect brought by processing the host with the hypochlorous acid concentration is unpredictable for the technical personnel in the field.
Example 5
The host is the fruit tip spotted borer; the gradient temperature changing mode is as follows: and putting the alternate host pupae after the bee inoculation for 36 hours into a container with the temperature of 25 ℃ and the illumination of 16L:8D, culturing in an incubator with humidity of 50% or in a greenhouse for 8 days, transferring into a greenhouse with temperature of 15 ℃, and irradiating with light of 12L: and 12D, culturing in an incubator with humidity of 50% for 3 days, and then transferring to a room or an incubator with temperature of 4 ℃ and full darkness illumination for preservation. The comparison example is that the alternate host pupae after 36 hours of bee inoculation are put into a chamber with the temperature of 15 ℃ and the illumination of 12L: 12D, performing low-temperature refrigeration after propagation and preservation in an incubator with the humidity of 50%; and temperature 25 ℃, illumination 16L: and 8D, performing low-temperature refrigeration after propagation and storage in a constant temperature box with the humidity of 50%, wherein other conditions of the comparative example are the same as those of the gradient temperature changing mode.
Comparing the data in table 6, it can be seen that on one hand, the parasitism capability of the red tip pinworm nibalism walskii to the fruit tip pinworm can be maintained, and the parasitism rate is hardly reduced; on the other hand, the shelf life of the Chouioia cunea Yang can be prolonged, and the Chouioia cunea Yang can be stored for 180 days at most.
TABLE 6 influence of different propagation and preservation conditions on bee emergence
Claims (4)
1. A method for breeding Chouioia cunea Yang of Pinus koraiensis Chun is characterized by comprising the following steps: step one, bee species collection and low-temperature preservation: collecting pupa of Pinus koraiensis Chun in forest at pupa stage, placing into finger-shaped glass tube, culturing the finger-shaped tube containing pupa in room or incubator at 25-28 deg.C under 16L/8D, identifying bee species after pupa apis cerana Humifera emergence, and storing at low temperature; step two, propagation and preservation of bee species: taking out the red pine tip borer wasps stored at low temperature by adopting a finger tube bee breeding method or an open bee breeding method, inoculating 2-30 heads of the red pine tip borer wasps into each host pupa, and then putting the host pupas inoculated with the red pine tip borer wasps into a thermostat or a greenhouse for culturing and storing; wherein, before the second step of inoculating the bees, the host pupae is put into a hypochlorous acid aqueous solution with the concentration of 5 to 25 percent for soaking, then taken out and washed clean by distilled water, and naturally dried at room temperature;
in the second step, the host pupae inoculated with the bees for 36-48 hours are placed at the temperature of 25-28 ℃ and the illumination is 16L:8D, culturing for 7-10 days in a thermostat with humidity of 45% -60% or a greenhouse, then transferring into a container with temperature of 15 ℃ and illumination of 12L: 12D, culturing in an incubator with the humidity of 45-60% for 3 days, and then transferring to a room or an incubator with the temperature of 4 ℃ and the illumination of full darkness for preservation;
the diapause of the bee species is relieved after the step two: the immigration temperature of the preserved breeding bees is 15 ℃, and the illumination is 16L:8D, culturing in a room or an incubator with humidity of 45% -60% for 3 days, then transferring to a room or an incubator with temperature of 25 ℃ again, and illuminating for 16L:8D, culturing the seeds in a room or an incubator with the humidity of 45-60% until emergence, and then starting propagation or releasing the seeds in the field after 2 weeks of culture; in the second step, the effective accumulated temperature of the bred Chouioia cunea Yang is 342.50 +/-7.48 days, and the humidity is 50-75%; the Chouioia cunea Yang is a female bee which emerges on the same day or 1-3 days after the emergence;
in the second step, the host pupa is one or more of the fruit tip striped rice borer, the tussah, the corn borer, the wax moth, the rice borer or the silkworm.
2. The method for rearing the Chouioia punctiferalis as recited in claim 1, wherein in the first step, the temperature for cryogenically preserving the Chouia punctiferalis is-10 ℃ to 10 ℃.
3. The method for cultivating the Chouioia cunea Yang as claimed in claim 1, wherein the method further comprises rejuvenation of bee species after indoor breeding of the Chouioia cunea Yang for 10 generations, and comprises the following specific operations: after the Chouioia cunea Yang is bred indoors for 10 generations, putting the alternative host pupae into a wide-mouth bottle, hanging the wide-mouth bottle in a forest, and recovering for feeding after several days; or covering the plants which substitute the hosts and are favored with gauze, putting a plurality of head substitute host larvae into the plants, taking back the larvae to be bred after the larvae are pupated and the larvae of the red pine tip borer in nature are parasitized in the pupae, and then rejuvenating once every 10 generations of breeding.
4. The method for cultivating Chouioia punctiferalis Guenee according to claim 1, wherein the temperature of the hypochlorous acid aqueous solution in the second step is 20-30 ℃, and the host pupae are soaked in the hypochlorous acid aqueous solution for 50-70 s.
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| CN103650697A (en) * | 2012-09-21 | 2014-03-26 | 张勇 | Method for promoting cynomorium songaricum seeds to germinate |
| CN103141443B (en) * | 2013-03-08 | 2015-09-16 | 中国农业科学院草原研究所 | A kind of regulate and control the method for beet webworm Ah lattice ichneumon wasp diapause |
| CN105145487B (en) * | 2014-07-23 | 2017-12-08 | 中国科学院动物研究所 | The artificial breeding production method of Trichogramma |
| CN105028333B (en) * | 2015-06-25 | 2017-07-21 | 吉林省林业科学研究院 | A kind of method of artificial room's mass rearing Berry size |
| CN105900748A (en) * | 2016-05-05 | 2016-08-31 | 四川农业大学 | Method for biologically preventing and treating Clytus validus Fairmaire larvae through Scleroderma sichuanensis Xiao |
| CN105850897B (en) * | 2016-05-09 | 2018-10-26 | 漯河市森林病虫害防治检疫站 | A kind of artificial fecundation method of Tetrastichus septentrionalis |
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- 2018-05-30 CN CN202011116128.6A patent/CN112219796B/en not_active Expired - Fee Related
- 2018-05-30 CN CN201810540470.5A patent/CN108575918B/en not_active Expired - Fee Related
- 2018-05-30 CN CN202011116133.7A patent/CN112219797B/en not_active Expired - Fee Related
- 2018-05-30 CN CN202011116146.4A patent/CN112219798B/en not_active Expired - Fee Related
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Also Published As
| Publication number | Publication date |
|---|---|
| CN112219798B (en) | 2022-01-11 |
| CN112219796A (en) | 2021-01-15 |
| CN112219798A (en) | 2021-01-15 |
| CN112219797B (en) | 2022-02-01 |
| CN108575918A (en) | 2018-09-28 |
| CN112219796B (en) | 2022-01-14 |
| CN112219797A (en) | 2021-01-15 |
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