Low-temperature storage method for dominant parasitic wasps in mulberry field

Abstract

A low-temperature storage method for dominant parasitic wasps in a mulberry field comprises the following steps: by using female polyneomyzigus marmoreus as parasitic female wasps, enabling the female wasps to parasitize greater wax moths to obtain offspring wasps for low-temperature storage; selecting pupae and adults of the offspring bees at different growth and development stages for low-temperature storage; placing the bee pupae and the adults fed with the honey in a low-temperature environment of 4 DEG C, and performing low-temperature storage; after the low-temperature storage is finished, the bee pupae and the adults are taken out and subjected to transition treatment in a transition temperature environment; and feeding the treated bee pupae and adults for later use. The low-temperature storage method is easy and convenient to operate and low in cost, effective low-temperature storage of the parasitic wasps is achieved, the shelf life of the parasitic wasps is prolonged, and a good foundation is laid for field large-scale release.

Description

Technical Field

[0001] The invention belongs to the technical field of biological control of agricultural pests, and particularly relates to a low-temperature storage method for dominant parasitic wasps in mulberry gardens. Background Art

[0002] There are many kinds of mulberry pests. Lepidoptera pests such as Glyphodes pyloalis, Phthonandria atrineata, and Rondotia menciana occur in mulberry gardens all year round. In the year of repeated outbreaks, mulberry leaves may be completely lost, which seriously threatens the development of the sericulture industry (Wu Fuan, 2015). Due to long-term irrational use, the problem of mulberry pest resistance caused by chemical pesticides has become increasingly prominent (Xu Jinsong, 2006). In particular, the commonly used pesticides used in mulberry gardens are very likely to cause frequent poisoning of silkworms. It is urgent to develop new methods to control mulberry pests.

[0003] Biological control of parasitic wasps is an important part of the green pest management system and a key development direction for mulberry pest control. Although mulberry pests have abundant natural enemy resources, to date, biological control practices of parasitic wasps against mulberry pests have been rarely carried out. The parasitic wasps that are being promoted for use are still in the large-scale breeding stage, and the long-term stable supply of parasitic wasps is still under exploration. Low-temperature refrigeration is currently a more common method for preserving parasitic wasps, which can effectively extend the shelf life of parasitic wasp products, reduce production costs, and improve the efficiency of field control. At present, research has been carried out on parasitic wasp groups such as trichogrammatids, aphid wasps, aphid wasps, and gnawing wasps, and certain results have been achieved, but relevant work has not yet been carried out on parasitic wasps of mulberry garden pests, which greatly limits the promotion and application of biological control of mulberry pests.

[0004] Iseropus kuwanae (Vlereck), belonging to the family Ichneumonidae of the order Hymenoptera, is the main parasitic wasp of the mulberry pest mulberry leeches. Iseropus kuwanae has a wide range of hosts, including important Lepidoptera pests such as mulberry leeches, mulberry borers, striped stem borers, rice leaf folders, and wax moths. Under laboratory conditions, Iseropus kuwanae can lay eggs all year round. Under field conditions, it overwinters as a pupa in the third generation of mulberry leeches in late November. At present, the indoor parasitism and propagation techniques of this parasitic wasp have been gradually mastered, but there has been no breakthrough in long-term storage, resulting in high breeding costs and limited release in the wild. Therefore, it is urgent to develop an effective low-temperature storage method for Iseropus kuwanae, which can extend the shelf life without changing the main parasitic characteristics of the parasitic wasp, and provide a solution for giving full play to the biological control potential of the parasitic wasp. Summary of the invention

[0005] Technical problem solved: In view of the above technical problems, the present invention provides a low-temperature storage method for dominant parasitic bees in mulberry gardens, which prolongs the shelf life of parasitic bee products, ensures the parasitic indicators of parasitic bees, and provides a solution for the biological control of natural enemies of mulberry pests.

[0006] Technical solution: The application of feeding honey to prolong the shelf life of female Mylotrichum fasciatus wasps stored at low temperatures and improve their parasitism rate.

[0007] Preferably, the low-temperature storage temperature of the female Mylotrichum sphaerocephalus is 4° C., and the storage time is 5-15 days.

[0008] The female Mylotrichum sphaerocephalus is a female adult obtained from a 5-10 day old mother bee.

[0009] Preferably, the female Mylotrichum mulberryii tubercle-forming wasp is a female adult obtained from a 5-day-old mother bee.

[0010] The female adult mentioned above is a female Myriophyllum sphaerocephalum which has emerged for 1-12 hours.

[0011] The honey is honey water with a concentration of 5wt.%-30wt.%.

[0012] Preferably, the concentration of the honey water is 5wt.%.

[0013] The application also includes the step of placing the product at 8-24°C for 6 hours after low temperature storage.

[0014] The above-mentioned storage temperature is 16°C.

[0015] A method for low-temperature storage of dominant parasitic bees in mulberry gardens comprises: (1) using 5-10 day old female bees of the mulberry lily polypterus as parasitic mother bees, parasitizing wax moths, and obtaining offspring bees that have emerged for 1-12 hours for low-temperature storage; (2) selecting 5-7 day old female pupae and adults that have emerged for 1-12 hours for storage at 4°C for no more than 15 days; and (3) after the low-temperature storage is completed, taking out the pupae and adults and placing them at 8-24°C for 6 hours for standby use.

[0016] Beneficial effects: The present invention selects different developmental stages of the mulberry lily polyplastid suitable for low-temperature storage according to the biological characteristics of the parasitic wasps of mulberry garden pests, and realizes low-temperature storage of multiple stages of parasitic wasps using one parasitic wasp. The parasitic wasps stored at low temperatures can smoothly resume development, and their lifespan and reproductive capacity are not affected. The low-temperature storage method used in the present invention is simple to operate, low in cost, and realizes effective low-temperature storage of parasitic wasps, prolongs the shelf life of parasitic wasps, and lays a good foundation for large-scale release in the wild. DETAILED DESCRIPTION

[0017] The present invention will be further illustrated below in conjunction with specific embodiments. It should be understood that these embodiments are only used to illustrate the present invention and are not intended to limit the scope of the present invention. In addition, it should be understood that after reading the content of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms fall within the scope limited by the appended claims of the application equally.

[0018] Example 1 Low temperature storage of Mytilus sphaerocephalus

[0019] The 5-day-old female pupae, 5-day-old male pupae, female adults 12 hours after eclosion, and male adults 12 hours after eclosion of the mulberry tuber were used as the low-temperature storage objects, and the storage time was 5d, 15d, and 30d, respectively, and they were stored in a 4℃ refrigerator. The pupae were placed in 1.5mL centrifuge tubes (5 heads / tube), and the adults were placed in 50mL centrifuge tubes (20 heads / tube). After the low-temperature storage was completed, the pupae or adults of the parasitic bees were taken out and placed in a 20℃ incubator for 6h. After that, the survival rate of pupae and adults and the parasitism rate of female bees were investigated, and the parasitism rate of the refrigerated female pupae was investigated after eclosion. The survey results are shown in Table 1. The survival rate of female pupae, male pupae, female adults, and male adults was the highest after 5d of low-temperature storage, and the survival rate of female adults refrigerated for 15d was slightly lower than that of the 5d treatment. The parasitism rate of female adults after 15d of storage was lower than that of the 5d storage treatment, but it was within an acceptable range. Therefore, under the above conditions, female adults can still maintain a high survival rate and parasitism rate within 15 days of low-temperature storage.

[0020] Table 1 Effects of different low temperature storage time on the survival rate and parasitism rate of Ichneumonella spp.

[0021]

[0022] Different lowercase letters indicate significant differences (p<0.05)

[0023] Example 2 Low temperature storage of Myriophyllum sphaerocephalum

[0024] 5, 10 and 15-day-old female bees of the mulberry tuber were used as parasitoid mother bees to obtain bee adults and bee pupae for low-temperature storage. 5-day-old female and male pupae, and female and male adults 12 hours after eclosion of the mulberry tuber were used as low-temperature storage objects, and the storage time was 15 days, stored in a 4°C refrigerator. Bee pupae were placed in 1.5mL centrifuge tubes (5 heads / tube), and adults were placed in 50mL centrifuge tubes (20 heads / tube). After the low-temperature storage was completed, the pupae or adults of the parasitic bee were taken out and placed in a 20°C incubator for 6 hours. After that, the survival rate of pupae and adults and the parasitism rate of female bees were investigated, and the parasitism rate of the refrigerated female pupae was investigated after eclosion. The results of the survey are shown in Table 2. There was no significant difference in the survival rate of male pupae parasitized by 5-day-old and 10-day-old mother bees after low-temperature storage. The survival rates of female pupae, female adults and male adults decreased slightly, which were significantly higher than those of male and female adults obtained from 15-day-old mother bees. The parasitism rate of female bees obtained from 5-day-old mother bees after refrigeration was slightly different from that of 10-day-old mother bees, but significantly higher than that of 15-day-old mother bees. The parasitism rate of female pupae obtained from 5-day-old mother bees was significantly higher than that of female pupae obtained from 10-day-old and 15-day-old mother bees after low-temperature storage and emergence.

[0025] Table 2 Effects of different-age queen bees on the survival rate and parasitism rate of Ichneumonella mulberrye after low-temperature storage

[0026]

[0027] Different lowercase letters indicate significant differences (p<0.05)

[0028] Example 3 Low temperature storage of Mytilus sphaerocephalus

[0029] The 5-day-old female bees of the mulberry tuber-growth wasp were used as parasitoid mother bees to obtain bee pupae for low-temperature storage. The 5d, 7d and 9-day-old male and female pupae of the mulberry tuber-growth wasp were used as low-temperature storage objects, and the storage time was 15d, stored in a 4℃ refrigerator. The pupae were placed in 1.5mL centrifuge tubes (5 heads / tube), and the adults were placed in 50mL centrifuge tubes (20 heads / tube). After the low-temperature storage was completed, the pupae or adults of the parasitic bees were taken out and placed in a 20℃ incubator for 6h. After that, the survival rate of the pupae and the parasitism rate of the female bees were investigated. The survey results are shown in Table 3. The survival rate of male and female pupae aged 5 to 7 days was slightly different, which was significantly higher than that of male and female pupae aged 9 days. After the 5d and 7-day-old female pupae emerged into adults, the parasitism rate was slightly different, which was significantly higher than that of female pupae aged 9 days.

[0030] Table 3 Effects of different ages on the survival rate and parasitism rate of Pleurotus eryngii pupae after low temperature storage

[0031]

[0032] Different lowercase letters indicate significant differences (p<0.05)

[0033] Example 4 Low temperature storage of Mytilus sphaerocephalus

[0034] 5-day-old female bees of the mulberry leech gall-shaped ichneumon was used as parasitoid mother bees to obtain bee pupae for low-temperature storage. Male and female adults of the mulberry leech gall-shaped ichneumon was used as low-temperature storage objects at 1h, 12h, 24h, and 48h after eclosion for 15d in a 4℃ refrigerator. Bee pupae were placed in 1.5mL centrifuge tubes (5 heads / tube), and adults were placed in 50mL centrifuge tubes (20 heads / tube). After the low-temperature storage was completed, the pupae or adults of the parasitic bee were taken out and placed in a 20℃ incubator for 6h. Thereafter, the survival rate of adults and the parasitism rate of female bees were investigated. The results of the investigation are shown in Table 4. There was no significant difference in the survival rate of male and female adults 1h and 12h after eclosion in low-temperature storage, which was significantly higher than that of male and female adults 24h and 48h after eclosion. There was no significant difference in the parasitism rate of female adults stored at low temperature for 1 hour and 12 hours after emergence after refrigeration, but it was significantly higher than that of female bees stored at low temperature for 24 hours and 48 hours.

[0035] Table 4 Effects of storage of adults at different times after emergence on the survival rate and parasitism rate of adults of the mulberry larvae

[0036]

[0037] Different lowercase letters indicate significant differences (p<0.05)

[0038] Example 5 Low temperature storage of Myriophyllum spp.

[0039] The 5-day-old female bees of the mulberry tuber-growthed ichneumon was used as the parasitoid mother bees to obtain the bee pupae for low-temperature storage. The male and female adults of the mulberry tuber-growthed ichneumon were used as the objects of low-temperature storage 12 hours after emergence. Before the adults were stored, they were fed with honey water with a mass fraction of 5%, 15% and 30%, respectively. The storage time was 15 days and stored in a refrigerator at 4°C. The adults were placed in 50mL centrifuge tubes (20 heads / tube). After the low-temperature storage was completed, the adults of the parasitic bees were taken out and placed in a 20°C incubator for 6 hours. After that, the survival rate of the adults and the parasitism rate of the female bees were investigated. The survey results are shown in Table 5. Feeding different concentrations of honey water before low-temperature storage had no significant effect on the survival rate of male and female adults. After feeding 30% honey water for storage, the survival rate of female bees and male bees was the highest. There was no significant difference in the parasitism rate of female bees after the cold storage after being fed with honey water of different concentrations before low-temperature storage. The parasitism rate of female bees fed with 5% honey water and then refrigerated was the highest.

[0040] Table 5 Effects of feeding different concentrations of honey water before low-temperature storage on the survival rate and parasitism rate of Ichneumonella mulberrye

[0041]

[0042] Different lowercase letters indicate significant differences (p<0.05)

[0043] Example 6 Low temperature storage of Myriophyllum sphaerocephalum

[0044] The 5-day-old female bees of the mulberry tuber-growthed ichneumon was used as the parasitoid mother bees to obtain the bee pupae for low-temperature storage. The male and female adults of the mulberry tuber-growthed ichneumon were used as the objects of low-temperature storage 12 hours after emergence. They were fed with 5% honey water by mass before storage. The storage time was 15 days and stored in a refrigerator at 4°C. The pupae were placed in 1.5mL centrifuge tubes (5 heads / tube), and the adults were placed in 50mL centrifuge tubes (20 heads / tube). After the low-temperature storage was completed, the pupae or adults of the parasitic bees were taken out and placed in the incubators at 8°C, 16°C and 24°C for 6 hours. After that, the survival rate of adults and pupae and the parasitism rate of female bees were investigated. The survey results are shown in Table 6. The survival rate of male and female adults and male and female pupae treated at 16°C after low-temperature storage was the highest. The parasitism rate of female adults and adults that emerged from female pupae treated at 16°C after low-temperature storage was also the highest.

[0045] Table 6 Effects of different transition temperatures on the survival rate and parasitism rate of Ichneumonella mulberrye after low-temperature storage

[0046]

[0047] Different lowercase letters indicate significant differences (p<0.05).

Claims

1. Application of feeding honey to prolong the shelf life of female Myxocybe spp. stored at low temperature and improve its parasitism rate.

2. The use according to claim 1, characterized in that: The low-temperature storage temperature of the female Mylotrichum sphaerocephalus is 4° C., and the storage time is 5-15 days.

3. The use according to claim 1, characterized in that: The female mulberry lily tumor-forming wasp is a female adult obtained from a 5-10 day old mother wasp.

4. The use according to claim 3, characterized in that: The female Mylotrichum sphaerocephalus is a female adult obtained from a 5-day-old mother bee.

5. The use according to claim 4, characterized in that: The female adult is a female Myriophyllum mulberrye-like gall wasp that has emerged for 1-12 hours.

6. The use according to claim 1, characterized in that: The honey is honey water with a concentration of 5wt.%-30wt.%.

7. The use according to claim 5, characterized in that: The concentration of the honey water is 5wt.%.

8. The use according to claim 1, characterized in that: The method also includes the step of placing the mixture at 8-24°C for 6 hours after low temperature storage.

9. The use according to claim 7, characterized in that: The storage temperature is 16°C.

10. A method for low temperature storage of dominant parasitic wasps in mulberry gardens, characterized in that: (1) Use 5-10 day old female bees of the polypida as parasitoid queens, parasitize the wax moth, and obtain offspring bees that have emerged for 1-12 hours for low temperature storage; (2) Select 5-7 day old female pupae and 1-12 hour old adults for storage at 4°C for no more than 15 days; (3) After the low-temperature storage is completed, take out the pupae and adults and place them at 8-24℃ for 6 hours for later use.

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