Method for mass rearing of aphidius gifuensis and its application in preventing and treating of calophya japonica

Abstract

The application discloses a method for breeding of the Tamarixia radiata and application of the Tamarixia radiata in prevention and treatment of the Sibicides brouni, wherein the culture tube containing the paired Tamarixia radiata adult is vertically placed in a culture box for culture, the culture tube is placed in a low-temperature storage environment and is supplemented with food 1-2 times per week, a green bean weevil host is selected, the Tamarixia radiata in the tube is taken out from the low-temperature environment, 30-40 green bean weevils with the green bean weevil hosts are put into each culture tube after the tube cover is opened, the culture tube is closed, the culture tube is placed horizontally in the culture box for mating, the host is regularly replaced after oviposition, the replaced host is placed in a new tube together with the green bean and is placed horizontally in the culture box or in a room temperature environment until the next generation is eclosed. The method can prepare a large number of parasitic wasps, further test host types, select other coleopteran weevil hosts and has potential natural enemy prevention and treatment application value, and is a new measure for prevention and treatment of natural enemies of garden pests.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of agriculture and forestry industry, in particular to a method for breeding Anisequispina sp. and application thereof in preventing and treating Adoxophyes sp. BACKGROUND

[0002] Anisequispina sp. is a parasitic insect of Tenthredinidae, which moves agilely and jumps well, is heterogonous, mainly parasitizes on Acanthoscelides obtectus in Shandong province, and a small amount of it can parasitize on broadshouldered small bees. It has 3 generations in a year, goes through multiple hosts, and has unbalanced generations. The old mature larvae hibernate in the seeds parasitized by the host. The male bees are earlier than the female bees when they are hatched, and the male bees can survive for 5-8 days after feeding 10% honey, while the female bees can survive for 13-20 days. The adult life is positively correlated with humidity. The female bees can continuously lay eggs for 6-8 days, lay 1-5 eggs in each host, and lay about 60 eggs in total. Only one adult can be hatched from each host, and the development cannot be completed on the young host. In the investigation in Wuhan area, it is found that the overwintering generation widely parasitizes on Acanthoscelides obtectus, and the occurrence varies in different places. In the subsequent host test, it is found that it has good parasitic characteristics on common warehouse pests Callosobruchus chinensis and common garden leaf-eating pests Adoxophyes sp., and has great development potential.

[0003] At present, Anisequispina sp. lacks an applicable breeding method. At the same time, Adoxophyes sp. is a Coleoptera Curculionidae insect, which is an important leaf-eating pest of Ligustrum lucidum, Ligustrum sinense and Ligustrum delavayanum. It occurs in the Yangtze River Basin every spring, and the occurrence is large in local areas. The leaves can be eaten up in the peak period. The common prevention and treatment method is chemical prevention and treatment, but the plants are urban greening tree species, long-term pesticide spraying pollutes the environment, endangers the human population, and easily makes the pest population produce drug resistance. Since Adoxophyes sp. only has one generation in the Yangtze River Basin in a year, the cycle is long, and it is not suitable for laboratory breeding. At present, there are few special parasitic enemies of Curculionidae hosts in the research reports, and there is no effective controllable breeding technology. The research on the natural enemies of Adoxophyes sp. has been blank. SUMMARY

[0004] The present application aims at overcoming the shortcomings of the prior art, and provides a method for breeding Anisequispina sp. and application thereof in preventing and treating Adoxophyes sp. The present application can breed Anisequispina sp. (parasitic bee) in large quantities, which has the characteristics of short intergenerational time, long female survival time and strong parasitic ability. The Anisequispina sp. prefers to parasitize Curculionidae pests (especially Adoxophyes sp.), and has great significance for preventing and treating the garden leaf-eating pest Adoxophyes sp.

[0005] To achieve the above-mentioned purpose, the technical scheme designed by the present application is as follows:

[0006] The present application provides a method for breeding Anisequispina sp., which comprises the following steps:

[0007] 1) Dip the cotton wool with honey water on the gauze of the tube cover of the culture tube, then place the culture tube with pairs of bean weevil parasitoid adult in the incubator vertically for 5-24 hours, then place the culture tube in the low temperature storage for 4-5 weeks and feed 1-2 times per week, wherein, each time when feeding, take the culture tube out of the low temperature environment, supply honey water nutrition by dropping on the cotton wool of the tube opening with a dropper, and put it back after 2 hours of feeding (low temperature storage can significantly prolong the survival life of male and female parasitoids, female parasitoids can survive for 20-35 days, and male parasitoids can survive for 15-20 days);

[0008] 2) Select the green bean weevil host, then take the tube-mounted bean weevil parasitoid out of the low temperature environment, open the tube cover, put 30-40 green beans with green bean weevil hosts into each culture tube, cover it, and after the temperature rises to above 25 °, supply honey water nutrition by dropping on the cotton wool of the tube opening with a dropper, and place the culture tube horizontally in the above-mentioned incubator (to facilitate female parasitoids to find a suitable position for mating and oviposition), and 2-3 days after mating, oviposition behavior can be observed, and 15-20 hosts per female can be replaced every 6-7 days for repeated mating and oviposition (after oviposition, pay attention to supplement honey water and maintain humidity. One female parasitoid can mate and oviposit several times, and can survive for 3-4 months at the longest. If the male parasitoid dies, a newly emerged male parasitoid can be supplemented. If not remated, the female: male ratio gradually transitions from all females to similar numbers of males and females, and then all males peak. If the host is not replaced for a long time or is small, small individual males may peak, with a body length of about 1 / 2 and a volume of about 1 / 3 of the ordinary male, which can normally mate with female parasitoids, but have a shorter lifespan).

[0009] 3) Replace the host regularly after oviposition, and place the replaced host together with the green beans in a new tube horizontally in the above-mentioned incubator or at room temperature until the next generation emerges (the period from female oviposition to the completion of the emergence of the next generation is about 23-35 days).

[0010] Further, in step 1), the specific steps of the collection and culture method of the bean weevil parasitoid adult are as follows:

[0011] a. Collect the Alnus heterophylla pods in the overwintering generation (autumn and winter seasons), collect the bean weevil parasitoid adults in the first emergence peak, peel off the parasitized Alnus heterophylla seeds, and place them flat in a sealed transparent self-sealing bag. Raise the temperature to above 25 °C at room temperature or above 25 °C to promote parasitoid emergence (room temperature or above 25 °C can effectively promote parasitoid emergence), take out the emerged adults with a wormer, and place them in a 20-40 mesh gauze covered transparent culture tube, with two female and two male adults per tube;

[0012] b. The remaining acacia seeds after autumn emergence are pupae in hibernation state, the seeds are stored in cold and dry condition, the low temperature should be maintained at 7-8 DEG C (the high temperature is not more than 15 DEG C), and the temperature is increased to above 25 DEG C after entering spring, so that the remaining overwintering pupae are emerged, and the emerged insects are collected in the same way.

[0013] Further, in the step 1), the mesh size of the gauze is 20-40 mesh, and the low-temperature storage temperature is 8-10 DEG C.

[0014] Further, the concentration of the honey water is 10%.

[0015] Further, in the step 2), the Callosobruchus chinensis is the Callosobruchus chinensis in the old larva stage or the pupa stage.

[0016] Further, in the steps 1) and 2), the optimal ratio is two to three pairs of the Callosobruchus sp.

[0017] Further, the temperature of the incubator is 20-30 DEG C.

[0018] The application further provides application of the Callosobruchus sp. multiplied by the method to the prevention and treatment of the Callosobruchus chinensis.

[0019] Further, the application method comprises the following steps:

[0020] The Callosobruchus chinensis with a plurality of shelled pupae is placed in a transparent tube with a cover, a plurality of pairs of the Callosobruchus sp. are transferred into the tube, and the tube is horizontally placed in an incubator with a temperature of 20-30 DEG C and a humidity of 60-80% to mate and lay eggs; the egg-laying behavior can be observed 2-3 days after mating, white larvae appear in the pupal shell of the Callosobruchus chinensis 15 days after egg-laying, and the Callosobruchus chinensis is emerged 30-35 days after egg-laying.

[0021] Further, the Callosobruchus sp. is two to three pairs, and the parasitic rate of the Callosobruchus sp. is 40-60%.

[0022] The application has the following beneficial effects:

[0023] The application explores an artificial cultivation, preservation and multiplication method of the parasitic bee which has not been applied, so that the parasitic bee can successfully complete the life history in an artificial environment by using an alternative host which is easy to obtain, and the cultivation condition is easy to achieve. The female parasitic bee can survive for a long time with food supply, can lay eggs in batches, and is easy to preserve the species source. The parasitic rate of the parasitic bee on the leaf-eating pest Callosobruchus chinensis is good, and the parasitic bee can provide a selection of natural enemy control, and the multi-host parasitic characteristics have potential subsequent natural enemy development capacity.

[0024] The application can guide the collection of Anisea spinulata parasitic wasps, and expand the new type of parasitic wasps in the laboratory, and apply them in the prevention and control of garden pests Anisea spinulata. The common parasitic wasps are mostly parasitic on Lepidoptera pests, and the parasitic wasp species of Coleoptera is less. The parasitic wasp shows preference for parasitizing Curculionoidea pests in host testing, has shorter intergenerational time, longer female survival time, and great potential in parasitic ability, and can be developed as a new type of biocontrol natural enemy species. Therefore, the host of the parasitic wasp is more, and a large number of parasitic wasps can be prepared by the method, and the host species are further tested, and other Coleoptera Curculionoidea hosts are screened out, which has potential natural enemy prevention and control application value, and is a new measure for garden pest natural enemy prevention and control. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 Figure 1 is a diagram of Anisea spinulata parasitic wasps laying eggs on Anisea spinulata pupae;

[0026] Figure 2 Figure 2 is a diagram of white parasitic wasp larvae in Anisea spinulata pupae after Anisea spinulata parasitic wasps parasitize the pupae;

[0027] Figure 3 Figure 3 is a diagram of parasitic wasp larvae and hosts taken out of the pupal case;

[0028] Figure 4 Figure 4 is a diagram of Anisea spinulata pupal cases after parasitic wasps emerge. DETAILED DESCRIPTION

[0029] The application will be further described in detail below with specific examples, so that those skilled in the art can understand.

[0030] Example 1

[0031] A method for expanding Anisea spinulata parasitic wasps, comprising the following steps:

[0032] 1) The specific steps of the collection and culture method of Anisea spinulata adult insects are as follows:

[0033] a. In the overwintering generation (autumn and winter seasons), Anisea spinulata parasitic wasps are collected, Anisea spinulata adult insects are collected at the first emergence peak, parasitized Albizia lebbeck seeds are peeled off, and are placed flat in a sealed transparent self-sealing bag. Room temperature or temperature above 25 DEG C is used to promote the emergence of parasitic wasps (room temperature or temperature above 25 DEG C can effectively promote the emergence of parasitic wasps), and the emerged adult insects are taken out with a wormer and are placed in a transparent culture tube with a cover and a mesh size not less than 20 meshes, two male and two female adult insects are placed in each tube;

[0034] b. The remaining Albizia lebbeck seeds after emergence in autumn are in a state of entering hibernation, the seeds are stored in a cold and dry state, the low temperature should be maintained at 7-8 DEG C (the high temperature should not be higher than 15 DEG C), and the temperature is increased to above 25 DEG C after entering spring, so that the remaining overwintering pupae emerge, and the emerged adult insects are collected in the same way.

[0035] 2) Dip the cotton wool in honey water (10% concentration) and fix it on the gauze (20-40 mesh) of the tube cover of the culture tube, then place the culture tube containing pairs (2-3 pairs) of Tetrastichus giffardii adults vertically in the incubator (temperature 20-30°C) for 5-24 h, then store the culture tube in a low temperature environment (8-10°C) and feed the insects 1-2 times a week, wherein, each time the culture tube is taken out from the low temperature environment, honey water is supplied to the insects by dripping the honey water on the cotton wool at the tube opening with a dropper, and the culture tube is put back after 2 h of feeding, for 4-5 weeks (low temperature storage can significantly prolong the survival life of male and female wasps, and the female wasps can survive for 20-35 d and the male wasps can survive for 15-20 d).

[0036] 3) Select green gram bruchids in the older larval stage or in the pupal stage, then take out the Tetrastichus giffardii adults from the low temperature environment, open the tube cover, and put 30-40 green gram bruchids with green gram bruchid hosts into each culture tube, then close the tube cover, and after the temperature rises to above 25°C, supply honey water to the insects by dripping the honey water on the cotton wool at the tube opening with a dropper, and place the culture tube horizontally in the incubator (this is convenient for the female wasps to find a suitable position for mating and laying eggs), and mating can be observed 2-3 d later, and the hosts can be replaced every 6-7 d to repeat the mating and egg laying of the female wasps (15-20 hosts per female wasp) (after laying eggs, pay attention to supplement the honey water and maintain the humidity. One female wasp can mate and lay eggs for several times, and the longest survival time is 3-4 months. If the male wasps die, new male wasps can be supplemented. If the female wasps are not re-mated, the female: male ratio gradually changes from all female wasps to a similar number of male and female wasps, and then all male wasps appear. If the hosts are not replaced for a long time or the hosts are small, small male wasps can appear, which are about 1 / 2 of the size of ordinary male wasps and about 1 / 3 of the volume, and can mate with female wasps normally, but have a shorter life span).

[0037] 4) Replace the hosts regularly after the female wasps lay eggs, and place the replaced hosts together with green gram in a new tube, and place the tube horizontally in the incubator or in a room temperature environment, until the next generation of wasps hatches (the period from when the female wasps lay eggs to when the next generation of wasps hatches is about 23-35 d).

[0038] Based on the above method steps, green gram bruchid hosts are collected and screened, and the hosts are placed in culture tubes in proportion to allow Tetrastichus giffardii wasps to parasitize. The parasitic wasps are removed every 4-8 days, and the green gram hosts on which eggs have been laid are left to culture. The first batch of hosts hatches 8 male wasps. The second batch of hosts hatches 25 female wasps and 18 male wasps in 6 batches. The female: male ratio in the 6 batches is 100:0, 58.8:41.2, 50:50, 50:50, 0:100 and 0:100, respectively.

[0039] Table 1 Life cycle and hatching sequence of Tetrastichus giffardii on alternative host Callosobruchus chinensis

[0040]

[0041] Note: the first and second batches are different parents, a generation apart, the second batch is all eggs laid by the same generation of small bees in succession, divided into different time periods.

[0042] As shown in Table 1: Callosa rotundipennis can complete the life cycle on the alternative host Callosa oryzae, the life cycle period is 23-35 days; female bees emerge earlier, male bees emerge later, and female bees are more active, can mate multiple times, and survive longer.

[0043] Example 2

[0044] The application of Callosa rotundipennis propagated based on the method of Example 1 in the control of Ligustrum boll weevil has the following specific steps:

[0045] Put multiple shelled pupae of Ligustrum boll weevil into a transparent tube with a lid, and transfer multiple pairs of Callosa rotundipennis into the tube, and place it horizontally in a incubator with a temperature of 25-28°C and a humidity of 60-80%. After mating for 2-3 days, oviposition behavior can be observed, and 15 days after oviposition, white larvae can be observed in the pupal shell of Ligustrum boll weevil, and 30-35 days after oviposition, emergence can be observed.

[0046] Based on the above method, 18 pupae of Ligustrum boll weevil were collected for Callosa rotundipennis parasitism. On April 6, oviposition behavior was observed on the pupae of Ligustrum boll weevil, and on April 11, the bees were removed. On April 20, white larvae appeared in the pupal shell of Ligustrum boll weevil, and on May 5, the first batch of 6 male bees emerged, and another 2(5.7)+2(5.9) emerged. On May 9, the last two males emerged, with a large difference in size, with the small males being about 1 / 2 the size and 1 / 3 the volume of the average male, but the small males could mate with females normally. A total of 10 male bees emerged, with a parasitism rate of 55%. The results of subsequent tests on the parasitism of Callosa rotundipennis on the pupae of Ligustrum boll weevil are as follows.

[0047] As shown in Table 2: the parasitism rate of Callosa rotundipennis on the pupae of Ligustrum boll weevil is between 55-85%, and when the host is continuously supplied, the parasitism rate of the middle batch can reach more than 60%. Conclusion: Callosa rotundipennis can quickly recognize the pupae of Ligustrum boll weevil, oviposit actively (see Figure 1 ), has a similar length of life cycle on the alternative host Callosa oryzae, has good parasitism (see Figure 2 , Figure 3 ), has a high parasitism rate, and the overall parasitism rate can reach more than 80%, which can effectively inhibit the emergence of Ligustrum boll weevil (see Figure 4 ).

[0048] Table 2 Parasitism of Callosa rotundipennis on the pupae of Ligustrum boll weevil

[0049] Number of pupae of Adoxophyes spp. Initial oviposition day Final oviposition day Incubation period Total number of chalcid emergence Dead pupae Adoxophyes spp. emergence Parasitism rate 18 April 6 April 11 4.6-5.5 10 0 8 55.56％ 41 March 20 March 31 3.20-4.25 22 12 7 82.93％ 23 March 31 April 7 3.31-4.25 0 14 9 60.87％ 20 April 3 April 11 4.3-5.1 10 7 3 85.00％

[0050] Other parts not described in detail are prior art. Although the above embodiment has made a detailed description of the present application, it is only a part of the embodiment of the present application, not all the embodiments, and people can also obtain other embodiments according to the present embodiment without creativity, which all belong to the protection scope of the present application.

Claims

1. A method of propagating the parasitic wasp Diglyphus begini, characterized in that: It comprises the following steps: 1) dip the cotton wool with honey water on the gauze of the tube cover of the culture tube, then vertically place the culture tube containing pairs of bean weevil parasitoid adult in the incubator for 5-24 hours, then place the culture tube in the low temperature storage for 4-5 weeks and feed 1-2 times per week, wherein, each time when feeding, take the culture tube out of the low temperature environment, drop the honey water nutrition on the cotton wool at the tube opening with a dropper, and place it back in the culture tube after feeding for 2 hours; the specific steps of the collection and culture method of the bean weevil parasitoid adult are as follows: a. collect the bean weevil parasitoid adult in the first emergence peak after collecting the albizia lebbeck seed in the overwintering generation, peel off the parasitized albizia lebbeck seed, place it in a sealed transparent self-sealing bag, and promote the emergence of the parasitic bee at room temperature or by increasing the temperature to above 25°C. The emerged adult is taken out with a sucking device and placed in a 20-40 mesh gauze transparent culture tube with a cover, with two male and two female adults in each tube; b. the remaining albizia lebbeck seed after autumn emergence is in the state of hibernating bee pupa, which is stored in a dry and cold environment. The low temperature should be maintained at 7-8°C, and the temperature should be increased to above 25°C after entering spring to make the remaining overwintering bee pupa emerge. The emerged adult is collected in the same way; 2) select green bean weevil host, then take the tube containing bean weevil parasitoid out of the low temperature environment, open the tube cover, put 30-40 green bean weevils with green bean weevil host into each culture tube, cover it, and after the temperature rises to above 25°C, drop the honey water nutrition on the cotton wool at the tube opening with a dropper, and place it in the above-mentioned incubator. The mating behavior can be observed 2-3 days after mating, and the host can be replaced every 6-7 days to repeat mating and oviposition, with 15-20 hosts per female bee; 3) replace the host regularly after oviposition, and place the replaced host together with the green bean in a new tube, which is placed in the above-mentioned incubator or at room temperature until the next generation emerges.

2. The method of claim 1 wherein the mass-reared parasitoids are T. podagrica. In step 1), the mesh size of the gauze is 20-40 mesh, and the low temperature storage temperature is 8-10°C.

3. The method of claim 1, wherein the mass-reared parasitoid is: The concentration of the honey water is 10%.

4. The method of claim 1 wherein the mass-reared parasitoids are T. podagrica. In step 2), the green bean weevil is a large larva or a pupa of green bean weevil.

5. The method of claim 1 wherein the mass-reared parasitoids are T. podagrica. In steps 1) and 2), the optimal ratio is two to three pairs of bean weevil parasitoids per tube.

6. The method of claim 1 wherein the mass-reared parasitoids are T. podagrica. The temperature of the incubator is 20-30°C.

7. The application of the bean weevil parasitoid propagated by the method of claim 1 in the prevention and control of ligustrum borer.

8. Use according to claim 7, characterized in that: The specific steps of the method are as follows: Put a plurality of shelled pupae of ligustrum borer into a transparent tube with a cover, transfer a plurality of pairs of bean weevil parasitoids into the tube, and place it horizontally in an incubator with a temperature of 20-30°C and a humidity of 60-80% to mate and oviposit. The oviposition behavior can be observed 2-3 days after mating, and white larvae can be observed in the pupal case of ligustrum borer 15 days after oviposition, and the ligustrum borer emerges 30-35 days after oviposition.

9. Use according to claim 7, characterized in that: The bean weevil parasitoids are 2-3 pairs, and the parasitization rate of the bean weevil parasitoids is 40-60%.

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