Microsatellite primers, applications and methods for identifying the sex of the pupae of the fly pupae, the egg of the wasp

A kind of technology of fly pupa and golden wasp, which is applied in the field of insect evolution and achieves the effects of high polymorphism and stable PCR amplification results

Active Publication Date: 2020-01-10
ANHUI NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The current sex identification of the chrysalis chrysalis can only be identified by appearance in the adult stage

Method used

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  • Microsatellite primers, applications and methods for identifying the sex of the pupae of the fly pupae, the egg of the wasp
  • Microsatellite primers, applications and methods for identifying the sex of the pupae of the fly pupae, the egg of the wasp
  • Microsatellite primers, applications and methods for identifying the sex of the pupae of the fly pupae, the egg of the wasp

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] A kind of microsatellite primer for identification of the sex of the fly chrysalis chrysalis ovum, the parameters such as the nucleic acid sequence position and the repeating unit of the microsatellite primer are as shown in table 1 below:

[0057] Table 1

[0058]

[0059] The design method of the microsatellite primers for the identification of the sex of the pupae of the fly chrysalis:

[0060] Breeding and establishing housefly breeding strains, obtaining housefly pupae, and then luring parasitic wasps in different geographical locations, through morphological identification, obtaining fly pupae golden wasps, and constructing culture strains of different geographical strains fly pupae golden wasps; The DNA of the chrysalis chrysalis from different regions was extracted, randomly fragmented, purified and then repaired at the end, "A" was added, and the sequencing adapter was connected, and then the 350bp fragment was selected by agarose gel electrophoresis, and fi...

Embodiment 2

[0121] The application of the microsatellite primer for identifying the sex of the chrysalis chrysalis egg is used to identify the sex of the chrysalis chrysalis egg.

Embodiment 3

[0123] The method for identifying the sex of the fly chrysalis chrysalis ovum by using microsatellite primers for identifying the sex of the chrysalis chrysalis egg comprises the following steps:

[0124] 1) Raising and establishing housefly breeding strains in the laboratory to obtain housefly pupae; using housefly pupae to lure parasitic wasps in three different geographical regions in China (Wuhu in Anhui, Urumqi in Xinjiang and Dunhuang in Gansu), Through the morphological identification, the chrysalis chrysalis was obtained; the culture strains of various geographical strains of the chrysalis chrysalis were established in the laboratory;

[0125] 2) After the parasitoid wasps parasitize the housefly pupae (host) for 15 days, use a cryopreservation tube with a capacity of 5ml to pack individual housefly pupae to ensure that each parasitoid that emerges from the bee is in an unmated state. The unmated parasitoids after bee emergence were raised for one day, and mated with t...

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Abstract

The invention provides a microsatellite primer and application for identifying fly pupa nasonia eggs sex and a method for identifying fly pupa nasonia eggs sex. The method includes the steps of making female fly pupa nasonias and male fly pupa nasonias from different strains mate to obtain parasitic wasp eggs with discriminability; extracting DNA from the obtained parasitic wasp eggs with discriminability; taking the extracted DNA as a template, using a designed primer to conduct PCR amplification, and conducting strip running on an obtained product through agarose gel electrophoresis; judging the sex of the eggs according to the strips , wherein if single strips are unfertilized haploid eggs, the eggs are male, and if the double strips are fertilized diploid eggs, the eggs are female. Compared with the prior art, the method uses the characteristics of a microsatellite to design the microsatellite primer to be combined with the sex determination mode of parasitic wasps, and therefore the fly pupa nasonia eggs sex can be identified successfully.

Description

technical field [0001] The invention belongs to the field of insect evolution, and in particular relates to a microsatellite primer for identifying the gender of a fly pupae, an egg of the chrysalis, its application, and a method for identifying the sex of a fly pupae of the egg of the wasp. Background technique [0002] Pachycrepoideus vindemmiae Rondani belongs to Chalcidoidea, Pteromalidae and Pteromalinae, and is a parasitic wasp in the pupal stage of many insects such as Diptera. It is one of the most common species of parasitic wasps in the pupal stage of the housefly. The golden wasp is widely distributed in different regions of the country and is the dominant species among many parasitic wasps. [0003] The current sex identification of the chrysalis chrysalis can only be identified by appearance in the adult stage. The more obvious ovipositing needles on the tail of female parasitoids become the main basis for sex identification of parasitoid adults. [0004] Micr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12N15/11
CPCC12Q1/686C12Q1/6888C12Q2600/156C12Q2525/151
Inventor 李镜胡好远李琴朱承节
Owner ANHUI NORMAL UNIV
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