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Method for obtaining nilaparvata lugens new strains stably infected with Cardinium by artificial transfection

A technology of brown planthopper and a new strain, applied in the biological field, can solve the problems of inability to transfect new hosts, low infection rate, and inability to stably infect

Active Publication Date: 2019-09-27
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Brown planthopper can only infect Wolbachia at present, but the infection rate is extremely low, and it cannot infect stably, and Wolbachia cannot induce CI, while the infection of Cardinium in brown planthopper has not been reported
[0005] In summary, Cardinium has excellent characteristics such as strong CI and complete maternal transmission in some insects, but it has been unable to transfect to new hosts, especially In brown planthopper, it cannot be further applied, so we urgently need to establish a method to obtain a new strain of brown planthopper infected with Cardinium

Method used

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  • Method for obtaining nilaparvata lugens new strains stably infected with Cardinium by artificial transfection
  • Method for obtaining nilaparvata lugens new strains stably infected with Cardinium by artificial transfection
  • Method for obtaining nilaparvata lugens new strains stably infected with Cardinium by artificial transfection

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Embodiment Construction

[0062] The following examples are further descriptions of the present invention, rather than limitations of the present invention.

[0063] (1) Collection of planthopper eggs

[0064] 30 mature brown planthoppers infected with Cardinium and 30 mature brown planthoppers not infected with symbiotic bacteria were respectively taken to lay eggs in two cups of rice seedlings for 4-5 hours.

[0065] Place the oviposited rice seedlings on a clean petri dish, remove the roots and leaves, carefully dissect the seedlings under a stereomicroscope with dissecting tweezers, and then transfer their eggs to two pieces of wet filter paper, These embryos were then placed on nitrocellulose membranes and transferred to glass slides fixed with double-sided tape.

[0066] The dissected eggs of white-backed planthopper and brown planthopper were used as donors and recipients of artificial transfection, among which eggs of white-backed planthopper infected with Cardinium were used as donors of arti...

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Abstract

The invention discloses a method for obtaining nilaparvata lugens new strains stably infected with Cardinium by artificial transfection and belongs to the technical field of agricultural biology. According to the method, eggs newly laid by sogatella furcifera and nilaparvata lugens are used as a donor and a receptor for artificial transfection respectively, and a cytoplasm component of the donor is sucked by a micro-injection instrument and rapidly injected into receptor eggs. The receptor eggs are cultured and developed into primary female adults (G0), the primary female adults are mated with non-infected males to produce a first generation (G1), and infected positive females are screened from the G1 to be continuously mated with non-infected males to produce a second generation (G2). The G2 and the G1 are detected to be positive, so that the nilaparvata lugens new strains stably infected with Cardinium are obtained successfully. The method can be used for rapidly establishing the nilaparvata lugens new strains infected with Cardinium, the fertility and hatching rate of the new strains are significantly reduced, so that harm of the nilaparvata lugens is controlled. The invention also provides models for transfection of other symbiotic bacteria.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for obtaining a new strain of brown planthopper stably infecting Cardinium by artificial transfection Background technique [0002] Cardinium is a class of intracellular Gram-negative commensal bacteria that can be transmitted maternally. They are widely found in many arthropods, and Cardinium has been found in some important agricultural pests such as planthoppers, spider mites, whitefly and parasitic wasps. Similar to Cardinium, Wolbachia is another class of intracellular symbionts widely found in arthropods and nematodes. These symbiotic bacteria can regulate the reproduction and development of the host. Among them, the most important reproductive phenotype induced by Cardinium is Cytoplasmic incompatibility (CI). The occurrence of CI is related to the infection of symbiotic bacteria, when uninfected females mate with infected males, the offspring produced ...

Claims

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Application Information

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IPC IPC(8): C12N15/89A01K67/033C12Q1/6841
CPCC12N15/89A01K67/0339C12Q1/6841A01K2207/12A01K2227/706C12Q2563/107Y02A50/30
Inventor 洪晓月李同浦龚君淘周迎春查思思
Owner NANJING AGRICULTURAL UNIVERSITY
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