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Method for locating distribution of symbiotic bacteria Cardinium in different tissues of rice planthopper

A rice planthopper and tissue technology, applied in the biological field, can solve the problem that the distribution of Cardinium is not accurately located, and achieve the effects of high specificity, cost saving, and good interaction relationship

Active Publication Date: 2019-09-17
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the tissue distribution of Cardinium in two species of rice planthoppers has not been precisely mapped

Method used

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  • Method for locating distribution of symbiotic bacteria Cardinium in different tissues of rice planthopper
  • Method for locating distribution of symbiotic bacteria Cardinium in different tissues of rice planthopper
  • Method for locating distribution of symbiotic bacteria Cardinium in different tissues of rice planthopper

Examples

Experimental program
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Effect test

Embodiment 1

[0042] Example 1: Localization of the distribution of the symbiotic bacterium Cardinium in different tissues of the white-backed planthopper.

[0043] (1) Dissection and isolation of white-backed planthopper tissues

[0044] Collect 20 mature white-backed planthopper females and males, place the planthoppers to be dissected in a clean petri dish, carefully dissect out salivary glands, ovaries, testes, intestines and other tissues with dissecting forceps under a microscope, and dissect them Transfer to a tube containing 1x PBS.

[0045] (2) Design of oligonucleotide probes

[0046] According to the 16S rRNA sequence of Cardinium, three specific oligonucleotide probes with fluorescent labels were designed, and Cy5 was used to label the 5' end of the probe sequence, which were SEQ ID NO.1 and SEQ ID NO.2 and SEQ ID NO.3.

[0047] (3) Hybridization of oligonucleotide probes with Cardinium in white-backed planthopper

[0048] The hybridization of the oligonucleotide probe with ...

Embodiment 2

[0070] Example 2: Localization of the distribution of the symbiotic bacterium Cardinium in different tissues of the brown planthopper.

[0071] (1) Dissection and isolation of N. lugens tissue

[0072] Collect 20 mature brown planthopper females and males, place the planthoppers to be dissected in a clean petri dish, carefully dissect out salivary glands, ovaries, testes, intestines and other tissues with dissecting forceps under a microscope, and transfer them to a culture medium containing in a test tube of PBS.

[0073] (2) Design of oligonucleotide probes

[0074] According to the 16S rRNA sequence of Cardinium, three fluorescently labeled oligonucleotide probes with specificity were designed, and Cy5 was used to label the 5' end of the probe sequence, which are SEQ ID NO.1 and SEQ ID NO.2 and SEQ ID NO.3.

[0075] (3) Hybridization of oligonucleotide probes with Cardinium in brown planthopper

[0076] The hybridization of oligonucleotide probes and Cardinium in brown ...

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Abstract

The invention discloses a method for locating the distribution of symbiotic bacteria Cardinium in different tissues of rice planthopper, and belongs to the field of agricultural biotechnology. The method includes the steps that firstly, dissection is carried out to separate out different tissues of the rice planthopper; then, three specific oligonucleotide probes with fluorescent markers are designed according to the 16S rRNA sequence of the Cardinium; finally, hybridization is carried out on the Cardinium in the rice planthopper through the specific probes, and the Cardinium in the tissues of the rice planthopper can be accurately located through scanning by a laser scanning confocal microscope. The location method has high specificity, sensitivity, feasibility and effectiveness and can save cost. The method is beneficial to better research on the interaction relationship between the Cardinium and the host and also facilitates research on a strategy for suppressing pest populations and a mechanism for resisting viral transmission.

Description

technical field [0001] The invention belongs to the field of biological technology, and in particular relates to a method for locating the distribution of symbiotic bacteria Cardinium in different tissues of rice planthopper. Background technique [0002] Cardinium is a class of intracellular Gram-negative endosymbiont 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 rice planthopper, spider mite, whitefly and parasitic wasps. They can regulate host reproduction and development, including cytoplasmic incompatibility (Cytoplasmic incompatibility, CI), parthenogenesis (parthenogenesis inducing), feminization (Feminization) and male-killing (Male-killing). Among them, the most important reproductive phenotype induced by Cardinium is CI. The occurrence of CI is related to the infection of symbiotic bacteria, when uninfected females mate with infected males, the offsp...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6816C12Q1/689C12R1/01
CPCC12Q1/6816C12Q1/689C12Q2563/107Y02A50/30
Inventor 洪晓月李同浦周迎春查思思
Owner NANJING AGRICULTURAL UNIVERSITY
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