Method for improving killing of German cockroaches by metarhizium anisopliae through interfering genes and application of method

A technology of Blattella germanica and Metarhizium anisopliae, applied in application, genetic engineering, plant gene improvement, etc., can solve the problems of slow insecticidal speed, low lethality, poor host selectivity, etc., and achieve increased sensitivity and improved poison killing effect of effect

Pending Publication Date: 2022-05-17
SHANDONG NORMAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] Yet the contriver finds, only use Metarhizium anisopliae as insecticide, also has shortcoming, and its shortcoming mainly is that effect is slow, desinsection speed is slow, and desinsection period is too long, and possible toxicity is not ideal, and lethal rate is low, and The poor selectivity to the host leads to the inconspicuous effect of killing cockroaches, etc.

Method used

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  • Method for improving killing of German cockroaches by metarhizium anisopliae through interfering genes and application of method
  • Method for improving killing of German cockroaches by metarhizium anisopliae through interfering genes and application of method
  • Method for improving killing of German cockroaches by metarhizium anisopliae through interfering genes and application of method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] 1. Observe the changes of Akirin gene expression in different time periods after the German cockroach sensitive strain (S) and German cockroach resistant strain (R) are infected with Metarhizium anisopliae, and verify the immune function of the Akirin gene.

[0079] After Blattella germanica was infected with Metarhizium anisopliae, collect the whole worms of S and R strains of Blattella germanica at 12h, 36h, 48h, 72h, 84h, 108h after the infection of the control group and Metarhizium anisopliae, and treat Blattella germanica with 75% alcohol. Disinfect. The treated worms were cut off with sterilized scissors and tweezers to remove the wings and antennae, and rinsed three times in PBS to remove impurities. The obtained tissues were used for the extraction of total RNA. Reverse transcription and cDNA synthesis were performed on the RNA samples extracted above, and the RNA concentration was adjusted to be consistent before inversion, and the samples of the two strains w...

Embodiment 2

[0122] 1. Detect the changes of Bgchi gene expression at different time periods after the German cockroach sensitive strain (S) and German cockroach resistant strain (R) are infected with Metarhizium anisopliae, and verify the immune effect of the Bgchi gene (GenBank accession number: KJ789158).

[0123] (1) Primer design

[0124] In this example, the Bgchi gene, an immune gene related to the German cockroach's resistance to pathogenic fungi, was selected, and Primer5.0 was used to design primers to obtain the following primer sequences:

[0125]

[0126] (2) RNA extraction

[0127] After Blattella germanica was infected with Metarhizium anisopliae, the whole worms of S and R strains of Blattella germanica were collected from the control group and 12h, 36h, 48h, 72h, 84h, and 108h after infection with Metarhizium anisopliae. The german cockroaches were disinfected with 75% alcohol. The treated worms were cut off with sterilized scissors and tweezers for wings and antennae...

Embodiment 3

[0169] 1. Changes of CYP4G19 gene expression after Blattella germanica infection with Metarhizium anisopliae

[0170] (1) Primer design

[0171] The CYP4G19 gene (GenBank accession number: AY176056.1) selected in this embodiment is an immune gene related to the defense against pathogenic fungi by Blattella germanica. Primers were designed with Primer5.0 to obtain the following primer sequences:

[0172]

[0173] (2) RNA extraction

[0174] After Blattella germanica was infected with Metarhizium anisopliae, the whole worms of S and R strains of Blattella germanica were collected from the control group and 12h, 36h, 48h, 72h, 84h, and 108h after infection with Metarhizium anisopliae. The german cockroaches were disinfected with 75% alcohol. The treated worms were cut off with sterilized scissors and tweezers for wings and antennae, and rinsed three times in PBS to remove impurities. The obtained tissues were used for the extraction of total RNA.

[0175] RNA extraction ki...

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Abstract

The invention provides a method for improving killing of German cockroaches by metarhizium anisopliae through interfering genes and application of the method, and belongs to the technical field of biological control. According to the application disclosed by the invention, the research finds that the resistance of German cockroach to pathogenic fungi (such as metarhizium anisopliae) can be effectively reduced by down-regulating the expression of German cockroach Akirin, Bgchi or CYP4G19 genes by adopting an RNAi technology, so that the sensitivity of German cockroach, especially anti-beta-cypermethrin strain German cockroach to pathogenic fungi (such as metarhizium anisopliae) is increased, and the resistance of German cockroach to pathogenic fungi (such as metarhizium anisopliae is improved. Therefore, the poison killing effect of the biological insecticide is effectively improved, and the biological insecticide has good practical application value.

Description

technical field [0001] The invention belongs to the technical field of biological control, and in particular relates to a method for improving the killing of Blattella germanica by interfering genes and its application. Background technique [0002] The information disclosed in the Background of the Invention is only intended to increase the understanding of the general background of the invention, and is not necessarily to be taken as an acknowledgment or any form of suggestion that the information constitutes the prior art that is already known to those skilled in the art. [0003] German cockroach, commonly known as cockroach, is one of the most widely distributed indoor pests in the world, causing certain harm to human life and health. At present, the control of German cockroaches is still dominated by chemical control. However, there are many problems in the process of using chemical insecticides, such as overdosage and improper use methods, which gradually increase the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/54C12N15/53A01K67/033A01N63/30A01P7/04
CPCC12N9/1081C12N9/0081C12Y204/01016C12Y114/15006C07K14/43563A01K67/0339A01N63/30
Inventor 张凡刘宝睿赵海政刘彩霞张宪翠姜萌臧雅南王学军
Owner SHANDONG NORMAL UNIV
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