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The brown planthopper nlcdc2 gene and its encoded product and application

A brown planthopper, gene technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve problems such as difficulty in adopting targeted strategies and lack of target information

Inactive Publication Date: 2017-04-26
CHINA JILIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is difficult to adopt targeted strategies in the control of brown planthopper due to the lack of ideal target information

Method used

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  • The brown planthopper nlcdc2 gene and its encoded product and application
  • The brown planthopper nlcdc2 gene and its encoded product and application
  • The brown planthopper nlcdc2 gene and its encoded product and application

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Embodiment 1: brown planthopper Nlcdc 2 Gene cloning and sequence analysis

[0027] 1, Nlcdc2 RACE PCR and PCR amplification of core fragments

[0028] The N. lugens cDNA was used as a template, and the core sequence was amplified by PCR with primers Nlcdc2-F1 and Nlcdc2-R1 (Table 1), and the target fragment of the expected size was obtained. The sequencing results showed that the fragment was about 500 bp in size, and compared with the transcriptome Provides sequence consensus ( figure 1 ). After NCBI Blastp analysis, it was found that the deduced amino acid sequence of the gene was similar to that of Monarch butterfly Danaus plexippus (EHJ71170) homologous gene coded amino acid sequence homology reached 71%, as shown in Table 2. According to the core sequence sequencing results, RACE primers (Table 1) were designed to clone the full-length cDNA. 5'RACE obtained a specific sequence of about 750 bp, and 3'RACE obtained a specific sequence of about 250 bp ( f...

Embodiment 2

[0040] Example 2: Different developmental stages Nlcdc 2 mRNA expression levels

[0041] 1. Brown planthopper Nlcdc2 with β-actin Real-time PCR primer amplification efficiency

[0042] The N. lugens cDNA template was diluted 4 times, and 6 gradients were diluted. The abscissa of the standard curve is the logarithmic value of the dilution factor, and the ordinate is the CT value. Nlcdc2 with β-actin Fluorescent quantitative PCR standard curve ( Figure 4 ).

[0043] Real-Time PCR judges the specificity of primers by melting curves. Nlcdc2 with β-actin The melting curve shows that the product is single and there is no non-specific amplification ( Figure 5 ), and the results can be used for later data analysis.

[0044] 2. Different stages of development Nlcdc2 expression analysis of

[0045] Developmental stages of brown planthopper nymphs and brachypterous females using fluorescent quantitative PCR Nlcdc2 Quantification of transcript levels showed that Nlcdc2...

Embodiment 3

[0046] Embodiment 3: Synthesis and RNA interference of brown planthopper dsRNA

[0047] 1. Synthesis of dsRNA from brown planthopper

[0048] Use primers dsNlcdc2-F, dsNlcdc2-R and dsGFP-F, dsGFP-R (Table 1) to synthesize the fragment dsNlcdc2 for Nlcdc2 RNA interference and the non-target control interference fragment dsGFP respectively, and the results of RNA electrophoresis obtained fragments of the expected size, among which the dsNlcdc2 fragment The length is about 500 bp, and the length of the dsGFP fragment is 350 bp ( Figure 7 ).

[0049] 2. The effect of feeding dsRNA on the survival rate of brown planthopper

[0050] The artificial diet with a final concentration of 0.5 μg / μL dsGFP was used as the non-target control group (dsGFP), and the artificial diet with a final concentration of 0.5 μg / μL dsNlcdc2 was used as the test group (dsNlcdc2-1). Survival rates were counted daily. The results showed that there were significant differences between dsNlcdc2 test group...

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Abstract

The invention discloses a brown planthopper N1cdc2 gene as well as an encoding product and an application of the brown planthopper N1cdc2 gene, and belongs to the technical field of biology. On the one hand, the invention discloses a brown planthopper N1cdc2 gene sequence and an encoding protein amino acid sequence of the brown planthopper N1cdc2 gene, and on the other hand, the invention also discloses an application of the brown planthopper N1cdc2 gene in control of the survival rate and the reproductive capacity of brown planthopper. Research is carried out mainly aiming at a cell division cycle gene N1cdc2 which is related to brown planthopper reproduction, the expression pattern and related function of the gene in brown planthopper reproduction expression are ascertained, the potential of the brown planthopper N1cdc2 gene as a new target for preventing an injurious insect is defined, and a foundation is established for development of an N1cdc2-based pest control new strategy.

Description

technical field [0001] The invention belongs to the field of biological technology, in particular to the cell division cycle gene of brown planthopper Nlcdc2 And its coding products and applications. Background technique [0002] Brown planthopper ( Nilaparvata lugens Stål) is a rice pest with strong fecundity and serious damage. It mainly damages rice plants by sucking phloem sap and spreading rice viruses. In recent years, brown planthoppers have frequently broken out and become disasters, and have developed into an important pest in Asian rice regions. At present, the control of BPH mainly relies on measures such as spraying chemical pesticides and promoting rice resistant varieties. However, practice has shown that brown planthoppers can develop strong resistance to pesticides, making chemical control impossible; similarly, brown planthoppers can gradually develop into insect-resistant rice species that can adapt to the resistance of rice varieties in the process of...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/12C07K14/435A01N57/16A01P7/04
Inventor 俞晓平郝培应马艳申屠旭萍陆潮峰
Owner CHINA JILIANG UNIV
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