Pest control constructor and method thereof

A technology for constructs and pests, applied in the directions of botanical equipment and methods, biochemical equipment and methods, pesticides, etc., can solve problems such as hindering the promotion of genetically modified resistant materials, increasing production costs, and low resistance.

Active Publication Date: 2014-04-30
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In order to control plant pests, people usually use broad-spectrum chemical insecticides and biological insecticides, but both have limitations in practical application: chemical insecticides will cause environmental pollution problems and lead to the emergence of resistant insects. However, biopesticides are easily degraded in the environment and need to be repeatedly applied in production, which greatly increases the production cost
[0004] At present, most of the Cry2Aa protein and Cry1Ab / Ac protein are used in plants in the form of a single gene, especially the Cry2Aa protein. When it exists in a single gene form, it has low resistance to Chilorrhizae and S. The emergence of resistance to these two genes alone by the borer and the large borer hampers the promotion of transgenic resistant materials

Method used

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  • Pest control constructor and method thereof
  • Pest control constructor and method thereof
  • Pest control constructor and method thereof

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0082] The first embodiment, the acquisition and synthesis of Cry2Aa gene and Cry1Ab / Ac gene

[0083] 1. Obtain Cry2Aa and Cry1Ab / Ac nucleotide sequences

[0084] The amino acid sequence (633 amino acids) of the Cry2Aa insecticidal protein, as shown in SEQ ID NO: 1 in the sequence listing; the Cry2Aa nucleotide sequence (1902) encoding the amino acid sequence (633 amino acids) corresponding to the Cry2Aa insecticidal protein nucleotides), as shown in SEQ ID NO:3 in the sequence listing.

[0085] The amino acid sequence (609 amino acids) of the Cry1Ab / Ac insecticidal protein, as shown in SEQ ID NO: 2 in the sequence listing; Cry1Ab / Ac encoding the amino acid sequence (609 amino acids) corresponding to the Cry1Ab / Ac insecticidal protein Nucleotide sequence (1830 nucleotides), as shown in SEQ ID NO:4 in the sequence listing.

[0086] 2. Synthesize the above nucleotide sequence

[0087]The Cry2Aa nucleotide sequence (as shown in SEQ ID NO: 3 in the sequence listing) and the Cry...

no. 2 example

[0088] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0089] 1. Construction of a recombinant cloning vector containing the Cry2Aa gene and the Cry1Ab / Ac gene

[0090] The synthesized Cry2Aa nucleotide sequence was connected to the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and the operation steps were carried out according to the instructions of the pGEM-T vector produced by Promega Company to obtain the recombinant cloning vector DBN01-T, which Build process such as figure 1 Shown (wherein, Amp represents the ampicillin resistance gene; f1 represents the replication origin of phage f1; LacZ is the LacZ start codon; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; Cry2Aa is the Cry2Aa nucleoside acid sequence (SEQ ID NO: 3); MCS is a multiple cloning site).

[0091] Then, the recombinant cloning vector DBN01-T was transformed into Esche...

no. 3 example

[0101] The third embodiment, the acquisition and verification of corn plants transferred to Cry2Aa and Cry1Ab / Ac genes

[0102] 1. Obtaining corn plants transferred to Cry2Aa and Cry1Ab / Ac genes

[0103] According to the commonly used Agrobacterium infection method, the immature embryos of the aseptically cultured maize variety Zong 31 (Z31) were co-cultured with the Agrobacterium described in 3 in the second example, so that the The T-DNA in recombinant expression vectors DBN100074, DBN100056 and DBN100058 (including the promoter sequence of maize Ubiquitin gene, Cry2Aa nucleotide sequence, Cry1Ab / Ac nucleotide sequence, PMI gene and Nos terminator sequence) were transferred to maize chromosome In the group, the maize plant transferred into the Cry2Aa nucleotide sequence, the maize plant transferred into the Cry1Ab / Ac nucleotide sequence and the maize plant transferred into the Cry2Aa-Cry1Ab / Ac nucleotide sequence were obtained; as comparison.

[0104] For Agrobacterium-med...

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Abstract

The invention relates to a pest control constructor and a method thereof. The constructor includes Cry2Aa protein and Cry1A protein. The pest control constructor and the method disclosed by the invention control pests through the Cry2Aa protein and the Cry1Ab / Ac protein generated in plants; the constructor and the method, compared to an agricultural control method, a chemical control method and a biological control method in the prior art, are not only strong in toxicity and thorough in effect, but also capable of preventing and controlling pests through protecting the whole plants in the whole growth period; and the constructor is free of pollution and residues, stable in effect, simple, convenient and economical.

Description

technical field [0001] The present invention relates to a construct and method for controlling pests, in particular to a method for controlling pests to damage plants by using a construct expressing Cry2Aa protein and Cry1Ab / Ac protein in plants. Background technique [0002] Corn and rice are important food crops in China. Every year, plant pests cause huge food losses, and even affect the living conditions of the local population. In order to control plant pests, people usually use broad-spectrum chemical insecticides and biological insecticides, but both have limitations in practical application: chemical insecticides will cause environmental pollution problems and lead to the emergence of resistant insects. However, biopesticides are easily degraded in the environment and require repeated application in production, which greatly increases production costs. [0003] In order to solve the limitations in the practical application of chemical insecticides and biological ins...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/84C12P21/02A01H5/00A01N47/44A01N57/16A01P7/04
Inventor 韩超庞洁丁德荣李胜兵
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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