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Application of insecticidal proteins

A protein and pest technology, applied in the field of insecticidal protein

Active Publication Date: 2019-05-28
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] It has been proved that plants transgenic for Cry1A can resist the attack of Lepidoptera pests such as corn borer, cotton bollworm, fall armyworm (Fall Armyworm), etc. Reports on Controlling the Harm to Plants by the Spodoptera moth

Method used

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  • Application of insecticidal proteins
  • Application of insecticidal proteins
  • Application of insecticidal proteins

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0086] The first embodiment, the acquisition and synthesis of genes

[0087] 1. Obtain the nucleotide sequence

[0088] The amino acid sequence (818 amino acids) of Cry1Ab-01 insecticidal protein, as shown in SEQ ID NO:1 in the sequence listing; Cry1Ab-01 nucleotide sequence ( 2457 nucleotides), as shown in SEQ ID NO:2 in the sequence listing. The amino acid sequence (615 amino acids) of Cry1Ab-02 insecticidal protein, as shown in SEQ ID NO:3 in the sequence listing; Cry1Ab-02 nucleotide sequence ( 1848 nucleotides), as shown in SEQ ID NO:4 in the sequence listing.

[0089] The amino acid sequence (1178 amino acids) of Cry1Ac-01 insecticidal protein, as shown in SEQ ID NO:5 in the sequence listing; Cry1Ac-01 nucleotide sequence ( 3537 nucleotides), as shown in SEQ ID NO:6 in the sequence listing. The amino acid sequence (1156 amino acids) of Cry1Ac-02 insecticidal protein, as shown in SEQ ID NO: 7 in the sequence listing; Cry1Ac-02 nucleotide sequence ( 3471 nucleotides),...

no. 2 example

[0095] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0096] 1. Construction of a recombinant cloning vector containing the Cry1A gene

[0097] The synthesized Cry1Ab-01 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 , its construction process is as follows figure 1 Shown (wherein, Amp represents the ampicillin resistance gene; fiori represents the replication origin of phage f1; LacZ is the LacZ initiation codon; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; Cry1Ab-01 is the Cry1Ab -01 nucleotide sequence (SEQ ID NO: 2); MCS is the multiple cloning site).

[0098] Then, the recombinant cloning vector DBN01-T was transformed into Es...

no. 3 example

[0117] The third embodiment, the acquisition of transgenic plants

[0118] 1. Obtaining transgenic soybean plants

[0119] According to the routinely used Agrobacterium infection method, the cotyledon node tissue of the aseptically cultivated soybean variety Zhonghuang 13 was co-cultured with the Agrobacterium described in 3 in the second embodiment, so that the soybean constructed in 2 in the second embodiment T-DNA of recombinant expression vectors DBN100125, DBN100744, DBN100645, DBN100175, DBN100656, DBN100757 and DBN100176 (including Cry1Ab-01 nucleotide sequence, Cry1Ab-02 nucleotide sequence, Cry1Ac-01 nucleotide sequence, Cry1Ac-02 core nucleotide sequence, Cry1A.105 nucleotide sequence, Cry2Ab nucleotide sequence, Cry1Fa nucleotide sequence and PAT gene) were transferred into the soybean genome, and soybean plants transferred to the Cry1Ab-01 nucleotide sequence were obtained, Soybean plant transferred to Cry1Ab-02 nucleotide sequence, soybean plant transferred to Cr...

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Abstract

The invention relates to an application of insecticidal proteins. The application of the insecticidal proteins comprises the following steps that argyrogramma agnata pests are at least in contact withCry1A proteins. According to the application of the insecticidal proteins, the Cry1A proteins capable of killing argyrogramma agnata are generated in plant bodies to control the argyrogramma agnata pests. Compared with an agricultural control method, a chemical control method, a physical control method and a biological control method used in the prior art, full-growth period and full-plant protection on plants are adopted in the application of the insecticidal proteins to control the invasion of the argyrogramma agnata pests, and the application of the insecticidal proteins has no pollution and residue, has stable and thorough effects, and is simple, convenient and economic.

Description

technical field [0001] The present invention relates to the use of an insecticidal protein, in particular to the use of a Cry1A protein to control plant damage to plants by expressing the protein Cry1A. Background technique [0002] Argyrogramma agnata, belonging to Lepidoptera Noctuidae, is mainly distributed in the Yangtze River Basin and the Yellow River Basin in my country. It is one of the main soybean pests in the main soybean producing areas of China. Silver print moth is a polyphagous pest, mainly harming beans, rapeseed, cabbage, cauliflower, cabbage, radish and other cruciferous vegetables and other crops. Eat up the leaves and affect the yield. [0003] Cultivated soybean (Glycine max (L.) Merri) is an important economic crop grown globally as the main source of vegetable oil and vegetable protein, and is an important food crop in China. Soybean is one of the plants that silver-leaved moths like to feed on. Every year, silver-leafed moths will cause different de...

Claims

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Application Information

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IPC IPC(8): A01G13/00A01G22/15A01G22/25A01G22/40A01H1/02A01H4/00C12N15/32C12N15/82A01H5/00A01H6/20A01H6/54
Inventor 张爱红杨淑靖任振涛贺建波
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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