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Use of insecticidal protein

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

Active Publication Date: 2019-04-26
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] Plants transgenic for Vip3Aa have been shown to be resistant to pests of the order Lepidoptera such as cutworms, cotton bollworms, and fall armyworm. Reports on Moth Harms to Plants

Method used

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Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0084] The first embodiment, the acquisition and synthesis of genes

[0085] 1. Obtain the nucleotide sequence

[0086] The amino acid sequence (789 amino acids) of Vip3Aa-01 insecticidal protein, as shown in SEQ ID NO: 1 in the sequence listing; The Vip3Aa nucleotide sequence (2370 nucleosides) of encoding corresponding to the amino acid sequence of said Vip3Aa insecticidal protein acid), as shown in SEQ ID NO:2 in the sequence listing.

[0087] Amino acid sequence (789 amino acids) of Vip3Aa-02 insecticidal protein, as shown in SEQ ID NO:3 in the sequence listing; Vip3Aa-02 nucleotide sequence ( 2370 nucleotides), as shown in SEQ ID NO:4 in the sequence listing.

[0088] The amino acid sequence (615 amino acids) of Cry1Ab insecticidal protein, as shown in SEQ ID NO:5 in the sequence listing; Cry1Ab nucleotide sequence (1848 nucleotides) of encoding corresponding to the amino acid sequence of said Cry1Ab insecticidal protein , as shown in SEQ ID NO:6 in the sequence listin...

no. 2 example

[0092] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0093] 1. Construction of a recombinant cloning vector containing the Vip3Aa gene

[0094]The synthetic Vip3Aa-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, whose construction process is as follows figure 1 Shown (among them, Amp represents the ampicillin resistance gene; f1ori represents the replication origin of phage f1; LacZ is the actual codon of LacZ; SP6 is the promoter of SP6 RNA polymerase; T7 is the promoter of T7 RNA polymerase; Vip3Aa-01 is the promoter of Vip3Aa- 01 nucleotide sequence (SEQ ID NO: 2); MCS is a multiple cloning site).

[0095] Then, the recombinant cloning vector DBN01-T was t...

no. 3 example

[0108] The third embodiment, the acquisition of transgenic plants

[0109] 1. Obtaining transgenic soybean plants

[0110] 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 example, so that the recombinant gene constructed in 2 in the second example T-DNA of expression vectors DBN100002, DBN100741, DBN100003 and DBN100370 (including the promoter sequence of Arabidopsis ubiquitin gene, Vip3Aa-01 nucleotide sequence, Vip3Aa-02 nucleotide sequence, Vip3Aa-02-Cry1Ab nucleotide sequence sequence, Vip3Aa-02-Cry2Ab nucleotide sequence, PAT gene and tNos terminator sequence) were transferred into the soybean genome, and the soybean plants transferred to the Vip3Aa-01 nucleotide sequence and the Vip3Aa-02 nucleoside Acid sequence soybean plants, soybean plants transformed with Vip3Aa-02-Cry1Ab nucleotide sequence, and s...

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Abstract

The invention relates to use of an insecticidal protein. According to the use, argyrogramma agnata pests are enabled to be at least in contact with a Vip3Aa protein. The argyrogramma agnata pests arecontrolled by causing the Vip3Aa protein, capable of killing the argyrogramma agnata pests, to generate in plant bodies. Compared with the agricultural control method, physical control method, biological control method and chemical control method in the prior art, the whole plants are protected in a whole growth period, so that the invasion of the argyrogramma agnata pests can be prevented and controlled; furthermore, the insecticidal protein is free from pollution and residues, stable and thorough in effect, simple, convenient and economical.

Description

technical field [0001] The present invention relates to the use of an insecticidal protein, in particular to the use of a Vip3Aa protein to control plant damage to plants by expressing the plant. 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. 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 degrees of food lo...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12N15/29C07K14/415A01H5/00A01H6/54A01N37/46A01P7/04
CPCA01N37/46C12N15/8286C07K14/415Y02A40/146
Inventor 谢香庭杨淑靖任振涛
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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