Application of insecticidal protein

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

Active Publication Date: 2018-10-02
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] Plants transgenic for the Vip3Aa gene have been shown to be resistant to pests of the order Lepidoptera such as cutworms, cotton bollworms, and fall armyworms,

Method used

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  • Application of insecticidal protein
  • Application of insecticidal protein
  • Application of insecticidal protein

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0085] The first embodiment, the acquisition and synthesis of genes

[0086] 1. Obtain the nucleotide sequence

[0087] 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 nucleotides) of coding corresponding to the amino acid sequence of described Vip3Aa insecticidal protein ), as shown in SEQ ID NO:2 in the sequence listing.

[0088] 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.

[0089] The amino acid sequence (615 amino acids) of Cry1Ab-01 insecticidal protein, as shown in SEQ ID NO:5 in the sequence listing; Cry1Ab-01 nucleotide sequence (1848 nucleotides), as shown in SEQ ID NO:6 in the sequence listing.

[0090] The amino acid sequence (634 amino acids) of Cry2Ab-01 insecticidal prote...

no. 2 example

[0093] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0094] 1. Construction of a recombinant cloning vector containing the Vip3Aa gene

[0095]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; f1 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).

[0096] Then, the recombinant cloning vector DBN01-T was tran...

no. 3 example

[0109] The third embodiment, the acquisition of transgenic plants

[0110] 1. Obtaining transgenic soybean plants

[0111] 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 promoter sequence of Arabidopsis ubiquitin gene, Vip3Aa-01 nucleotide sequence, Vip3Aa-02 nucleotide sequence, Vip3Aa-02-Cry1Ab-01 core Nucleotide sequence, Vip3Aa-02-Cry2Ab-01 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, the transferred Vip3Aa The soybean plant of -02 nucleotide sequence, the soybean plant transferred to the Vip3Aa-02-Cry1Ab-01 nucleotid...

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Abstract

The invention relates to an application of an insecticidal protein, which includes a step of at least contacting an injurious insect, clanis bilineata tsingtauica, with Vip3Aa protein. In the invention, the clanis bilineata tsingtauica can be controlled by means of the Vip3Aa protein, produced in plants, which can kill the clanis bilineata tsingtauica; compared with agricultural, physical, biological and chemical prevention and treatment methods in the prior art, the insecticidal protein, can protect plants in all growth periods and on whole plant for preventing and treating the clanis bilineata tsingtauica. The insecticidal protein is free of pollution and residue, has stable and complete effect, and is simple, convenient and economical.

Description

technical field [0001] The invention relates to an application of an insecticidal protein, in particular to an application of a Vip3Aa protein expressed in a plant to control the plant damage caused by the bean hornworm. Background technique [0002] Bean horn moth Clanis bilineata (walker), whose larvae are commonly known as bean worm, bean dan, and bean cicada, belong to the genus Bean hawk moth of the family Scutellaridae Scutellaria subfamily of Lepidoptera, and are mainly distributed in the Huanghuai River Basin and the Yangtze River Basin and South China, the main soybean producing area in my country, is one of the main soybean pests. The bean hornworm is an explosive pest, and its damage characteristics are: the larvae overeat the leaves, the light ones are bitten into holes and nicks, and the severe ones become polished stems, and then the petioles remain, and they cannot form pods, which seriously affects the yield. [0003] Cultivated soybean (Glycine max (L.) Merr...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00
CPCC07K14/32C12N15/8261C12N15/8286
Inventor 韩超杨淑靖任振涛吴竹筠
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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