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Application of insecticidal protein

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

Active Publication Date: 2018-08-24
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 plant damage caused by bean hornworm

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

[0092] The first embodiment, the acquisition and synthesis of genes

[0093] 1. Obtain the nucleotide sequence

[0094] 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 coding corresponding to the amino acid sequence (818 amino acids) of said Cry1Ab-01 insecticidal protein 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.

[0095] The amino acid sequence (1178 amino acids) of the Cry1Ac-01 insecticidal protein, as shown in SEQ ID NO: 5 in the sequence listing; Cry1Ac-01 encoding the amino acid sequence (1178 amino acids) corresponding to the Cry1Ac-01 insecticidal protein Nucleotide sequence (3537 nucleotides), as shown in SEQ ...

no. 2 example

[0101] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0102] 1. Construction of a recombinant cloning vector containing the Cry1A gene

[0103] 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; f1 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).

[0104] Then, the recombinant cloning vector DBN01-T was transformed into Esche...

no. 3 example

[0123] The third embodiment, the acquisition of transgenic plants

[0124] 1. Obtaining transgenic soybean plants

[0125] 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 DBN100125, DBN100744, DBN100745, DBN100075, DBN100756, DBN100757 and DBN100076 (including promoter sequence of Arabidopsis ubiquitin gene, Cry1Ab-01 nucleotide sequence, Cry1Ab-02 nucleotide sequence, Cry1Ac- 01 nucleotide sequence, Cry1Ac-02 nucleotide sequence, Cry1A.105 nucleotide sequence, Cry2Ab nucleotide sequence, Cry1Fa nucleotide sequence, PAT gene and tNos terminator sequence) into the soybean genome, Obtained soybean plants transferred to Cry1Ab-01 nucleotide sequence, soybean plants transferred to Cry1Ab-02 nucleotide sequence...

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Abstract

The invention relates to application of an insecticidal protein. The application of the insecticidal protein comprises enabling a clanis bilineata tsingtauica pest to be in contact with Cry1A protein.The Cry1A protein which is generated in a plant body and is capable of killing clanis bilineata tsingtauica is used for controlling the clanis bilineata tsingtauica pest; compared with the agricultural control method, the chemical control method, the physical control method and the biological control method used in the prior art, the insecticidal protein is capable of protecting whole plants in awhole growth period and controlling the infestation of the clanis bilineata tsingtauica, is free of pollution, is free of residues, has a stable and thorough effect, and is 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 Cry1A protein to control plant damage to plants by Cry1A protein expressed in plants. Background technique [0002] Bean horn moth Clanis bilineata (walker), its larvae are commonly known as bean worm, bean dan, bean cicada, belongs to the genus Bean horn moth of Lepidoptera Scutellaridae Scutellaria subfamily, 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, enter the residual petioles, and cannot set pods, seriously affecting the yield. [0003] Cultivated soybean (Glycine max (L.) Merri) is an important economic crop grown globally as ...

Claims

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Application Information

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IPC IPC(8): A01N37/46A01P7/04A01H1/02A01H4/00
CPCA01H1/02A01H4/008A01N37/46
Inventor 吴竹筠韩超杨淑靖任振涛
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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