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The use of insecticidal protein

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

Active Publication Date: 2019-10-18
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] It has been proved that the plants transgenic for Cry1Fa can resist the attack of lepidoptera (Lepidoptera) pests such as the small cutworm. However, there is no report on the control of plant damage by the bean hornworm by producing transgenic plants expressing the Cry1Fa protein.

Method used

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  • The use of insecticidal protein
  • The use of insecticidal protein
  • The use of insecticidal protein

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

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

[0085] 1. Obtain the Cry1Fa nucleotide sequence

[0086] The amino acid sequence (605 amino acids) of Cry1Fa-01 insecticidal protein, as shown in SEQ ID NO:1 in the sequence listing; Cry1Fa-01 nucleotide sequence ( 1818 nucleotides), as shown in SEQ ID NO: 3 in the sequence listing; the amino acid sequence (1148 amino acids) of Cry1Fa-02 insecticidal protein, as shown in SEQ ID NO: 2 in the sequence listing; the encoding corresponds to the The Cry1Fa-02 nucleotide sequence (3447 nucleotides) of the amino acid sequence of the Cry1Fa-02 insecticidal protein is shown in SEQ ID NO:4 in the sequence listing.

[0087] 2. Obtain the Cry1Ac nucleotide sequence

[0088] The amino acid sequence (1156 amino acids) of Cry1Ac insecticidal protein, as shown in SEQ ID NO:5 in the sequence listing; Cry1Ac nucleotide sequence (3471 nucleotides) of encoding corresponding to the amino acid sequence of said Cry1Ac insecticida...

no. 2 example

[0091] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0092] 1. Construction of a recombinant cloning vector containing the Cry1Fa gene

[0093] The synthetic Cry1Fa-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 start codon; SP6 is the promoter of SP6 RNA polymerase; T7 is the promoter of T7 RNA polymerase; Cry1Fa-01 is the promoter of Cry1Fa -01 nucleotide sequence (SEQ ID NO: 3); MCS is the multiple cloning site).

[0094]Then, the recombinant cloning vector DBN01-T was transformed...

no. 3 example

[0105] The third embodiment, the acquisition of transgenic plants

[0106] 1. Obtaining transgenic soybean plants

[0107] 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 DBN100014, DBN100013 and DBN100756 (including the promoter sequence of Arabidopsis ubiquitin gene, Cry1Fa-01 nucleotide sequence, Cry1Fa-02 nucleotide sequence, Cry1Ac nucleotide sequence, PAT gene and tNos terminator sequence) into the soybean genome, obtained the soybean plant transferred to the Cry1Fa-01 nucleotide sequence, the soybean plant transferred to the Cry1Fa-02 nucleotide sequence and the Cry1Fa-02-Cry1Ac nucleotide sequence sequenced soybean plants, and wild-type soybean plants were used as controls.

[0108] For Agrobacter...

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Abstract

The invention relates to application of an insecticidal protein. The application includes that at least greenish brown hawk moth pests are in contact with a Cry1Fa protein. The greenish brown hawk moth pests are controlled by producing the Cry1Fa protein capable of killing the greenish brown hawk moths in plant bodies. Compared with agricultural prevention and control methods, chemical preventionand control methods, physical prevention and control methods and biological prevention and control methods in the prior art, whole plants are protected in a whole growing period to prevent and controlattack of greenish brown hawk moth pests, and the insecticidal protein is free of pollution and residues, stable and thorough in effect, 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 Cry1Fa protein expressed in a plant to control the bean hornworm from injuring 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 globa...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82A01H5/00A01H6/54A01N37/46A01P7/04A01H1/02A01H4/00A01H1/06
Inventor 杨淑靖任振涛吴竹筠韩超
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD