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Use of insecticidal proteins

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

Active Publication Date: 2019-01-18
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 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 meadow borer damage to plants by producing transgenic plants expressing the Cry1Fa protein.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0087] The first embodiment, the acquisition and synthesis of genes

[0088] 1. Obtain the nucleotide sequence

[0089] 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: 2 in the sequence listing; the amino acid sequence (1148 amino acids) of Cry1Fa-02 insecticidal protein, as shown in SEQ ID NO: 3 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.

[0090] 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 insecticidal protein , as shown in SEQ ID NO:6 in the sequence listi...

no. 2 example

[0094] The second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0095] 1. Construction of a recombinant cloning vector containing the Cry1Fa gene

[0096]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: 2); MCS is the multiple cloning site).

[0097] Then, the recombinant cloning vector DBN01-T was transformed...

no. 3 example

[0112] The third embodiment, the acquisition of transgenic plants

[0113] 1. Obtaining transgenic soybean plants

[0114] According to the commonly used Agrobacterium infection method, the cotyledon node tissue of the aseptically cultivated soybean variety Zhonghuang 13 was co-cultured with the Agrobacterium described in 4 in the second embodiment, so that the soybean constructed in 2 in the second embodiment The T-DNA of recombinant expression vectors DBN100656 and DBN100003 (including Cry1Fa-02 nucleotide sequence, Cry1Ac nucleotide sequence, Vip3A nucleotide sequence and PAT gene) were transferred into the soybean genome, and the transferred Cry1Ac-Cry1Fa -02 nucleotide sequence soybean plants and soybean plants transformed into Vip3A nucleotide sequence, and wild-type soybean plants were used as controls.

[0115] For Agrobacterium-mediated soybean transformation, briefly, mature soybean seeds were germinated in soybean germination medium (B5 salt 3.1 g / L, B5 vitamin, su...

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PUM

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Abstract

The invention relates to the use of an insecticidal protein, comprising: contacting a grass moth pest with at least Cry1Fa protein. Cry1Fa protein capable of killing meadow moth is generated in plantto control meadow moth pest; Compared with the agricultural prevention and control method, chemical prevention and control method, physical prevention and control method and biological prevention andcontrol method used in the prior art, the invention protects the whole plant during the whole growth period of the plant to prevent and control the invasion of the grass moth pest, and has no pollution, no residue, stable effect, thoroughness, simplicity, convenience and economy.

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 plants to control the damage of plants by the meadow borer. Background technique [0002] The meadow moth Loxostege sticticalis, belonging to the Lepidoptera Boreridae, is mainly distributed in the Northeast, Northwest, and North China, namely Jilin, Inner Mongolia, Heilongjiang, Ningxia, Gansu, Qinghai, Hebei, Shanxi, Shaanxi, Jiangsu and other provinces . Meadow borer is a polyphagous large pest, which can feed on 35 families and more than 200 kinds of plants, mainly sugar beets, soybeans, sunflowers, potatoes, hemp, vegetables, medicinal materials and other crops; suffer from it. The meadow moth usually has a major outbreak every 10 years or so. The characteristics of its damage are: the larvae mostly gather on the branches and form nets to hide, eat the mesophyll, and leave the epidermis and veins. In severe cases, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82A01H5/00A01H6/54A01N37/46A01P7/04A01H1/02A01H4/00A01H1/06
CPCA01H1/02A01H1/06A01H4/008A01N37/46C12N15/8286
Inventor 韩超杨淑靖任振涛陶青
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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