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Application of insecticidal proteins

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

Active Publication Date: 2019-05-28
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 damage of Lepidoptera pests such as the small cutworm. However, there is no report about controlling the damage to plants by producing transgenic plants expressing the Cry1Fa protein.

Method used

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  • Application of insecticidal proteins
  • Application of insecticidal proteins
  • Application of insecticidal proteins

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

[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 Cry1Fa gene

[0095] 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; f1ori 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).

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

no. 3 example

[0107] The third embodiment, the acquisition of transgenic plants

[0108] 1. Obtaining transgenic soybean plants

[0109] 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 embodiment, so that the soybean constructed in 2 in the second embodiment The T-DNA of recombinant expression vectors DBN100015, DBN100013 and DBN100656 (including Cry1Fa-01 nucleotide sequence, Cry1Fa-02 nucleotide sequence, Cry1Ac nucleotide sequence and PAT gene) were transferred into the soybean genome, and the transgenic Soybean plants with Cry1Fa-01 nucleotide sequence, soybean plants with Cry1Fa-02 nucleotide sequence, soybean plants with Cry1Ac-Cry1Fa-02 nucleotide sequence, and wild-type soybean plants as controls.

[0110] For Agrobacterium-mediated soybean transformation, briefly, mature soybean seeds were germinated in soybean ge...

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Abstract

The invention relates to an application of insecticidal proteins. The application of the insecticidal proteins comprises the following steps that argyrogramma agnata pests are at least in contact withCry1Fa proteins. According to the application of the insecticidal proteins, the Cry1Fa proteins capable of killing argyrogramma agnata are generated in plant bodies to control the argyrogramma agnatapests. Compared with an agricultural control method, a chemical control method, a physical control method and a biological control method used in the prior art, full-growth period and full-plant protection on plants are adopted in the application of the insecticidal proteins to control the invasion of the argyrogramma agnata pests, and the application of the insecticidal proteins has no pollutionand residue, has stable and thorough effects, and is simple, convenient and economic.

Description

technical field [0001] The present invention relates to the use of an insecticidal protein, in particular to the use of a Cry1Fa protein to control plant damage to plants by expressing the protein in plants. 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. [0003] 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 differe...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01G13/00A01G22/15A01G22/40A01H1/02A01H4/00A01H5/00A01H6/20A01H6/54C12N15/32C12N15/82
Inventor 韩超杨淑靖任振涛
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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