Insecticide protein composition and method thereof for managing insect resistance

An insect resistance and protein technology, applied in the fields of pesticides, chemical instruments and methods, and botanical equipment and methods, can solve problems such as poor sustainability, lower total yield, and cumbersome farmers' actual operation.

Active Publication Date: 2017-04-26
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The purpose of setting up shelters is to maintain a certain proportion of non-resistant alleles to delay the emergence of insects resistant to high doses of insect-resistant proteins, but this strategy will make farmers more cumbersome in practice and to some extent reduce total output
[0006] 2) Increase the dosage of a single anti-insect protein. This strategy is less sustainable, and insects will develop higher resistance to a single protein under continuous selection pressure
In view of the fact that Bt proteins specifically bind to the receptors of sensitive insects, it is necessary to assess whether insects have cross-resistance to different insect-resistant proteins, that is, whether they share or compete for the binding site, so the superimposed use of insect-resistant proteins has high uncertainty. So far, there is no report on whether Cry2Ab protein has cross-resistance to O. corn Borer and Cry1Ab protein

Method used

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  • Insecticide protein composition and method thereof for managing insect resistance
  • Insecticide protein composition and method thereof for managing insect resistance
  • Insecticide protein composition and method thereof for managing insect resistance

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0088] The first embodiment, expression, purification and activation of Cry2Ab protein

[0089] 1. Construction of recombinant expression vector containing Cry2Ab nucleotide sequence

[0090] The amino acid sequence (634 amino acids) of the Cry2Ab insecticidal protein is shown in SEQ ID NO: 1 in the sequence listing; the nucleotide sequence of the Cry2Ab insecticidal protein is optimized according to the codon preference of Escherichia coli, and the code corresponding to The Cry2Ab nucleotide sequence (1905 nucleotides) of the amino acid sequence of the Cry2Ab insecticidal protein is shown in SEQ ID NO:3 in the sequence listing.

[0091] The Cry2Ab nucleotide sequence (shown as SEQ ID NO: 3 in the sequence listing) was synthesized by Nanjing GenScript Biotechnology Co., Ltd., and the structural representation of the recombinant expression vector Cry2Ab-pET30 constructed therefrom is shown in figure 1 Shown (wherein, Kan represents the kanamycin resistance gene; f1 represents ...

no. 2 example

[0112] Second embodiment, bioassay of Bt-sensitive corn borer strain and corn borer-resistant strain (Cry1Ab protein)

[0113] 1. The source of the tested insects

[0114] The tested insects were the Bt-sensitive strain of Ostrinia furnacalis (hereinafter referred to as ACB-BtS to represent the Bt-sensitive strain of Ostrinia furnacalis) and the Bt-resistant strain of Asian corn borer (Ostrinia furnacalis) (hereinafter referred to as ACB-BtS). AbR represents the Asian corn borer Bt-resistant strain Ab0.8, that is, the Asian corn borer Bt-resistant strain subcultured with feed with a Cry1Ab concentration of 0.8 μg / g), and the insects of the above two strains are from China Institute of Plant Protection, Academy of Agricultural Sciences.

[0115] 2. Bioassay method for Asian corn borer

[0116]Bioassays were performed with different concentrations for each Bt protein, the concentration range of Cry1Ab protein was 0 μg / g to 50 μg / g, and the concentration range of Cry2Ab protein...

no. 3 example

[0127] The third embodiment, the application of Cry2Ab protein and Cry1Ab protein in the production of insect-resistant transgenic plants

[0128] According to the experimental results of the second embodiment, the Cry2Ab protein and the Cry1Ab protein are expected to be used for combined expression in transgenic plants such as corn plants to delay or prevent O. corn borer from developing resistance to the plant.

[0129] The first method is sequential transformation: where a plant transformed with a first gene (eg the Cry1Ab gene) is retransformed to introduce a second gene (eg the Cry2Ab gene). This sequential transformation preferably uses two different selectable marker genes, eg a kanamycin resistance gene and a phosphinothricin acetyltransferase gene (eg pat or bar gene) conferring resistance to the glufosinate herbicide.

[0130] The second method is the co-transformation method: the nucleotide sequence encoding the Cry1Ab protein is expressed in plants together with th...

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Abstract

The invention relates to an insecticide protein composition and a method thereof for managing insect resistance. The method for managing insect resistance comprises the steps that Asiatic corn borer is in contact with at least of Cry2Ab protein and Cry1Ab protein. The Asiatic corn borer is effectively delayed or prevented from producing resistance by the two kinds of insecticide protein Cry2Ab protein and Cry1Ab protein, the Asiatic corn borer is controlled or prevented and cured, the plant can be protected to the greater extent, and the yield is stabilized.

Description

technical field [0001] The present invention relates to an insecticidal protein combination and a method for managing insect resistance thereof, in particular to a method for managing resistance to Ostrinia sativa by combining Cry1Ab protein and Cry2Ab protein. Background technique [0002] Insect pests cause huge economic losses in crop production worldwide, and farmers face the threat of yield loss due to insect pests every year. The Asian corn borer (Ostrinia furnacalis), commonly known as the borer, belongs to the family Lepidoptera and is the main pest in corn production in my country. This kind of insect eats the leaves of corn and eats into the main stem or ear of corn, reducing photosynthesis and affecting Nutrient transportation can also lead to various secondary diseases, resulting in reduced yield and quality of corn. In recent years, with the change of climatic conditions, the change of farming system, the increase of corn planting density, and the improvement of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/70C07K14/325C12N15/32A01N47/44A01P7/04
CPCA01N47/44C12N15/70C12N15/8286C07K14/325A01N2300/00Y02A40/146
Inventor 陶青杨旭李建勇
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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