Method for managing resistance of ostrinia nubilalis to Bt insecticidal protein

A corn borer, resistance technology, applied in the direction of instruments, hybridization, sequence analysis, etc., can solve the problems of corn borer no insecticidal activity, corn borer resistance management has no effect, etc., to prolong the service life and alleviate Bt toxin resistance the effect of

Pending Publication Date: 2021-12-14
HANGZHOU RUIFENG BIOTECH LIMITED
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, the widely used insecticidal protein Vip3Aa has no insecticidal activity against corn borer
Therefore, the use of Vip3Aa has no effect on the resistance management of the corn borer

Method used

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  • Method for managing resistance of ostrinia nubilalis to Bt insecticidal protein
  • Method for managing resistance of ostrinia nubilalis to Bt insecticidal protein
  • Method for managing resistance of ostrinia nubilalis to Bt insecticidal protein

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Embodiment 1, the preparation of Bt recombinant protein

[0031] The coding genes involved in the present invention are all artificially synthesized by Shanghai Sangong Company, wherein the amino acid sequence of the Cry1Ab protein is shown in SEQ ID NO: 1, and the nucleotide sequence of the coding gene cry1Ab is shown in SEQ ID NO: 4; Cry1Fa The amino acid sequence of the protein is shown in SEQ ID NO: 2, the nucleotide sequence of the coding gene cry1Fa is shown in SEQ ID NO: 5; the amino acid sequence of the Cry2Ab protein is shown in SEQ ID NO: 3, the nucleotide sequence of the coding gene cry1Fa The acid sequence is shown in SEQ ID NO: 6; the amino acid sequence of the Vip3Af protein is shown in SEQ ID NO: 7, and the nucleotide sequence of the coding gene vip3Af is shown in SEQ ID NO: 10; the amino acid sequence of the Vip3Ag protein is shown in SEQ ID NO Shown in: 8, the nucleotide sequence of coding gene vip3Ag is shown in SEQ ID NO: 11; The aminoacid sequence of...

Embodiment 2

[0039] Embodiment 2, the biological activity assay of recombinant protein to corn borer

[0040] Experimental objects: The newly hatched larvae of common corn borer (collected from the field of Sanya Experimental Base in Hainan), Cry1Ab-resistant corn borer and Cry1Fa-resistant corn borer within 2 hours of birth were selected as the tested insects. Resistance to corn borer (anti-Cry1Ab corn borer and anti-Cry1Fa corn borer) was obtained by artificial breeding and screening, the specific method refers to the literature (Kain, Wendy C., et al. "Inheritance of resistance to Bacillus thuringiensis Cry1Ac toxin in greenhouse-derived strain of cabbage looper(Lepidoptera: Noctuidae). "Journal of economic entomology 97.6(2004):2073-2078.; Huang, Fangneng, et al."Baseline susceptibility and changes in susceptibility to Bacillus thuringiensis opept. Pyralidae). "Journal of Economic Entomology 90.5(1997): 1137-1143.) to screen. The resistant corn borer was not sensitive to Bt insecticid...

Embodiment 3

[0051] Embodiment 3, the construction of transformation vector

[0052] In this example, the transformation vector is constructed based on the pCambia1300 (NCBI sequence number AF234296) vector. Readers of the present invention can achieve the same effect on other plant genetic transformation vectors according to the method provided in this example, such as pCambia1301, pCambia2301 et al. This example takes the construction of an expression cassette containing two genes cry1Fa and vip3Af as an example to illustrate the construction method of the transformation vector, replace the cry1Fa gene with any of the Cry proteins described in the present invention, and replace the vip3Af gene with any of the Cry proteins described in the present invention The Vip3 protein can realize the construction of the transformation vector. In addition, the promoter used in the expression cassette of the insect-resistant gene can be any constitutive promoter, such as the cauliflower mosaic virus ...

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Abstract

The invention discloses a method for managing resistance of ostrinia nubilalis to Bt insecticidal protein. The method is realized by simultaneously expressing at least one Bt insecticidal crystal protein Cry and at least one Bt vegetative insecticidal protein Vip3 in the same plant cell. According to the invention, the coding gene of the Bt insecticidal crystal protein Cry and the coding gene of the Bt vegetative insecticidal protein Vip3 are simultaneously introduced into plant cells so that the obtained transgenic plant can simultaneously express two Bt toxins which have an efficient insecticidal effect on ostrinia nubilalis and do not have cross resistance, and the resistance of the ostrinia nubilalis to the Bt toxins can be effectively managed; the generation of Bt toxin resistance of corn borers can be greatly relieved, the service life of transgenic plants is greatly prolonged, important germplasm resource reserve is provided for achieving the national biological breeding strategy, and the method is creative improvement of an existing pest resistance control technology.

Description

(1) Technical field [0001] The present invention relates to a method for managing the resistance of the Lepidoptera pest corn borer, including European corn borer (Ostrinia nubilalis) and Asian corn borer (Ostrinia furnacalis), to Bt toxin from Bacillus thuringiensis. (2) Background technology [0002] Pests cost billions of dollars annually in global agricultural production. The control of pests currently mainly relies on the use of chemical pesticides and the promotion of genetically modified crops. Residues of chemical pesticides can have adverse effects on human health and the environment. The use of biotechnology to cultivate transgenic insect-resistant crops containing insect-resistant genes can greatly reduce the use of chemical pesticides and effectively protect crops from pests. There are multiple insecticidal proteins, and the most used insecticidal crystal proteins from Bacillus thuringiensis (Bt) at present, such as Cry1A, Cry1F, etc.; al. 1996, Adang, Crickmo...

Claims

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Application Information

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IPC IPC(8): G16B25/10G16B30/00
CPCG16B25/10G16B30/00
Inventor 许超沈志成
Owner HANGZHOU RUIFENG BIOTECH LIMITED
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