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Method for controlling pests

A technology for pests and Spodoptera frugiperda, applied in the field of pest control

Active Publication Date: 2013-03-20
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] It has been proved that plants transgenic for Cry1F can resist damage from Lepidoptera pests such as cutworms. However, there is no report on controlling the damage of plants by Cry1F protein by producing transgenic plants expressing Cry1F protein

Method used

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Examples

Experimental program
Comparison scheme
Effect test

no. 1 example

[0084] The first embodiment, the acquisition and synthesis of Cry1Fa gene

[0085] 1. Obtain the Cry1Fa nucleotide sequence

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

[0087] 2. Obtain the nucleotide sequences of Cry1A and Vip3A

[0088] The Cry1Ab nucleotide sequence (1848 nucleotides) encoding the amino acid sequence (615 amino acids) of the Cry1Ab ...

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 Cry1F gene

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

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

no. 3 example

[0106] The third embodiment, the acquisition and verification of corn plants transferred to the Cry1F gene

[0107] 1. Obtaining corn plants transferred to the Cry1F gene

[0108] According to the commonly used Agrobacterium infection method, the immature embryos of the aseptically cultured maize variety Zong 31 (Z31) were co-cultured with the Agrobacterium described in 3 in the second example, so that the T-DNA in recombinant expression vectors DBN100014, DBN100012 and DBN100276 (including the promoter sequence of corn Ubiquitin gene, Cry1Fa-01 nucleotide sequence, Cry1Fa-02 nucleotide sequence, Cry1Ab nucleotide sequence, Vip3A nucleotide sequence , PMI gene and Nos terminator sequence) were transferred into the maize genome, and the maize plants transferred to the Cry1Fa-01 nucleotide sequence, the maize plants transferred to the Cry1Fa-01-Cry1Ab nucleotide sequence and the transferred Cry1Fa - the maize plant of 02-Vip3A nucleotide sequence; meanwhile, the wild type maize...

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Abstract

The invention relates to a method for controlling athetis lepigones, which comprises the steps of getting the athetis lepigones to be contacted with protein of CrylF. According to the method, the athetis lepigones are controlled by the protein of CrylF which is capable of killing the athetis lepigones and is generated in a plant. Compared with the agricultural control method and the chemical control method used in the prior art, the method for controlling pests can be used for protecting the whole plant from being damaged by the athetis lepigones in the whole growth period, and the method has the advantages of being pollution-fee, stable in effect, thorough in killing pests, simple, convenient and economic, and having no residue.

Description

technical field [0001] The invention relates to a method for controlling pests, in particular to a method for using Cry1F protein expressed in plants to control the damage of plants by Spodoptera spp. Background technique [0002] Athetis lepigone belongs to Lepidoptera Noctuidae and is an omnivorous pest. At present, it is found that it mainly damages corn crops. It is mainly distributed in the summer corn planting areas of Huanghuaihai in my country, and it is mainly distributed in Japan, North Korea, Russia, Europe and other places. The two-spot moth mainly damages the corn roots at the soil surface of the aerial roots of corn, and bites off the corn stems or superficial roots of the corn. A large area of ​​blank land appeared, and the plots were seriously damaged or even destroyed. [0003] Corn is the main food crop in China. On July 9, 2011, CCTV News Network reported for the first time the occurrence of Noctuid moth in China. From the autumn of 2011 to May 31, 2012, ...

Claims

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Application Information

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IPC IPC(8): A01N47/44A01P7/04
CPCC07K14/325C12N15/8286A01N63/02Y02A40/146
Inventor 程鹏丁德荣庞洁李胜兵王利君宋金岭张迪李凯丽田康乐汤勇
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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