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Pest control method

A technology for pests, Spodoptera litura, applied in the directions of botanical equipment and methods, biochemical equipment and methods, pesticides, etc.

Active Publication Date: 2014-04-16
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] It has been proved that the plants transgenic for Cry1F gene can resist the attack of Lepidoptera pests such as the small cutworm, however, there is no report on the control of plant damage by Spodoptera litura by producing transgenic plants expressing Cry1F protein

Method used

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Examples

Experimental program
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Effect test

no. 1 example

[0087] The first embodiment, the acquisition and synthesis of Cry1Fa gene

[0088] 1. Obtain the Cry1Fa nucleotide sequence

[0089] 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; amino acid sequence (1148 amino acids) of Cry1Fa-02 insecticidal protein, as shown in SEQ ID NO:2 in the sequence listing ; Cry1Fa-02 nucleotide sequence (3447 nucleotides) encoding the amino acid sequence (1148 amino acids) corresponding to the Cry1Fa-02 insecticidal protein, as shown in SEQ ID NO:4 in the sequence listing.

[0090] 2. Obtain the nucleotide sequences of Cry1Ab and Vip3A

[0091] The Cry1Ab nucleotide sequence (2457 nucleotides) encoding the amino acid sequence (818 amino acids) of the Cry1Ab insectic...

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 Cry1F gene

[0096] 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 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: 3); MCS is a multiple cloning site).

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

no. 3 example

[0114] The third embodiment, the acquisition and verification of corn plants transferred to the Cry1F gene

[0115] 1. Obtaining corn plants transferred to the Cry1F gene

[0116] 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, DBN100013, DBN100075 and DBN100276 (including the promoter sequence of corn Ubiquitin gene, Cry1Fa-01 nucleotide sequence, Cry1Fa-02 nucleotide sequence, Cry1Ab nucleotide sequence, Vip3A nucleotide sequence acid sequence, PMI gene and Nos terminator sequence) into the maize genome, and obtained the maize plant with the Cry1Fa-01 nucleotide sequence, the maize plant with the Cry1Fa-02 nucleotide sequence, and the Cry1Fa -01-Cry1Ab nucleotide sequence maize plant and Cry1Fa-01-Vip3A nucleotide sequence-transferred maize plan...

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Abstract

The invention relates to a method for controlling prodenia litura pests. The method comprises the step that the prodenia litura pests are contacted with Cry1F protein. According to the invention, the prodenia litura pests are controlled by Cry1F protein which is generated in plants and can kill prodenia litura; and compared with an agricultural control method, a chemical control method and a physical control method used in the prior art, the pest control method can be used for protecting whole plants at the whole growth periods so as to prevent and control attack of the prodenia litura pests, and is further free of pollution and residue, stable and thorough in effect, simple, convenient and economical.

Description

technical field [0001] The invention relates to a method for controlling pests, in particular to a method for controlling Spodoptera litura from damaging plants by using Cry1F protein expressed in plants. Background technique [0002] Prodenia litura belongs to Lepidoptera Noctuidae. It is an omnivorous and gluttonous pest. It damages a wide range of hosts. In addition to corn and soybeans, it can also damage melons, eggplants, beans, green onions, leeks, spinach and There are nearly 100 families and more than 300 kinds of plants including cruciferous vegetables, grains, and cash crops. Spodoptera litura is distributed worldwide and occurs in all parts of the country, mainly in the Yangtze River Basin and the Yellow River Basin. Spodoptera litura mainly damages the whole plant with larvae, and gathers and eats on the back of leaves when it is young; after the third instar, it disperses and damages leaves, tender stems, and old larvae can eat fruits. [0003] Corn and soybea...

Claims

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Application Information

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IPC IPC(8): A01N47/44A01N57/16A01P7/04C12N15/84A01H5/00
CPCA01N37/46C07K14/325C12N15/8286Y02A40/146
Inventor 张爱红鲍晓明刘海利裴艳新张云珠
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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