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Methods of Controlling Pests

By expressing Cry1F protein in plants, the pollution and instability problems of giant borer control in the existing technology are solved, the control effect of the whole growth period and the whole plant is achieved, and the effective control of giant borer is ensured.

Active Publication Date: 2017-05-17
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 can resist the attack of Lepidoptera pests such as the small cutworm. However, there is no report on the control of the plant damage caused by the large borer by producing transgenic plants expressing the Cry1F protein.

Method used

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  • Methods of Controlling Pests
  • Methods of Controlling Pests
  • Methods of Controlling Pests

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 is shown in SEQ ID NO: 1 in the sequence listing; Cry1Fa-01 encoding the corresponding amino acid sequence (605 amino acids) of 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 the Cry1Fa-02 insecticidal protein, as shown in SEQ ID NO: 2 in the sequence listing ; Encoding the Cry1Fa-02 nucleotide sequence (3447 nucleotides) corresponding to the amino acid sequence (1148 amino acids) of the Cry1Fa-02 insecticidal protein, as shown in SEQID NO:4 in the sequence listing.

[0087] 2. Obtain the nucleotide sequences of Cry1Ab and Vip3A

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

no. 2 example

[0091] Second embodiment, construction of recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0092] 1. Construction of recombinant cloning vector containing Cry1F gene

[0093] The synthesized Cry1Fa-01 nucleotide sequence was ligated into 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 , the construction process is as follows figure 1 (wherein, Amp is the ampicillin resistance gene; f1 is the origin of replication of phage f1; LacZ is the LacZ initiation 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 E. coli T1 competent cells (Transgen...

no. 3 example

[0106] The third embodiment, the acquisition and verification of the corn plant transformed into the Cry1F gene

[0107] 1. Obtain maize plants transformed with Cry1F gene

[0108] According to the conventional 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 embryos constructed in 2 in the second example were co-cultured. T-DNA in recombinant expression vectors DBN100014, DBN100012 and DBN100276 (including the promoter sequence of maize Ubiquitin gene, Cry1Fa-01 nucleotide sequence, Cry1Fa-02 nucleotide sequence, Cry1Ab nucleotide sequence, Vip3A nucleotide sequence , PMI gene and Nos terminator sequence) into the maize genome, and obtained maize plants with Cry1Fa-01 nucleotide sequences, Cry1Fa-01-Cry1Ab nucleotide sequences, and Cry1Fa -02-Vip3A nucleotide sequence of maize plants; wild-type maize plants were also used as co...

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Abstract

The invention relates to a method for controlling the pest of Sinophila spp., comprising: contacting the pest of C. chinensis with Cry1F protein. The present invention controls the large borer pest by producing the Cry1F protein capable of killing the large borer in the plant; compared with the agricultural control methods, chemical control methods and biological control methods used in the prior art, the present invention controls the plants during the whole growth period and the whole growth period. The protection of plants is to prevent and control the damage of the large borer pest, and there is no pollution and no residue, the effect is stable, thorough, simple, convenient and economical.

Description

technical field [0001] The present invention relates to a method for controlling pests, in particular to a method for controlling giant borers to damage plants by using Cry1F protein expressed in plants. Background technique [0002] The giant borer (Sesamia inferens) belongs to the Lepidoptera Noctuidae, and is an omnivorous pest. Besides maize, it also damages rice, sugarcane, sorghum and other gramineous crops. It is widely distributed in central and southeastern my country, especially in Shaanxi and Henan. Most of the rice area to the south. Large borer larvae bore into the stems of crops to cause damage, which can cause dead seedlings or the death of the whole plant. The boreholes are generally large, and a large amount of insect excrement is discharged from the stems. Occurrence is heavier than spring corn. [0003] Maize and sorghum are important food crops in China. The annual food loss caused by the giant borer is huge, and it even affects the living conditions of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01N63/50
CPCC12N15/8286Y02A40/146A01N63/50A01N25/00
Inventor 韩超庞洁康越景刘海利张云珠张成伟许春苹魏玫黄金存田康乐王乾钦
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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