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Polynucleotide of insect resistant plant resistant to herbicide, expression cassette including polynucleotide and application thereof

A technology of polynucleotides and insect-resistant plants, applied in the field of genetic engineering, can solve the problems of pests in cotton fields, achieve high expression levels, reduce demand, and reduce the chance of resistance

Inactive Publication Date: 2016-10-26
达尼亚·贾德·库雷西·索·贾德·萨利姆·库雷西 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although Bt cotton varieties Bollguard-II (B G-2) and Roundup Ready Flex (RRF) cotton are available in the southern Punjab and Sindh provinces, farmers have surprisingly reported Low levels of Bt toxins in crops have resulted in constant pest infestation in cotton fields (GMWatch, 2014)

Method used

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  • Polynucleotide of insect resistant plant resistant to herbicide, expression cassette including polynucleotide and application thereof
  • Polynucleotide of insect resistant plant resistant to herbicide, expression cassette including polynucleotide and application thereof
  • Polynucleotide of insect resistant plant resistant to herbicide, expression cassette including polynucleotide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] An expression cassette for a herbicide-resistant insect-resistant plant: comprising a first expression cassette, a second expression cassette, a third expression cassette and a fourth expression cassette;

[0093] The first expression cassette comprises a first promoter, a 141bp long first 5' non-coding region, a first polynucleotide and a first terminator; the first polynucleotide comprises a coding region of EPSP synthase and a 108bp long the coding region of the first CTP;

[0094] The first promoter is connected to the 5' end of the first 5' non-coding region; the coding region of the first CTP is connected to the 3' end of the first 5' non-coding region, and the first 5' non-coding region and the first CTP The sequence connected to the coding region is shown in SEQ ID No.17; the coding region of the EPSP synthase is connected at the 3' end of the coding region of the first CTP; the first terminator is connected at the 3' end of the coding region of the EPSP synthas...

Embodiment 2

[0116] Each expression cassette in Example 1 was prepared by the following process:

[0117] The coding sequence of CTP is operably combined at the 3' end of the 5' non-coding region including the corresponding promoter; then further tagged to the coding region of EPSP synthase, the coding region of the vegetative insecticidal protein VIP3A or the insecticidal protein Cry2A The 5' end of the DNA sequence of the coding region; the corresponding terminator is further connected to the 3' end of the DNA sequence encoding the protein.

Embodiment 3

[0119] This example is used to illustrate the process of obtaining the carrier:

[0120] 1) The first expression cassette, the second expression cassette, the third expression cassette and the fourth expression cassette obtained in Example 1 and Example 2 are cloned between the left border sequence and the right border sequence of the same T-DNA of the plasmid vector , the sequence of the T-DNA is shown in SEQ ID No.26; obtain the vector inserted into the expression cassette;

[0121] 2) Using the Agrobacterium electroporation transformation technique, the plasmid vector obtained in step 1) is transformed into Agrobacterium tumefaciens LB4404 strain, and plants are infected by Agrobacterium tumefaciens.

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Abstract

The invention relates to polynucleotide of an insect resistant plant resistant to herbicide, an expression cassette including polynucleotide and application thereof. The polynucleotide of an insect resistant plant resistant to herbicide includes first polynucleotide, second polynucleotide and third polynucleotide. The first polynucleotide includes a coding area of EPSP synthase; the second polynucleotide includes a coding area of a vegetative period insecticide protein VIP3A; and the third polynucleotide includes a coding area of an insecticide protein Cry2A. The polynucleotide can code the EPSP synthase, the vegetative period insecticide protein VIP3A, and bacillus thuringiensis Delta-endotoxin, i.e., the insecticide protein Cry2A. The invention further relates to a DNA builder formed by a first expression cassette, a second expression cassette, a third expression cassette and a fourth expression cassette, and a method for transforming a plant by using the DNA builder. The transgenic plant has stronger resistance to lepidoptera insects than a first generation single-gene Bt plant through enhanced coordinate expression, and has a herbicide resistance character, thereby facilitating farmland weeding.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, more specifically, to a polynucleotide of a herbicide-resistant insect-resistant plant, an expression cassette comprising the polynucleotide and applications thereof. Background technique [0002] Cotton is an important source of raw material in the textile industry and accounts for 1.4% of GDP and 6.7% of Pakistan's agricultural output. During the period from July 2013 to March 2014, the textile industry received US$10.385 billion in foreign exchange earnings. During 2013-2014, 2,806,000 hectares were planted to the cotton crop, 2.5 percent less than the 2,879,000 hectares in 2012-2013. During 2013-2014, cotton production stood at 12.8 million bales against the target of 14.1 million bales, showing a 9.2% decline from the target and a 2.0 % decline (Pakistan Economic Survey, 2014). [0003] Insect pests are a significant problem for cotton crops in cotton growing regions of the wo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/54C12N15/32C12N15/82C12N9/10C07K14/325
CPCC07K14/325C12N9/1092C12N15/8274C12N15/8286C12Y205/01019C07K2319/55C12N15/1003C12N15/8275C12Q1/6806C12Q1/6895C12N15/10G01N33/6854Y02A40/146
Inventor 达尼亚·贾德·库雷西·索·贾德·萨利姆·库雷西哈姆萨·纳迪姆·库雷西·索·穆罕默德·纳迪姆·库雷西
Owner 达尼亚·贾德·库雷西·索·贾德·萨利姆·库雷西
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