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Screening method for eliminating transgenic maize gene instable plants

A technology for transgenic corn and plants, applied in botany equipment and methods, fertilization methods, plant gene improvement, etc., can solve problems such as inability to meet the needs of breeding work, achieve stable and reliable screening results, improve genetic stability, and improve operating procedures clear and simple effect

Inactive Publication Date: 2017-07-07
TIANJIN UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the methods used to screen the resistance of transgenic maize mainly include leaf spraying, smearing, seed germination, and root absorption. Experiments have proved that leaf spraying method has the best screening effect. Lost, the effect of using the same spraying intensity can no longer meet the needs of breeding work

Method used

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  • Screening method for eliminating transgenic maize gene instable plants
  • Screening method for eliminating transgenic maize gene instable plants
  • Screening method for eliminating transgenic maize gene instable plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] A screening method for eliminating transgenic corn genetically unstable plants, comprising the steps of:

[0030] (1) Acquisition of transgenic corn plants:

[0031] The in situ transformation method of mature embryos was used to transform the vector containing the BAR gene ( figure 1 The vector pCambia2300-BAR) was introduced into the non-transgenic corn material Tianta No. 5 to obtain the transformed plant seedlings of the T0 generation. After they grew robustly, they were transplanted into the isolated experimental field of the Institute of Genetic Engineering, Tianjin University, and the transformed plants were subjected to conventional agricultural management and self-pollination. , the T1 generation seeds were harvested at the maturity stage, dried and threshed, and stored at -20°C;

[0032] (2) Spray screening of the herbicide glufosinate-ammonium aqueous solution:

[0033] Select the plump and disease-free T1 generation seeds, plant them in the field and spray...

Embodiment 2

[0056] A screening method for eliminating transgenic corn genetically unstable plants, comprising the steps of:

[0057] (1) Acquisition of transgenic corn plants:

[0058] The vector pCAMBIA2300-BAR containing the BAR gene was introduced into the non-transgenic corn material Fe 7922 by the pollen tube passage method, and the transformed plant seedlings of the T0 generation were obtained. After growing vigorously, they were transplanted into the field for conventional agricultural management and self-pollination. Harvest T1 generation seeds;

[0059] (2) Spray screening of herbicide glufosinate-ammonium aqueous solution

[0060] Select plump and disease-free T1 generation seeds, plant them in the field and spray and water them; when the T1 generation plants grow to the three-leaf stage, spray the first glufosinate-ammonium aqueous solution: the spraying rate is 0.25L / m 2 ; The initial spraying treatment was 9 days, and the second glufosinate-ammonium aqueous solution sprayin...

Embodiment 3

[0066] A screening method for eliminating transgenic corn genetically unstable plants, comprising the steps of:

[0067] (1) Acquisition of transgenic corn plants:

[0068] The vector pCAMBIA2300-BAR containing the Bar gene was introduced into the non-transgenic corn material Zong 31 by the Agrobacterium-mediated method, and the transformed plant seedlings of the T0 generation were obtained. After growing vigorously, they were transplanted into the field for conventional agricultural management and self-pollination. Harvest the T1 generation seeds;

[0069] (2) Spray screening of the herbicide glufosinate-ammonium aqueous solution:

[0070] Select plump and disease-free T1 generation seeds, plant them in the field and spray and water them; when the T1 generation plants grow to the three-leaf stage, spray the first glufosinate-ammonium aqueous solution: the spraying rate is 0.3L / m 2 ; The initial spraying treatment was 7 days, and the second glufosinate-ammonium aqueous solut...

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Abstract

The invention discloses a screening method for eliminating transgenic maize gene instable plants. The screening method comprises the steps of guiding a carrier pCAMBIA2300-BAR containing a BAR gene into non-GMO (non-Genetically Modified Organism) maize materials, obtaining T0 generation of transformation plant seedlings, transplanting into a land for growing field crops, carrying out conventional farming management and self-pollination, and harvesting T1 generation of seeds in a mature period; spraying herbicide glufosinate-ammonium water solutions with different concentrations on the T1 generation of plants for three times, and screening to eliminating dead plants; carrying out self-pollination on survived plants and harvesting seeds. The method provided by the invention is clear and simple in operation flow, fast and efficient in screening process, stable and reliable in screening result, capable of effectively avoiding the phenomena of gene transposition, silence or loss and other gene instable phenomena existed widely in maize transgenosis events and improving gene stability of transgenic maize progeny, provides technical assistance and support for culturing high-quality maize new species by utilizing a gene engineering method, and has higher field application, production and popularization values.

Description

technical field [0001] The invention belongs to the field of corn transgenics, and relates to a screening method for eliminating genetically unstable transgenic corn plants. Background technique [0002] In recent years, problems such as increasing world population, decreasing arable land, environmental pollution, global warming, and increasing area of ​​salinized land have seriously affected the sustainable development of agriculture, resulting in land crisis, food crisis and energy crisis. It has become a global problem that needs to be solved urgently. Traditional breeding methods can no longer meet the needs of human social development. As an emerging biotechnology, transgenic technology has shown great potential and application prospects. As the main food crop, corn's quality and yield are of great significance to the sustainable development of all human beings. Therefore, it has become a huge opportunity and challenge to cultivate new plant varieties to adapt to cli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/02A01H1/04A01C21/00
CPCA01C21/005A01H1/02A01H1/04
Inventor 季静尹日晖王罡金超张松皓安婷
Owner TIANJIN UNIV
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