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Glyphosate-resistant transgenic soybean as well as preparation method and application thereof

A technology of genetically modified soybeans and soybeans, which is applied in the field of plant genetic engineering technology or breeding, and can solve the problems of not being able to prevent and control large areas in time, time-consuming and labor-intensive, and limiting the use of herbicides, etc.

Active Publication Date: 2016-04-20
INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional manual weeding includes manual weeding and weeding with simple farm tools, etc., which is time-consuming and labor-intensive, with low efficacy and cannot prevent and control large areas in time
The use of chemical herbicides has greatly improved the control efficiency of weeds in farmland, but traditional herbicides such as chlorimuron-methyl and metsulfuron-methyl have serious residues in the soil and pollute the environment, which limits the scope of use of these herbicides

Method used

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  • Glyphosate-resistant transgenic soybean as well as preparation method and application thereof
  • Glyphosate-resistant transgenic soybean as well as preparation method and application thereof
  • Glyphosate-resistant transgenic soybean as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0071] Embodiment 1, genetic transformation of transgenic soybean

[0072] 1. Obtaining the recombinant vector

[0073] (1) According to the optimized synthetic sequence of A.thaliana-rbcS plus G2-EPSPS (G2-aroA) gene, the PCR primers were redesigned. The rbcS-G2-EPSPS(aroA) fragment (nucleotides 8204-9784 from the 5' end of Sequence 1) was added and connected to the T-A vector to obtain the plasmid prbcS-G2-EPSPS(aroA), which was digested and sequenced for verification.

[0074] (2) pBI121 (Wang Huaxin, Cao Jiashu, Xiang Xun et al. Construction of pBI121 expression vector and rapid identification of transformed plants. Journal of Zhejiang University (Agriculture and Life Sciences), 2008, 34(2): 137-142; public available Obtained from the Institute of Crop Science, Chinese Academy of Agricultural Sciences) and pCAMBIA2300 (Gong Yuanyong, Feng Yongkun, Ni Wanchao, etc. Construction and verification of plant expression vector pCAMBIA2300-35S-GUS-CaMVterm. China Biotechnology Jo...

Embodiment 2

[0094] Embodiment 2, glyphosate resistance analysis of transgenic soybean

[0095] To the T obtained in Example 1 0 Glyphosate resistance analysis of successive generations of transgenic soybean GE-J16 found that it was more resistant to glyphosate than the non-transgenic wild-type control Jack, as follows:

[0096] collect T 0 Genetically modified soybean GE-J16 seeds were sown to obtain T 1 The transgenic soybean GE-J16 was sprayed with 3L / ha (200ml / mu) glyphosate isopropylamine salt (Roundup) solution during the first to third compound leaf stages, and the phytotoxicity was investigated after 2 weeks.

[0097] The phenotype of non-resistant plants is dehydration and chlorosis of leaves, curling and shriveling of leaves, and necrosis of top meristem until the whole plant dies; the phenotype of resistant plants is vigorous growth, leaves do not chlorosis, curl or shrink .

[0098] T 1 The results of transgenic soybean GE-J16 are as follows: image 3 Shown, A: negative c...

Embodiment 3

[0105] Example 3. Analysis of exogenous DNA copy number and molecular integration stability in transgenic soybean

[0106] 1. Copy number of integrated exogenous T-DNA

[0107] Determination of T by Southern blotting analysis of restriction fragments extending beyond the left and right border sequences of the transformation vector used 0 Copy numbers of exogenous DNA fragments integrated in the genome of transgenic soybean GE-J16.

[0108] The probe for Southernblotting analysis is to select a 338bp carrier DNA sequence design probe (sequence 6) from the carrier T-DNA region, and prepare a high Sim-labeled probes.

[0109] Referring to the instruction manual of the digoxin hybridization kit produced by Beijing Meilaibo Medical Technology Co., Ltd., the recombinant plasmid pKT-rGE used was used as a positive control, and the genomic DNA of the non-transgenic wild-type genetically transformed recipient Jack was used as a negative control.

[0110] Extract the T obtained in th...

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Abstract

The invention discloses glyphosate-resistant transgenic soybean as well as a preparation method and application thereof. The invention provides a method for cultivating transgenic soybean. In the method, an exogenous DNA fragment is inserted between the 50543767th site-50543792nd site of the chromosome No.19 of the target soybean genome to obtain transgenic soybean. The transgenic soybean disclosed by the invention is tolerant to high-dose glyphosate, and the transgenic soybean can be further hybridized with a good soybean strain so as to improve and optimize other agronomic characters such as yield and quality. A detection method provided by the invention can be used for identifying the binding domain between the inserted T-DNA and plant genome DNA and further identifying the cells, tissues, organs, products and the like of the transgenic soybean and progenies thereof.

Description

technical field [0001] The present invention relates to the field of plant genetic engineering technology or breeding. Specifically, the present invention relates to transgenic soybeans and applications thereof. More specifically, the present invention relates to glyphosate-resistant transgenic soybeans, a preparation method and application thereof. Background technique [0002] Soybean (Glycinemax (L.).Merr) originated in China, is an important oil crop and economic crop in the world, and is also the main source of edible vegetable oil and vegetable protein. As an important part of the agricultural ecosystem, weeds can compete with crops for resources such as light, water, and nutrients, thereby reducing crop yields. Therefore, the effective control of weeds in soybean field is one of the key factors for stable and high yield of soybean. Traditional manual weeding includes weeding by hand and weeding with simple agricultural tools, which is time-consuming and labor-intensi...

Claims

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Application Information

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IPC IPC(8): C12N15/82A01H5/00C12Q1/68
CPCC12N9/1029C12N9/1092C12N15/8271C12N15/8275C12Q1/6895C12Y205/01019C12N15/82C12Q1/68
Inventor 邱丽娟郭兵福郭勇张丽娟洪慧龙金龙国王爽杨慧
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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