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Transgenic glyphosate-resistant soybeans as well as preparation method and application thereof

A technology for transgenic soybeans and soybeans, applied in the field of plant genetic engineering technology or breeding, can solve the problems of polluting the environment, small scope of use, and low efficacy.

Active Publication Date: 2015-03-25
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 herbicides has greatly improved the control efficiency of weeds in farmland. Traditional herbicides such as chlorimuron-methyl and metsulfuron-methyl have serious residues in the soil, pollute the environment, and have a small range of use.

Method used

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  • Transgenic glyphosate-resistant soybeans as well as preparation method and application thereof
  • Transgenic glyphosate-resistant soybeans as well as preparation method and application thereof
  • Transgenic glyphosate-resistant soybeans as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1, genetic transformation of transgenic soybean

[0064] 1. Obtaining the recombinant vector

[0065] (1) According to the optimized synthetic sequence of A.thaliana-rbcS plus G2-EPSPS gene, redesign the PCR primers, with the restriction site XbaI in the upstream and SacI in the downstream, and PCR amplify rbcS-G2- The EPSPS fragment (nucleotides 8204-9784 from the 5' end of Sequence 1) was connected to the T-A vector to obtain the plasmid prbcS-G2-EPSPS, which was digested and sequenced for verification.

[0066] (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...

Embodiment 2

[0085] Embodiment 2, glyphosate resistance analysis of transgenic soybean

[0086] The glyphosate resistance analysis of successive generations of the T0 transgenic soybean ZH10-6 obtained in Example 1 found that it has significantly more resistance to glyphosate than the non-transgenic control Zhonghuang No. 10, as follows:

[0087] Collect T0 generation transgenic soybean ZH10-6 seeds, sow, and obtain T1 generation transgenic soybean ZH10-6, which is smeared with 1 μl glyphosate isopropylamine salt (Roundup) per plant at the seedling stage (cotyledons unearthed and true leaves are not fully expanded), 2 Investigation of drug damage after a week.

[0088] 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 .

[0089] The result is as image 3 As shown, 1: non-resist...

Embodiment 3

[0097] Example 3. Analysis of exogenous DNA copy number and molecular integration stability in transgenic soybean

[0098] 1. Copy number of integrated exogenous T-DNA

[0099] The copy numbers of exogenous DNA fragments integrated in the genome of T1 transgenic soybean ZH10-6 were determined by Southern blotting analysis of restriction fragments extending beyond the left and right border sequences of the transformation vectors used.

[0100] The probe for Southern blotting analysis is to select the carrier DNA sequence design probe (sequence 6) of 338bp from the carrier T-DNA region, and refer to the instruction manual of the PCR method DIG labeling kit produced by Beijing Meilaibo Medical Technology Co., Ltd., the place of preparation Goxigenin-labeled probes.

[0101] 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 gen...

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Abstract

The invention discloses transgenic glyphosate-resistant soybeans as well as a preparation method and application thereof. The invention provides a method of cultivating transgenic soybeans. The method comprises the following step: inserting a foreign DNA fragment in a range from a 7980527th position to a 7982541st position of chromosome 17 of a target soybean genome to obtain the transgenic soybeans. The transgenic soybeans have tolerance to high-dose glyphosate and can be further improved by being hybridized with an excellent soybean strain so as to optimize other agronomic characters including quality and yield. The detection method is capable of identifying a combined region of an inserted T-DNA and a plant genome DNA and further identifying cells, tissues, organs, products and the like of the transgenic soybeans and progenies of the transgenic soybeans.

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 and their preparation methods and applications. Background technique [0002] Soybean (Glycine max (L.).Merr) originated in China, is an important oil crop and economic crop, and is also the main source of edible vegetable oil and vegetable protein. Weeds are an important part of the agricultural ecosystem. Weeds compete with crops for resources such as light, water, and nutrients, 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-intensive, with low eff...

Claims

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Application Information

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IPC IPC(8): C12N15/82C12Q1/68A01H5/00
CPCC12N15/82C12Q1/68
Inventor 邱丽娟郭兵福郭勇张丽娟金龙国洪慧龙化宿南
Owner INST OF CROP SCI CHINESE ACAD OF AGRI SCI
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