Degradation type glyphosate-resistant gene, plant expression vector, and cultivation method and application of degradation type glyphosate-resistant transgenic rice

A plant expression vector and a technology for transgenic rice, applied in the field of plant genetic engineering, can solve the problems of restricting the application of transgenic technology, consuming a lot of manpower, material resources and time, etc., reducing glyphosate residues, reducing identification sites, and avoiding abnormality expressive effect

Active Publication Date: 2021-10-01
HUAZHONG AGRI UNIV
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  • Abstract
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  • Claims
  • Application Information

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Problems solved by technology

[0006] Some rice varieties can obtain a large number of transgenic rice plants through the method of genetic transformation, but there will be factors such as somatic cell variation and foreign gene insertion site effects in the tissue culture process, and it will take a lot of manpower, material resources and time to develop ideal transgenic rice plants. Screening of strains limits the application of transgenic technology in agricultural production

Method used

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  • Degradation type glyphosate-resistant gene, plant expression vector, and cultivation method and application of degradation type glyphosate-resistant transgenic rice
  • Degradation type glyphosate-resistant gene, plant expression vector, and cultivation method and application of degradation type glyphosate-resistant transgenic rice
  • Degradation type glyphosate-resistant gene, plant expression vector, and cultivation method and application of degradation type glyphosate-resistant transgenic rice

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Embodiment 1

[0051] Codon optimization of BceGO-B3S1 sequence

[0052] The original sequence of the target glyphosate-resistant gene (hereinafter referred to as WBceGO-B3S1) used in the present invention was obtained by the research group of Professor Liu Ziduo of Huazhong Agricultural University by cloning it from a microorganism, namely the mutant B3S1 of the bacterium Bacillus cereus (gene function verification is also verified in microorganisms It has not been verified in plants, especially in rice transformation, see literature: Zhan, T, et al. (2013). Improving Glyphosate Oxidation Activity of GlycineOxidase from Bacillus cereus by Directed Evolution. PLOS ONE.), the original gene Named BceGO-B3S1, its nucleotide sequence is shown in SEQ ID No.6, and its encoded amino acid sequence is shown in SEQ ID No.7.

[0053] The analysis of the original BceGO-B3S1 gene and its application to the breeding of glyphosate-resistant rice may have the following problems. (1) The content of base A i...

Embodiment 2

[0061] Construction of plant expression vectors

[0062] The original transformation vector used in the present invention is PYNU vector. Degradable glyphosate-resistant gene and UTR-CTP * Inserted into the PYNU vector to obtain a plant expression vector. PYNU-WBceGO-B3S1 was transformed into Agrobacterium EHA105, and the transformed Agrobacterium strain was stored at -80°C until use.

Embodiment 3

[0064] Genetic transformation of rice ZH11 mediated by Agrobacterium

[0065]The genetic transformation mainly refers to the operating instructions of the State Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University (see 3.1-3.8 for the operation steps). It mainly includes callus induction, callus subculture, Agrobacterium culture, Agrobacterium infection, callus washing and selection culture, differentiation, rooting and transplanting.

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Abstract

The invention provides a degradation type glyphosate-resistant gene, a plant expression vector, and a cultivation method and the application of degradation type glyphosate-resistant transgenic rice, and belongs to the technical field of plant genetic engineering. The nucleotide sequence of the degradation type glyphosate-resistant gene is as shown in SEQ ID No.1. The flanking sequence of the insertion site of the exogenous gene of the degraded glyphosate-resistant rice obtained by cultivation is shown as SEQ ID No.2. The degradation type glyphosate-resistant gene provided by the invention has the effect of degrading glyphosate. The degradation type glyphosate-resistant gene provided by the invention has an amino acid sequence which is completely the same as that coded by an original gene BceGO-B3S1 (SEQ ID No.6); the optimal codon use frequency of amino acids such as leucine and glutamic acid is improved.

Description

technical field [0001] The invention belongs to the technical field of plant genetic engineering, and in particular relates to a degradable glyphosate-resistant gene, a plant expression vector, and a cultivation method and application of a degradable glyphosate-resistant transgenic rice. Background technique [0002] Rice is one of the most important food crops in my country and even in the world. It is the basic food that more than half of the world's population depends on for survival, and plays a very important role in agricultural production. According to statistics from the Food and Agriculture Organization of the United Nations, the global population will reach 9 billion by 2050, and food production needs to increase by 60%. Although traditional genetic breeding methods have made important contributions to improving rice production, resistance and quality, but Due to the reduction of arable land, the limited germplasm resources of Oryza genus and the limitations of con...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12N15/82A01H5/00A01H5/10A01H6/46
CPCC07K14/32C12N15/8275
Inventor 林拥军温莉娴钟珏段镇淳李昌焱
Owner HUAZHONG AGRI UNIV
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