A degradable glyphosate-resistant gene, a plant expression vector, and a cultivation method and application of degradable 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, and achieving reduction of glyphosate residues, reduction of identification sites, and accelerated cultivation. Effect

Active Publication Date: 2022-08-09
HUAZHONG AGRI UNIV
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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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  • A degradable glyphosate-resistant gene, a plant expression vector, and a cultivation method and application of degradable glyphosate-resistant transgenic rice
  • A degradable glyphosate-resistant gene, a plant expression vector, and a cultivation method and application of degradable glyphosate-resistant transgenic rice
  • A degradable glyphosate-resistant gene, a plant expression vector, and a cultivation method and application of degradable glyphosate-resistant transgenic rice

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Experimental program
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Effect test

Embodiment 1

[0051] Codon optimization of the BceGO-B3S1 sequence

[0052] The original sequence of the target gene glyphosate-resistant gene (hereinafter referred to as WBceGO-B3S1) used in the present invention was cloned from the microorganism, Bacillus cereus mutant B3S1 by the research group of Professor Liu Ziduo of Huazhong Agricultural University (the gene function verification was also verified in the microorganism). 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 It is named BceGO-B3S1, its nucleotide sequence is shown in SEQ ID No.6, and its amino acid sequence is shown in SEQ ID No.7.

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

Embodiment 2

[0061] Construction of plant expression vector

[0062] The original transformation vector used in the present invention is the PYNU vector. Degradable glyphosate-resistant gene and UTR-CTP * Insert into PYNU vector to obtain 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] Agrobacterium-mediated genetic transformation of rice ZH11

[0065]Genetic transformation mainly refers to the operation guidelines 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 selective culture, differentiation, rooting and transplanting.

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Abstract

The invention provides a degradable glyphosate-resistant gene, a plant expression vector, and a cultivation method and application of degradable glyphosate-resistant transgenic rice, belonging to the technical field of plant genetic engineering. The nucleotide sequence is shown in SEQ ID No.1. In the degraded glyphosate-resistant rice obtained by cultivation, the flanking sequence of the foreign gene insertion site is shown in SEQ ID No. 2. The degradable glyphosate-resistant gene provided by the invention has the effect of degrading glyphosate. The degradable glyphosate-resistant gene provided by the present invention is identical to the amino acid sequence encoded by the original gene BceGO-B3S1 (SEQ ID No. 6); the optimal codon usage frequency of amino acids such as leucine and glutamic acid is increased.

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 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, and occupies a very important position in agricultural production. According to the United Nations Food and Agriculture Organization, 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 yield, resistance and quality, but Due to the reduction of arable land, limited rice germplasm resources and limitations of conventional breeding methods, traditional agricultural t...

Claims

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Application Information

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Patent Type & Authority Patents(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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