Creation of winter rapeseed Brassica napus L. commercial line of biotechnological plants, resistant to the glyphosate action

Q4 Biochemistry, Genetics and Molecular Biology Biopolymers and Cell Pub Date : 2020-12-31 DOI:10.7124/BC.000A40

I. Hnatiuk, O. Varchenko, M. Kuchuk, M. Parii, Y. Symonenko

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Abstract

Aim. To create biotechnological plants of a commercial line winter rapeseed, which carry the glyphosate and phosphinostricin resistance genes. Methods. In vitro tissue culture method, Agrobacterium -mediated genetic transformation, polymerase chain reaction and χ 2 methods were used. Results. The PCR analysis results are presented for the biotechnological winter rapeseed plants obtained after genetic transformation to confirm the presence of CP4 epsps gene insert in 11 of 12 plants, as well as the presence of bar gene integration in 6 of 7 plants. All obtained regenerant plants were adapted to in vivo conditions, treated with glyphosate and vernalized. The obtained seeds were sterile germinated and the segregation was determined based on the resistance to the selective agent. Conclusion. 11 biotechnological plants with the glyphosate ( epsps ) and kanamycin ( nptII ) resistance genes were obtained, as well as 6 biotechnological plants, which carry phosphinothricin ( bar ) resistance genes in addition to the epsps gene. The presence of the CP4 EPSPS protein in transgenic rapeseed plants has been proven by the molecular analysis of protein expression using an immunoassay system. The expression of selective agents resistance genes was confirmed in T 1 generation.

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创建了甘蓝型冬季油菜商业系的生物技术植物，抗草甘膦作用

的目标。培育具有抗草甘膦和抗磷抑菌素基因的商品品系冬油菜籽生物技术植株。方法。采用离体组织培养法、农杆菌介导的遗传转化法、聚合酶链反应法和χ 2法。结果。对转基因后获得的生物技术冬油菜植株进行PCR分析，证实12株中有11株存在CP4 epsps基因插入，7株中有6株存在bar基因整合。所有获得的再生植株都适应了体内条件，用草甘膦处理并春化。获得的种子无菌发芽，并根据对选择剂的抗性来确定分离。结论:共获得了11株具有草甘膦(epsps)和卡那霉素(nptII)抗性基因的生物技术植物，以及6株除epsps基因外还具有磷丙氨酸(bar)抗性基因的生物技术植物。利用免疫分析系统对CP4 EPSPS蛋白的表达进行了分子分析，证实了转基因油菜中存在CP4 EPSPS蛋白。在t1代中证实了选择性耐药基因的表达。

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来源期刊

Biopolymers and Cell Biochemistry, Genetics and Molecular Biology-Biochemistry, Genetics and Molecular Biology (all)

CiteScore

1.10

自引率

0.00%

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期刊介绍： “Biopolymer and cell” is published since 1985 at the Institute of Molecular Biology and Genetics NAS of Ukraine under the supervision of the National Academy of Sciences of Ukraine. Our journal covers a wide scope of problems related to molecular biology and genetics including structural and functional genomics, transcriptomics, proteomics, bioinformatics, biomedicine, molecular enzymology, molecular virology and immunology, theoretical bases of biotechnology, physics and physical chemistry of proteins and nucleic acids and bioorganic chemistry.