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Rapeseed Transformation with aroA Bacterial Gene Containing P101S Mutation Confers Glyphosate Resistance

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Abstract

Field weed infestations can cause serious problems and require regular and planned programs to control them. Glyphosate is a broad-spectrum herbicide that inactivates the 5-enolpyruvylshikimate 3-phosphate synthase (EPSPS) enzyme and causes plant death. It has been reported that the mutation of proline 101 to serine in EPSPS is one of the effective mutations to reduce the affinity of glyphosate to EPSPS enzyme. In this study, we investigated the effect of the bacterial P101S mutant aromatic acid (aroA) gene on glyphosate resistance in transgenic rapeseeds. For this purpose, the mutant gene was synthesized and cloned into the pUC18 and pBI121 vectors. The gene was transferred to rapeseed by the Agrobacterium-mediated method. In this experiment, three generations of transgenic plants (T0, T1, and T2) were studied under in vitro and in vivo conditions. After the treatment of 75 putative transgenic plants with 10 mM glyphosate in T0 generation, resistant plants were identified and their seeds were harvested. In the T1 generation, out of 200 cultivated plants, 141 showed resistance. After the plants were treated with herbicides and resistance was determined, the seeds were harvested when they mature. In the T2 generation, most plants (162 plants of 200) were resistant to glyphosate. Therefore, the inheritance of resistance followed Mendel's first law, which is a sign of the monogenic character of resistance. Purification and increasing the percentage of resistant plants will be carried out in the next generations. It is concluded that P101S mutation guarantees glyphosate resistance of rapeseed and is recommended to study it in other plants.

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Abbreviations

P101S:

Proline 101 to serine

EPSPS:

5-Enolpyruvylshikimate 3-phosphate synthase

E. coli :

Escherichia coli

aroA:

Aromatic acid

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Authors and Affiliations

  1. Department of Agronomy and Plant Breeding, Kermanshah Branch, Islamic Azad University, Kermanshah, Iran

    Amir Roeintan & Seyed Mehdi Safavi

  2. Department of Agronomy and Plant Breeding, Razi University, Kermanshah, Iran

    Danial Kahrizi

Authors
  1. Amir Roeintan
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  2. Seyed Mehdi Safavi
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  3. Danial Kahrizi
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Contributions

AR: Execution research project, data analysis, manuscript preparation. SMS: Data analysis, proofreading of the article, manuscript preparation. DK: Experimental design, data analysis, gene expression analysis.

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Correspondence to Danial Kahrizi.

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This study was approved by the Ethics Committee of Razi University, Kermanshah, Iran.

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Roeintan, A., Safavi, S.M. & Kahrizi, D. Rapeseed Transformation with aroA Bacterial Gene Containing P101S Mutation Confers Glyphosate Resistance. Biochem Genet 60, 953–968 (2022). https://doi.org/10.1007/s10528-021-10136-w

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