SUBSTITUTION OF AMINO ACID ALA-183 TO THR IN 5-ENOLPYRUVYLSHIKIMATE 3-PHOSHPATE SYNTHASE IN ORDER TO MAKE TOLERANCE TO GLYPHOSATE HERBICIDE IN TRANSGENIC RAPESEED PLANT (BRASSICA NAPUS L.).

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RAZI KAHRIZI D.; SALMANIAN A.H.; MOUSAVI AMIR; MOEINI AHMAD; KARIMZADEH GHASEM

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Journal: Year:2008 | Volume:21 | Issue:2 (79 IN AGRONOMY AND HORTICULTURE) Page(s): 151-159

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Title

SUBSTITUTION OF AMINO ACID ALA-183 TO THR IN 5-ENOLPYRUVYLSHIKIMATE 3-PHOSHPATE SYNTHASE IN ORDER TO MAKE TOLERANCE TO GLYPHOSATE HERBICIDE IN TRANSGENIC RAPESEED PLANT (BRASSICA NAPUS L.).

Author(s)

Keywords

RAPESEED (BRASSICA NAPUS L.)Q3

HERBICIDE RESISTANCEQ3

GLYPHOSATEQ2

MANIPULATED EPSPS AND GENE TRANSFORMATIONQ3

Abstract

Rapeseed (Brassica napus L.) is an important oil seed crop in the world. The application of genetic engineering to B. napus will lead to the generation of plant varieties possessing more agriculturally and economically viable genetic traits. The manipulation of bacterial EPSPS gene in order to reduce its affinity to GLYPHOSATE is one of the most effective methods for production of GLYPHOSATE tolerant plants. GLYPHOSATE is a non-selective and broad-spectrum herbicide that inhibits 5-enolpyruvylshikimate 3-phoshpate synthase (EPSPS), a key enzyme in the aromatic amino acid biosynthesis in microorganisms and plants. In this research, we study on alanine183 of E. coli (k12) EPSPS enzyme. This amino acid is an important residue for attachment of GLYPHOSATE herbicide to EPSPS enzyme. We used site directed mutagenesis method to inducing a point mutation in E. coli EPSPS gene to convert alanine 183 to threonine. The manipulated EPSPS gene was cloned in pUC18 as a universal cloning and pBI121 as a plant expression vectors. The results of molecular analyses and sequencing showed that the genes has been changed and cloned in correct orientation in both plasmids. Recombinant pBI121 containing altered EPSPS gene were transferred to rapeseed (B. napus L.), PF-7045-91 cv, via Agrobacterium tumefaciens mediated transformation method. The target tissue for transformation was the cut end of cotyledonery petioles. At the first step the transformed explants were screened in kanamycin containing media. Results showed that plant regeneration and transformation frequency was 74% and 28٪ respectively. Molecular analyses and GLYPHOSATE tolerance test, confirm the gene presence and it.s effective function in transgenic plant.

Cites

INVESTIGATION ON INHERITANCE OF GLYPHOSATE TOLERANCE IN T1 GENERATION OF TRANSGENIC RAPESEED (BRASSICA NAPUS L.) WITH THE MUTATED BACTERIAL EPSPS GENE

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APA: Copy

RAZI KAHRIZI, D., SALMANIAN, A.H., MOUSAVI, AMIR, MOEINI, AHMAD, & KARIMZADEH, GHASEM. (2008). SUBSTITUTION OF AMINO ACID ALA-183 TO THR IN 5-ENOLPYRUVYLSHIKIMATE 3-PHOSHPATE SYNTHASE IN ORDER TO MAKE TOLERANCE TO GLYPHOSATE HERBICIDE IN TRANSGENIC RAPESEED PLANT (BRASSICA NAPUS L.).. PAJOUHESH-VA-SAZANDEGI, 21(2 (79 IN AGRONOMY AND HORTICULTURE)), 151-159. SID. https://sid.ir/paper/18720/en

Vancouver: Copy

RAZI KAHRIZI D., SALMANIAN A.H., MOUSAVI AMIR, MOEINI AHMAD, KARIMZADEH GHASEM. SUBSTITUTION OF AMINO ACID ALA-183 TO THR IN 5-ENOLPYRUVYLSHIKIMATE 3-PHOSHPATE SYNTHASE IN ORDER TO MAKE TOLERANCE TO GLYPHOSATE HERBICIDE IN TRANSGENIC RAPESEED PLANT (BRASSICA NAPUS L.).. PAJOUHESH-VA-SAZANDEGI[Internet]. 2008;21(2 (79 IN AGRONOMY AND HORTICULTURE)):151-159. Available from: https://sid.ir/paper/18720/en

IEEE: Copy

D. RAZI KAHRIZI, A.H. SALMANIAN, AMIR MOUSAVI, AHMAD MOEINI, and GHASEM KARIMZADEH, “SUBSTITUTION OF AMINO ACID ALA-183 TO THR IN 5-ENOLPYRUVYLSHIKIMATE 3-PHOSHPATE SYNTHASE IN ORDER TO MAKE TOLERANCE TO GLYPHOSATE HERBICIDE IN TRANSGENIC RAPESEED PLANT (BRASSICA NAPUS L.).,” PAJOUHESH-VA-SAZANDEGI, vol. 21, no. 2 (79 IN AGRONOMY AND HORTICULTURE), pp. 151–159, 2008, [Online]. Available: https://sid.ir/paper/18720/en

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