Transgenic induction in Sesamum indicum with recombinant pBI121 expression construct containing CYP81Q1 and aroA genes using Agrobacterium tomfacensis

Document Type : Research Paper

Authors

1 Assistant Professor, Department of Agricultural Engineering, Payame Noor University, Tehran, Iran.

2 MSc Student, Department of Cell and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.

Abstract

Objective
Oilseeds are the second-largest source of calories for human societies after cereals. Sesamum indicum has been considered recently due to its oil quality and content of 43-46% unsaturated fatty acids. To investigate the possibility of transferring the recombinant aroA-CYP81Q1 gene through Agrobacterium tumefaciens to the Sesamum indicum plant (reported for the first time in the country), after optimizing tissue culture and selecting the best hormonal combination, a factorial experiment was performed. 
Materials and methods
In this experiment, the recombinant aroA-CYP81Q1 gene was synthesized and transformed into an Agrobacterium strain (LB4404). Gene cloning was confirmed by PCR and then confirmed by sequencing. The efficacy and frequency of transgenic Sesamum indicum were evaluated in the selected medium containing 50 mg/l kanamycin. Finally, to confirm the transgenicity of the regenerated plants, PCR analyzes were performed with specific primers on selected plants. Also, the expression of the sesamin-producing gene (CYP81Q1) in control groups and transgenic groups was measured at a significant level of P <0.01.   
Results
Statistical analysis showed that the percentage of regeneration of transgenic seedlings using Agrobacterium (LB4404) in the selected medium containing kanamycin was 33%. Also, PCR analysis of transgenic plants showed a prevalence of transgenics of 33%. In addition, amplification of the bp fragment indicated the accuracy of cloning in transgenic plants. The results indicate the successful transfer of this gene to the sesame plant to increase its industrial properties. The results of the analysis of variance showed that there was a significant difference in the expression of sesamin-producing genes between transgenic cultivar and control groups (P <0.01). 
Conclusions
Since pBI121-based expression vectors are more efficient than other expression vectors and are widely used in the transfer of recombinant genes to plants, the recombinant vector constructed in this study indicates successful gene transfer. Sesame to sesame plant to increase its industrial properties.

Keywords

References

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Agricultural Biotechnology Journal
Volume 14, Issue 3
September 2022
Pages 223-242

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