Cloning and Agrobacterium-mediated transfer of SoBADH gene to Brassica napus L. to confer osmotic stress tolerance

Document Type : Original Article

Authors

1 Department of Plant Sciences, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran

2 National Institute of Genetic Engineering and Biotechnology

3 Kharazmi University

4 Central Branch of Islamic Azad University

10.22034/nbr.2026.132149.0
Abstract
Objective: Canola (Brassica napus L.) is a major source of edible oil, but its production is severely affected by osmotic stresses such as drought and salinity. Glycine betaine (GB) plays a key role in stress tolerance, yet many crops including canola cannot accumulate it. This study aimed to transfer the SoBADH gene from spinach into canola to enhance GB biosynthesis and improve osmotic stress tolerance.

Methods: The synthesized SoBADH gene was cloned into pUC57, digested with SalI and NdeI, and ligated into pRI201-AN using T4 ligase. The recombinant vector was transferred into Agrobacterium tumefaciens via freeze-thaw method. Canola cotyledonary explants were inoculated with Agrobacterium, and transgenic plants were regenerated on kanamycin-containing medium.

Results: The presence of the SoBADH gene was confirmed by PCR analysis, and its expression was verified by RT-PCR. Osmotic stress tolerance was evaluated using 20% PEG6000 ( 0.7 MPa). The results showed that transgenic plants exhibited a 60% increase in glycine betaine content compared to control plants and displayed greater leaf development and greener color under osmotic stress.

Conclusion: Transfer of the SoBADH gene into canola effectively enhances GB accumulation and osmotic stress tolerance, offering a practical approach for developing drought- and salt-tolerant cultivars for sustainable agriculture.

Keywords

Subjects


Articles in Press, Accepted Manuscript
Available Online from 07 September 2026

  • Receive Date 16 May 2026
  • Revise Date 02 September 2026
  • Accept Date 07 September 2026
  • Publish Date 07 September 2026