Efficient Agrobacterium-mediated transformation and genome editing of Fagopyrum tataricum

Artur Pinski; Alexander Betekhtin

doi:10.3389/fpls.2023.1270150

Efficient Agrobacterium-mediated transformation and genome editing of Fagopyrum tataricum

Front Plant Sci. 2023 Sep 8:14:1270150. doi: 10.3389/fpls.2023.1270150. eCollection 2023.

Authors

Artur Pinski¹, Alexander Betekhtin¹

Affiliation

¹ Institute of Biology, Biotechnology and Environmental Protection, Faculty of Natural Sciences, University of Silesia in Katowice, Katowice, Poland.

Abstract

Fagopyrum tataricum (L.) Gaertn. is an exceptional crop known for its remarkable health benefits, high levels of beneficial polyphenols and gluten-free properties, making it highly sought-after as a functional food. Its self-fertilisation capability and adaptability to challenging environments further contribute to its potential as a sustainable agricultural option. To harness its unique traits, genetic transformation in F. tataricum is crucial. In this study, we optimised the Agrobacterium-mediated transformation protocol for F. tataricum callus, resulting in a transformation rate of regenerated plants of approximately 20%. The protocol's effectiveness was confirmed through successful GUS staining, GFP expression, and the generation of albino plants via FtPDS gene inactivation. These results validate the feasibility of genetic manipulation and highlight the potential for trait enhancement in F. tataricum.

Keywords: Agrobacterium-mediated transformation; CRISPR/Cas9 system; Green Fluorescent Protein (GFP); GreenGate system; buckwheat; callus; phytoene desaturase (PDS); β-glucuronidase (GUS).

Grants and funding

The authors declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by the National Science Centre, Poland. Research project OPUS-19 (No. Reg. 2020/37/B/NZ9/01499 awarded to AB) and SONATA BIS 10 (No. Reg. 2020/38/E/NZ9/00033 awarded to AB).