An established protocol for generating transgenic wheat for wheat functional genomics via particle bombardment

Yaqiong Wang; Jian Zeng; Peipei Su; Hongyan Zhao; Li Li; Xiaoxue Xie; Qian Zhang; Ya'nan Wu; Ruibin Wang; Yufan Zhang; Boju Yu; Mingjie Chen; Yuesheng Wang; Guangxiao Yang; Guangyuan He; Junli Chang; Yin Li

doi:10.3389/fpls.2022.979540

An established protocol for generating transgenic wheat for wheat functional genomics via particle bombardment

Front Plant Sci. 2022 Dec 8:13:979540. doi: 10.3389/fpls.2022.979540. eCollection 2022.

Authors

Yaqiong Wang¹, Jian Zeng², Peipei Su¹, Hongyan Zhao¹, Li Li¹, Xiaoxue Xie¹, Qian Zhang¹, Ya'nan Wu¹, Ruibin Wang¹, Yufan Zhang¹, Boju Yu¹, Mingjie Chen¹, Yuesheng Wang¹, Guangxiao Yang¹, Guangyuan He¹, Junli Chang¹, Yin Li¹

Affiliations

¹ The Genetic Engineering International Cooperation Base of Chinese Ministry of Science and Technology, The Key Laboratory of Molecular Biophysics of Chinese Ministry of Education, College of Life Science and Technology, Huazhong University of Science & Technology, Wuhan, China.
² Guangdong Provincial Key Laboratory of Utilization and Conservation of Food and Medicinal Resources in Northern Region, Shaoguan University, Shaoguan, Guangdong, China.

Abstract

Wheat is one of the most important food crops in the world and is considered one of the top targets in crop biotechnology. With the high-quality reference genomes of wheat and its relative species and the recent burst of genomic resources in Triticeae, demands to perform gene functional studies in wheat and genetic improvement have been rapidly increasing, requiring that production of transgenic wheat should become a routine technique. While established for more than 20 years, the particle bombardment-mediated wheat transformation has not become routine yet, with only a handful of labs being proficient in this technique. This could be due to, at least partly, the low transformation efficiency and the technical difficulties. Here, we describe the current version of this method through adaptation and optimization. We report the detailed protocol of producing transgenic wheat by the particle gun, including several critical steps, from the selection of appropriate explants (i.e., immature scutella), the preparation of DNA-coated gold particles, and several established strategies of tissue culture. More importantly, with over 20 years of experience in wheat transformation in our lab, we share the many technical details and recommendations and emphasize that the particle bombardment-mediated approach has fewer limitations in genotype dependency and vector construction when compared with the Agrobacterium-mediated methods. The particle bombardment-mediated method has been successful for over 30 wheat genotypes, from the tetraploid durum wheat to the hexaploid common wheat, from modern elite varieties to landraces. In conclusion, the particle bombardment-mediated wheat transformation has demonstrated its potential and wide applications, and the full set of protocol, experience, and successful reports in many wheat genotypes described here will further its impacts, making it a routine and robust technique in crop research labs worldwide.

Keywords: functional genomics; genetic improvement; genetic transformation; particle bombardment; particle bombardment-mediated wheat transformation research topic: women in plant biotechnology 2022 wheat; transgenic plants.