Abstract
Transgene expression and genome editing can help improve cucumber varieties to better respond to climate change. This study aimed to evaluate the applicability of the CsUBL5 promoter in transgene expression and genome editing in cucumber. The CsUBL5 promoter was cloned and analyzed to identify cis-elements that respond to abiotic signals, hormones, signal molecules, and nutrient treatments. 5′ deletion constructs of the promoter were tested for their ability to drive GUS reporter expression in cucumber cotyledons, Arabidopsis seedlings, and tobacco leaves, and their response to various treatments including SA, light, drought, IAA, and GA was determined. The results showed that the CsUBL5 promoter effectively drove transgene expression in these plants, and their expressions under treatments were consistent with the predicted cis-elements, with some exceptions. Furthermore, the pCsUBL5-749 deletion construct can improve genome editing efficiency in cucumber when driving Cas9 expression. The editing efficiency of two sgRNAs targeting the ATG6 gene in cucumber was up to 4.6-fold higher using pCsUBL5-749 compared to a rice UBI promoter, although the effects of changing promoter on the editing efficiency is sgRNA specific. These findings highlight the potential utility of the CsUBL5 promoter for improving cucumber varieties through genetic engineering and genome editing. It also demonstrates the importance of modulating Cas9 expression to increase genome editing efficiency in cucumbers.
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Acknowledgements
This research is supported by the Graduate Program Scholarship from The Graduate School, Kasetsart University for KT. ID was supported by Science Achievement Scholarship of Thailand (SAST). AS was supported by Agricultural Research Development Agency (CRP6405030740). SV was supported by the National Research Council of Thailand (NRCT5-RSA63002-02) and The Office of the Ministry of Higher Education, Science, Research and Innovation; and the Thailand Science Research and Innovation through the Kasetsart University Reinventing University Program 2021. Dr. Thidaporn Supapakorn, Dr. Passorn Wonnapinij, and Kunnika Thongrak provided assistance in data and statistical analysis.
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AS, KT, and SV conceived the project and designed experiments. KT and ID performed the experiments. KT, ID and AS analyzed the data. KT, ID, SV, and AS prepared Figures and Tables and wrote the manuscript. All authors have reviewed the manuscript and approved the final version.
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Taway, K., Dachphun, I., Vuttipongchaikij, S. et al. Evaluation of cucumber UBL5 promoter as a tool for transgene expression and genome editing in plants. Transgenic Res 32, 437–449 (2023). https://doi.org/10.1007/s11248-023-00359-5
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DOI: https://doi.org/10.1007/s11248-023-00359-5