Abstract
Objective
In this study, we established an efficient and rapid transient expression system in the protoplasts of Pinellia ternata (Thunb.) Breit. (P. ternata).
Results
The protoplasts of P. ternata were prepared from plant leaves as the source material by digesting them with the combination of 20 g·l−1 cellulase and 15 g·l−1 macerozyme for 6 h. Based on the screening of PEG concentration, the conditions for PEG-mediated protoplast transformation were improved, and the highest transformation efficiency was found for 40% PEG 4000. Furthermore, we used the subcellular protein localization technique in P. ternata protoplasts to allow further validation of transient expression system.
Conclusions
We present the method that can be applicable for studying both gene verification and expression in P. ternata protoplasts, thus allowing for engineering the improved varieties of P. ternata through molecular plant breeding techniques. This method can also be widely applicable for analyzing protein interactions, detecting promoter activity, for somatic cell fusion in plant breeding, as well as for other related studies.
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Data availability
The data that support the findings of this study are available from the corresponding author upon reasonable request.
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Acknowledgements
The authors are grateful to Hong Yi from the Instrument and Equipment Sharing Platform of Large Scale Instruments & Equipments of Guizhou University, China. The authors would like to express their gratitude to EditSprings for the expert linguistic services provided.
Funding
This work was supported by the Major Special Project of Science and Technology Program in Guizhou (2017-5411-06); the Science and Technology Support Project of Guizhou (2018-2797); the Guizhou University Cultivation Project (2017-5788 & 702029201101); the Construction Project of Modern Industry Technology system of traditional Chinese Medicinal Materials in Guizhou (GZCYTX-02); the Expert Team Construction Project of Pinellia ternata in Guizhou (2019-175); the Construction Program of Biology First-Class Discipline in Guizhou (GNYL-2017-009-FX1KT02).
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MZ: Conceptualization, methodology and funding acquisition. YT and ML: Investigation, validation, and writing-original draft preparation. LT, YZ, YH, LS and YZ: Discussion, writing-reviewing and editing. All authors read and approved the final manuscript.
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Tian, Yh., Liu, M., Tang, L. et al. Establishment of protoplasts transient expression system in Pinellia ternata (Thunb.) Breit. Biotechnol Lett 45, 1381–1391 (2023). https://doi.org/10.1007/s10529-023-03420-9
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DOI: https://doi.org/10.1007/s10529-023-03420-9