Publication:
Transfecting Taxus x media protoplasts to study transcription factors BIS2 and TSAR2 as activators of taxane-related genes

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Date
2020-03
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Authors
Sánchez Muñoz, Raul ; Cusido, Rosa M. ; Bonfill, Mercedes ; Palazón, Javier ; Moyano, Elisabeth ; Almagro Romero, Lorena
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Publisher
Oxford University Press
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Biología Vegetal
DOI
https://doi.org/10.1093/pcp/pcz225
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info:eu-repo/semantics/article
Description
© The Author(s) 2019. This document is the Published Manuscript version of a Published Work that appeared in final form in Plant & Cell Physiology. To access the final edited and published work see https://doi.org/10.1093/pcp/pcz225
Abstract
Taxane diterpenes are secondary metabolites with an im-portant pharmacological role in the treatment of cancer. Taxus spp. biofactories have been used for taxane produc-tion, but the lack of knowledge about the taxane biosynthet-ic pathway and its molecular regulation hinders their optimal function. The difficulties in introducing foreign genes in Taxus spp. genomes hinder the study of the mo-lecular mechanisms involved in taxane production, and a new approach is required to overcome them. In this study, a reliable, simple and fast method to obtain Taxus 􀀁 media protoplasts was developed, allowing their manipulation in downstream assays for the study of physiological changes in Taxus spp. cells. Using this method, Taxus protoplasts were transiently transfected for the first time, corroborating their suitability for transfection assays and the study of specific physiological responses. The two assayed transcription fac-tors (BIS2 and TSAR2) had a positive effect on the expression of several taxane-related genes, suggesting their potential use for the improvement of taxane yields. Furthermore, the results indicate that the developed method is suitable for obtaining T. 􀀁 media protoplasts for transfection with the aim of unraveling regulatory mechanisms in tax-ane production.
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Metabolic engineering , Protoplasts , Taxanes , Taxus x media , Transcription factors , Transient transfection
Citation
Plant and Cell Physiology, 2020, Vol. 61, Issue 3, pp. 576–583
URI
http://hdl.handle.net/10201/149643
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