Agrobacterium-mediated transient gene expression in developing Ricinus communis seeds: a first step in making the castor oil plant a chemical biofactory
- 1Instituto de la Grasa (IG), Spain
- 2University of Málaga, Spain
The castor oil plant represents a promising platform to produce oils with industrial applications. However, its use in biotechnology is limited by the absence of a well-established procedure to transform it, and a poor understanding of gene regulation and promoter use in this species. As such, a quick method has been developed to express proteins or hairpin-RNA in this plant, a method based on the direct injection of Agrobacterium into the developing endosperm of castor oil fruit, enabling different constructs and promoters to be tested. This method produces a high rate of transformation and a good proportion of viable seeds that express reporter genes for up to 20 days after imbibition (DAI). Gene expression under the control of different promoters was tested by RT-qPCR and by directly assaying the activity of the GUS reporter gene, which proved to be strongest when driven by the glycinin promoter. Constructs expressing a fatty acid elongase from Lesquerella fendlery were tested, the expression of which provoked an important increase in the lesquerolic acid in the castor oil endosperm at 5 and 10 DAI, although this fatty acid did not accumulate significantly in the final mature seeds. The nature of this response could reflect the poor availability of substrates for this enzyme. In the light of this data, the potential of this technique to test promoters and different constructs in castor oil plants and other oilseeds is discussed.
Keywords: Ricinus communis, transient expression, Promoter, Lesquerella fedleri, elongase, Acyl-CoA pool
Received: 23 Jan 2019;
Accepted: 11 Oct 2019.
Copyright: © 2019 Sánchez-Álvarez, Ruiz-Lopez, Moreno-Perez, Martinez Force, Garcés and Salas. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
* Correspondence: Dr. Joaquín J. Salas, Instituto de la Grasa (IG), Seville, 41013, Spain, firstname.lastname@example.org