Insights into enhancing Centella asiatica organ cell biofactories via hairy root protein profiling

Miguel Angel Alcalde; Diego Hidalgo-Martinez; Roque Bru Martínez; Susana Sellés-Marchart; Mercedes Bonfill; Javier Palazon

doi:10.3389/fpls.2023.1274767

Insights into enhancing Centella asiatica organ cell biofactories via hairy root protein profiling

Front Plant Sci. 2023 Oct 30:14:1274767. doi: 10.3389/fpls.2023.1274767. eCollection 2023.

Authors

Miguel Angel Alcalde^{1

2}, Diego Hidalgo-Martinez², Roque Bru Martínez³, Susana Sellés-Marchart³, Mercedes Bonfill², Javier Palazon²

Affiliations

¹ Biotechnology, Health and Education Research Group, Posgraduate School, Cesar Vallejo University, Trujillo, Peru.
² Department of Biology, Healthcare and the Environment, Faculty of Pharmacy and Food Sciences, University of Barcelona, Barcelona, Spain.
³ Plant Proteomics and Functional Genomics Group, Department of Biochemistry and Molecular Biology, Soil Science and Agricultural Chemistry, Faculty of Science, University of Alicante, Alicante, Spain.

Abstract

Recent advancements in plant biotechnology have highlighted the potential of hairy roots as a biotechnological platform, primarily due to their rapid growth and ability to produce specialized metabolites. This study aimed to delve deeper into hairy root development in C. asiatica and explore the optimization of genetic transformation for enhanced bioactive compound production. Previously established hairy root lines of C. asiatica were categorized based on their centelloside production capacity into HIGH, MID, or LOW groups. These lines were then subjected to a meticulous label-free proteomic analysis to identify and quantify proteins. Subsequent multivariate and protein network analyses were conducted to discern proteome differences and commonalities. Additionally, the quantification of rol gene copy numbers was undertaken using qPCR, followed by gene expression measurements. From the proteomic analysis, 213 proteins were identified. Distinct proteome differences, especially between the LOW line and other lines, were observed. Key proteins related to essential processes like photosynthesis and specialized metabolism were identified. Notably, potential biomarkers, such as the Tr-type G domain-containing protein and alcohol dehydrogenase, were found in the HIGH group. The presence of ornithine cyclodeaminase in the hairy roots emerged as a significant biomarker linked with centelloside production capacity lines, indicating successful Rhizobium-mediated genetic transformation. However, qPCR results showed an inconsistency with rol gene expression levels, with the HIGH line displaying notably higher expression, particularly of the rolD gene. The study unveiled the importance of ornithine cyclodeaminase as a traceable biomarker for centelloside production capacity. The strong correlation between this biomarker and the rolD gene emphasizes its potential role in optimizing genetic transformation processes in C. asiatica.

Keywords: Rhizobium rhizogenes; biomarkers; centelloside production; molecular farming; plant biotechnology.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This research was funded by Agencia Estatal de Investigación. REF: PID2020-113438RBI00/AEI/10.13039/501100011033, by the Agència de Gestió d’Ajuts Universitaris i de Recerca (AGAUR) del Departament de Recerca i Universitats de la Generalitat de Catalunya 2021 SGR00693 and Valencian Conselleria d’Innovació, Universitats, Ciencia y Societat Digital grant CIAICO/2021/167. DH-M is a Postdoctoral researcher at Maria Zambrano at the University of Barcelona. His contract is financed by the Ministry of Universities, the European Union Next GenerationEU/PRTRi, and the Recovery, Transformation and Resilience Plan.