Dataset Information

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

ABSTRACT: The objectives of this study encompassed a thorough exploration of the potential implications of protein profiling in hairy roots, specifically focusing on optimizing and enhancing C. asiatica organ cell biofactories. In this pursuit, we categorized established C. asiatica hairy root lines according to their capacity for centelloside production, classifying them into HIGH, MID, or LOW categories. For comparative analysis, wild adventitious (Adv) roots were extracted from in vitro C. asiatica seedlings and cultivated in solid MS medium at 25°C in complete darkness, serving as control specimens. This meticulous, label-free proteomic analysis enabled the successful identification of several proteins. Our research substantially builds upon and extends the findings presented by Alcalde et al. (2022) (DOI: 10.3389/fpls.2022.1001023). In their study, distinctive morphological and metabolic variations were noted among different C. asiatica hairy root lines. Such differences are presumably attributable to the random incorporation of a selective set of genes from the T-DNA, with particular emphasis on the rol and aux genes.

INSTRUMENT(S): 6550 iFunnel Q-TOF LC/MS

ORGANISM(S): Centella Asiatica

TISSUE(S): Hairy Root Culture

SUBMITTER: Diego Alberto Hidalgo Martinez

LAB HEAD: Diego A. Hidalgo

PROVIDER: PXD045705 | Pride | 2024-01-26

REPOSITORIES: Pride

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

Alcalde Miguel Angel MA Hidalgo-Martinez Diego D Bru Martínez Roque R Sellés-Marchart Susana S Bonfill Mercedes M Palazon Javier J

Frontiers in plant science 20231030

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 produc ...[more]

PMID: 37965024

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Project description:Multigene families encoding diverse secreted peptide hormones play important roles in plant development. A need exists to efficiently elucidate the structures and post-translational-modifications of these difficult-to-isolate peptide hormones in planta so that their biological functions can be determined. A mass spectrometry and bioinformatics approach was developed to comprehensively analyse the secreted peptidome of Medicago hairy root cultures and xylem sap. We identified 759 spectra corresponding to the secreted products of twelve peptide hormones including four CEP (C-TERMINALLY ENCODED PEPTIDE), two CLE (CLV3/ENDOSPERM SURROUNDING REGION RELATED) and six XAP (XYLEM SAP ASSOCIATED PEPTIDE) peptides. The MtCEP1, MtCEP2, MtCEP5 and MtCEP8 peptides identified differed in post-translational-modifications. Most were hydroxylated at conserved proline residues but some MtCEP1 derivatives were tri-arabinosylated. In addition, many CEP peptides possessed unexpected N- and C-terminal extensions. The pattern of these extensions suggested roles for endo- and exoproteases in CEP peptide maturation. Longer -than -expected, hydroxylated and homogeneously modified mono- and tri-arabinosylated CEP peptides corresponding to their in vivo structures were chemically synthesised to probe the effect of these post-translational-modifications on function. The ability of CEP peptides to elevate root nodule number was increased by hydroxylation at key positions. MtCEP1 peptides with N-terminal extensions or with tri-arabinosylation modification, however, were unable to impart increased nodulation. The MtCLE5 and MtCLE17 peptides identified were of precise size, and inhibited main root growth and increased lateral root number. Six XAP peptides, each beginning with a conserved DY sulfation motif, were identified including MtXAP1a, MtXAP1b, MtXAP1c, MtXAP3, MtXAP5 and MtXAP7. MtXAP1a and MtXAP5 inhibited lateral root emergence. Transcriptional analyses demonstrated peptide hormone gene expression in the root vasculature and tip. Since hairy roots can be induced on many plants, their corresponding root cultures may represent ideal source materials to efficiently identify diverse peptide hormones in vivo in a broad range of species.

Dataset Information

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

Publications

Insights into enhancing <i>Centella asiatica</i> organ cell biofactories via hairy root protein profiling.

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