Abstract
Key message
Extensive leaf transcriptome profiling and differential gene expression analysis of field grown and elicited shoot cultures of L. speciosa suggest that differential synthesis of CRA is mediated primarily by CYP and TS genes, showing functional diversity.
Abstract
Lagerstroemia speciosa L. is a tree species with medicinal and horticultural attributes. The pentacyclic triterpene, Corosolic acid (CRA) obtained from this species is widely used for the management of diabetes mellitus in traditional medicine. The high mercantile value of the compound and limited availability of innate resources entail exploration of alternative sources for CRA production. Metabolic pathway engineering for enhanced bioproduction of plant secondary metabolites is an attractive proposition for which, candidate genes in the pathway need to be identified and characterized. Therefore, in the present investigation, we focused on the identification of cytochrome P450 (CYP450) and oxidosqualene cyclases (OSC) genes and their differential expression during biosynthesis of CRA. The pattern of differential expression of these genes in the shoot cultures of L. speciosa, elicited with different epigenetic modifiers (azacytidine (AzaC), sodium butyrate (NaBu) and anacardic acid (AA)), was studied in comparison with field grown plant. Further, in vitro cultures with varying (low to high) concentrations of CRA were systematically assessed for the expression of CYP-TS and associated genes involved in CRA biosynthesis by transcriptome sequencing. The sequenced samples were de novo assembled into 180,290 transcripts of which, 92,983 transcripts were further annotated by UniProt. The results are collectively given in co-occurrence heat maps to identify the differentially expressed genes. The combined transcript and metabolite profiles along with RT-qPCR analysis resulted in the identification of CYP-TS genes with high sequence variation. Further, instances of concordant/discordant relation between CRA biosynthesis and CYP-TS gene expression were observed, indicating functional diversity in genes.
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Abbreviations
- AA:
-
Anacardic acid
- AzaC:
-
Azacytidine
- BAP:
-
6-Benzylaminopurine
- CYP450:
-
Cytochrome P450
- GO:
-
Gene ontology
- HPLC:
-
High-performance liquid chromatography
- NaBu:
-
Sodium butyrate
- OSC:
-
Oxidosqualene cyclase
- TPS:
-
Terpene synthase
- TS:
-
Terpenoid synthase
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Acknowledgements
We greatly acknowledge the Vice Chancellor of Central University of Kerala, Kasaragod, for all the supports rendered throughout this study. K.S acknowledges with thanks the Department of Science and Technology, Govt. of India for providing the DST-INSPIRE fellowship (DST/INSPIRE Fellowship/2017/IF170450).
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The work was partly supported by funding from Central University of Kerala.
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K.S performed all analyses and experiments concerning tissue culture, CYP450 enzymes and TS. K.S and S.P conducted the validation studies of transcriptome data. P.P conceptualized the research problem and designed the experiments. K.S prepared the draft and P.P refined and revised the manuscript. All authors read and approved the manuscript.
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Surendran, K., Pradeep, S. & Pillai, P.P. Comparative transcriptome and metabolite profiling reveal diverse pattern of CYP-TS gene expression during corosolic acid biosynthesis in Lagerstroemia speciosa (L.) Pers. Plant Cell Rep 43, 122 (2024). https://doi.org/10.1007/s00299-024-03203-0
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DOI: https://doi.org/10.1007/s00299-024-03203-0