Abstract
Bixa orellana L. seeds are an important source of the liposoluble apocarotenoid bixin and the natural hydrosoluble dye norbixin. These natural dyes are used for food, pharmaceutical, and cosmetic applications. The participation of methyltransferases has already been demonstrated in the bixin biosynthesis pathway, and the alteration of these genes expression can lead to changes in the production of bixin. Treatment with demethylating agents, such as 5-azacytidine (5-AzaC), was previously related to an increased production of secondary compounds and serves for the study of secondary metabolism pathways in plants. Here, we evaluated for the first time the effect of 5-AzaC on the expression of SABATH methyltransferase genes as well as some bixin pathway candidate genes, global DNA methylation, and bixin, chlorophylls, and total carotenoid content in the leaves of B. orellana plants cultured in vitro. Exposing shoot apexes to 5-AzaC for 48 h affected global genome methylation but did not alter chlorophylls and total carotenoid content in the leaves of B. orellana. Importantly, plants submitted to this demethylating agent exhibited downregulation of the SABATH and CCD4 families of genes, upregulation of the ALDH3H1 gene, and increased bixin content in the leaves, suggesting that this methyltransferase inhibitor can selectively regulate the biosynthesis and production of bixin in B. orellana by an unknown mechanism. Moreover, here we demonstrate the possibility to use 5-AzaC to increase bixin contents in B. orellana plants.
Key message
The methyltransferase inhibitor, 5-Azacytidine, can selectively regulate the biosynthesis and production of bixin in annatto plant leaves, by upregulating BoALDH3H1 gene and downregulating BoSABATH and BoCCD4 gene families.
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Abbreviations
- 5-AzaC:
-
5-Azacytidine
- ALDH :
-
Aldehyde dehydrogenase
- BoADH :
-
Bixin aldehyde dehydrogenase
- BoLCD :
-
Bixin carotene cleavage dioxygenase
- BonBMT :
-
Norbixin methyltransferase
- CCD4 :
-
Carotenoid cleavage dioxygenase 4
- COMT :
-
Caffeic acid O-methyltransferase
- DEPC:
-
Diethyl pyrocarbonate
- DW:
-
Dry weight
- DXS :
-
1-Deoxy-d-xylulose-5-phosphate synthase
- IBA:
-
Indole-3-butyric acid
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Acknowledgements
Fundação de Amparo à Pesquisa do Estado de Minas Gerais—FAPEMIG (Grant number APQ-02372-17, Belo Horizonte, MG), Conselho Nacional de Desenvolvimento Científico e Tecnológico—CNPq (Brasília, DF), and Coordenação de Aperfeiçoamento de Pessoal de Ensino Superior—CAPES (Finance code 01, Brasília, DF) are acknowledged for their financial support. Christian Hansen Ind. Com. Ltda. (Valinhos, SP, Brazil) is acknowledged for kindly donating the seeds of Bixa orellana ‘Piave Vermelha’. The authors are in debt to the Laboratory for Molecular Genetics of Bacteria (BIOAGRO/UFV) for making available the facilities for quantitative RT-PCR analyses. DVF was a recipient of a postdoctoral scholarship from the CNPq (Grant number 155615/2018-1). Editage (www.editage.com) is also acknowledged for English language editing.
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DFV and LNFC cultivated the plants in vitro and performed the experiments. DSB and DFV performed statistical analysis. CEV performed bixin analysis. ASH performed DNA methylation analysis. DFV, WCO, DSB, MGCC, LNFC, MPG and CD-l-P contributed to the design and interpretation of the data and to the writing of the paper.
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Communicated by Manoj Prasad.
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Faria, D.V., de Freitas Correia, L.N., Batista, D.S. et al. 5-Azacytidine downregulates the SABATH methyltransferase genes and augments bixin content in Bixa orellana L. leaves. Plant Cell Tiss Organ Cult 142, 425–434 (2020). https://doi.org/10.1007/s11240-020-01857-8
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DOI: https://doi.org/10.1007/s11240-020-01857-8