CO2 enrichment and supporting material impact the primary metabolism and 20-hydroxyecdysone levels in Brazilian ginseng grown under photoautotrophy
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In vitro photoautotrophic propagation system has been successfully established for Pfaffia glomerata, a medicinal species that produces the phytoecdysteroid 20-hydroxyecdysone (20E) under forced ventilation and CO2 enrichment. For that, an adequate supporting material with high porosity in place of agar is required. In this study, we investigated metabolic and morpho-anatomical alterations of two accessions (Ac) of P. glomerata (Ac 22 and Ac 43) under photoautotrophy conditions, using two supporting materials (agar and Florialite®) and two CO2 concentrations (360 and 1000 µL CO2 L−1). High CO2 concentration and the use of Florialite® as supporting material enhanced the production of 20E, and influenced on the levels of amino acids, sugars, tricarboxylic acid cycle intermediates, stress related metabolites (aromatic amines and shikimate), and osmotic adjustment-related compounds (hydroxyproline, aspartate and myo-inositol). Interestingly, Ac 22 displayed less tolerance to stress caused by low photoautotrophy compared to Ac 43, as indicated by the higher total polyamines in Ac 22. Moreover, the accessions showed different metabolic responses under photoautotrophy. These findings provide a better understanding of how the supporting material and CO2 enrichment influence in vitro metabolism under photoautotrophic system in P. glomerata, wherein higher levels of gas exchange, enabled by use of Florialite® and CO2 enrichment, increased total sugars as well as the levels of 20E in the plants. This information will be fundamental to optimize the in vitro culture systems of P. glomerata for the production of 20E.
Photoautotrophic propagation has the unique advantage of combining biological and environmental perspectives, resulting in morpho-physiologically well-adjusted plants grown in vitro or ex vitro. Our results revealed that the combined use of CO2 enrichment (1000 μL CO2 L−1) and supporting material (Florialite®) had great influence in the in vitro performance of P. glomerata and increased 20-hydroxyecdysone content. This combination also led to increased levels of amino acids, sugars, TCA cycle intermediates, and stress-related metabolites such as aromatic amines and shikimate, and osmotic adjustment-related compounds (hydroxyproline, aspartate, and myo-inositol).
KeywordsAmaranthaceae CO2 enrichment Metabolomics Pfaffia glomerata Photoautotrophic growth
Prof. Takeshi Kamada (Universidade de Rio Verde, Rio Verde, GO, Brazil), Dr. Roberto Fontes Vieira and Dr. Rosa Belém das Neves Alves (National Center for Genetic Resources and Biotechnology—Embrapa/Cenargen, Brasília, DF, Brazil) are acknowledged for providing P. glomerata accessions. The authors are also grateful to the Núcleo de Análise de Biomoléculas (NuBiomol) of the Universidade Federal de Viçosa for providing the facilities for the metabolite analysis. PRBF was recipient of a scholarship from CAPES.
PRBF and WCO planned and designed the research. PRBF, DSB, ACFC, ADK, and SHF conducted experiments and analyzed the data. IGA conducted and photographed part of the experiments. LAN and DIR contributed with anatomy analysis and manuscript writing. PRBF wrote the manuscript, WCO, DSB and AN-N reviewed and corrected it.
This work was supported by the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) (Brasília, DF, Brazil) [Grants MCT/CNPq 480675/2009-0; PDJ 500874/2012-3; PQ 303201/2010-10 to WCO], Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) (Belo Horizonte, MG, Brazil) [Grants PRONEX-CAG-APQ-01036-09; CRA-APQ-01651-13; CRA-BPD-00046-14; CBB-BPD-00020-16; RED-00053-16]; and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) (Brasília, DF, Brazil) Finance Code 001 and CAPES—PNPD. RPBS was recipient of a scholarship from CAPES.
Compliance with ethical standards
Conflict of interest
The authors declare that there are no conflict of interest.
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