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.
- Badr A, Angers P, Desjardins Y (2015) Comprehensive analysis of in vitro to ex vitro transition of tissue cultured potato plantlets grown with or without sucrose using metabolic profiling technique. Plant Cell Tiss Organ Cult 122:491–508. https://doi.org/10.1007/s11240-015-0786-3 CrossRefGoogle Scholar
- Batista DS, Castro KM, Koehler AD, Porto BN, Silva AR, Souza VC, Teixeira ML, Cardoso MG, Santos MO, Viccini LF, Otoni WC (2017) Elevated CO2 improves growth, modifies anatomy, and modulates essential oil qualitative production and gene expression in Lippia alba (Verbenaceae). Plant Cell Tiss Organ Cult 128:357–368. https://doi.org/10.1007/s11240-016-1115-1 CrossRefGoogle Scholar
- Batista DS, Felipe SHS, Silva TD, Castro KM, Mamedes-Rodrigues TC, Miranda NA, Ríos-Ríos AM, Faria DV, Fortini EA, Chagas K, Torres-Silva G, Xavier A, Arencibia AD, Otoni WC (2018) Light quality in plant tissue culture: does it matter? In Vitro Cell Dev Biol Plant 54:195–215. https://doi.org/10.1007/s11627-018-9902-5 CrossRefGoogle Scholar
- Batista DS, Koehler AD, Romanel E, Souza VC, Silva TD, Almeida MC, Maciel TEF, Ferreira PRB, Felipe SHS, Saldanha CW, Maldaner J, Dias LLC, Festucci-Buselli RA, Otoni WC (2019) De novo assembly and transcriptome of Pfaffia glomerata uncovers the role of photoautotrophy and the P450 family genes in 20-hydroxyecdysone production. Protoplasma 256:601–614. https://doi.org/10.1007/s00709-018-1322-1 CrossRefGoogle Scholar
- Bernard B, Gautier B (2005) Use of ecdysteroids for preparing dermatological or cosmetological anti-hair loss compositions. United States Patent, US 0137175 A1Google Scholar
- Cha-Um S, Kirdmanee C (2008) Effect of osmotic stress on proline accumulation, photosynthetic abilities and growth of sugarcane plantlets (Saccharum officinarum L.). Pak J Bot 40:2541–2552Google Scholar
- Correa JPO, Viibtal CE, Pinheiro MVM, Batista DS, Azevedo JFL, Saldanha CW, Cruz ACF, DaMatta FM, Otoni WC (2015) In vitro photoautotrophic potential and ex vitro photosynthetic competence of Pfaffia glomerata (Spreng.) Pedersen accessions. Plant Cell Tiss Organ Cult 121:289–300. https://doi.org/10.1007/s11240-014-0700-4 CrossRefGoogle Scholar
- Correa JPO, Vital CE, Pinheiro MVM, Batista DS, Saldanha CW, Cruz ACF, Notoni MM, Freitas DMS, DaMatta FM, Otoni WC (2016) Induced polyploidization increases 20-hydroxyecdysone content, in vitro photoautotrophic growth, and ex vitro biomass accumulation in Pfaffia glomerata (Spreng.) Pedersen. In Vitro Cell Dev Biol Plant 52:45–55. https://doi.org/10.1007/s11627-016-9746-9 CrossRefGoogle Scholar
- Dias FCR, Martins ALP, Melo FCSA, Cupertino MC, Gomes MLM, Oliveira JM, Damasceno EM, Silva J, Otoni WC, Matta SLP (2019) Hydroalcoholic extract of Pfaffia glomerata alters the organization of the seminiferous tubules by modulating the oxidative state and the microstructural reorganization of the mice testes. J Ethnopharmacol 233:179–189. https://doi.org/10.1016/j.jep.2018.12.047 CrossRefGoogle Scholar
- Gomes ACMM, Nicole M, Mattos JK, Pereira SIV, Pereira P, Silva DB, Vieira R, Capeville G, Moita AW, Carneiro RMDG (2010) Concentration of β-ecydisone (20E) in susceptible and resistant accessions of Pfaffia glomerata infected with Meloidogyne incognita and histological characterisation of resistance. Nematology 12:701–709. https://doi.org/10.1163/138855409X12597616267771 CrossRefGoogle Scholar
- Jozwiak A, Ples M, Skorupinka-Tudek K, Kania M, Dydak M, Danikiewicz W, Swiezewska E (2013) Sugar availability modulates polyisoprenoid and phytosterol profiles in Arabidopsis thaliana hairy root culture. Biochim Biophys Acta 1831:438–447. https://doi.org/10.1016/j.bbalip.2012.11.006 CrossRefGoogle Scholar
- Kamada T (2006) Avaliação da diversidade genética de populações de fáfia (Pfaffia glomerata (Spreng.) Pedersen) por RAPD, caracteres morfológicos e teor de beta-ecdisona. Tese de doutorado, Universidade Federal de Viçosa, Viçosa, MG, Brazil. http://locus.ufv.br/handle/123456789/1298. Accessed 16 Jan 2017
- Kamada T, Picoli EAT, Vieira RF, Barbosa LCA, Cruz CD, Otoni WC (2009) Variação dos caracteres morfológicos e fisiológicos de populações naturais de Pfaffia glomerata (Spreng.) Pederson e correlação com a produção de β-ecdisona. Rev Bras Plantas Med 11:247–256. https://doi.org/10.1007/s11240-016-1089-z CrossRefGoogle Scholar
- Karnovsky MJ (1965) A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron microscopy. J Cell Biol 10: 188–204. http://www.jstor.org/stable/1604673. Accessed 16 Jan 2017
- Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x CrossRefGoogle Scholar
- Ngyuen QT, Xiao Y, Kozai T (2016) Photoautotrophic propagation. In: Kozai T, Niu G, Takagaki M (eds) Plant factory. An indoor vertical farming system for efficient quality food production. Academic Press, Cambridge, p 432Google Scholar
- O’Brien TP, McCully ME (1981) The study of plant structure principles and selected methods, vol 14, 1st edn. Termarcarphi Pty, Melbourne, p 357Google Scholar
- Oh M-M, Seo JH, Park JS, Son JE (2012) Physicochemical properties of mixtures of inorganic supporting materials affect growth of potato (Solanum tuberosum L.) plantlets cultured photoautotrophically in a nutrient-circulated micropropagation system. Hortic Environ Biotechnol 53:497–504. https://doi.org/10.1007/s13580-012-0043-1 CrossRefGoogle Scholar
- Olalde JA (2008) Multiple sclerosis synergistic phyto-nutraceutical composition. United States Patent, US 0081046 A1Google Scholar
- Paudel JR, Amirizian A, Krosse S, Giddings J, Ismail SAA, Xia J, Gloer JB, Dam NM, Bede JC (2016) Effect of atmospheric carbon dioxide levels and nitrate fertilization on glucosinolate biosynthesis in mechanically damaged Arabidopsis plants. BMC Plant Biol 16:68. https://doi.org/10.1186/s12870-016-0752-1 CrossRefGoogle Scholar
- Pérez-Jímenez M, López-Perez AJ, Otalora-Alcon G, Marin-Nicolas D, Piñero MC, Amor FM (2015) A regime of high CO2 concentration improves the acclimatization process and increases plant quality and survival. Plant Cell Tiss Organ Cult 121:547–557. https://doi.org/10.1007/s11240-015-0724-4 CrossRefGoogle Scholar
- Pott A, Pott VS (1994) Plantas do pantanal. Embrapa-SPI, CorumbáGoogle Scholar
- Saldanha CW, Otoni CG, Azevedo JLF, Dias LLC, Rêgo MM, Otoni WC (2012) A low-cost alternative membrane system that promotes growth in nodal cultures of Brazilian ginseng [Pfaffia glomerata (Spreng.) Pedersen]. Plant Cell Tiss Organ Cult 110:413–422. https://doi.org/10.1007/s11240-012-0162-5 CrossRefGoogle Scholar
- Saldanha CW, Otoni CG, Notini MN, Kuki KN, Cruz ACF, Neto AR, Dias LLC, Otoni WC (2013) A CO2-enriched atmosphere improves in vitro growth of Brazilian ginseng [Pfaffia glomerata (Spreng.) Pedersen]. In Vitro Cell Dev Biol Plant 49:433–444. https://doi.org/10.1007/s11627-013-9529-5 CrossRefGoogle Scholar
- Saldanha CW, Otoni CG, Rocha DI, Cavatte PC, Detmann KSC, Tanaka FAO, Dias LLC, DaMatta FM, Otoni WC (2014) A CO2-enriched atmosphere and supporting material impact the growth morphophysiology and ultrastructure of in vitro Brazilian ginseng [Pfaffia glomerata (Spreng.) Pedersen] plantlets. Plant Cell Tiss Organ Cult 118:87–99. https://doi.org/10.1007/s11240-014-0464-x CrossRefGoogle Scholar
- Silva TD, Chagas K, Batista DS, Felipe SHS, Louback E, Machado LT, Fernandes AM, Buttrós VHT, Koehler AD, Farias LM, Santos AF, Silva PO, Otoni WC (2019) Morpho-physiological in vitro performance of Brazilian ginseng [Pfaffia glomerata (Spreng.) Pedersen] based on culture media formulations. In Vitro Cell Dev Biol Plant. https://doi.org/10.1007/s11627-019-10003-9 Google Scholar
- Thorpe T, Stasolla C, Yeung EC, De Klerk GJ, Roberts A, George EF (2008) The components of plant tissue culture media II: organic additions, osmotic and pH effects, and support systems. In: George EF, Hall MA, De Klerk GJ (eds) Plant propagation by tissue culture, vol 1, 3rd edn. The background. Springer, Dordrecht, pp 115–173Google Scholar
- Vasconcelos JM, Saldanha CW, Dias LLC, Maldaner J, Rêgo MM, Silva LC, Otoni WC (2014) In vitro propagation of Brazilian ginseng [Pfaffia glomerata (Spreng.) Pedersen] as affected by carbon sources. In Vitro Cell Dev Biol Plant 50:746–751. https://doi.org/10.1007/s11627-014-9651-z CrossRefGoogle Scholar