Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 136, Issue 2, pp 255–267 | Cite as

Indirect somatic embryogenesis in Coffea with different ploidy levels: a revisiting and updating study

  • Natália Arruda Sanglard
  • Paulo Marcos Amaral-Silva
  • Mariana Cansian Sattler
  • Stéfanie Cristina de Oliveira
  • Letícia Miranda Cesário
  • Adésio Ferreira
  • Carlos Roberto Carvalho
  • Wellington Ronildo ClarindoEmail author
Original Article


Indirect somatic embryogenesis (ISE) is required for plant propagation and a prerequisite for applications that may provide new germplasms. Genetic, epigenetic and physiological features of the explant donor are barriers for ISE establishment, hindering its wide use. Despite the identification and/or expression analysis of genes during ISE, no approach to establish the karyotype aspects has been performed so far. So, this study aims to establish the ISE and compare the in vitro responses between diploid (Coffea canephora and Coffea eugenioides), allotriploid (“Híbrido de Timor”—HT) and true allotetraploid (Coffea arabica) Coffea in a taxonomic and evolutive scenario. Under the same in vitro conditions, the four Coffea differed from each other during ISE. Leaf explants of the true allopolyploids yielded the highest mean number of friable calli (FC) in relative short time and visually exhibiting more pronounced length. FC of the allotetraploid C. arabica presented the highest mean number of mature cotyledonary somatic embryos (MCSE), which were also recovered faster in this species. However, MCSE mean number in HT was the same or lower than diploid Coffea. Besides, intraspecific variation related to the ISE responses was observed in each Coffea, mainly the mean number of FC obtained from ex vitro and in vitro C. arabica and C. eugenioides explants. So, epigenetic and physiologic features may also have influenced the ISE responses. The findings provide the basis for performing other approaches considering the ploidy level, epigenetic and physiological backgrounds. Besides, the data also contributed for understanding about the consequences of polyploidy.


Coffee Plant tissue culture In vitro somatic embryogenesis Euploidy Chromosome number Nuclear 2C value 



2,4-Dichlorophenoxyacetic acid




Friable calli


Gibberellic acid


“Híbrido de Timor”


Indirect somatic embryogenesis


Mature cotyledonary somatic embryo


Secondary constriction


Somatic embryos



We would like to thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq, Brasília—DF, Brazil, Grant: 443801/2014-2), Fundação de Amparo à Pesquisa do Espírito Santo (FAPES, Vitória—ES, Brazil, Grant: 65942604/2014 and 82/2017), and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES, Brasília—DF, Brazil) for financial support. We also thank Dr. Eveline Teixeira Caixeta (Embrapa Café, Empresa Brasileira de Pesquisa Agropecuária, BIOAGRO, Laboratório BioCafé, Universidade Federal de Viçosa, MG, Brazil) for providing leaves in 2015 of the C. eugenioides and C. arabica ‘Catuaí Vermelho’ plants.

Author contributions

The authors NAS, PMA, LMC and WRC conceived, designed and conducted the tissue culture experiments. MCS, SCDO and WRC carried out the cytogenetic analyses. WRC and CRC contributed with flow cytometry analysis. The authors NAS and AF performed the statistical analysis.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  • Natália Arruda Sanglard
    • 1
  • Paulo Marcos Amaral-Silva
    • 1
  • Mariana Cansian Sattler
    • 2
  • Stéfanie Cristina de Oliveira
    • 1
  • Letícia Miranda Cesário
    • 1
  • Adésio Ferreira
    • 3
  • Carlos Roberto Carvalho
    • 2
  • Wellington Ronildo Clarindo
    • 2
    Email author
  1. 1.Laboratório de Citogenética e Cultura de Tecidos Vegetais, Departamento de BiologiaUniversidade Federal do Espírito SantoAlegreBrazil
  2. 2.Laboratório de Citogenética e Citometria, Departamento de Biologia Geral, Centro de Ciências Biológicas e da SaúdeUniversidade Federal de ViçosaViçosaBrazil
  3. 3.Laboratório de Biometria, Departamento de AgronomiaUniversidade Federal do Espírito SantoAlegreBrazil

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