Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Species delimitation within Campomanesia (Myrtaceae) using morphology and amplification profiles of microsatellite markers


Campomanesia is a well-circumscribed genus within Myrtaceae, but the boundaries among some of its species remain poorly understood. Microsatellite amplification profiles have proven useful DNA fingerprinting for plant species identification, but have never been applied to the taxonomy of this genus. Here we studied 12 species of Campomanesia, including a set of related and non-related species, as well as species with doubtful boundaries. Forty-eight morphological characters and 50 primers were used to test the species. A cluster analysis followed by a group sharpness test and a heat map analysis was performed with both sets of data. A regression tree was constructed using only on the molecular data. Seven groups based on morphological characters and five groups based on molecular characters were revealed by cluster analysis and the sharpness group test. The microsatellite amplification profiles were efficient at distinguishing the studied species, despite the smaller number of distinct groups formed by this marker compared with the morphological data. Among the 50 primers included, 13 were informative for species distinction, and 10 were indicated by the regression tree to species identification. Microsatellite amplification profiles proved to be an informative tool for the taxonomy of Campomanesia.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4


  1. Borcard D, Gillet F, Legendre P (2011) Numerical ecology with R. Springer, New York

  2. Chae WB, Hong SJ, Gifford JM, Rayburn AL, Sacks EJ, Juvik JA (2014) Plant morphology, genome size, and SSR markers differentiate five distinct taxonomic groups among accessions in the genus Miscanthus. GCB Bioenergy 6:646–660.

  3. Christ JA, Hollunder RK, Carvalho MS, Ferreira MF, Garbin ML, Carrijo TT (2018) DNA fingerprinting based on SSR amplification profiles for Piper species identification (Piperaceae). Acta Bot Brasil 32:511–520.

  4. Crawley MJ (2007) The R Book. Wiley, West Sussex

  5. Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

  6. Duminil J, Di Michele M (2009) Plant species delimitation: a comparison of morphological and molecular markers. Plant Biosyst 143:528–542.

  7. Galili T, Mitelpunkt A, Shachar N, Marcus-Kalish M, Benjamini Y (2014) Categorize, cluster and classify: a 3-C strategy for scientific discovery in the Medical Informatics platform of the Human Brain Project. In: Discovery science, pp 73–86

  8. Landrum LR (1986) Campomanesia, Pimenta, Blepharocalyx, Legrandia, Acca, Myrrhinium and Luma (Myrtaceae). Flora Neotropica Monograph 45:7–72

  9. Landrum LR (2001) Two new species of Campomanesia (Myrtaceae) from Espírito Santo and Bahia, Brazil. Brittonia 53:534–538

  10. Legendre P, Legendre L (2012) Numerical ecology, 3rd edn. Elsevier, Amsterdam

  11. Lima DF, Mauad AVS, Silva-Pereira V, Smidt EC, Goldenberg R (2015) Species boundaries inferred from ISSR markers in the Myrcia laruotteana complex (Myrtaceae). Plant Syst Evol 301:353–363.

  12. Luber J, Oliveira MIU, Ferreira MFS, Carrijo TT (2017) Flora do Espírito Santo: Campomanesia (Myrtaceae). Rodriguésia 68:1767–1790.

  13. Maechler M, Rousseeuw P, Struyf A, Hubert M, Hornik K (2016) Cluster: cluster analysis basics and extensions. R package version 2.0.4

  14. Nogueira AM, Ferreira A, da Silva-Ferreira MF (2016) Transferability of Microsatellites from Psidium guajava to Eugenia, Myrciaria, Campomanesia, and Syzygium Species (Myrtaceae). Plant Mol Bio Rep 34:249–256

  15. Oksanen J, Blanchet FG, Kindt R (2016) Vegan: community ecology package. R package version 1.17-1. 2010. Accessed 20 Oct 2018

  16. Oliveira MIU (2013) Estudos taxonômicos e populacionais em Campomanesia Ruiz & Pavón (Myrtaceae, Myrteae), com ênfase no “Complexo Campomanesia xanthocarpa”. Thesis. Universidade Estadual de Feira de Santana, Bahia

  17. Oliveira MIU, Funch LS, Landrum LR (2012) Flora da Bahia: Campomanesia (Myrtaceae). Sitientibus 12:91–107

  18. Pillar VD (1999) How sharp are classifications? Ecology 80:2508–2516.

  19. Pillar VD (2006) MULTIV, multivariate exploratory analysis, randomization testing and bootstrap resampling. User’s Guide v. 2.4. Accessed 20 Oct 2018

  20. R Core Team (2016) R: a language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. Accessed 20 Oct 2018

  21. Ripley B (2016) tree: Classification and Regression Trees. R package version 1.0-37. Accessed 20 Oct 2018

  22. Samah S, De Pardo CVT, Cruz MAS, Valadez-Moctezuma E (2016) Genetic diversity, genotype discrimination, and population structure of Mexican Opuntia sp., determined by SSR markers. Plant Mol Bio Rep 34:146–159.

  23. Satya P, Paswan PK, Ghosh S, Majumdar S, Ali N (2016) Confamiliar transferability of simple sequence repeat (SSR) markers from cotton (Gossypium hirsutum L.) and jute (Corchorus olitorius L.) to twenty-two Malvaceous species. 3 Biotech 6:65.

  24. Sharawy S, Karakish E (2015) Taxonomic relationships of some species of Orobanche L. evidence from RAPD-PCR and ISSR markers. Pak J Bot 47:437–452

  25. Spooner DM (2016) Species delimitations in plants: lessons learned from potato taxonomy by a practicing taxonomist. J Syst Evol 54(3):191–203.

  26. Tóth G, Gáspari Z, Jurka J (2000) Microsatellites in different eukaryotic genomes: survey and analysis. Genome Res 10:967–981

  27. Tuler AC, Carrijo TT, Nóia LR, Ferreira A, Peixoto AL, da Silva Ferreira MF (2015) SSR markers: a tool for species identification in Psidium (Myrtaceae). Mol Biol Rep 42:1501–1513.

  28. Weber JL, May PE (1989) Abundant class of human DNA polymorphisms which can be typed using the Polymerase Chain Reaction. Am J Hum Genet 44:388–396

  29. Zhang Y, Yuan X, Teng W, Chen C, Wu J (2016) Identification and Phylogenetic Classification of Pennisetum (Poaceae) Ornamental Grasses Based on SSR Locus Polymorphisms. Plant Mol Biol Rep 34:1181–1192.

Download references


We thank Universidade Federal do Espírito Santo for logistical support; the managers and employees of the Conservation Units for their support during visits; the owners of private areas for permission to collect on their properties; Fundação de Amparo à Pesquisa e Inovação do Espírito Santo - FAPES for the fellowship granted to the first author; and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for the grants to T.T. Carrijo and M.F.S. Ferreira. This study was financed in part by the Coordernação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) - Finance Code 001 to the first and second authors.

Author information

JL and TTC designed and conducted the study and wrote the article. JL, JAC, and MFS Ferreira conceived and conducted the molecular marker analysis. All authors equally contributed to the editing and revision of the manuscript and approved the final version for submission.

Correspondence to Tatiana Tavares Carrijo.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (DOCX 46 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Luber, J., Christ, J.A., Ferreira, M.F.d. et al. Species delimitation within Campomanesia (Myrtaceae) using morphology and amplification profiles of microsatellite markers. Braz. J. Bot (2020).

Download citation


  • Pimenta Group
  • SRR markers
  • Taxonomy
  • Transferability