Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 125, Issue 3, pp 445–456 | Cite as

Accumulation of loganin by genotypes of Palicourea rigida and related differential gene expression as determined by cDNA-SRAP

  • Mayara Valdevite
  • Bianca Waléria Bertoni
  • Silvia Helena Taleb Contini
  • Suzelei de Castro França
  • Ana Maria Soares Pereira
Original Article


Palicourea rigida, an endangered medicinal plant from the Brazilian Cerrado, produces the iridoid glucoside loganin as an active principle. The objectives of this study were to establish a micropropagation method for the species, to evaluate loganin accumulation in different genotypes cultivated in vitro, and to perform cDNA-sequence-related amplified polymorphism (cDNA-SRAP) analysis to discriminate the genotypes. Shoot cultures were initiated from apical explants derived from 7-day-old seedlings and incubated on agar medium without growth regulators. Nodal segments were excised from established shoots and cultured on agar medium supplemented with 6-benzylaminopurine (BAP), kinetin or 2-isopentenyl adenine. Rooting experiments in vitro were conducted using agar medium supplemented with 1-naphthaleneacetic acid or indole-3-butyric acid as well as common substrates (sand, sandy soil and clay soil), while rooting experiments ex vitro were performed using sandy soil or commercial Bioplant® substrate. Multiplication of shoots in vitro was improved by the presence of 0.1 µM BAP, while rooting only occurred in clay soil. Rooting under ex vitro conditions was improved when sandy soil was employed as substrate. High-performance liquid chromatographic analysis revealed that shoots cultured in the presence of 0.1 µM kinetin exhibited maximum levels of loganin (32.6 mg/g dry weight). cDNA-SRAP analysis was efficient in discriminating seven genotypes generated from seeds collected in the Cerrado. The genotypes were clustered into five groups, four of which had similar chemical profiles while the most genetically distant group accumulated no detectable amounts of loganin and could be classified as a chemotype.


In vitro cultures Micropropagation Iridoid glucoside Differential gene expression Rubiaceae 



2-Isopentenyl adenine


Analysis of variance




Differentially expressed genes


High-performance liquid chromatography


Indole-3-acetic acid


Indole-3-butyric acid (IBA)


Limit of detection


Limit of quantification


Murashige and Skoog


Naphthalene acetic acid (NAA)


Principal coordinates analysis


Reverse transcription-polymerase chain reaction


Signal to noise ratio


Sequence-related amplified polymorphism


Tris–borate-EDTA buffer



The authors wish to thank the Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP) for financial support for the study (Grant No. 07/58503-3).

Authors’ contribution

Study conception and design: AMSP, MV; Acquisition of data: MV, AMSP; Analysis and interpretation of data: MV, AMSP, BWB, SHTC; Drafting of manuscript: AMSP; Critical revision: BWB, SCF.


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Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Mayara Valdevite
    • 1
  • Bianca Waléria Bertoni
    • 1
  • Silvia Helena Taleb Contini
    • 1
  • Suzelei de Castro França
    • 1
  • Ana Maria Soares Pereira
    • 1
  1. 1.Departamento de Biotecnologia em Plantas MedicinaisUniversidade de Ribeirão PretoRibeirão PrêtoBrazil

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