Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 125, Issue 3, pp 445–456

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

DOI: 10.1007/s11240-016-0959-8

Cite this article as:
Valdevite, M., Bertoni, B.W., Contini, S.H.T. et al. Plant Cell Tiss Organ Cult (2016) 125: 445. doi:10.1007/s11240-016-0959-8
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Abstract

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.

Keywords

In vitro cultures Micropropagation Iridoid glucoside Differential gene expression Rubiaceae 

Abbreviations

2iP

2-Isopentenyl adenine

ANOVA

Analysis of variance

BAP

6-Benzylaminopurine

DEG

Differentially expressed genes

HPLC

High-performance liquid chromatography

IAA

Indole-3-acetic acid

IBA

Indole-3-butyric acid (IBA)

LOD

Limit of detection

LOQ

Limit of quantification

MS

Murashige and Skoog

NAA

Naphthalene acetic acid (NAA)

PCoA

Principal coordinates analysis

RT-PCR

Reverse transcription-polymerase chain reaction

S/N

Signal to noise ratio

SRAP

Sequence-related amplified polymorphism

TBE

Tris–borate-EDTA buffer

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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