Explant type and natural ventilation systems influence growth and content of carvacrol and thymol of Lippia gracilis Schauer

  • Luiz Eduardo Santos Lazzarini
  • Suzan Kelly Vilela Bertolucci
  • Alexandre Alves de Carvalho
  • Alexsandro Carvalho Santiago
  • Fernanda Ventorim Pacheco
  • Buhara Yucesan
  • José Eduardo Brasil Pereira PintoEmail author
Original Article


The objective of this study was to evaluate the effects of different natural ventilation systems and explant types on the growth and volatile compound content of Lippia gracilis cultured in vitro. The treatments consisted of four membrane systems (without membrane, with one, two, and four porous membranes) and two explant types (nodal segments with and without a pair of leaves). The evaluation of growth, photosynthetic pigments and chemical analysis of the volatile fraction were performed at 35 days of cultivation in half strength MS basal medium. Natural ventilation systems significantly influenced the in vitro growth and volatile fraction of L. gracilis. Explants with a pair of leaves obtained the best experimental responses. The natural ventilation system with four membranes provided the best growth parameters and leaf area response of L. gracilis explants with leaves. The photosynthetic pigments decreased with an increase in the number of porous membranes in the culture flask. Variations in the number, content, and profile of volatile compounds under the influence of natural ventilation systems were observed. Major constituents such as ρ-cymene, γ-terpinene, thymol, carvacrol, and E-caryophyllene, regardless of experimental conditions, were identified. The highest carvacrol and thymol contents were observed in plantlets grown in culture flasks with four porous membranes. To maximize the content of carvacrol and thymol from the in vitro culture of L. gracilis, explants with a pair of leaves and four porous membranes in culture flasks are recommended for use.


Alternative membrane Medicinal plant Micropropagation Volatile fraction 



Gas chromatography/mass spectrometry


No membrane system


Alternative membrane system with one filter


Alternative membrane system with two filters


Alternative membrane system with four filters




Murashige and Skoog


Principal component analysis



This study was financed in part by the National Council for Scientific and Technological Development (CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico), the Minas Gerais State Research Foundation (FAPEMIG - Fundação de Pesquisa do Estado de Minas Gerais), and the Coordination for the Improvement of Higher Education Personnel (CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brasil) (CAPES – Finance Code 001).

Author contribution statement

LESL, ACS and FVP performed the laboratory work and the experimental work. SKVB performed the chemical analyses. AAC participated in the statistical analysis. BY participated in the correction of the manuscript. JEBPP wrote and corrected the manuscript. The authors accepted the final version of our manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest.


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Luiz Eduardo Santos Lazzarini
    • 1
  • Suzan Kelly Vilela Bertolucci
    • 1
  • Alexandre Alves de Carvalho
    • 1
  • Alexsandro Carvalho Santiago
    • 1
  • Fernanda Ventorim Pacheco
    • 1
  • Buhara Yucesan
    • 2
  • José Eduardo Brasil Pereira Pinto
    • 1
    Email author
  1. 1.Laboratory of Tissue Culture and Medicinal Plants, Department of AgricultureFederal University of Lavras, University CampusLavrasBrazil
  2. 2.Faculty of Natural and Agricultural Sciences, Department of Seed Science and TechnologyAbant Izzet Baysal UniversityBoluTurkey

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