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Establishment of an in vitro propagation protocol for Thymus lotocephalus, a rare aromatic species of the Algarve (Portugal)

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Abstract

The aim of this work was to develop an in vitro propagation protocol for the endangered species Thymus lotocephalus using seedlings as explants. Several macronutrient concentrations of Murashige and Skoog medium (MS), cytokinin types and concentrations, and cytokinin/auxin combinations were tested to assess the shoots’ proliferation capacity. Although the best proliferation results were obtained with 6-benzyladenine, high percentages of hyperhidric shoots were observed. Because high proliferation of healthy shoots was observed in MS medium that was free of plant growth regulators, this medium was chosen for proliferation studies. The best rooting results were achieved in ¼MS medium without auxins (92.00 ± 6.11%, 6.54 ± 0.52 and 1.60 ± 0.10 cm regarding rooting frequency, number of roots per shoot and longest roots, respectively) or supplemented with 0.5 mg l−1 indole-3-acetic acid (98.00 ± 2.11%, 11.14 ± 0.75 and 2.40 ± 0.24 cm, respectively). Plantlets were successfully acclimatised to ex vitro conditions with a survival rate of 93.33%. With the development of this micropropagation protocol, an important contribution has been made to the conservation of the endangered species T. lotocephalus.

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Abbreviations

2,4-D:

2,4-Dichlorophenoxyacetic

BA:

6-Benzyladenine

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butyric acid

KIN:

Kinetin

MS:

Murashige and Skoog medium (1962)

NAA:

1-Naphthalene-acetic acid

PGRs:

Plant growth regulators

ZEA:

Zeatin

References

  • Andrade LB, Echeverrigaray S, Fracaro F, Pauletti GF, Rota L (1999) The effect of growth regulators on shoot propagation and rooting of common lavender (Lavandula vera DC). Plant Cell Tissue Organ Cult 56:79–83

    Article  CAS  Google Scholar 

  • Benson EE, Danaher JE, Pimbley IM, Anderson CT, Wake JE, Daley S, Adams LK (2000) In vitro micropropagation of Primula scotica: a rare Scottish plant. Biodiv Conserv 9:711–726

    Article  Google Scholar 

  • Bouhouche N, Ksiksi T (2007) An efficient in vitro plant regeneration system for the medicinal plant Teucrium stocksianum Boiss. Plant Biotechnol Rep 1:179–184

    Article  Google Scholar 

  • Cosentino S, Tuberoso CIG, Pisano B, Satta M, Mascia V, Arzedi E, Palmas F (1999) In vitro antimicrobial activity and chemical composition of Sardinian Thymus essential oils. Lett Appl Microbiol 29:130–135

    Article  PubMed  CAS  Google Scholar 

  • Cuenca S, Amo-Marco JB (2000) In vitro propagation of two Spanish endemic species of Salvia through bud multiplication. In Vitro Cell Dev Biol Plant 36:225–229

    Article  CAS  Google Scholar 

  • de Klerk G-J (2002) Rooting of microcuttings: theory and practice. In Vitro Cell Dev Biol Plant 38:415–422

    Article  Google Scholar 

  • de Klerk G-J, van der Krieken W, de Jong JC (1999) The formation of adventitious roots: new concepts, new possibilities. In Vitro Cell Dev Biol Plant 35:189–199

    Article  Google Scholar 

  • Dias MC, Almeida R, Romano A (2002) Rapid clonal multiplication of Lavandula viridis L’Hér through in vitro axillary shoot proliferation. Plant Cell Tissue Organ Cult 68:99–102

    Article  CAS  Google Scholar 

  • Faleiro ML, Miguel MG, Ladeiro F, Venâncio F, Tavares R, Brito JC, Figueiredo AC, Barroso JG, Pedro LG (2003) Antimicrobial activity of essential oils isolated from Portuguese endemic species of Thymus. Lett Appl Microbiol 36:35–40

    Article  PubMed  CAS  Google Scholar 

  • Fay M (1992) Conservation of rare and endangered plants using in vitro methods. In Vitro Cell Dev Biol Plant 28:1–4

    Google Scholar 

  • Figueiredo AC, Miguel MG, Duarte AMF, Barroso JG, Pedro LG (2001) Essential oil composition of Thymus lotocephalus G. López & R. Morales, collected during flowering and vegetative phases. Flavour Frag J 16:417–421

    Article  CAS  Google Scholar 

  • Furmanowa M, Olszowska O (1992) Micropropagation of Thyme (Thymus vulgaris L.). In: Stahl-Biskup E, Saéz F (eds) Thyme: the genus Thymus, medicinal and aromatic plants—industrial profiles. Taylor & Francis, London, pp 177–196

    Google Scholar 

  • Gonçalves S, Romano A (2005) Micropropagation of Drosophyllum lusitanicum (L.) Link. an endangered West Mediterranean endemic insectivorous plant. Biodiv Conserv 14:1071–1081

    Article  Google Scholar 

  • Gonçalves S, Fernandes L, Romano A (2010) High-frequency in vitro propagation of the endangered species Tuberaria major. Plant Cell Tissue Organ Cult 101:359–363

    Article  Google Scholar 

  • Hazarika BN (2006) Morpho-physiological disorders in in vitro culture of plants. Sci Hortic 108:105–120

    Article  CAS  Google Scholar 

  • ICN (Instituto da Conservação da Natureza) (2006) Plano Sectorial da Rede Natura. Flora: Thymus lotocephalus G. López & R. Morales. http://www.icn.pt/psrn2000/caracterizacao_valores_naturais/flora/Thymus%20lotocephalus.pdf. Accessed 4 April 2011

  • Ioio RD, Nakamura K, Moubayidin L, Perilli S, Taniguchi M, Morita MT, Aoyama T, Costantino P, Sabatini S (2008) Root system architecture: opportunities and constraints for genetic improvement of crops. Science 322:1380–1384

    Article  Google Scholar 

  • Lê CL (1989) Microbouturage in vitro du thym (Thymus vulgaris L.). In: Stahl-Biskup E, Saéz F (eds) Thyme: the genus Thymus, medicinal and aromatic plants—industrial profiles. Taylor & Francis, London, pp 177–196

    Google Scholar 

  • Mendes ML, Romano A (1999) In vitro cloning of Thymus mastichina L. field-grown plants. Acta Hort 502:303–306

    CAS  Google Scholar 

  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Ozudogru EA, Kaya E, Kirdok E, Issever-Ozturk S (2011) In vitro propagation from young and mature explants of thyme (Thymus vulgaris and T. longicaulis) resulting in genetically stable shoots. In Vitro Cell Dev Biol Plant 47:309–320

    Article  Google Scholar 

  • Paek K-Y, Hahn E-J (2000) Cytokinins, auxins and activated charcoal affected organogenesis and anatomical characteristics of shoot-tip cultures of Lisianthus [Eustoma grandiflorum (raf.) Shinn]. In Vitro Cell Dev Biol Plant 36:128–132

    Article  CAS  Google Scholar 

  • Saéz F, Sánchez P, Piqueras A (1994) Micropropagation of Thymus piperella. Plant Cell Tissue Organ Cult 39:269–272

    Article  Google Scholar 

  • Salgueiro LR, Vila R, Tomàs X, Cañigueral S, Paiva J, Cunha A-P, Adzet T (2000) Essential oil composition and variability of Thymus lotocephalus and Thymus × mourae. Biochem Syst Ecol 28:457–470

    Article  PubMed  CAS  Google Scholar 

  • Sorin C, Bussell JD, Camus I, Ljung K, Kowalczyk M, Geiss G, McKhann H, Garcion C, Vaucheret H, Sandberg G, Bellinia C (2005) Auxin and light control of adventitious rooting in Arabidopsis require ARGONAUTE1. Plant Cell 17:1–17

    Article  Google Scholar 

  • Stahl-Biskup E, Saéz F (2002) Thyme: the genus Thymus, medicinal and aromatic plants—industrial profiles. Taylor & Francis, London

    Google Scholar 

Download references

Acknowledgments

N Coelho and S Gonçalves acknowledge a grant from the Portuguese Science and Technology Foundation (FCT, grants SFRH/BD/63501/2009 and SFRH/BPD/31534/2006, respectively).

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Authors and Affiliations

  1. Institute for Biotechnology and Bioengineering, Centre of Genomics and Biotechnology (IBB/CGB), Faculty of Sciences and Technology, University of Algarve, Campus de Gambelas, Ed. 8, 8005-139, Faro, Portugal

    Natacha Coelho, Sandra Gonçalves & Anabela Romano

  2. Departamento de Biología Vegetal, Escuela Universitaria de Ingeniería Técnica Agrícola, Universidad Politécnica de Madrid, Ciudad Universitaria s/n, 28040, Madrid, Spain

    María Elena González-Benito

Authors
  1. Natacha Coelho
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  2. Sandra Gonçalves
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  3. María Elena González-Benito
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  4. Anabela Romano
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Correspondence to Anabela Romano.

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Coelho, N., Gonçalves, S., González-Benito, M.E. et al. Establishment of an in vitro propagation protocol for Thymus lotocephalus, a rare aromatic species of the Algarve (Portugal). Plant Growth Regul 66, 69–74 (2012). https://doi.org/10.1007/s10725-011-9630-x

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  • DOI: https://doi.org/10.1007/s10725-011-9630-x

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