MICROPROPAGATION AND PLOIDY STABILITY OF Lippia lacunosa Mart. & Schauer: AN ENDANGERED BRAZILIAN MEDICINAL PLANT

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Authors

  • Diego Pandeló José Universidade Federal do Triângulo Mineiro, campus Universitário de Iturama
  • José Marcello Salabert de Campos Universidade Federal de Juiz de Fora, Instituto de Ciências Biológicas, Depto de Biologia, Laboratório de Genética
  • Lyderson Facio Viccini Universidade Federal de Juiz de Fora, Instituto de Ciências Biológicas, Depto de Biologia, Laboratório de Genética
  • Emilly Ruas Alkimim Universidade Federal do Triângulo Mineiro, campus Universitário de Iturama
  • Marcelo de Oliveira Santos Universidade Federal de Juiz de Fora, Instituto de Ciências Biológicas, Depto de Biologia, Laboratório de Genética

DOI:

https://doi.org/10.32404/rean.v6i1.3203

Abstract

Lippia lacunosa is a Brazilian savanna plant that belongs to the Verbenaceae family. It has been used in folk medicine as a treatment for different diseases. This species represents an endangered Brazilian medicinal plant, and this is the first report documenting a reliable protocol for the in vitro propagation and regeneration of L. lacunosa. Axenic explants were cultivated in MS medium containing different concentrations of naphthalene acetic acid (NAA) to induce root growth. The mean shoot length and the number of roots were highest with 0.06 mg·L-1 NAA. The highest number of buds in shoot regeneration was induced with 2 mg·L-1 6-benzylaminopurine (BA). To obtain a long-term culture, the dwarf shoots were elongated on MS media containing 0.5 mg·L-1 BA alternated with MS containing 2 mg·L-1 BA every 40 days. In the present protocol, the long-term shoots retained the ability to root even after long periods of BA treatment. In addition, we evaluated the nuclear DNA content and ploidy levels, including the occurrence of endopolyploidy, in long-term micropropagated plant leaves using flow cytometry analysis. The plants propagated in vitro over several years possessed nuclear DNA contents ranging from 2.940 to 3.095 pg, and no differences in DNA content were found among in vitro plants or between these plants and the control (L. lacunosa from a greenhouse with a DNA content of 3.08 pg). The flow cytometry analysis also demonstrated that there was no polyploidization. The present study will be useful for biotechnological approaches and provides the first estimate of the nuclear DNA content of this species using flow cytometry.

Author Biography

Diego Pandeló José, Universidade Federal do Triângulo Mineiro, campus Universitário de Iturama

Atua nas áreas de Genética, Biologia Molecular e Biotecnologia

References

(I) Alan, A.R., Zeng, H., Assani, A., Shi, W.L., McRae, H.E., Murch, S.J., Saxena, P.K., 2007. Assessment of genetic stability of the germplasm lines of medicinal plant Scutellaria baicalensis Georgi (Huang-qin) in long-term, in vitro maintained cultures. Plant Cell Reports, 26, 1345–1355.

(II) Campos, J.M.S., Sousa, S.M., Silva, P.S., Pinheiro, L.C., Sampaio, F., Viccini, L.F., 2011. Chromosome numbers and DNA C values in the genus Lippia (Verbenaceae). Plant Systematics and Evolution, 291, 133–140.

(III) Canter, P.H., Thomas, H., Ernst, E., 2005. Bringing medicinal plants into cultivation: opportunities and challenges for biotechnology. Trends in Biotechnology, 23, 180–185.

(IV) Dolezel, J., 1997. Application of flow cytometry for the study of plant genomes. Journal of Applied Genetics, 38, 285–302.

(V) Dolezel, J., Greilhuber, J., Suda, J., 2007. Flow Cytometry with Plant Cells, first ed. Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

(VI) Galbraith, D.W., Lambert, G.M., Macas, J., Dolezel, J., 2001. Analysis of Nuclear DNA Content and Ploidy in Higher Plants, in: Robinson, J.P., (Ed.), Current Protocols in Cytometry. John Wiley & Sons, New York, pp 7.6.1-7.6.22.

(VII) Gomide, M., Lemos, F., Reis, D., José, G., Lopes, M., Machadoc, M.A., Alves, T., Coelho, C.M., 2016. Identification of dysregulated microRNA expression and their potential role in the antiproliferative effect of the essential oils from four different Lippia species against the CT26.WT colon tumor cell line. Revista Brasileira de Farmacognosia, 26, 627–633.

(VIII) Jain, S.M., 2001. Tissue culture-derived variation in crop improvement. Euphytica, 118, 153–166.

(IX) Leitão, S.G., Castro, O., Fonseca, E.N., Julião, L.S., Tavares, E.S., Leo, R.R.T., Vieira, R.C., Oliveira, D.R., Leitão, G.G., Martino, V., Sulsen, V., Barbosa, Y.A.G., Pinheiro, D.P.G., Silva, P.E.A., Teixeira, D.F., Junior, I.N., Lourenço, M.C.S., 2006. Screening of Central and South American plant extracts for antimycobacterial activity by the Alamar Blue test. Revista Brasileira de Farmacognosia, 16, 6–11.

(X) Loureiro, J., Capelo, A., Brito, G., Rodriguez, E., Silva, S., Pinto, G., Santos, C., 2007. Micropropagation of Juniperus phoenicea from adult plant explants and analysis of ploidy stability using flow cytometry. Biologia Plantarum, 51, 7–14.

(XI) Marie, D., Brown, S.C., 1993. A cytometric exercise in plant DNA histograms, with 2C values for 70 species. Biology of the cell, 78, 41–51.

(XII) Murashige, T., Skoog, F., 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiologia Plantarum, 15, 473–497.

(XIII) Palomino, G., Dolezel, J., Cid, R., Brunnera, I., Méndez, I., Rubluo, A., 1999. Nuclear genome stability of Mammillaria san-angelensis (Cactaceae) regenerants induced by auxins in long-term in vitro culture. Plant Science, 141, 191–200.

(XIV) Pascual, M.E., Slowing, K., Carretero, E., Mata, D.S., Villara, A., 2001. Lippia: traditional uses, chemistry and pharmacology: a review. Journal of Ethnopharmacology, 76, 201–214.

(XV) Peixoto, P.H.P., Salimena, F.R.G., Santos, M.O., Garcia, L.S., Pierre, P.M.O., Viccini, L.F., Otoni, W.C., 2006. In vitro propagation of endangered Lippia filifolia mart. and schauer ex schauer. In Vitro Cellular & Developmental Biology - Plant, 42, 558–561.

(XVI) Phillips, R.L., Kaepples, S.M., Olhoft, P., 1994. Genetic instability of plant tissue cultures: breakdown of normal controls. Proceedings of the National Academy of Sciences of the United States of America, 91, 5222–5226.

(XVII) Salimena, F.R.G., Múlgura, M.E., 2015. Notas taxonômicas em Verbenaceae do Brasil. Rodriguésia, 66, 191–197.

(XVIII) Sousa, S.M., Torres, G.A., Viccini, L.F., 2012. Karyological studies in Brazilian species of Lippia L. (Verbenaceae). Anais da Academia Brasileira de Ciências, 84, 1029–1037.

(XIX) Troncoso, N.S., 1974. Los Géneros de Verbenáceas de Sudamérica extratropical (Argentina, Chile, Bolivia, Paraguay, Uruguay y Sur de Brasil). Darwiniana, 18, 295–412.

(XX) Viccini, L.F., Costa, D.C.S., Machado, M.A., Campos, A.L., 2004. Genetic diversity among nine species of Lippia (Verbenaceae) based on RAPD Markers. Plant Systematics and Evolution, 246, 1–8.

(XXI) Viccini, L.F., Praça, M.M., Romanel, E., Peixoto, P.H.P., Pierre, P., Souza, D., Sousa, S.M., Salimena, F.R.G., 2005. Chromosome numbers in the genus Lippia (Verbenaceae). Plant Systematics and Evolution, 256, 171–178.

(XXII) Vieira, R. F., 1999. Conservation of medicinal and aromatic plants in Brazil, in: Janick, J. (Ed.), Perspectives on new crops and new uses. VA: ASHS Press, Alexandria, pp 152–159.

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Published

2019-03-08

How to Cite

José, D. P., de Campos, J. M. S., Viccini, L. F., Alkimim, E. R., & Santos, M. de O. (2019). MICROPROPAGATION AND PLOIDY STABILITY OF Lippia lacunosa Mart. & Schauer: AN ENDANGERED BRAZILIAN MEDICINAL PLANT. REVISTA DE AGRICULTURA NEOTROPICAL, 6(1), 1–7. https://doi.org/10.32404/rean.v6i1.3203

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Vol. 6 No. 1 (2019): REVISTA DE AGRICULTURA NEOTROPICAL

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