Wounding and medium formulation affect de novo shoot organogenic responses in hypocotyl-derived explants of annatto (Bixa orellana L.)
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Bixa orellana L. (annatto) is the unique exploited source for the production of bixin and norbixin natural dyes, which are widely used in the food, pharmaceutical, cosmetic, and textile industries. In tissue culture, adequate culture conditions are essential for the development of the plant. Among them, the saline composition of the culture medium and the type of explants are limiting factors. In this study, the effect of MS (Murashige and Skoog), Correia and colleagues (JADS) medium, Woody Plant Medium (WPM), and Driver Kuniyuki Walnut (DKW) culture media and wounding of hypocotyl explants on de novo shoot organogenesis (DNSO) of annatto were evaluated. It was demonstrated that JADS medium improved the nutritional status of shoots, which promoted the best development and quality of the plants generated after acclimatization, when compared to MS medium. The use of JADS medium for annatto DNSO during 45 d of culture was sufficient to regenerate and stimulate the growth of shoots, which had adequate elongation and rhizogenic responses. In addition, enlargement of the wound surface, using longitudinal sections of the hypocotyl explant cut surface down on JADS medium, significantly improved the frequency and number of newly formed shoots. This protocol enabled production of an average of 17 30-mm-long shoots per explant after 90 d of culture. These results will be useful for future studies related to annatto biotechnology.
KeywordsNutritional status Media formulation Hypocotyl explants Regeneration wounding response
Chr. Hansen Ind. Com. Ltda. (Valinhos, SP, Brazil) for kindly donating the seeds of Bixa orellana ‘Piave Vermelha’, and the Laboratory of Plant Anatomy-Plant Biology Department/UFV for the facilities and support of the anatomical analyses.
DFV, EMM, LNFC, and MVCS raised the in vitro plants for the experiments and performed the experiments; DSB, EMM, and DFV performed statistical analysis; and DFV, EMM, WCO, MGCC, LNFC, VBPN, MR, and AX contributed to the design and interpretation of the research and to the writing of the paper.
Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG, Grant number APQ-02372-17, Belo Horizonte, MG, Brazil), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior-Brasil (CAPES)–Finance Code 001 (Brasília, DF, Brazil) and CNPq (Brasília, DF, Brazil) provided financial support.
Compliance with ethical standards
Conflicts of interest
The authors declare that they have no conflicts of interest.
- Correia D, Gonçalves AN, Couto HYZ, Ribeiro MC (1995) Efeito do meio de cultura líquido e sólido no crescimento e desenvolvimento de gemas de Eucalyptus grandis x Eucalyptus urophylla na multiplicação in vitro. IPEF 48/49:107–116Google Scholar
- Driver JA, Kuniyuki AH (1984) In vitro propagation of paradox walnut root stock. HortiScience 19:507–509Google Scholar
- Empresa Brasileira de Pesquisa Agropecuária – EMBRAPA (2009) Manual de análises químicas de solos,plantas e fertilizantes. In: Silva FC (ed) 2nd ed. Embrapa Informação Tecnológica, Brasília, p 627Google Scholar
- George EF, de Klerk G-J (2008) The components of plant tissue culture media I: macro- and micro-nutrients. In: George EF, Hall MA, de Klerk G-J (eds) Plant propagation by tissue culture, 3rd ed, v. 1. The background. Springer-Verlag, DordrechtGoogle Scholar
- Iwase A, Harashima H, Ikeuchi M, Rymen B, Ohnuma M, Komaki S, Morohashi K, Kurata T, Nakata M, Ohme-Takagi M, Grotewold E, Grotewold E, Sugimoto K (2017) WIND1 promotes shoot regeneration through transcriptional activation of ENHANCER OF SHOOT REGENERATION1 in Arabidopsis. Plant Cell 29:54–69CrossRefGoogle Scholar
- Joseph N, Siril EA (2013) Multiplication of annatto (Bixa orellana L.) using cotyledons and leaf explants. Res Plant Biol 3:24–32Google Scholar
- Karnovsky MJ (1965) A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron microscopy. J Cell Biol 27:137–138Google Scholar
- Lewandowska M, Sirko A (2008) Recent advances in understanding plant response to sulfur-deficiency stress. Acta Biochimic Polonica 55:457–471Google Scholar
- Lloyd G, McCown BH (1980) Commercially-feasible micropropagation of mountain Laurel, Kalmia latifolia, by shoot tip culture. Int Plant Prop Soc Proc 30:421–427Google Scholar
- Lorenzi H, Matos FJA (2002) Plantas medicinais no Brasil: nativas e exóticas. Jardim Botânico Plantarum, Instituto Plantarum, Nova OdessaGoogle Scholar
- O’Brien TP, McCully ME (1981) The study of plant structure principles and selected methods. Termarcarphi Pty Ltd., MelbourneGoogle Scholar
- Parimalan R, Giridhar P, Gururaj HB, Ravishankar GA (2007) Organogenesis from cotyledon and hypocotyl-derived explants of japhara (Bixa orellana L.). Acta Bot Croatica 66:153–160Google Scholar
- Porra RJ, Thompson WA, Kriedemann PE (1989) Determination of accurate extinction coefficients and simultaneous equations for assaying chlorophylls a and b extracted with four different solvents: verification of the concentration of chlorophyll standards by atomic absorption spectroscopy. BBA-Bioenergetics 975:384–394Google Scholar
- Venugopalan A, Giridhar P, Ravishankar GA (2011) Food, ethanobotanical and diversified applications of Bixa orellana L.: a scope for its improvement through biotechnological mediation. Indian J fund Appl. Life Sci 1:9–31Google Scholar