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Multiplication and cryopreservation of adventitious roots of Cleome rosea Vahl

  • Developmental Biology/Morphogenesis
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Abstract

Adventitious root cultures of Cleome rosea were established using root segments from proximal and middle root regions of in vitro-propagated plants. The root explants were cultured in liquid MS medium supplemented with the following growth regulators: naphthaleneacetic acid (NAA), indole-3-acetic acid (IAA), indole-3-butyric acid (IBA), 2,4-dichlorophenoxyacetic acid (2,4-D), and N6-furfuryladenine (KIN). The highest biomass accumulation was obtained with roots, cultured in media with NAA. The use of 2,4-D resulted in callus formation, while explants cultured in the presence of KIN developed buds. Protocols for long-term maintenance of in vitro roots through cryopreservation by vitrification were also evaluated using plant vitrification solution 2 (PVS2). The use of NAA during pre- and post-culture steps was essential to recovery after exposure to liquid nitrogen. On the other hand, the presence or absence of light during culture maintenance had little effect on the process. The highest recovery frequency after cooling in liquid nitrogen (63.6 ± 14.4%) was reached after exposure to PVS2 for 30 min. The cryopreserved roots showed excellent ability to multiply, which remained stable for up to five subcultures. Moreover, these roots also displayed the capacity for shoot induction when cultivated on solid MS medium supplemented with 0.50 mg/L 6-benzyladenine (BA), reaching regeneration frequencies of 60–82% over five subcultures.

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Acknowledgments

This work was supported by the Fundação Carlos Chagas Filho de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) and the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq). The authors are grateful to Adriana M. Lanziotti for the valuable technical assistance.

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Authors and Affiliations

  1. Núcleo de Biotecnologia Vegetal, Instituto de Biologia Roberto Alcantara Gomes, Universidade do Estado do Rio de Janeiro (UERJ), Rio de Janeiro, RJ, Brazil

    Lívia da Silva Cordeiro, Claudia Simões-Gurgel & Norma Albarello

  2. Instituto de Biologia Roberto Alcantara Gomes, Núcleo de Biotecnologia Vegetal—Laboratório de Biotecnologia de Plantas (LABPLAN), Universidade do Estado do Rio de Janeiro—Centro Biomédico, Rua São Francisco Xavier, 524—PHLC, sala 509, Maracanã, 20550-013, Rio de Janeiro, RJ, Brazil

    Lívia da Silva Cordeiro

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  1. Lívia da Silva Cordeiro
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  2. Claudia Simões-Gurgel
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  3. Norma Albarello
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Correspondence to Lívia da Silva Cordeiro.

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Editor: Rida Shibli

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da Silva Cordeiro, L., Simões-Gurgel, C. & Albarello, N. Multiplication and cryopreservation of adventitious roots of Cleome rosea Vahl. In Vitro Cell.Dev.Biol.-Plant 51, 249–257 (2015). https://doi.org/10.1007/s11627-015-9688-7

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  • DOI: https://doi.org/10.1007/s11627-015-9688-7

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