New Forests

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In vitro serial subculture to improve rooting of Eucalyptus urophylla

  • Evânia Galvão MendonçaEmail author
  • Tânia Regina Batista
  • Vanessa Cristina Stein
  • Flávia Pereira Balieiro
  • José Renato de Abreu
  • Marinês Ferreira Pires
  • Patrícia Aparecida de Souza
  • Luciano Vilela Paiva


The aim of this study was to improve the rooting efficiency of Eucalyptus urophylla clones by in vitro reinvigoration/rejuvenation in two clones (02 and 04) from the breeding program of the V&M Florestal company. An in vitro culture began with 200 meristems of each clone, which were excised, disinfected, and inoculated in culture medium. When shoots from these first meristems inoculated reached a height of 3 cm, 100 new meristematic regions of 0.5 cm were isolated and inoculated in culture medium. The other shoots from were inoculated in a rooting medium, where they remained for 30 days. After this period, the plants were acclimatized and used as stock plants for shoot production in a commercial nursery. This process was repeated until the shoots attained an ex vitro rooting rate of more than 80%. After reinvigoration/rejuvenation of clones 02 and 04, the relationship between rooting and the presence of starch and phenolic compounds at the base of the minicuttings was histochemically analyzed. For clone 02, three in vitro subcultures were needed to increase the rooting rate, and for clone 04, only one in vitro subculture was required. In vitro reinvigoration/rejuvenation is a determining factor for greater rooting efficiency of minicuttings of 02 and 04 clones. Production of sclerenchyma fibers around the root vascular cylinder and starch and phenolic compound production are directly related to rooting efficiency.


Phenolic compounds Rooting Anatomical barriers Rejuvenation Starch Clonal forestry 



Our thanks to the V&M Florestal company for donating the eucalypt plants used in this study and to the National Research Council (CNPq), Coordination for the Improvement of Personnel in Higher Education (Capes), and the Research Support Foundation of the State of Minas Gerais (FAPEMIG) for financial support.

Author contributions

All authors contributed equally to all parts of the development and revision of this work.

Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest.

Supplementary material

11056_2019_9761_MOESM1_ESM.jpg (112 kb)
Fig. 1Rejuvenation of clone 02 from natural Eucalyptus urophylla, 21 days after pruning. Nursery plants (1), G1 (2), G2 (3), G3 (4), and G4 (5). (a) Tubette plants. (b) Plants without tubette. Scale bar – 2 cm. (JPEG 111 kb)
11056_2019_9761_MOESM2_ESM.jpg (107 kb)
Fig. 2Rejuvenation of clone 04 from natural Eucalyptus urophylla, 21 days after pruning. Nursery plants (1), G1 (2), G2 (3), and G3 (4). (a) Tubette plants. (b) Plants without tubette. Scale bar – 2 cm. (JPEG 107 kb)
11056_2019_9761_MOESM3_ESM.jpg (373 kb)
Fig. 3Cross sections of the base of Eucalyptus urophylla stem minicuttings from clone 02. (a) Nursery plants. (b) First subculture generation. (c) Second subculture generation. (d) Third subculture generation stained with safrablau solution. ep – epidermis. cx - cortex. sc - sclerenchyma. ph - phloema. xl - xylem. ss - secretory structure. mp - medullary parenchyma. (JPEG 372 kb)
11056_2019_9761_MOESM4_ESM.jpg (372 kb)
Fig. 4Cross sections of the base of Eucalyptus urophylla stem minicuttings from clone 04. (a) Nursery plants. (b) First subculture generation. (c) Second subculture generation. (d) Third subculture generation. ep – epidermis. cx - cortex. sc - sclerenchyma. ph - phloema. xl - xylem. mp - medullary parenchyma. (JPEG 372 kb)


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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Evânia Galvão Mendonça
    • 1
    Email author
  • Tânia Regina Batista
    • 2
  • Vanessa Cristina Stein
    • 3
  • Flávia Pereira Balieiro
    • 4
  • José Renato de Abreu
    • 5
  • Marinês Ferreira Pires
    • 6
  • Patrícia Aparecida de Souza
    • 1
  • Luciano Vilela Paiva
    • 6
  1. 1.Departamento de Ciências FlorestaisUniversidade Federal de São João del-ReiSete LagoasBrazil
  2. 2.Instituto de Pesquisas e Estudos FlorestaisPiracicabaBrazil
  3. 3.Laboratório de FarmacobotânicaUniversidade Federal de São João del-ReiDivinópolisBrazil
  4. 4.Centro de Tecnologia CanavieiraPiracicabaBrazil
  5. 5.Denpasa- Dendê do BrasilSanta Bárbara do ParáBrazil
  6. 6.Departamento de BiologiaUniversidade Federal de LavrasLavrasBrazil

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