Physiology and Molecular Biology of Plants

, Volume 24, Issue 4, pp 693–702 | Cite as

Analysis of nutrient deficiencies affecting in vitro growth and development of Eucalyptus dunnii Maiden

  • Gustavo Pedro Javier OberschelpEmail author
  • Antônio Natal Gonçalves
Research Article


Although basal medium optimization is a key factor in the success of tissue culture, its mineral composition is frequently disregarded when optimizing in vitro propagation protocols. A previous work on Eucalyptus dunnii micropropagation suggests that excessive callus formation and leaf chlorosis are related to specific nutritional conditions of the basal media. Recently, a novel basal medium based on the mineral nutrient analysis of E. dunnii young stump shoots was developed and successfully tested in plant regeneration and micropropagation of E. dunnii, avoiding all these issues. Considering this basal medium as an ideal growth condition, a mild deprivation of each macro and micronutrient and of the total organic fraction was imposed to E. dunnii in vitro cultures for 30 d. As a result, K, Mg, Mn, Cl, Zn, Mo, Ni or Co deprivation quantitatively affected growth and development of axillary shoots. Moreover, leaf chlorosis and the development of organogenic callus under Fe deficiency, and leaf drop along with shoot tip necrosis under N deficiency were observed. These symptoms suggest that nutrient content in E. dunnii tissues needs to be above 420.3 mg kg−1 for Fe and 27.7 g kg−1 for N to avoid the symptoms of leaf chlorosis and shoot tip necrosis. Additionally, the main role of Mn in quantitative responses and the antagonism between ions, especially for Mg/K and Mg/Zn, were denoted by the multivariate analysis. Overall, these results make a relevant contribution to the optimization of in vitro propagation of E. dunnii and other hard-to-propagate related species.


Mineral nutrition Deficiency symptoms Micropropagation Eucalypts Chlorosis 



The authors thank the Instituto Nacional de Tecnología Agropecuaria (INTA) and the Escola Superior de Agricultura Luiz de Queiroz, University of São Paulo (ESALQ/USP) for the support.

Author contributions

Author 1 (corresponding author): Design of the work, data collection, data analysis and interpretation, drafting the article and critical revision of the article.

Author 2: Conception of the work, support, data interpretation and critical revision of the article.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2018

Authors and Affiliations

  1. 1.Instituto Nacional de Tecnología Agropecuaria (INTA), EEA ConcordiaConcordiaArgentina
  2. 2.Escola Superior de Agricultura “Luiz de Queiroz” (ESALQ), Depto. de Ciências Florestais, PPG em Recursos FlorestaisUniversidade de São Paulo (USP)PiracicabaBrazil

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