Micropropagation of Eucalyptus benthamii to form a clonal micro-garden

• Published in: In vitro cellular & developmental biology. Plant Vol. 48; no. 5; pp. 478 - 487
• Main Authors: Brondani, Gilvano Ebling, de Wit Ondas, Heron Wilhelmus, Baccarin, Francisco José Benedini, Gonçalves, Antonio Natal, de Almeida, Marcilio
• Format: Journal Article
• Language: English
• Published: New York Springer-Verlag 01.10.2012; Springer Science + Business Media; Springer Nature B.V
• Subjects: acclimation; Acclimatization; Adventitious roots; Adventitious shoots; benzyladenine; Biomedical and Life Sciences; buds; cambium; Cell Biology; clones; Cloning; Cold regions; cold zones; Culture media; Developmental Biology; Eucalyptus benthamii; explants; Forestry; Gardens & gardening; genotype; Genotypes; gibberellic acid; Growth regulators; Histology; Immunization; Life Sciences; Micropropagation; Multiplication & division; naphthaleneacetic acid; Plant Breeding/Biotechnology; Plant cells; Plant Genetics and Genomics; Plant growth; Plant growth regulators; Plant Sciences; Plant species; plantation forestry; Plants; Rooting; Seeds; Shoots; trees; wood; Micro-stumps; Acclimatization; Multiplication; rooting; and; Elongation
• ISSN: 1054-5476; 1475-2689
• DOI: 10.1007/s11627-012-9449-9

Eucalyptus benthamii is an important component of forestry plantations in cold regions, but it is difficult to obtain clonal plants of this species, especially by low rooting. In this study, we developed a method for cloning selected genotypes of E. benthamii using a micropropagation technique, enabling the formation of a clonal micro-garden. Nodal segments from sprouts of mini-stumps in the clonal mini-garden were used as explants. After in vitro establishment of the explants, we tested two selected clones (BP101 and BP118), three culture media (Wood Plant Medium (WPM), Correia and colleagues JADS medium, and Murashige and Skoog medium), and two plant growth regulators (6-benzylaminopurine (BAP) and α-naphthaleneacetic acid (NAA)) for the multiplication of adventitious buds. Additionally, combinations of two other plant growth regulators (BAP and gibberellic acid (GA3)) were tested for the elongation of shoots. The in vitro and ex vitro rooting of micro-plantlets prior to acclimatization were compared. The in vitro bud multiplication of E. benthamii depended on the clone, culture medium, and concentration of plant growth regulators. The best results were obtained with WPM supplemented with 0.5 mg L−1 BAP and 0.05 mg L−1 NAA. The elongation of shoots depended on the clone and plant growth regulator, and the best results were obtained with nutrient medium free of GA3 and BAP. Histological analysis showed that both in vitro and ex vitro rooting were successful, resulting in normal development of adventitious roots showing a vascular connection with the vascular cambium. The new protocol is efficient for micro-plantlet production of E. benthamii and can be used for the formation of a clonal micro-garden for other Eucalyptus or tree species.