Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 125, Issue 3, pp 457–470 | Cite as

Establishment of an in vitro propagation and transformation system of Balanites aegyptiaca

  • Galal Khamis
  • Traud Winkelmann
  • Frank Schaarschmidt
  • Jutta PapenbrockEmail author
Original Article


Balanites aegyptiaca (Balanitaceae) is a drought-tolerant but salt-sensitive tree species distributed in the tropical and arid lands in Africa and Asia. The tree contains many secondary metabolites and a high percentage of oil in the kernels that can be used for biodiesel production. This study aimed to establish an in vitro propagation system of two B. aegyptiaca provenances (El-Kharga and Wadi El-Alaqi) from nodal and cotyledon explants. The explants were placed on Murashige and Skoog medium supplemented with different concentrations of 6-benzyladenine (BA) and thidiazuron (TDZ) for shoot induction. BA was significantly more effective in shoot induction from nodal explants and treatment with BA also resulted in higher regeneration rates of about 40–60 % of adventitious shoots on cotyledon explants, whereas on TDZ-containing medium slightly higher shoot numbers per explant but a negative effect on shoot length were recorded. Rooting was achieved in 40–60 % of the shoots on medium containing between 1.2 and 4.8 µM indole-3-butyric acid. Three different Agrobacterium tumefaciens strains (EHA105, GV3101, and LBA4404) harboring the plasmid pCAMBIA2301 containing the nptII marker and gus reporter genes were used to establish a transformation system in B. aegyptiaca. Strain GV3101 resulted in the highest survival rates and highest number of explants positive in the GUS assay. This selected A. tumefaciens strain was used to introduce pBinAR containing the sequence encoding ERD10 (early responsive to dehydration 10) to produce salt-tolerant B. aegyptiaca plants. The presence of the ERD10 and the nptII gene were detected by PCR in transformed B. aegyptiaca plants.


Agrobacterium tumefaciens Balanites aegyptiaca ERD10 Regeneration Rooting Salt stress 



We would like to thank Prof. Usama Radwan (Environmental Studies and Development Unit, Faculty of Sciences, Aswan University, Egypt) for the kind help to collect fruits of two genotypes (El-Kharga, Wadi El-Alaqi). We would like to thank the German Academic Exchange Service (DAAD) and the Ministry of Higher Education (MoHE) of the Arab Republic of Egypt cooperation agreement for the support and providing the PhD scholarship to Galal Khamis.

Author contributions

Galal Khamis has conducted all experiments. Jutta Papenbrock and Traud Winkelmann have planned, organized and supervised all experiments. Frank Schaarschmidt supported planning the experiments and evaluated the data statistically. The manuscript was written by all authors.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.


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Copyright information

© Springer Science+Business Media Dordrecht 2016

Authors and Affiliations

  • Galal Khamis
    • 1
    • 2
  • Traud Winkelmann
    • 3
  • Frank Schaarschmidt
    • 4
  • Jutta Papenbrock
    • 1
    Email author
  1. 1.Institute of BotanyLeibniz Universität HannoverHannoverGermany
  2. 2.Department of Laser Applications in Metrology, Photochemistry and Agriculture (LAMPA), National Institute of Laser Enhanced SciencesCairo UniversityGizaEgypt
  3. 3.Institute of Horticultural Production SystemsLeibniz Universität HannoverHannoverGermany
  4. 4.Institute of BiostatisticsLeibniz Universität Hannover30419 HannoverGermany

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