Development of a new culture medium and efficient protocol for in vitro micropropagation of Ceratonia siliqua L.

  • Assia LozziEmail author
  • Rabha Abdelwahd
  • Rachid Mentag
  • Abdelhadi Abousalim
Plant Tissue Culture


A new basal culture medium was developed and tested using a rapid and efficient protocol of in vitro axillary shoot bud proliferation of Ceratonia siliqua L., an important Mediterranean Fabaceae plant species. In a first experiment, the new formulated ‘LA’ mineral composition significantly improved shoot growth and proliferation as compared with Murashige and Skoog medium (MS, 1962) in both solid and liquid culture media. However, the liquid culture system proved to be the most suitable for shoot induction, shoot length (about fourfold higher), and multiplication rate (about two-fold higher), the difference being significant. The measured growth and proliferation parameters were further improved when LA mineral composition was optimized, in a second experiment. The highest multiplication rate (6.3) was achieved during the second subculture using the optimized ‘LAC’ medium. Noticeably, hyperhydricity and shoot-tip necrosis symptoms were absent in both formulated LA and LAC compositions when using the liquid culture system. In vitro rooting in solid medium showed 41.7 to 46.3% response on a solid medium which was more suitable than the liquid culture system, the difference being significant. In contrast, pretreated microcuttings with 3 μM IBA (indole-3-butyric acid) were successfully rooted ex vitro, showing significantly higher response (91.7%), average root number (8.3), and root length (31.5 mm). The plantlets were successfully acclimatized showing more than 90% survivability and normal morphology. The present study is a first cost-effective protocol for carob micropropagation combining the use of the newly formulated LAC basal medium, a liquid culture system, and ex vitro rooting.


Ceratonia siliqua Ex vitro rooting Micropropagation Liquid medium New basal medium 



The author acknowledges the assistance provided by Miss S. Sawabi in explants mineral assessment. Thanks are also due to Prof. Z. El Akkaoui for statistical analysis advice.

Compliance with ethical standards

Conflict of interest

A provisional patent application based on the technology described in this work has been filed.


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© The Society for In Vitro Biology 2019

Authors and Affiliations

  1. 1.Plant Tissue Culture Laboratory, Horticultural and Local Products Unit, Plant Production, Protection and Biotechnology DepartmentHassan II Institute of Agronomy and Veterinary MedicineRabat-InstitutsMorocco
  2. 2.Biotechnology Unit. Regional Center of Agricultural ResearchNational Institute of Agricultural ResearchRabatMorocco

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