Abstract
Trachyspermum ammi (L.) Sprague (Ajowan) is an endangered medicinal plant with useful pharmaceutical properties. Ex situ conservation of this medicinal plant needs the development of an in vitro regeneration protocol using somatic embryogenesis. In the present study, a high-precision image-processing approach was successfully applied to measure physical properties of embryogenic callus. Explant age and the concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), kinetin (Kin), and sucrose were used as inputs, and an artificial intelligence technique was applied to predict physical properties of embryogenic callus, and the number of somatic embryos produced. Artificial neural network (ANN) models were tested to find the best combinations of input variables that affected output variables. The lower values of root mean square error, and mean absolute error, and the highest values of determination coefficient, were achieved when all four input variables were applied to predict the number of somatic embryos, the area of the callus, the perimeter of the callus, the Feret diameter of the callus, the roundness of the callus, and the true density of the callus in ANN models. The highest measured and predicted number of somatic embryos were achieved from the interaction of 15-d-old explants × 1.5 mg L−1 2,4-D × 0.5 mg L−1 Kin × 2.5% (w/v) sucrose. Based on sensitivity analysis, the 2,4-D concentration was the most important component in the culture medium that affected the number of somatic embryos and physical properties of the embryogenic callus tissue.
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Acknowledgements
The authors are thankful to the Research Institute of Forests and Rangelands of Iran for procuring the seeds, and also grateful to Dr. M.H Asare, the secretary of science and technological development staff of medicinal plants and traditional medicine of Islamic Republic of Iran, for his kind support of ajowan project. The first author is thankful to Piama Svoboda for her kind assistance in the English language editing of the manuscript.
Funding
This work was financially supported by the Biotechnology Development Council of the Islamic Republic of Iran [grant number 950620].
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Editor: Randall P. Niedz
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Niazian, M., Sadat-Noori, S.A., Abdipour, M. et al. Image Processing and Artificial Neural Network-Based Models to Measure and Predict Physical Properties of Embryogenic Callus and Number of Somatic Embryos in Ajowan (Trachyspermum ammi (L.) Sprague). In Vitro Cell.Dev.Biol.-Plant 54, 54–68 (2018). https://doi.org/10.1007/s11627-017-9877-7
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DOI: https://doi.org/10.1007/s11627-017-9877-7