Abstract
The present investigation was carried out to establish a simple and efficient in vitro method for rapid propagation of Daucus carota L for different applications. Root, stem, leaf and seed explants were tested to induce callus on Gamborg medium supplemented with 2,4-Dichlorophenoxyacetic acid (2,4-D) for cell suspension culture establishment. The leaf, stem, seed explants induced calluses were more embryogenic and viable when compared to callus derived from tap root explants. Plant regeneration potential of embryogenic callus from tap root showed very slow response producing only fewer shoots, whereas the response of other explants derived calluses like stem, leaves and seeds exhibited rapid multiple shoot formation within three weeks’ time. Tap root derived suspension culture showed more clumps with less viable embryogenic cells, in contrast to the stem and seed derived suspension cultures, where cells were more embryogenic and proliferative. Explant, concentration of carbohydrates, 2,4-D and amino acids were tested for embryo induction response and it was found that 3% sucrose, 1.0 mg/L 2,4-D and amino acid alanine had great influence on embryo induction. The somatic embryos were subjected to desiccation prior to plating on the regeneration medium. It was found that 1 h desiccation increased the regeneration percentage of the embryos and the conversion frequency of embryos to plants decreased on increased desiccation time. These optimized conditions will be very useful for studies involving genetic transformation, cell cycle, cell proliferation, synthetic seed production and other physiological studies of D. carota species.
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Acknowledgments
Author Sathish. S acknowledges Indian Council for Medical Research, New Delhi, India (No.3/1/2/102/2018-Nut.) for fellowship support. Safia N thanks University Grants Commission- Basic Science Research, New Delhi for fellowship support (UGC BSR No. F.25.1/2014-15). Dr. Sivakumar. S thanks National Post-Doctoral Fellowship (Sanction Order No: PDF/2016/002258), Science and Engineering Research Board, Department of Science and Technology, Government of India. We would also like to thank, University Grants Commission-Special Assistance Programme and Department of Science & Technology-Fund for Improvement of S&T Infrastructure for the financial support to carry out this research.
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Figure. S1
In vitro grown carrot plants used for explant preparation a b seedling grown in MS medium at 4 and 6 weeks c d individual plantlets used for steam and leaf explant preparation e lateral and tap root explants after excision Scale bar = 5 cm;d = 3cm (PDF 11662 kb)
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Sundararajan, S., Nayeem, S., Subiramani, S. et al. Optimizing culture conditions for high frequency somatic embryogenesis and plantlet conversion in Daucus carota L. Biologia 74, 695–707 (2019). https://doi.org/10.2478/s11756-019-00223-0
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DOI: https://doi.org/10.2478/s11756-019-00223-0