Optimizing culture conditions for high frequency somatic embryogenesis and plantlet conversion in Daucus carota L
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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.
KeywordsDaucus carota L. Somatic embryogenesis Desiccation Regeneration Amino acids
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.
Compliance with ethical standards
Conflict of interest
The authors declare that there is no conflict of interest.
- Ammirato PV (1986) Carrot. In: Evans DA, Sharp WR, Ammirato PV (eds) Handbook of plant cell culture: techniques and application, vol 4. Macmillan, New York, pp 457–499Google Scholar
- Elhag HM, Whipkey A, Janick J (1987) Induction of somatic embryogenesis from callus in Theobroma cacao in response to carbon source and concentration. Rev Theobroma 17:153–162Google Scholar
- Gaj MD (2004) Factors influencing somatic embryogenesis induction and plant regeneration with particular reference to Arabidopsis thaliana (L.) Heynh. Plant Growth Regul 43:27–47. https://doi.org/10.1023/B:GROW.0000038275.29262.fb CrossRefGoogle Scholar
- Guy CL (1990) Cold acclimation and freezing stress tolerance: role of protein metabolism. Annu Rev Plant Physiol Plant Mol Biol 41:187–223. https://doi.org/10.1146/annurev.pp41.060190.001155 CrossRefGoogle Scholar
- Kochhar S L (Ed.) (1998) Economic botany in the tropics. Macmillan India Ltd, Delhi, p 608Google Scholar
- Komamine A, Kawahara R, Matsumoto M, Sunabori S, Toya T, Fujiwara A, Tsukuhara M, Smith J, Ito M, Fukuda H, Nomura K, Fujimura T (1992) Mechanisms of somatic embryogenesis in cell cultures: physiology, biochemistry, and molecular biology. In Vitro Cell Dev Biol-Plant 28:11–14. https://doi.org/10.1007/BF02632185 CrossRefGoogle Scholar
- Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x CrossRefGoogle Scholar
- Nomura K, Komamine A (1986) Molecular mechanisms of somatic embryogenesis. Oxford Surveys Plant Mol Cell Biol 3:456–466Google Scholar
- Pescador R, Kerbauy GB, De Melo Ferreira W, Purgatto E, Suzuki RM, Guerra MP (2012) A hormonal misunderstanding in Acca sellowiana embryogenesis: levels of zygotic embryogenesis do not match those of somatic embryogenesis. Plant Growth Regul 68:67–76. https://doi.org/10.1007/s10725-012-9694-2 CrossRefGoogle Scholar
- Raghavan V (2006) Can carrot and Arabidopsis serve as model systems to study the molecular biology of somatic embryogenesis? Curr Sci 90:1336–1343 Google Scholar
- Sotiropoulos TE, Molassiotis AN, Mouhtaridou GI, Papadakis I, Dimassi KN, Therios IN, Diamantidis G (2006) Sucrose and sorbitol effects on shoot growth and proliferation in vitro, nutritional status and peroxidase and catalase isoenzymes of M 9 and MM 106 apple (Malus domestica Borkh.) rootstocks. Eur J Hortic Sci 71:114–119Google Scholar
- Steward FC, Mapes MO, Mears K (1958) Growth and organized development of cultured cells. II. Organization in cultures grown from freely suspended cells. Am J Bot 45:705–708. https://doi.org/10.1002/j.1537-2197.1958.tb10599.x CrossRefGoogle Scholar