Embryogenesis from microinjected single cells in a carrot cell suspension culture |
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| Affiliation: | 1. Department of Mechanical Engineering, Federal University of Petroleum Resources, Effurun, Delta State P.M.B 1221, Nigeria;2. Department of Mechanical Engineering, Federal University of Agriculture, Abeokuta, Nigeria;3. Mechanical Engineering of Biosystems Department, Lorestan University, Khorramabad, Iran;4. Department of Mechanical Engineering, Maulana Azad National Institute of Technology, Bhopal, MP 462003, India;5. Department of Agricultural Engineering, Federal University of Agriculture, Abeokuta, Nigeria;6. Department of Mechanical Engineering, Faculty of Engineering, Ikole-Ekiti Campus, Federal University Oye-Ekiti, P.M.B. 373, Oye-Ekiti, Nigeria;7. Department of Mechanical Engineering, P. A. College of Engineering Affiliated to Visvesvaraya Technological University, Belagavi, Mangaluru, India;1. Santa Cruz Genomics Institute, MS CBSE, 1156 High Street, University of California, Santa Cruz, CA 95064, USA;2. Laboratoire Structure et Instabilité des Génomes, Inserm U1154, CNRS UMR7196, Muséum national d''Histoire naturelle, 43 rue Cuvier, 75005 Paris, France |
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| Abstract: | A micrroinjection method was established for intact single cells with cell walls using a carrot suspension culture system in which selected single cells differentiate to embryos at high frequency. A solution of a fluorescent dye, Lucifer Yellow CH, was microinjected into those single cells, using an inverted microscope and a hydraulic micro-manipulator. In order to hold cells with cell walls and to overcome their turgor pressure, certain modification to conventional microinjection methods for protoplasts were necessary. The microinjected cells could divide and differentiate to embryos at a frequency of about 50%. |
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