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Optimization of somatic embryogenesis in suspension cultures of horsegram [Macrotyloma uniflorum (Lam.) Verdc.]—A hardy grain legume
Institution:1. Laboratory of Molecular Genetics, Agronomy Division, Taiwan Agricultural Research Institute, Council of Agriculture, 189 ChungCheng Rd., Wufeng, Taichung 413, Taiwan, ROC;2. Department of Food Science and Nutrition, Hung Kuang University, Sha Lu, Taichung 433, Taiwan, ROC;1. Institute of Biology and Soil Science, Far East Branch of Russian Academy of Sciences, Vladivostok 690022, Russia;2. Far Eastern Federal University, Vladivostok 690950, Russia;1. Junior Research Group Cellular Immunobiology, Leibniz Institute for Natural Product Research and Infection Biology – Hans Knöll Institute, Jena, Germany;2. MTA-ELTE “Lendület” Complement Research Group, Department of Immunology, Eötvös Loránd University, Budapest, Hungary;3. MTA-ELTE Immunology Research Group, Department of Immunology, Eötvös Loránd University, Budapest, Hungary;4. Junior Research Group Fundamental Molecular Biology of Pathogenic Fungi, Leibniz Institute for Natural Product Research and Infection Biology – Hans Knöll Institute, Jena, Germany;5. Research and Development, Kneipp GmbH, Würzburg, Germany;6. Department of Immunology, Eötvös Loránd University, Budapest, Hungary;7. Institute for Transfusion Medicine, University Hospital of Jena, Germany;1. School of Chinese Pharmacy, Beijing University of Chinese Medicine, Beijing, China;2. Henan Province for Drug Evaluation Certification Center, Zhengzhou, China;1. Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot, 76100, Israel;2. Institute of Soil, Water and Environmental Sciences, Agricultural Research Organization, Volcani Center, POB 6, Bet Dagan, 50250, Israel;3. Institute of Plant Sciences, Agricultural Research Organization, Newe Ya''ar Research Center, Ramat Yishay, 30095, Israel;1. Department of Physics, Regional Centre, Anna University Tirunelveli Region, Tirunelveli 627 007, India;2. Department of Physics, University College of Engineering, Nagercoil, Anna University, Nagercoil 629 004, India;3. Department of Physics, Madurai Kamaraj University, Madurai 625 021, India
Abstract:Cell suspension cultures were established from immature cotyledon derived calli from drought tolerant legume horsegram Macrotyloma uniflorum (Lam.) Verdc.]. Embryogenic callus could be originated from cut slices of the immature cotyledons on MS solid medium Murashige, T. Skoog, K., 1962. A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15, 473–497] augmented with 1.0 μM zeatin and 4.5 μM NAA. Numerous somatic embryoids (26.4%) appeared on MS liquid basal nutrient medium with 5.6 μM NAA and with absence of zeatin during 3 weeks culture. Sustained cell division resulted in the formation of cell aggregates, and then progressed to globular, heart and further if they differentiate properly to torpedo and cotyledonary stages within 5 weeks. Transfer of individual embryos on to a fresh MS basal medium with no plant growth regulators was able to achieve complete maturation. Only a relatively few number of embryos developed into root/shoot when transferred to 0.9 μM GA3, 15 g/l−1 sucrose and 2.4 g/l−1 gelrite containing medium. Substitution of sucrose associated with the use of l-glutamine gave, in the range of concentrations tested, the strongest enhancement of the embryo growth and development. About 5% of somatic embryos were converted into true-to-type fertile plants.
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