Aluminum borate whisker-mediated production of transgenic tobacco plants |
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Authors: | Kiyotaka?Mizuno Wataru?Takahashi Toshio?Beppu Takiko?Shimada Email author" target="_blank">Osamu?TanakaEmail author |
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Institution: | (1) Department of Biology, Faculty of Science and Engineering, Konan University, Okamoto, Higashinada-ku, Kobe 658-8501, Japan;(2) Present address: Forage Crop Research Institute, Japan Grassland Farming and Forage Seed Association, 388-5 Higashiakada, Nishinasuno, Tochigi 329-2742, Japan;(3) Department of Bioscience, Faculty of Science and Engineering, Teikyou University of Science and Technology, 2525 Uenohara, Yatsuzawa, Yamanashi 409-0193, Japan;(4) Research Institute of Agricultural Resource, Ishikawa Agricultural College, 1-308 Suematsu, Nonoichi, Ishikawa 921-8836, Japan |
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Abstract: | An aluminum borate whiskers-mediated transformation system for calluses of tobacco (Nicotiana tabacum, cv. SR-1) has been developed. A total of 50 small pieces of calluses were vigorously agitated in a liquid medium containing aluminum borate whiskers, pBI221 plasmid carrying the -glucuronidase (GUS) gene, and pBI222 plasmid carrying the hygromycin phosphotransferase (HPT) gene. After treatment, calluses were cultured to select for hygromycin resistance, and three resistant calluses were obtained. Adventitious shoots were produced from each hygromycin-resistant callus and were transferred to rooting medium. A total of three plantlets obtained from each hygromycin-resistant callus were acclimatized and established in soil. Polymerase chain reaction analysis revealed that all the plantlets were cotransformed with both the GUS and HPT genes. Detached leaves of transgenic individuals showed clear hygromycin resistance when cultured in liquid medium. Histochemical assay for GUS revealed that one of these transgenic plants expressed the GUS gene, indicating coexpression of foreign genes. |
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Keywords: | callus -glucuronidase gene" target="_blank">gif" alt="beta" align="MIDDLE" BORDER="0">-glucuronidase gene whisker-mediated transformation |
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