Stable transformation ofArabidopsis with thebar gene using particle bombardment |
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Authors: | Tatsuya Sawasaki Motoaki Seki Hiroyuki Anzai Kohei Irifune Hiromichi Morikawa |
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Affiliation: | (1) Graduate Department of Gene Science, Faculty of Science, Hiroshima University, Kagamiyama, 724 Higashi-Hiroshima, Japan;(2) Pharmaceutical Research Center, Meiji Seika Kaisha Ltd., Kohuku-ku, 222 Yokohama, Japan |
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Abstract: | A plasmid pARK 22 harbouring thebar gene encoding phosphinothricin acetyltransferase (PAT) under the control of the cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase (NOS) terminator was constructed and introduced into root sections ofArabidopsis thaliana using the pneumatic particle gun. The root sections that had been bombarded with this plasmid gave four to eight times higher yield of drug-resistant calluses than those sections bombarded with pCaMVNEO or pCH, which respectively contain the neomycin phosphotransferase and hygromycin phosphotransferase genes. Among a number of primary transformant (T0) plants obtained from independent bialaphos-resistant calluses, three were studied by Southern blot hybridization and PAT enzyme activity analyses, confirming the stable integration of the foreign gene into theArabidopsis genome and its expression in plants. The progeny analysis showed transmission of the foreign gene and its expression in up to the T2 generation. Some of the T1 progeny showed morphological abnormalities. Thus, thebar gene can be used effectively to allow selection of transgenicA. thalianna plants. |
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Keywords: | Arabidopsis thaliana transgenic plants particle bombardment bar gene phosphinothricin acetyltransferase late-flowering mutant |
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