Efficient and Stable Transformation of Lactuca sativa L. cv. Cisco (lettuce) Plastids |
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Authors: | Hirosuke Kanamoto Atsushi Yamashita Hiroshi Asao Satoru Okumura Hisabumi Takase Masahira Hattori Akiho Yokota Ken-Ichi Tomizawa |
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Affiliation: | (1) Resarch Institute of Innovative Technology for the Earth, 9-2 Kizugawadai, Kizu-cho Soraku-gun, Kyoto 619-0292, Japan;(2) Kitasato Institute for Life Sciences, Kitasato University, 1-15-1 Kitasato, Sagamihara-shi, Kanagawa 228-8555, Japan;(3) Nara Prefectural Agricultural Experiment Station, 88 Shijyo-cho, Kashihara, Nara 634-0813, Japan;(4) Graduate School of Biological Science, Nara Institute of Science and Technology, Ikoma, Nara 630-0192, Japan |
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Abstract: | Transgenic plastids offer unique advantages in plant biotechnology, including high-level foreign protein expression. However, broad application of plastid genome engineering in biotechnology has been largely hampered by the lack of plastid transformation systems for major crops. Here we describe the development of a plastid transformation system for lettuce, Lactuca sativa L. cv. Cisco. The transforming DNA carries a spectinomycin-resistance gene (aadA) under the control of lettuce chloroplast regulatory expression elements, flanked by two adjacent lettuce plastid genome sequences allowing its targeted insertion between the rbcL and accD genes. On average, we obtained 1 transplastomic lettuce plant per bombardment. We show that lettuce leaf chloroplasts can express transgene-encoded GFP to ~36% of the total soluble protein. All transplastomic T0 plants were fertile and the T1 progeny uniformly showed stability of the transgene in the chloroplast genome. This system will open up new possibilities for the efficient production of edible vaccines, pharmaceuticals, and antibodies in plants. |
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Keywords: | lettuce plastid genome plastid transformation |
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