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Plant and Cell Physiology 2004 45(9):1176-1184; doi:10.1093/pcp/pch139
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© 2004 Oxford University Press

Production of Polyhydroxybutyrate by Polycistronic Expression of Bacterial Genes in Tobacco Plastid

Yuko Arai1,3,5,6, Toshiharu Shikanai4,7, Yoshiharu Doi3, Shigeo Yoshida2, Isamu Yamaguchi1,8 and Hideo Nakashita1,2,5,9

1 Microbial Toxicology Laboratory, RIKEN Institute, 2-1 Hirosawa, Wako-shi, Saitama, 351-0198 Japan
2 Plant Functions Laboratory, RIKEN Institute, 2-1 Hirosawa, Wako-shi, Saitama, 351-0198 Japan
3 Polymer Chemistry Laboratory, RIKEN Institute, 2-1 Hirosawa, Wako-shi, Saitama, 351-0198 Japan
4 Graduate School of Biological Sciences, Nara Institute of Technology, 8916-5 Takayama, Ikoma-shi, Nara, 630-0101 Japan

Transgenic techniques are used to enhance and improve crop production, and their application to the production of chemical resources in plants has been under investigation. To achieve this latter goal, multiple-gene transformation is required to improve or change plant metabolic pathways; when accomplished by plant nuclear transformation, however, this procedure is costly and time consuming. We succeeded in the metabolic engineering of the tobacco plant by introducing multiple genes within a bacteria-like operon into a plastid genome. A tobacco plastid was transformed with a polycistron consisting of the spectinomycin resistance gene and three bacterial genes for the biosynthesis of the biodegradable polyester, poly[(R)-3-hydroxybutyrate] (PHB), after modification of their ribosome binding sites. DNA and RNA analysis confirmed the insertion of the introduced genes into the plastid genome and their polycistronic expression. As the result, the transplastomic tobacco accumulated PHB in its leaves. The introduced genes and the PHB productivity were maternally inherited, avoiding genetic spread by pollen diffusion, and were maintained stably in the seed progeny. Despite the low PHB productivity, this report demonstrates the feasibility of transplastomic technology for metabolic engineering. This "phyto-fermentation" system can be applied to plant production of various chemical commodities and pharmaceuticals.

5 These authors contributed equally to this study.

6 Present address: Department of Cell Biology, National Institute for Basic Biology, Myodaijicho, Okazaki-shi, Aichi, 444-8585 Japan.

7 Present address: Department of Plant Resources, Kyushu University, 6-10-1 Hakozaki, Higashi-ku, Fukuoka-shi, Fukuoka, 812-8581 Japan.

8 Present address: Laboratory for Remediation Research, Plant Science Center, RIKEN Institute, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama-shi, Kanagawa, 230-0045 Japan.

9 Corresponding author: E-mail, nakashi@postman.riken.go.jp; Fax, +81-48-462-4959.


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[Abstract] [Full Text] [PDF]


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