Stable transformation of Lithospermum erythrorhizon by Agrobacterium rhizogenes and shikonin production of the transformants |
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Authors: | K Yazaki S Tanaka H Matsuoka F Sato |
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Institution: | (1) Laboratory of Molecular and Cellular Biology, Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kitashirakawa, Kyoto 606-8502, Japan Fax: +81-75-753-6398 e-mail: yazaki@kais.kyoto-u.ac.jp, JP;(2) Faculty of Pharmaceutical Sciences, Kyoto University, Yoshida, Kyoto 606-8501, Japan, JP |
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Abstract: | Seedling hypocotyls of Lithospermum erythrorhizon were infected with Agrobacterium rhizogenes (strain 15834) harboring a binary vector with an intron-bearing the β-glucuronidase (GUS) gene driven by cauliflower mosaic virus (CaMV) 35S promoter as well as the hygromycin phosphotransferase
(HPT) gene as the selection marker. About 20% of the hairy roots isolated were hygromycin resistant and had co-integrated
GUS and HPT genes in their Lithospermum genomic DNA. Because GUS activity was detected in almost all the hygromycin-resistant root tissues, the CaMV 35S promoter
seems to be ubiquitously active in L. erythrorhizon hairy roots. In pigment production medium M9, the hairy root cultures had shikonin productivity similar to that of cell suspension
cultures of Lithospermum. They also showed light-dependent inhibition of shikonin biosynthesis similar to that of Lithospermum cell cultures. These findings suggest that this hairy root system transformable with A. rhizogenes is a suitable model system for molecular characterization of shikonin biosynthesis via reverse genetics.
Received: 2 March 1998 / Revision received: 25 May 1998 / Accepted: 8 July 1998 |
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Keywords: | Agrobacterium rhizogenes β -Glucuronidase Hairy root Lithospermum erythrorhizon Stable transformation |
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