High-level expression of sucrose inducible sweet potato sporamin gene promoter: [beta]-glucuronidase fusion gene in transgenic Nicotiana plumbaginifolia hairy roots.

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Date: Oct. 2022
Publisher: Springer
Document Type: Report; Brief article
Length: 314 words

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Abstract :

Keywords: Agrobacterium rhizogenes; GUS; Sucrose inducible promoter; Hairy root Abstract We developed transgenic Nicotiana plumbaginifolia hairy roots with sucrose-inducible minimal promoter (Spo.sup.min)-[beta]-glucuronidase (GUS) gene--fused constructs with signal sequences for sorting to cytosol, apoplast, and ER, and we analyzed the GUS activities of hairy roots after sucrose treatment. Induced GUS activities by Spo.sup.min were about 10 times higher than those by the CaMV 35S promoter. GUS activities in hairy roots induced with a Spo.sup.min-UTR-GUS construct after 6% sucrose treatment were higher than those with either Spo.sup.min-UTR-GUS construct after 10% sucrose treatment, Spo.sup.min-apoplast or ER-GUS constructs after sucrose treatment. High GUS activities in hairy roots with Spo.sup.min constructs were induced during 20 wk by culturing hairy roots in LS liquid medium containing 6% sucrose in 300 ml conical flasks. Hairy roots in Linsmaier and Skoog (LS) liquid medium showed the increase of fresh weight (FW) for 16 wk. Total yield (µg) of GUS by the hairy roots with Spo.sup.min-UTR-GUS construct using 5 l culture container was calculated to about 6742 µg after 8-wk culture. Productive efficiencies of GUS were over 3.0% of total soluble protein after 1--2 wk 6% sucrose treatment. Productive efficiencies of GUS in transgenic N. plumbaginifolia hairy roots were higher than that in transgenic N. plumbaginifolia leaves, stems, and roots. And growth speeds of transgenic N. plumbaginifolia hairy roots were faster than transgenic N. plumbaginifolia plants. These results showed that N. plumbaginifolia hairy roots with Spo.sup.min-heterologous construct systems and sucrose treatment would be useful tools to develop protein production system. Author Affiliation: (1) Department of Global Agricultural Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, 113-8657, Tokyo, Japan (2) Institute for Horticultural Plant Breeding, 2-5-1 Kamishiki, Matsudo-shi, 270-2221, Chiba, Japan (3) Research Center for Food Safety, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, 113-8657, Tokyo, Japan (b) yamakawa@g.ecc.u-tokyo.ac.jp Article History: Registration Date: 06/14/2022 Received Date: 02/12/2022 Accepted Date: 06/07/2022 Online Date: 08/12/2022 Byline:

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Gale Document Number: GALE|A727538978