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A simple Agrobacterium-mediated stable transformation technique for the hornwort model Anthoceros agrestis


Frangedakis, Eftychios; Waller, Manuel; Nishiyama, Tomoaki; Tsukaya, Hirokazu; Xu, Xia; Yue, Yuling; Tjahjadi, Michelle; Gunadi, Andika; Van Eck, Joyce; Li, Fay-Wei; Szövényi, Péter; Sakakibara, Keiko (2021). A simple Agrobacterium-mediated stable transformation technique for the hornwort model Anthoceros agrestis. bioRxiv 425778, University of Zurich.

Abstract

We have developed a simple Agrobacterium-mediated method for the stable transformation of the hornwort Anthoceros agrestis, the fifth bryophyte species for which a genetic manipulation technique becomes available. High transformation efficiency was achieved by using thallus tissue grown under low-light conditions. We generated a total of 216 transgenic A. agrestis lines expressing the β-Glucuronidase (GUS), cyan, green, and yellow fluorescent proteins under the control of the CaMV 35S promoter and several endogenous promoters. Nuclear and plasma membrane localization with multiple color fluorescent proteins was also confirmed. The transformation technique described here should pave the way for detailed molecular and genetic studies of hornwort biology, providing much needed insight into the molecular mechanisms underlying symbiosis, carbon-concentrating mechanism, RNA editing, and land plant evolution in general.

Abstract

We have developed a simple Agrobacterium-mediated method for the stable transformation of the hornwort Anthoceros agrestis, the fifth bryophyte species for which a genetic manipulation technique becomes available. High transformation efficiency was achieved by using thallus tissue grown under low-light conditions. We generated a total of 216 transgenic A. agrestis lines expressing the β-Glucuronidase (GUS), cyan, green, and yellow fluorescent proteins under the control of the CaMV 35S promoter and several endogenous promoters. Nuclear and plasma membrane localization with multiple color fluorescent proteins was also confirmed. The transformation technique described here should pave the way for detailed molecular and genetic studies of hornwort biology, providing much needed insight into the molecular mechanisms underlying symbiosis, carbon-concentrating mechanism, RNA editing, and land plant evolution in general.

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Additional indexing

Item Type:Working Paper
Communities & Collections:07 Faculty of Science > Department of Systematic and Evolutionary Botany
07 Faculty of Science > Zurich-Basel Plant Science Center
Dewey Decimal Classification:580 Plants (Botany)
Language:English
Date:2021
Deposited On:01 Feb 2021 18:01
Last Modified:01 Feb 2021 18:01
Series Name:bioRxiv
Number of Pages:36
ISSN:2164-7844
OA Status:Green
Publisher DOI:https://doi.org/10.1101/2021.01.07.425778

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