Improved G-AgarTrap: A highly efficient transformation method for intact gemmalings of the liverwort Marchantia polymorpha

Scientific Reports
Shoko TsuboyamaYutaka Kodama

Abstract

Liverworts are key species for studies of plant evolution, occupying a basal position among the land plants. Marchantia polymorpha has emerged as a highly studied model liverwort, and many relevant techniques, including genetic transformation, have been established for this species. Agrobacterium-mediated transformation is widely used in many plant species because of its low cost. Recently, we developed a simplified Agrobacterium-mediated method for transforming M. polymorpha, known as AgarTrap (agar-utilized transformation with pouring solutions). The AgarTrap procedure, which involves culturing the liverwort tissue in various solutions on a single solid medium, yields up to a hundred independent transformants. AgarTrap is a simple procedure, requiring minimal expertise, cost, and time. Here, we investigated four factors that influence AgarTrap transformation efficiency: (1) humidity, (2) surfactant in the transformation buffer, (3) Agrobacterium strain, and (4) light/dark condition. We adapted the AgarTrap protocol for transforming intact gemmalings, achieving an exceptionally high transformation efficiency of 97%. The improved AgarTrap method will enhance the molecular biological study of M. polymorpha. Furthermore, this met...Continue Reading

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Citations

Aug 6, 2020·International Journal of Molecular Sciences·Felix Althoff, Sabine Zachgo
Jan 28, 2021·Trends in Plant Science·Shubhajit DasJan Willem Borst
Dec 9, 2020·Biotechnology Advances·Jonas De SaegerStephen Depuydt

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Methods Mentioned

BETA
transgenic
Fluorescence
phosphotransferase
fluorescence microscopy

Software Mentioned

AgarTrap

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