High resolution imaging and characterisation of a simple plant system for synthetic biology

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Nuri Purswani, Tim Rudge, Fernan Federici, Lihua Robertson and Jim Haseloff

University of Cambridge, United Kingdom

Synthetic biology of multicellular organisms is extremely challenging due to their high genetic complexity and the difficulty of manipulation of the tissues. The liverwort Marchantia polymorpha (Marchantia) is a simple plant with a haploid genome and low genetic redundancy. The early embryonic stages of this organism are accessible for live imaging and there is no need to perforate a seed coat to visualise them. Marchantia offers the added benefit that the cells on the surface and their progeny can be easily observed and tracked without the need for clonal markers during early embryogenesis. The procedure for genetic transformation [1] of this plant is much faster than that of Arabidopsis thaliana (Arabidopsis), as it only takes 7 days to obtain transgenic plants and an additional 5-10 days to screen transient mutants with fluorescent reporters and antibiotic selection. This period of time is much shorter than the minimum 2 months required to obtain transgenic Arabidopsis plants. This organism is also very easy to manipulate and ideal for labs that wish to start to do synthetic biology in plants without prior experience. Therefore, it has the advantages of being a multicellular chassis that has an analogous simplicity to that of microbes. This project provides a detailed cellular characterisation of the organism through the application of high throughput imaging (laser scanning confocal microscopy) with fluorescent proteins to mark subcellular structures; combined with segmentation techniques for detailed analysis of cell geometry [2]. We are also transferring existing genetic markers from Arabidopsis into Marchantia for the purpose of future mapping and characterisation of cellular domains for engineering. [1] Ishizaki K, Chiyoda S, Yamato K, Kohchi T, Plant and Cell Physiology, 49:1084-1091 (2008) [2] Federici F, Dupuy L, Laplaze L, Heisler M and Haseloff J, Nature Methods, 9:483-485 (2012)