Development of synthetic biology tools and methods for plant systems
dc.contributor.advisor
Nakayama, Naomi
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dc.contributor.advisor
French, Chris
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dc.contributor.author
Andreou, Andreas
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dc.date.accessioned
2020-03-09T13:05:46Z
dc.date.available
2020-03-09T13:05:46Z
dc.date.issued
2020-07-04
dc.description.abstract
Plant synthetic biology is a fast-evolving field in which standardised tools and
methods are developed to empower research and commercial bioproduction in
plant systems. Nevertheless, in the whole synthetic biology landscape, plant
systems lag compared to microbial and mammalian systems. Plant cell cultures
are becoming a popular chassis for bioproduction, which can combine the
metabolic diversity and capacity of the plants with the benefits of cell cultures as
in microbial systems, such as fast growth, resource- and energy-efficient
cultivation, and secure containment. My PhD was focused on the establishment
of multiple enabling tools and methods to confer complex yet predictive genetic
engineering in plant systems. These plant synthetic biology resources are being
implemented to development cell-type-specific plant cell biofactories particularly
competent in the biosynthesis of specific compounds.
Firstly, I have developed Mobius Assembly, a user-friendly Golden Gate
Assembly system for fast and easy generation of complex DNA constructs.
Mobius Assembly toolkit was adapted for Plant Systems (MAPS). I devised a new
category of compact binary vectors, which can be employed for the whole plant,
plant cell culture, and plant protoplast transformations. I found that the
combination of binary vectors and Agrobacterial strains determine the efficiency
of plant cell culture transformation.
Secondly, I developed a high-throughput, protoplast expression protocol. Using
Mobius Assembly and the new protoplast expression analysis platform, I
characterised a library of short promoters and terminators for their regulatory
effect on reporter gene expression. The results highlighted the strong influence of
terminators in gene expression depending on different promoters; this observation
implies a synergistic interaction between terminators and promoters. I further
made and optimised standardised parts of three commonly used chemically
inducible gene expression systems in plant science and tested them for possible
crosstalks when used in conjunction.
Lastly, I employed MAPS to create a sequential plant expression system that can
be used to activate multiple gene expression at differential levels and timings. As
a pilot study, this sequential system was used to express master transcription
factors for the differentiation of plant cell cultures into xylem vessels, a cell type
that produces a high level of phenolic compounds and thus may serve as
biofactories for production of anthocyanin and other phenolics.
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dc.identifier.uri
https://hdl.handle.net/1842/36842
dc.identifier.uri
http://dx.doi.org/10.7488/era/144
dc.language.iso
en
dc.publisher
The University of Edinburgh
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dc.relation.hasversion
Andreou,A.I. and Nakayama,N. (2018) Mobius Assembly: A versatile Golden- Gate framework towards universal DNA assembly. PLoS One, 13, e0189892.
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dc.subject
synthetic biology
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dc.subject
bioproduction
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dc.subject
Mobius Assembly
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dc.subject
protoplast expression protocols
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dc.subject
xylem vessels
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dc.title
Development of synthetic biology tools and methods for plant systems
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dc.type
Thesis or Dissertation
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dc.type.qualificationlevel
Doctoral
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dc.type.qualificationname
PhD Doctor of Philosophy
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