Random mutagenesis of Phaeodactylum tricornutum using ultraviolet, chemical, and X-radiation demonstrates the need for temporal analysis of phenotype stability.

Macdonald Miller, S; Abbriano, RM; Herdean, A; Banati, R; Ralph, PJ; Pernice, M

Random mutagenesis of Phaeodactylum tricornutum using ultraviolet, chemical, and X-radiation demonstrates the need for temporal analysis of phenotype stability.

Macdonald Miller, S Abbriano, RM Herdean, A

Banati, R Ralph, PJ Pernice, M

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Publisher:: Springer Science and Business Media LLC
Publication Type:: Journal Article
Citation:: Sci Rep, 2023, 13, (1), pp. 22385
Issue Date:: 2023-12-16

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Field	Value	Language
dc.contributor.author	Macdonald Miller, S
dc.contributor.author	Abbriano, RM
dc.contributor.author	Herdean, A https://orcid.org/0000-0003-2143-0213
dc.contributor.author	Banati, R
dc.contributor.author	Ralph, PJ
dc.contributor.author	Pernice, M https://orcid.org/0000-0002-3431-2104
dc.date.accessioned	2024-01-17T22:45:13Z
dc.date.available	2023-10-25
dc.date.available	2024-01-17T22:45:13Z
dc.date.issued	2023-12-16
dc.identifier.citation	Sci Rep, 2023, 13, (1), pp. 22385
dc.identifier.issn	2045-2322
dc.identifier.issn	2045-2322
dc.identifier.uri	http://hdl.handle.net/10453/174736
dc.description.abstract	We investigated two non-ionising mutagens in the form of ultraviolet radiation (UV) and ethyl methanosulfonate (EMS) and an ionising mutagen (X-ray) as methods to increase fucoxanthin content in the model diatom Phaeodactylum tricornutum. We implemented an ultra-high throughput method using fluorescence-activated cell sorting (FACS) and live culture spectral deconvolution for isolation and screening of potential pigment mutants, and assessed phenotype stability by measuring pigment content over 6 months using high-performance liquid chromatography (HPLC) to investigate the viability of long-term mutants. Both UV and EMS resulted in significantly higher fucoxanthin within the 6 month period after treatment, likely as a result of phenotype instability. A maximum fucoxanthin content of 135 ± 10% wild-type found in the EMS strain, a 35% increase. We found mutants generated using all methods underwent reversion to the wild-type phenotype within a 6 month time period. X-ray treatments produced a consistently unstable phenotype even at the maximum treatment of 1000 Grays, while a UV mutant and an EMS mutant reverted to wild-type after 4 months and 6 months, respectively, despite showing previously higher fucoxanthin than wild-type. This work provides new insights into key areas of microalgal biotechnology, by (i) demonstrating the use of an ionising mutagen (X-ray) on a biotechnologically relevant microalga, and by (ii) introducing temporal analysis of mutants which has substantial implications for strain creation and utility for industrial applications.
dc.format	Electronic
dc.language	eng
dc.publisher	Springer Science and Business Media LLC
dc.relation.ispartof	Sci Rep
dc.relation.isbasedon	10.1038/s41598-023-45899-2
dc.rights	info:eu-repo/semantics/openAccess
dc.subject.mesh	X-Rays
dc.subject.mesh	Ultraviolet Rays
dc.subject.mesh	Diatoms
dc.subject.mesh	Mutagenesis
dc.subject.mesh	Mutagens
dc.subject.mesh	Phenotype
dc.subject.mesh	Diatoms
dc.subject.mesh	Mutagens
dc.subject.mesh	Ultraviolet Rays
dc.subject.mesh	Mutagenesis
dc.subject.mesh	Phenotype
dc.subject.mesh	X-Rays
dc.title	Random mutagenesis of Phaeodactylum tricornutum using ultraviolet, chemical, and X-radiation demonstrates the need for temporal analysis of phenotype stability.
dc.type	Journal Article
utslib.citation.volume	13
utslib.location.activity	England
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - C3 - Climate Change Cluster
utslib.copyright.status	open_access	*
dc.date.updated	2024-01-17T22:45:11Z
pubs.issue	1
pubs.publication-status	Published online
pubs.volume	13
utslib.citation.issue	1

Abstract:

We investigated two non-ionising mutagens in the form of ultraviolet radiation (UV) and ethyl methanosulfonate (EMS) and an ionising mutagen (X-ray) as methods to increase fucoxanthin content in the model diatom Phaeodactylum tricornutum. We implemented an ultra-high throughput method using fluorescence-activated cell sorting (FACS) and live culture spectral deconvolution for isolation and screening of potential pigment mutants, and assessed phenotype stability by measuring pigment content over 6 months using high-performance liquid chromatography (HPLC) to investigate the viability of long-term mutants. Both UV and EMS resulted in significantly higher fucoxanthin within the 6 month period after treatment, likely as a result of phenotype instability. A maximum fucoxanthin content of 135 ± 10% wild-type found in the EMS strain, a 35% increase. We found mutants generated using all methods underwent reversion to the wild-type phenotype within a 6 month time period. X-ray treatments produced a consistently unstable phenotype even at the maximum treatment of 1000 Grays, while a UV mutant and an EMS mutant reverted to wild-type after 4 months and 6 months, respectively, despite showing previously higher fucoxanthin than wild-type. This work provides new insights into key areas of microalgal biotechnology, by (i) demonstrating the use of an ionising mutagen (X-ray) on a biotechnologically relevant microalga, and by (ii) introducing temporal analysis of mutants which has substantial implications for strain creation and utility for industrial applications.

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http://hdl.handle.net/10453/174736