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    Rights statement: This is a pre-copy-editing, author-produced PDF of an article accepted for publication in Journal of Experimental Botany following peer review. The definitive publisher-authenticated version Panupon Khumsupan, Marta A Kozlowska, Douglas J Orr, Andreas I Andreou, Naomi Nakayama, Nicola Patron, Elizabete Carmo-Silva, Alistair J McCormick, Generating and characterising single- and multi-gene mutants of the Rubisco small subunit family in Arabidopsis, Journal of Experimental Botany, 71, (19), https://doi.org/10.1093/jxb/eraa316 is available online at: https://academic.oup.com/jxb/article/71/19/5963/5868035

    Accepted author manuscript, 1.86 MB, PDF document

    Embargo ends: 6/07/21

    Available under license: CC BY-NC: Creative Commons Attribution-NonCommercial 4.0 International License

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Generating and characterizing single- and multigene mutants of the Rubisco small subunit family in Arabidopsis

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<mark>Journal publication date</mark>1/10/2020
<mark>Journal</mark>Journal of Experimental Botany
Issue number19
Volume71
Number of pages13
Pages (from-to)5963–5975
Publication StatusPublished
Early online date6/07/20
<mark>Original language</mark>English

Abstract

The primary CO2-fixing enzyme Rubisco limits the productivity of plants. The small subunit of Rubisco (SSU) can influence overall Rubisco levels and catalytic efficiency, and is now receiving increasing attention as a potential engineering target to improve the performance of Rubisco. However, SSUs are encoded for by a family of nuclear rbcS genes in plants, which makes them challenging to engineer and study. Here we have used CRISPR/Cas9 and T-DNA insertion lines to generate a suite of single and multiple gene knockout mutants for the four members of the rbcS family in Arabidopsis, including two novel mutants 2b3b and 1a2b3b. 1a2b3b contained very low levels of Rubisco (ca. 3% relative to WT) and is the first example of a mutant with a homogenous Rubisco pool consisting of a single SSU isoform (1B). Growth under near-outdoor levels of light demonstrated Rubisco-limited growth phenotypes for several SSU mutants and the importance of the 1A and 3B isoforms. We also identified 1a1b as a likely lethal mutation, suggesting a key contributory role for the least expressed 1B isoform during early development. The successful use of CRISPR/Cas here suggests this is a viable approach for exploring the functional roles of SSU isoforms in plants.

Bibliographic note

This is a pre-copy-editing, author-produced PDF of an article accepted for publication in Journal of Experimental Botany following peer review. The definitive publisher-authenticated version Panupon Khumsupan, Marta A Kozlowska, Douglas J Orr, Andreas I Andreou, Naomi Nakayama, Nicola Patron, Elizabete Carmo-Silva, Alistair J McCormick, Generating and characterising single- and multi-gene mutants of the Rubisco small subunit family in Arabidopsis, Journal of Experimental Botany, 71, (19), https://doi.org/10.1093/jxb/eraa316 is available online at: https://academic.oup.com/jxb/article/71/19/5963/5868035