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    Rights statement: This is the peer reviewed version of the following article: Lin, M.T., Orr, D.J., Worrall, D., Parry, M.A.J., Carmo‐Silva, E. and Hanson, M.R. (2021), A procedure to introduce point mutations into the Rubisco large subunit gene in wild‐type plants. Plant J, 106: 876-887. https://doi.org/10.1111/tpj.15196 which has been published in final form at https://onlinelibrary.wiley.com/doi/10.1111/tpj.15196This article may be used for non-commercial purposes in accordance With Wiley Terms and Conditions for self-archiving.

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A procedure to introduce point mutations into the Rubisco large subunit gene in wild-type plants

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A procedure to introduce point mutations into the Rubisco large subunit gene in wild-type plants. / Lin, Myat T.; Orr, Douglas; Worrall, Dawn et al.

In: The Plant Journal, Vol. 106, No. 3, 31.05.2021, p. 876-887.

Research output: Contribution to Journal/MagazineJournal articlepeer-review

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Lin MT, Orr D, Worrall D, Parry M, Carmo-Silva E, Hanson MR. A procedure to introduce point mutations into the Rubisco large subunit gene in wild-type plants. The Plant Journal. 2021 May 31;106(3):876-887. Epub 2021 Feb 11. doi: 10.1111/tpj.15196

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@article{850983980bc6486bb6ca575fe119404a,
title = "A procedure to introduce point mutations into the Rubisco large subunit gene in wild-type plants",
abstract = "Photosynthetic inefficiencies limit the productivity and sustainability of crop production, and the resilience of agriculture to future societal and environmental challenges. Rubisco is a key target for improvement as it plays a central role in carbon fixation during photosynthesis and is remarkably inefficient. Introduction of mutations to the chloroplast-encoded Rubisco large subunit rbcL is of particular interest to improve the catalytic activity and efficiency of the enzyme. However, manipulation of rbcL is hampered by its location in the plastome, with many species recalcitrant to plastome transformation, and by the plastid{\textquoteright}s efficient repair system, which can prevent effective maintenance of mutations introduced with homologous recombination. Here we present a system where the introduction of a number of silent mutations into rbcL within the model plant Nicotiana tabacum facilitates simplified screening via additional restriction enzyme sites. This system was used to successfully generate a range of transplastomic lines from wild-type N. tabacum with stable point mutations within rbcL in 40% of the transformants, allowing assessment of the effect of these mutations on Rubisco assembly and activity. With further optimization, the approach offers a viable way forward for mutagenic testing of Rubisco function in planta within tobacco and modifying rbcL in other crops where chloroplast transformation is feasible. The transformation strategy could also be applied to introduce point mutations in other chloroplast-encoded genes.",
author = "Lin, {Myat T.} and Douglas Orr and Dawn Worrall and Martin Parry and Elizabete Carmo-Silva and Hanson, {Maureen R.}",
note = "This is the peer reviewed version of the following article: Lin, M.T., Orr, D.J., Worrall, D., Parry, M.A.J., Carmo‐Silva, E. and Hanson, M.R. (2021), A procedure to introduce point mutations into the Rubisco large subunit gene in wild‐type plants. Plant J, 106: 876-887. https://doi.org/10.1111/tpj.15196 which has been published in final form at https://onlinelibrary.wiley.com/doi/10.1111/tpj.15196This article may be used for non-commercial purposes in accordance With Wiley Terms and Conditions for self-archiving. ",
year = "2021",
month = may,
day = "31",
doi = "10.1111/tpj.15196",
language = "English",
volume = "106",
pages = "876--887",
journal = "The Plant Journal",
issn = "0960-7412",
publisher = "Blackwell Publishing Ltd",
number = "3",

}

RIS

TY - JOUR

T1 - A procedure to introduce point mutations into the Rubisco large subunit gene in wild-type plants

AU - Lin, Myat T.

AU - Orr, Douglas

AU - Worrall, Dawn

AU - Parry, Martin

AU - Carmo-Silva, Elizabete

AU - Hanson, Maureen R.

N1 - This is the peer reviewed version of the following article: Lin, M.T., Orr, D.J., Worrall, D., Parry, M.A.J., Carmo‐Silva, E. and Hanson, M.R. (2021), A procedure to introduce point mutations into the Rubisco large subunit gene in wild‐type plants. Plant J, 106: 876-887. https://doi.org/10.1111/tpj.15196 which has been published in final form at https://onlinelibrary.wiley.com/doi/10.1111/tpj.15196This article may be used for non-commercial purposes in accordance With Wiley Terms and Conditions for self-archiving.

PY - 2021/5/31

Y1 - 2021/5/31

N2 - Photosynthetic inefficiencies limit the productivity and sustainability of crop production, and the resilience of agriculture to future societal and environmental challenges. Rubisco is a key target for improvement as it plays a central role in carbon fixation during photosynthesis and is remarkably inefficient. Introduction of mutations to the chloroplast-encoded Rubisco large subunit rbcL is of particular interest to improve the catalytic activity and efficiency of the enzyme. However, manipulation of rbcL is hampered by its location in the plastome, with many species recalcitrant to plastome transformation, and by the plastid’s efficient repair system, which can prevent effective maintenance of mutations introduced with homologous recombination. Here we present a system where the introduction of a number of silent mutations into rbcL within the model plant Nicotiana tabacum facilitates simplified screening via additional restriction enzyme sites. This system was used to successfully generate a range of transplastomic lines from wild-type N. tabacum with stable point mutations within rbcL in 40% of the transformants, allowing assessment of the effect of these mutations on Rubisco assembly and activity. With further optimization, the approach offers a viable way forward for mutagenic testing of Rubisco function in planta within tobacco and modifying rbcL in other crops where chloroplast transformation is feasible. The transformation strategy could also be applied to introduce point mutations in other chloroplast-encoded genes.

AB - Photosynthetic inefficiencies limit the productivity and sustainability of crop production, and the resilience of agriculture to future societal and environmental challenges. Rubisco is a key target for improvement as it plays a central role in carbon fixation during photosynthesis and is remarkably inefficient. Introduction of mutations to the chloroplast-encoded Rubisco large subunit rbcL is of particular interest to improve the catalytic activity and efficiency of the enzyme. However, manipulation of rbcL is hampered by its location in the plastome, with many species recalcitrant to plastome transformation, and by the plastid’s efficient repair system, which can prevent effective maintenance of mutations introduced with homologous recombination. Here we present a system where the introduction of a number of silent mutations into rbcL within the model plant Nicotiana tabacum facilitates simplified screening via additional restriction enzyme sites. This system was used to successfully generate a range of transplastomic lines from wild-type N. tabacum with stable point mutations within rbcL in 40% of the transformants, allowing assessment of the effect of these mutations on Rubisco assembly and activity. With further optimization, the approach offers a viable way forward for mutagenic testing of Rubisco function in planta within tobacco and modifying rbcL in other crops where chloroplast transformation is feasible. The transformation strategy could also be applied to introduce point mutations in other chloroplast-encoded genes.

U2 - 10.1111/tpj.15196

DO - 10.1111/tpj.15196

M3 - Journal article

VL - 106

SP - 876

EP - 887

JO - The Plant Journal

JF - The Plant Journal

SN - 0960-7412

IS - 3

ER -