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Transplastomic integration of a cyanobacterial bicarbonate transporter into tobacco chloroplasts

Improving global yields of agricultural crops is a complex challenge with evidence indicating benefits in productivity are achieved by enhancing photosynthetic carbon assimilation. Towards improving rates of CO(2) capture within leaf chloroplasts, this study shows the versatility of plastome transfo...

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Autores principales: Pengelly, J. J. L., Förster, B., von Caemmerer, S., Badger, M. R., Price, G. D., Whitney, S. M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4071830/
https://www.ncbi.nlm.nih.gov/pubmed/24965541
http://dx.doi.org/10.1093/jxb/eru156
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author Pengelly, J. J. L.
Förster, B.
von Caemmerer, S.
Badger, M. R.
Price, G. D.
Whitney, S. M.
author_facet Pengelly, J. J. L.
Förster, B.
von Caemmerer, S.
Badger, M. R.
Price, G. D.
Whitney, S. M.
author_sort Pengelly, J. J. L.
collection PubMed
description Improving global yields of agricultural crops is a complex challenge with evidence indicating benefits in productivity are achieved by enhancing photosynthetic carbon assimilation. Towards improving rates of CO(2) capture within leaf chloroplasts, this study shows the versatility of plastome transformation for expressing the Synechococcus PCC7002 BicA bicarbonate transporter within tobacco plastids. Fractionation of chloroplast membranes from transplastomic tob(BicA) lines showed that ~75% of the BicA localized to the thylakoid membranes and ~25% to the chloroplast envelope. BicA levels were highest in young emerging tob(BicA) leaves (0.12 μmol m(–2), ≈7mg m(–2)) accounting for ~0.1% (w/w) of the leaf protein. In these leaves, the molar amount of BicA was 16-fold lower than the abundant thylakoid photosystem II D1 protein (~1.9 μmol m(–2)) which was comparable to the 9:1 molar ratio of D1:BicA measured in air-grown Synechococcus PCC7002 cells. The BicA produced had no discernible effect on chloroplast ultrastructure, photosynthetic CO(2)-assimilation rates, carbon isotope discrimination, or growth of the tob(BicA) plants, implying that the bicarbonate transporter had little or no activity. These findings demonstrate the utility of plastome transformation for targeting bicarbonate transporter proteins into the chloroplast membranes without impeding growth or plastid ultrastructure. This study establishes the span of experimental measurements required to verify heterologous bicarbonate transporter function and location in chloroplasts and underscores the need for more detailed understanding of BicA structure and function to identify solutions for enabling its activation and operation in leaf chloroplasts.
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spelling pubmed-40718302014-06-26 Transplastomic integration of a cyanobacterial bicarbonate transporter into tobacco chloroplasts Pengelly, J. J. L. Förster, B. von Caemmerer, S. Badger, M. R. Price, G. D. Whitney, S. M. J Exp Bot Research Paper Improving global yields of agricultural crops is a complex challenge with evidence indicating benefits in productivity are achieved by enhancing photosynthetic carbon assimilation. Towards improving rates of CO(2) capture within leaf chloroplasts, this study shows the versatility of plastome transformation for expressing the Synechococcus PCC7002 BicA bicarbonate transporter within tobacco plastids. Fractionation of chloroplast membranes from transplastomic tob(BicA) lines showed that ~75% of the BicA localized to the thylakoid membranes and ~25% to the chloroplast envelope. BicA levels were highest in young emerging tob(BicA) leaves (0.12 μmol m(–2), ≈7mg m(–2)) accounting for ~0.1% (w/w) of the leaf protein. In these leaves, the molar amount of BicA was 16-fold lower than the abundant thylakoid photosystem II D1 protein (~1.9 μmol m(–2)) which was comparable to the 9:1 molar ratio of D1:BicA measured in air-grown Synechococcus PCC7002 cells. The BicA produced had no discernible effect on chloroplast ultrastructure, photosynthetic CO(2)-assimilation rates, carbon isotope discrimination, or growth of the tob(BicA) plants, implying that the bicarbonate transporter had little or no activity. These findings demonstrate the utility of plastome transformation for targeting bicarbonate transporter proteins into the chloroplast membranes without impeding growth or plastid ultrastructure. This study establishes the span of experimental measurements required to verify heterologous bicarbonate transporter function and location in chloroplasts and underscores the need for more detailed understanding of BicA structure and function to identify solutions for enabling its activation and operation in leaf chloroplasts. Oxford University Press 2014-07 2014-04-18 /pmc/articles/PMC4071830/ /pubmed/24965541 http://dx.doi.org/10.1093/jxb/eru156 Text en © The Author 2014. Published by Oxford University Press on behalf of the Society for Experimental Biology. http://creativecommons.org/licenses/by/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Paper
Pengelly, J. J. L.
Förster, B.
von Caemmerer, S.
Badger, M. R.
Price, G. D.
Whitney, S. M.
Transplastomic integration of a cyanobacterial bicarbonate transporter into tobacco chloroplasts
title Transplastomic integration of a cyanobacterial bicarbonate transporter into tobacco chloroplasts
title_full Transplastomic integration of a cyanobacterial bicarbonate transporter into tobacco chloroplasts
title_fullStr Transplastomic integration of a cyanobacterial bicarbonate transporter into tobacco chloroplasts
title_full_unstemmed Transplastomic integration of a cyanobacterial bicarbonate transporter into tobacco chloroplasts
title_short Transplastomic integration of a cyanobacterial bicarbonate transporter into tobacco chloroplasts
title_sort transplastomic integration of a cyanobacterial bicarbonate transporter into tobacco chloroplasts
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4071830/
https://www.ncbi.nlm.nih.gov/pubmed/24965541
http://dx.doi.org/10.1093/jxb/eru156
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