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Agrobacterium tumefaciens-mediated transformation of Cleome gynandra L., a C(4) dicotyledon that is closely related to Arabidopsis thaliana

In leaves of most C(4) plants, the biochemistry of photosynthesis is partitioned between mesophyll and bundle sheath cells. In addition, their cell biology and development also differs from that in C(3) plants. We have a poor understanding of the mechanisms that generate the cell-specific accumulati...

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Autores principales: Newell, Christine A., Brown, Naomi J., Liu, Zheng, Pflug, Alexander, Gowik, Udo, Westhoff, Peter, Hibberd, Julian M.
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2837259/
https://www.ncbi.nlm.nih.gov/pubmed/20150516
http://dx.doi.org/10.1093/jxb/erq009
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author Newell, Christine A.
Brown, Naomi J.
Liu, Zheng
Pflug, Alexander
Gowik, Udo
Westhoff, Peter
Hibberd, Julian M.
author_facet Newell, Christine A.
Brown, Naomi J.
Liu, Zheng
Pflug, Alexander
Gowik, Udo
Westhoff, Peter
Hibberd, Julian M.
author_sort Newell, Christine A.
collection PubMed
description In leaves of most C(4) plants, the biochemistry of photosynthesis is partitioned between mesophyll and bundle sheath cells. In addition, their cell biology and development also differs from that in C(3) plants. We have a poor understanding of the mechanisms that generate the cell-specific accumulation of proteins used in the C(4) pathway, and there are few genes that have been shown to be important for the cell biology and development of C(4) leaves. To facilitate functional analysis of C(4) photosynthesis, and to enable knowledge from Arabidopsis thaliana to be translated to C(4) species, an Agrobacterium tumefaciens-mediated transformation protocol was developed for the C(4) species Cleome gynandra. A. tumefaciens, harbouring the binary vector SLJ1006, was used to transfer the uidA gene under the control of the CaMV 35S promoter into C. gynandra. Co-incubation of hypocotyls or cotyledons with SLJ1006 allowed efficient transfer of DNA into C. gynandra, and media that allowed callus production and then shoot regeneration were identified. Stable transformants of C. gynandra with detectable amounts of β-glucuronidase (GUS) were produced at an efficiency of 14%. When driven by the CaMV 35S promoter, GUS was visible in all leaf cells, whereas uidA translationally fused to a CgRbcS gene generated GUS accumulation specifically in bundle sheath cells. This transformation procedure is the first for an NAD-ME type C(4) plant and should significantly accelerate the analysis of mechanisms underlying C(4) photosynthesis.
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spelling pubmed-28372592010-03-15 Agrobacterium tumefaciens-mediated transformation of Cleome gynandra L., a C(4) dicotyledon that is closely related to Arabidopsis thaliana Newell, Christine A. Brown, Naomi J. Liu, Zheng Pflug, Alexander Gowik, Udo Westhoff, Peter Hibberd, Julian M. J Exp Bot Research Papers In leaves of most C(4) plants, the biochemistry of photosynthesis is partitioned between mesophyll and bundle sheath cells. In addition, their cell biology and development also differs from that in C(3) plants. We have a poor understanding of the mechanisms that generate the cell-specific accumulation of proteins used in the C(4) pathway, and there are few genes that have been shown to be important for the cell biology and development of C(4) leaves. To facilitate functional analysis of C(4) photosynthesis, and to enable knowledge from Arabidopsis thaliana to be translated to C(4) species, an Agrobacterium tumefaciens-mediated transformation protocol was developed for the C(4) species Cleome gynandra. A. tumefaciens, harbouring the binary vector SLJ1006, was used to transfer the uidA gene under the control of the CaMV 35S promoter into C. gynandra. Co-incubation of hypocotyls or cotyledons with SLJ1006 allowed efficient transfer of DNA into C. gynandra, and media that allowed callus production and then shoot regeneration were identified. Stable transformants of C. gynandra with detectable amounts of β-glucuronidase (GUS) were produced at an efficiency of 14%. When driven by the CaMV 35S promoter, GUS was visible in all leaf cells, whereas uidA translationally fused to a CgRbcS gene generated GUS accumulation specifically in bundle sheath cells. This transformation procedure is the first for an NAD-ME type C(4) plant and should significantly accelerate the analysis of mechanisms underlying C(4) photosynthesis. Oxford University Press 2010-03 2010-02-11 /pmc/articles/PMC2837259/ /pubmed/20150516 http://dx.doi.org/10.1093/jxb/erq009 Text en © 2010 The Author(s). This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. This paper is available online free of all access charges (see http://jxb.oxfordjournals.org/open_access.html for further details)
spellingShingle Research Papers
Newell, Christine A.
Brown, Naomi J.
Liu, Zheng
Pflug, Alexander
Gowik, Udo
Westhoff, Peter
Hibberd, Julian M.
Agrobacterium tumefaciens-mediated transformation of Cleome gynandra L., a C(4) dicotyledon that is closely related to Arabidopsis thaliana
title Agrobacterium tumefaciens-mediated transformation of Cleome gynandra L., a C(4) dicotyledon that is closely related to Arabidopsis thaliana
title_full Agrobacterium tumefaciens-mediated transformation of Cleome gynandra L., a C(4) dicotyledon that is closely related to Arabidopsis thaliana
title_fullStr Agrobacterium tumefaciens-mediated transformation of Cleome gynandra L., a C(4) dicotyledon that is closely related to Arabidopsis thaliana
title_full_unstemmed Agrobacterium tumefaciens-mediated transformation of Cleome gynandra L., a C(4) dicotyledon that is closely related to Arabidopsis thaliana
title_short Agrobacterium tumefaciens-mediated transformation of Cleome gynandra L., a C(4) dicotyledon that is closely related to Arabidopsis thaliana
title_sort agrobacterium tumefaciens-mediated transformation of cleome gynandra l., a c(4) dicotyledon that is closely related to arabidopsis thaliana
topic Research Papers
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2837259/
https://www.ncbi.nlm.nih.gov/pubmed/20150516
http://dx.doi.org/10.1093/jxb/erq009
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