Ectopic Transplastomic Expression of a Synthetic MatK Gene Leads to Cotyledon-Specific Leaf Variegation
Chloroplasts (and other plastids) harbor their own genetic material, with a bacterial-like gene-expression systems. Chloroplast RNA metabolism is complex and is predominantly mediated by nuclear-encoded RNA-binding proteins. In addition to these nuclear factors, the chloroplast-encoded intron matura...
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doaj-e98039705ff34a959f8a15a293fe7fef2020-11-24T20:56:59ZengFrontiers Media S.A.Frontiers in Plant Science1664-462X2018-10-01910.3389/fpls.2018.01453377404Ectopic Transplastomic Expression of a Synthetic MatK Gene Leads to Cotyledon-Specific Leaf VariegationYujiao Qu0Julia Legen1Jürgen Arndt2Stephanie Henkel3Galina Hoppe4Christopher Thieme5Giovanna Ranzini6Jose M. Muino7Andreas Weihe8Uwe Ohler9Gert Weber10Gert Weber11Oren Ostersetzer12Christian Schmitz-Linneweber13Institut für Biologie, Humboldt-Universität zu Berlin, Berlin, GermanyInstitut für Biologie, Humboldt-Universität zu Berlin, Berlin, GermanyInstitut für Biologie, Humboldt-Universität zu Berlin, Berlin, GermanyInstitut für Biologie, Humboldt-Universität zu Berlin, Berlin, GermanyInstitut für Biologie, Humboldt-Universität zu Berlin, Berlin, GermanyInstitut für Biologie, Humboldt-Universität zu Berlin, Berlin, GermanyInstitut für Biologie, Humboldt-Universität zu Berlin, Berlin, GermanyInstitut für Biologie, Humboldt-Universität zu Berlin, Berlin, GermanyInstitut für Biologie, Humboldt-Universität zu Berlin, Berlin, GermanyComputational Regulatory Genomics, Berlin Institute for Medical Systems Biology, Max Delbrück Center for Molecular Medicine, Berlin, GermanyInstitut für Biologie, Humboldt-Universität zu Berlin, Berlin, GermanyHelmholtz-Zentrum Berlin für Materialien und Energie, Joint Research Group Macromolecular Crystallography, Berlin, GermanyDepartment of Plant and Environmental Sciences, The Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem, Jerusalem, IsraelInstitut für Biologie, Humboldt-Universität zu Berlin, Berlin, GermanyChloroplasts (and other plastids) harbor their own genetic material, with a bacterial-like gene-expression systems. Chloroplast RNA metabolism is complex and is predominantly mediated by nuclear-encoded RNA-binding proteins. In addition to these nuclear factors, the chloroplast-encoded intron maturase MatK has been suggested to perform as a splicing factor for a subset of chloroplast introns. MatK is essential for plant cell survival in tobacco, and thus null mutants have not yet been isolated. We therefore attempted to over-express MatK from a neutral site in the chloroplast, placing it under the control of a theophylline-inducible riboswitch. This ectopic insertion of MatK lead to a variegated cotyledons phenotype. The addition of the inducer theophylline exacerbated the phenotype in a concentration-dependent manner. The extent of variegation was further modulated by light, sucrose and spectinomycin, suggesting that the function of MatK is intertwined with photosynthesis and plastid translation. Inhibiting translation in the transplastomic lines has a profound effect on the accumulation of several chloroplast mRNAs, including the accumulation of an RNA antisense to rpl33, a gene coding for an essential chloroplast ribosomal protein. Our study further supports the idea that MatK expression needs to be tightly regulated to prevent detrimental effects and establishes another link between leaf variegation and chloroplast translation.https://www.frontiersin.org/article/10.3389/fpls.2018.01453/fullchloroplastvariegationMatKsplicingtransplastomicNicotiana tabacum |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yujiao Qu Julia Legen Jürgen Arndt Stephanie Henkel Galina Hoppe Christopher Thieme Giovanna Ranzini Jose M. Muino Andreas Weihe Uwe Ohler Gert Weber Gert Weber Oren Ostersetzer Christian Schmitz-Linneweber |
spellingShingle |
Yujiao Qu Julia Legen Jürgen Arndt Stephanie Henkel Galina Hoppe Christopher Thieme Giovanna Ranzini Jose M. Muino Andreas Weihe Uwe Ohler Gert Weber Gert Weber Oren Ostersetzer Christian Schmitz-Linneweber Ectopic Transplastomic Expression of a Synthetic MatK Gene Leads to Cotyledon-Specific Leaf Variegation Frontiers in Plant Science chloroplast variegation MatK splicing transplastomic Nicotiana tabacum |
author_facet |
Yujiao Qu Julia Legen Jürgen Arndt Stephanie Henkel Galina Hoppe Christopher Thieme Giovanna Ranzini Jose M. Muino Andreas Weihe Uwe Ohler Gert Weber Gert Weber Oren Ostersetzer Christian Schmitz-Linneweber |
author_sort |
Yujiao Qu |
title |
Ectopic Transplastomic Expression of a Synthetic MatK Gene Leads to Cotyledon-Specific Leaf Variegation |
title_short |
Ectopic Transplastomic Expression of a Synthetic MatK Gene Leads to Cotyledon-Specific Leaf Variegation |
title_full |
Ectopic Transplastomic Expression of a Synthetic MatK Gene Leads to Cotyledon-Specific Leaf Variegation |
title_fullStr |
Ectopic Transplastomic Expression of a Synthetic MatK Gene Leads to Cotyledon-Specific Leaf Variegation |
title_full_unstemmed |
Ectopic Transplastomic Expression of a Synthetic MatK Gene Leads to Cotyledon-Specific Leaf Variegation |
title_sort |
ectopic transplastomic expression of a synthetic matk gene leads to cotyledon-specific leaf variegation |
publisher |
Frontiers Media S.A. |
series |
Frontiers in Plant Science |
issn |
1664-462X |
publishDate |
2018-10-01 |
description |
Chloroplasts (and other plastids) harbor their own genetic material, with a bacterial-like gene-expression systems. Chloroplast RNA metabolism is complex and is predominantly mediated by nuclear-encoded RNA-binding proteins. In addition to these nuclear factors, the chloroplast-encoded intron maturase MatK has been suggested to perform as a splicing factor for a subset of chloroplast introns. MatK is essential for plant cell survival in tobacco, and thus null mutants have not yet been isolated. We therefore attempted to over-express MatK from a neutral site in the chloroplast, placing it under the control of a theophylline-inducible riboswitch. This ectopic insertion of MatK lead to a variegated cotyledons phenotype. The addition of the inducer theophylline exacerbated the phenotype in a concentration-dependent manner. The extent of variegation was further modulated by light, sucrose and spectinomycin, suggesting that the function of MatK is intertwined with photosynthesis and plastid translation. Inhibiting translation in the transplastomic lines has a profound effect on the accumulation of several chloroplast mRNAs, including the accumulation of an RNA antisense to rpl33, a gene coding for an essential chloroplast ribosomal protein. Our study further supports the idea that MatK expression needs to be tightly regulated to prevent detrimental effects and establishes another link between leaf variegation and chloroplast translation. |
topic |
chloroplast variegation MatK splicing transplastomic Nicotiana tabacum |
url |
https://www.frontiersin.org/article/10.3389/fpls.2018.01453/full |
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