Plastid Transformation in Higher Plants

Maliga, Pál

doi:10.1146/annurev.arplant.55.031903.141633

Cited by 422 publications

(335 citation statements)

References 171 publications

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“…The lack of any known protein export mechanism from chloroplasts ensures the recombinant protein is localised to the target organelle, and obviates concerns over residual cytosolic expression due to inefficient chloroplast targeting. Furthermore, the importance of multi-component scavenging pathways, such as the ascorbate-glutathione cycle (Mittler 2002), together with the often limited effects of expressing individual components, suggest concerted expression of two or more genes might be more informative, something readily accomplished through chloroplast transformation (Bock 2001;Maliga 2004). This is illustrated by recent experiments in our laboratory (Le Martret et al 2011) in which we combine expression of dehydroascorbate reductase (DHAR) and glutathione reductase (GR) in tobacco chloroplasts.…”

Section: Discussionmentioning

confidence: 99%

Modification of reactive oxygen species scavenging capacity of chloroplasts through plastid transformation

et al. 2011

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Reactive oxygen species (ROS), including superoxide anions, hydrogen peroxide and hydroxyl radicals are generated through normal biochemical processes, but their production is increased by abiotic stresses. The prospects for enhancing ROS scavenging, and hence stress tolerance, by direct gene expression in a vulnerable cell compartment, the chloroplast, have been explored in tobacco. Several plastid transformants were generated which contained either a Nicotiana mitochondrial superoxide dismutase (MnSOD) or an Escherichia coli glutathione reductase (gor) gene. MnSOD lines had a three-fold increase in MnSOD activity, but interestingly a five to nine-fold increase in total chloroplast SOD activity. Gor transgenic lines had up to 6 times higher GR activity and up to 8 times total glutathione levels compared to wild type tobacco. Photosynthetic capacity of transplastomic plants, as measured by chlorophyll content and variable fluorescence of PSII was equivalent to non-transformed plants.The response of these transplastomic lines to several applied stresses was examined. In a number of cases improved stress tolerance was observed. Examples include enhanced methyl viologen (Paraquat)-induced oxidative stress tolerance in Mn-superoxidase dismutase overexpressing plants, improved heavy metal tolerance in glutathione reductase expressing lines, and improved tolerance to UV-B radiation in both sets of plants.

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Section: Discussionmentioning

confidence: 99%

Modification of reactive oxygen species scavenging capacity of chloroplasts through plastid transformation

et al. 2011

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“…Compared to nuclear transformation, the integration and expression of the transgene in the plastome has several potential advantages, as discussed in many original papers and literature reviews (Bock and Khan 2004;Maliga 2004;Bock 2007). However, plastid transformation has to date been applied mainly to the expression of heterologous genes involved in agronomic traits (e.g.…”

Section: Introductionmentioning

confidence: 99%

“…Although the generation of transplastomic plants became a reality with the ''binding-type'' vectors based on antibiotic-insensitive mutations in plastid genes (Svab et al 1990;Dix and Kavanagh 1995), agronomic trait manipulation by plastid transformation has been so far accomplished almost exclusively by using ''dominant-type'' vectors based on heterologous genes conferring antibiotic resistance (Maliga 2004;Bock 2007). Hence, to demonstrate the feasibility of using the ''binding-type'' vector strategy for manipulating the expression of agronomically relevant genes, the two D 9 desaturase genes were cloned in vectors containing only plant-derived sequences and inserted in transgenic plastids by PEG transformation of plant protoplasts.…”

Section: Introductionmentioning

confidence: 99%

Transplastomic tobacco plants expressing a fatty acid desaturase gene exhibit altered fatty acid profiles and improved cold tolerance

Craig¹,

Lenzi²,

Scotti³

et al. 2008

Transgenic Res

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The possibility of altering the unsaturation level of fatty acids in plant lipids by genetic transformation has implications for the stress tolerance of higher plants as well as for their nutritional value and industrial utilisation. While the integration and expression of transgenes in the plastome has several potential advantages over nuclear transformation, very few attempts have been made to manipulate fatty acid biosynthesis using plastid transformation. We produced transplastomic tobacco plants that express a D 9 desaturase gene from either the wild potato species Solanum commersonii or the cyanobacterium Anacystis nidulans, using PEG-mediated DNA uptake by protoplasts. Incorporation of chloroplast antibioticinsensitive point mutations in the transforming DNA was used to select transformants. The presence of the transcript and the D 9 desaturase protein in transplastomic plants was confirmed by northern and western blot analyses. In comparison with control plants, transplastomic plants showed altered fatty acid profiles and an increase in their unsaturation level both in leaves and seeds. The two transgenes produced comparable results. The results obtained demonstrate the feasibility of using plastid transformation to engineer lipid metabolic pathways in both vegetative and reproductive tissues and suggest an increase of cold tolerance in transplastomic plants showing altered leaf fatty acid profiles. This is the first example of transplastomic plants expressing an agronomically relevant gene produced with the ''binding-type'' vectors, which do not contain a heterologous marker gene. In fact, the transplastomic plants expressing the S. commersonii gene contain only plant-derived sequences, a clear attraction from a public acceptability perspective.

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“…Plastid transformation is a recently developed technique suitable for both fundamental studies on plastid gene function (Sugita et al 1997) and biotechnological applications (Bock 2001;Maliga 2003Maliga , 2004. The advantages of plastid transformation over nuclear transformation, including the potential for a high level of foreign protein production from plastomically inserted foreign transgenes, have been reviewed elsewhere (Heifetz 2000).…”

Section: Introductionmentioning

confidence: 99%

Plastid transformants of tomato selected using mutations affecting ribosome structure

Nugent

Have²,

Gulik³

et al. 2005

Plant Cell Rep

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Tomato plastid transformants were obtained using two vectors containing cloned plastid DNA of either Nicotiana tabacum or Solanum nigrum and including point mutations conferring resistance to spectinomycin and streptomycin. Transformants were recovered after PEGmediated direct DNA uptake into protoplasts, followed by selection on spectinomycin-containing medium. Sixteen lines contained the point mutation, as confirmed by mapping restriction enzyme sites. One line obtained with each vector was analysed in more detail, in comparison with a spontaneous spectinomycin-resistant mutant. Integration of the cloned Solanum or Nicotiana plastid DNA, by multiple recombination events, into the tomato plastome was confirmed by sequence analysis of the targeted region of plastid DNA in the inverted repeat region. Maternal inheritance of spectinomycin and streptomycin resistances or sensitivity in seedlings also confirmed the transplastomic status of the two transformants. The results demonstrate the efficacy in tomato of a selection strategy which avoids the integration of a dominant bacterial antibiotic resistance gene.Communicated by H. Lörz

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Plastid Transformation in Higher Plants

Cited by 422 publications

References 171 publications

Modification of reactive oxygen species scavenging capacity of chloroplasts through plastid transformation

Modification of reactive oxygen species scavenging capacity of chloroplasts through plastid transformation

Transplastomic tobacco plants expressing a fatty acid desaturase gene exhibit altered fatty acid profiles and improved cold tolerance

Plastid transformants of tomato selected using mutations affecting ribosome structure

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