The chloroplast transformation toolbox: selectable markers and marker removal

Day, Anil; Goldschmidt-Clermont, Michel

doi:10.1111/j.1467-7652.2011.00604.x

Cited by 172 publications

(141 citation statements)

References 117 publications

Supporting

Mentioning

139

Contrasting

Unclassified

Order By: Relevance

“…In contrast, genes introduced into the nuclear genome integrate randomly and are therefore prone to position effects and RNA silencing. (Day and Goldschmidt-Clermont, 2011).…”

Section: Introductionmentioning

confidence: 99%

Expression and membrane-targeting of an active plant cytochrome P450 in the chloroplast of the green alga Chlamydomonas reinhardtii

et al. 2015

View full text Add to dashboard Cite

The unicellular green alga Chlamydomonas reinhardtii has potential as a cell factory for the production of recombinant proteins and other novel compounds, but mainstream adoption has been hindered by a scarcity of genetic tools and a need to identify products that can be generated in a cost-effective manner. A promising strategy is to use algal chloroplasts as a site for synthesis of high value bioactive compounds such as diterpenoids since these are derived from metabolic building blocks that occur naturally within the organelle. However, synthesis of these complex plant metabolites requires the introduction of membraneassociated enzymes including cytochrome P450 enzymes (P450s). Here, we show that a gene (CYP79A1) encoding a model P450 can be introduced into the C. reinhardtii chloroplast genome using a simple transformation system. The gene is stably expressed and the P450 is efficiently targeted into chloroplast membranes, where it is active and accounts for 0.4% of total cell protein. These results provide proof of concept for the introduction of diterpenoid synthesis pathways into the chloroplast of Chlamydomonas reinhardtii.

show abstract

“…In contrast, genes introduced into the nuclear genome integrate randomly and are therefore prone to position effects and RNA silencing. (Day and Goldschmidt-Clermont, 2011).…”

Section: Introductionmentioning

confidence: 99%

Expression and membrane-targeting of an active plant cytochrome P450 in the chloroplast of the green alga Chlamydomonas reinhardtii

et al. 2015

View full text Add to dashboard Cite

show abstract

“…Although several selection markers and protocols have been developed, those based on aadA (encoding aminoglycoside 3″-adenylyltransferase, EC 2.7.7.47, and conferring spectinomycin and streptomycin resistance to bacteria by detoxification) are the most frequent ones (Table 1), most likely because they require low expression levels to confer phenotypic resistance. Detailed description and historical overview of the used vectors including promoters, effective selection markers, reporter genes, their insertion, and eventual removal are provided by recent reviews (Maliga 2003(Maliga , 2004Koop et al 2007;Verma and Daniell 2007;Ruhlman et al 2010;Day and Goldschmidt-Clermont 2011;Maliga and Bock 2011;Ahmad and Mukhtar 2013;Hanson et al 2013;Vafaee et al 2014).…”

Section: Genetic Transformation Of Plastidsmentioning

confidence: 99%

“…Several methods have been developed to remove the (antibiotic resistance) marker genes in order to facilitate the acceptance of transplastomic crops and also to allow multiple rounds of plastid transformation with the same marker gene (also called marker recycling) due to the low number of efficient selection systems available for plastid transformation (reviewed in Day et al 2005;Koop et al 2007;Day and Goldschmidt-Clermont 2011). In addition, constitutive and high level expression of a marker gene represents a metabolic burden to the transformed plant and may negatively impact its fitness and yield.…”

Section: Public Concerns Associated With Transplastomic Plantsmentioning

confidence: 99%

“…In order to obtain new and genetically uniform transplastomic crops, the transformed plastid DNA copies have to be maintained, while the plastids carrying nontransformed DNA have to be gradually eliminated on a selective medium (reviewed in Maliga 2003Maliga , 2004Day and Goldschmidt-Clermont 2011;Maliga and Bock 2011). Selective amplification of the transgenic DNA copies and elimination of the non-transformed ones first within the highly polyploid plastids, then in the plant cells, followed by subsequent identification and amplification of these so-called homoplasmic cells, and especially the reproducible regeneration protocol of genetically uniform and fertile plants containing a uniform population of transformed plastid genomes from these cells (and/or tissue cultures) ( Fig.…”

Section: Genetic Transformation Of Plastidsmentioning

confidence: 99%

“…This overview discusses data related to genetic modification of the plastid genome of higher plants with emphasis on crops. For details about plastid transformation protocols developed for eukaryotic (micro)algae, the readers are kindly directed to recent reviews (Koop et al 2007;Day and Goldschmidt-Clermont 2011;Purton et al 2013). …”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Transplastomic plants for innovations in agriculture. A review

Wani

Sah

Sági

et al. 2015

Agron. Sustain. Dev.

View full text Add to dashboard Cite

Food production has to be significantly increased in order to feed the fast growing global population estimated to be 9.1 billion by 2050. The Green Revolution and the development of advanced plant breeding tools have led to a significant increase in agricultural production since the 1960s. However, hundreds of millions of humans are still undernourished, while the area of total arable land is close to its maximum utilization and may even decrease due to climate change, urbanization, and pollution. All these issues necessitate a second Green Revolution, in which biotechnological engineering of economically and nutritionally important traits should be critically and carefully considered. Since the early 1990s, possible applications of plastid transformation in higher plants have been constantly developed. These represent viable alternatives to existing nuclear transgenic technologies, especially due to the better transgene containment of transplastomic plants. Here, we present an overview of plastid engineering techniques and their applications to improve crop quality and productivity under adverse growth conditions. These applications include (1) transplastomic plants producing insecticidal, antibacterial, and antifungal compounds. These plants are therefore resistant to pests and require less pesticides. (2) Transplastomic plants resistant to cold, drought, salt, chemical, and oxidative stress. Some pollution tolerant plants could even be used for phytoremediation. (3) Transplastomic plants having higher productivity as a result of improved photosynthesis. (4) Transplastomic plants with enhanced mineral, micronutrient, and macronutrient contents. We also evaluate field trials, biosafety issues, and public concerns on transplastomic plants. Nevertheless, the transplastomic technology is still unavailable for most staple crops, including cereals. Transplastomic plants have not been commercialized so far, but if this crop limitation were overcome, they could contribute to sustainable development in agriculture.

show abstract

Chloroplast Genome

Howe¹

2012

Encyclopedia of Life Sciences

View full text Add to dashboard Cite

Chloroplasts contain a genome that is a relic of the endosymbiont that gave rise to the organelle. These genomes typically contain 100–200 genes and encode proteins important for photosynthesis and other chloroplast functions, although dinoflagellate algae have a greatly reduced and fragmented chloroplast genome. Many nonphotosynthetic taxa retain a remnant chloroplast genome. In some organisms the chloroplast genome may be retained to allow redox‐mediated control of gene expression, whereas in others it may continue to exist because transfer of essential genes to the nucleus is no longer possible. Multiple ribonucleic acid (RNA) polymerases are involved in chloroplast transcription in land plants, and post‐transcriptional processing involving nuclear‐encoded RNA‐binding proteins also plays an important part in the expression of the chloroplast genome. Gene expression may be regulated at a number of levels, and redox poise in the organelle may be particularly important for this. Key Concepts: Chloroplasts originated in the ancestor of plants and red and green algae by endosymbiotic acquisition of a cyanobacterium, and then spread to many other eukaryotic lineages. There are other examples of stable acquisition of a cyanobacterium by a nonphotosynthetic host. Chloroplast genomes are typically 100–200 kbp in size, and include a set of genes for proteins essential to photosynthesis. Other genes present in the ancestral symbiont have been lost or relocated to the nucleus. Many nonphotosynthetic organisms retain remnant chloroplasts, with genomes, reflecting the fact that photosynthesis is not the only biochemical process that takes place in the chloroplast. In many organisms, gene transfer from chloroplast to nucleus can still take place, at an unexpectedly high frequency. Establishment of the chloroplast has resulted in the development of nuclear‐encoded RNA‐binding protein families and, in land plants, additional RNA polymerases that play a central role in chloroplast gene expression. Post‐transcriptional RNA processing plays an important role in chloroplast gene expression. In photosynthetic organisms, chloroplast gene expression is closely linked to the organelle's redox poise. The chloroplast can also influence the expression of nuclear genes.

show abstract

The chloroplast transformation toolbox: selectable markers and marker removal

Cited by 172 publications

References 117 publications

Expression and membrane-targeting of an active plant cytochrome P450 in the chloroplast of the green alga Chlamydomonas reinhardtii

Expression and membrane-targeting of an active plant cytochrome P450 in the chloroplast of the green alga Chlamydomonas reinhardtii

Transplastomic plants for innovations in agriculture. A review

Chloroplast Genome

Contact Info

Product

Resources

About