Long regions of homologous DNA are incorporated into the tobacco plastid genome by transformation.

Staub, Jeffrey M.; Maliga, Pál

doi:10.1105/tpc.4.1.39

Cited by 92 publications

(28 citation statements)

References 27 publications

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“…These authors also found that the transformation frequency in tomato was lower than that obtainable with tobacco. Staub and Maliga (1992) showed that almost all of the 6.2-kb homologous cloned ptDNA in the same ptDNA inverted repeat region as that targeted by pSSH1 integrated in two tobacco plastid transformants. However multiple recombinations were evident when a larger number of transformants were analysed in a homeologous (Solanum/Nicotiana) system with a 7.8-kb target with pSSH1 (Kavanagh et al 1999).…”

Section: Discussionmentioning

confidence: 99%

“…Selection based on binding-type markers, however, has the advantage that primary transformants are usually homoplasmic, where all plastome copies in cells contain the introduced DNA (Dix and Kavanagh 1995). The bindingtype markers are therefore a good alternative when using PEG-mediated transformation compared to biolistics, given that a large number of shots are needed to recover transformants with these plasmids (Svab et al 1990;Staub and Maliga 1992). An efficient protoplast regeneration system is required irrespective of the selection system used.…”

Section: Introductionmentioning

confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Plastid transformants of tomato selected using mutations affecting ribosome structure

Nugent

Have²,

Gulik³

et al. 2005

Plant Cell Rep

View full text Add to dashboard Cite

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“…The number of plastid genomes per organelle for Arabidopsis seedlings grown under our conditions is not known but is generally estimated to be between 5 and 100 copies per organelle (Herrmann, 1992;Staub and Maliga, 1992). Virtually all of the mitochondrial genes are expressed as monocistronic transcriptional units and do not present very large targets for UV-induced damage.…”

Section: Discussionmentioning

confidence: 99%

Little or No Repair of Cyclobutyl Pyrimidine Dimers Is Observed in the Organellar Genomes of the Young Arabidopsis Seedling

1996

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A Southern-blot-based, site-specific assay for ultraviolet (UV)-induced cyclobutyl pyrimidine dimers (CPDs), employing the CPDspecific enzyme 14 endonuclease V, was used to follow the repair of this lesion in particular DNA sequences in 5-to 6-d-old Arabidopsis fhaliana seedlings. CPDs, measured as enzyme-sensitive sites, in nuclear sequences were removed rapidly in the light but were repaired slowly, if at all, in the dark. This result was identical to that obtained in prior analyses of CPDs in total cellular DNA. Assay of representative chloroplast and mitochondrial sequences in the same DNA preparations revealed that, in contrast to nuclear sequences, enzyme-sensitive sites are inefficiently eliminated in both the presente and absence of visible light. These observations suggest that Arabidopsis seedlings possess little or no capacity for the repair of CPDs in the organellar genomes. Civen the fact that the UV dose employed only marginally affected the growth of the seedlings, we suggest that Arabidopsis seedlings must possess very efficient mechanism(s) for the tolerance of UV-induced DNA damage.Plant cells contain three distinct genomes encoded by the nucleus, the plastid, and the mitochondrion. A11 three genomes inevitably suffer damage due to the actions of UV light, oxidative damage, and spontaneous hydrolysis (Britt, 1996). Because damaged bases can act as blocks to both DNA replication and transcription, each of these three organelles must have one or more pathway(s) for the repair and/ or tolerance of DNA damage. UV-B induces severa1 different kinds of DNA damage products. Among these, CPDs and 6 4 photoproducts are the most abundant. Both of these lesions have been shown to block the progress of RNA polymerase in mammalian systems (Protic-Sabljic and Kraemer, 1986; Mitchell, et al., 1989). By assaying repair of total cellular DNA extracted from Arabidopsis thaliana seedlings, it has been established that this plant maintains two distinct photorepair systems that specifically and efficiently eliminate both classes of dimers (Pang and Hays, 1991;Chen, et al., 1994). At least some fraction of this repair must represent photorepair of nDNA, because more than 70% of these lesions are repaired within 2 h of exposure to visible light. Arabidopsis also has been shown to possess light-independent ("dark) repair pathways (Pang and Hays, 1991;Britt, et al., 1993). In the dark, Arabidopsis seedlings eliminate 6-4 photoproducts much more efficiently than CPDs from their total cellular DNA. This is also the case in mammalian, particularly rodent, cell lines (Mitchell, et al., 1985).Studies of repair in Arabidopsis have so far been limited to the analysis of total cellular DNA. Because no repair proteins are known to be encoded by the organellar genomes, and because any nuclear-encoded organellar proteins must be specifically targeted to the organelle, it is likely that each of the three plant genomes maintains a very different battery of DNA damage repair and tolerance pathways. To study DNA repair in a single ge...

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“…Although it is feasible to target gene interruptions or deletions to the chloroplast genome of plants (Svab et al, 1990;Staub and Maliga, 1992), such experiments can be performed more quickly and easily with the prokaryotic cyanobacteria. Most cyanobacterial and chloroplast genes show a remarkable degree of sequence homology, and in some cases, the same gene organization has also been reported (Steinmüller et al, 1989;Anderson and Mclntosh, 1991).…”

Section: Introductionmentioning

confidence: 99%

Inactivation of a Synechocystis sp strain PCC 6803 gene with homology to conserved chloroplast open reading frame 184 increases the photosystem II-to-photosystem I ratio.

et al. 1995

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A gene of the unicellular cyanobacterium Synechocystis sp strain PCC 6803 that is homologous to the conserved chloroplast open reading frame orf784 has been cloned and sequenced. The nucleotide sequence of the gene predicts a protein of 184 amino acids with a calculated molecular mass of 21.5 kD and two membrane-spanning regions. Amino acid sequence analysis showed 46 to 37% homology of the cyanobacterial orf784 with tobacco orf784, rice orf785, liverwort orf184, and Euglena gracilis orf206 sequences. Two orf784-specific mutants of Synechocystis sp PCC 6803 were constructed by insertion mutagenesis. Cells of mutants showed growth characteristics similar to those of the wild type. Their pigment composition was distinctly different from the wild type, as indicated by an increase in the phycocyanin-tochlorophyll ratio. In addition, mutants also had a two-to threefold increase in photosynthetic electron transfer rates as well as in photosystem Il-to-photosystem I ratio-a phenomenon hitherto not reported for mutants with altered photosynthetic characteristics. The observed alterations in the orfl84-specific mutants provide strong evidence for a functional role of the orf784 gene product in photosynthetic processes. INTRODUCTIONConsiderable sequence data on plastid genomes have been accumulated during recent years. The plastid DNA from severa1 sources has been completely sequenced, including liverwort (Ohyama et al., 1986), tobacco (Shinozaki et al., 1986, rice (Hiratsuka et ai., 1989), Epifagus virginiana (Wolfe et al., 1992a, 1992b, and Euglena gradis (Hallick et al., 1993). In addition to genes encoding rRNAs, tRNAs, and proteins that are mostly involved in housekeeping and photosynthesis, all plastid genomes have been found to contain several open reading frames (ORFs). Ten of these ORFs found in tobacco chloroplast DNA are also present in the genomes of liverwort and rice (Shimada and Sugiura, 1991). Euglefla gracilis (Hallick et al., 1993) and Epifagus virginiana (Wolfe et al., 1992a(Wolfe et al., , 1992b plastid DNAs each contain three of these conserved ORFs. The retained primary structure of the ORFs even from different phylogenetic groups indicates a functional role for these putative genes.Possible functions for some of these ORFs have been predicted on the basis of homology with already known genes (Maurizi et al., 1990; Li and Cronan, 1992;Wolfe, 1994). An alternative approach to gain new information about putative ORF proteins would be to examine the phenotypic 1 To whom correspondence should b e addressed.consequences of their inactivation. Although it is feasible to target gene interruptions or deletions to the chloroplast genome of plants (Svab et al., 1990;Staub and Maliga, 1992), such experiments can be performed more quickly and easily with the prokaryotic cyanobacteria. Most cyanobacterial and chloroplast genes show a remarkable degree of sequence homology, and in some cases, the same gene organization has also been reported (Steinmüller et al., 1989;Anderson and Mclntosh, 1991). Thus, cyanobact...

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Long regions of homologous DNA are incorporated into the tobacco plastid genome by transformation.

Cited by 92 publications

References 27 publications

Plastid transformants of tomato selected using mutations affecting ribosome structure

Plastid transformants of tomato selected using mutations affecting ribosome structure

Little or No Repair of Cyclobutyl Pyrimidine Dimers Is Observed in the Organellar Genomes of the Young Arabidopsis Seedling

Inactivation of a Synechocystis sp strain PCC 6803 gene with homology to conserved chloroplast open reading frame 184 increases the photosystem II-to-photosystem I ratio.

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