Genetic flexibility of plant chloroplasts

Frey, Jürg E.

doi:10.1038/18139

Cited by 33 publications

(21 citation statements)

References 1 publication

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“…Total leaf cellular DNA was extracted from plants using a method described previously [16]. Polymerase chain reactions (PCRs) were carried out using primers specific for the chimeric aadA or gfp genes and analysis for homoplasmy used a pair of primers (RBCL/ACCD) or (pSSH-rev/trnV-Solanum) flanking the transgene insertion site in the potato chloroplast genome.…”

Section: Pcr and Southern Blot Analyses Of Total Cellular Dnamentioning

confidence: 99%

Generation of homoplasmic plastid transformants of a commercial cultivar of potato (Solanum tuberosum L.)

et al. 2005

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This report describes the integration and expression of foreign genes into the plastid genome of a commercial cultivar of potato. Plastid transformation of potato was achieved using two tobacco specific plastid transformation vectors, pZS197 (Prrn/aadA/ psbA3 0 ) and pMSK18 (trc/gfp/Prrn/aadA/ psbA3 0 ). Selection was for spectinomycin resistance after biolistic delivery of plasmid DNA into leaf cells of Solanum tuberosum cv. Desiree. Ten transplastomic lines were obtained from 179 bombarded samples with vector pZS197 and four transplastomic lines selected out of 103 bombarded samples with vector pMSK18. Southern blot and PCR analyses confirmed homoplasmy in the primary regenerants, and incorporation of the aadA and gfp genes into the potato plastid genome by two homologous recombination events via the flanking plastid DNA sequences. Fluorometric measurements confirmed GFP expression in leaves and tubers of pMSK18 lines. No transformants were obtained with a third tobacco vector, pNtcZ7 (Prrn/gfp/ psbA3 0 /trc/aadA/rrnB-ter) in which the selectable marker gene is driven by a bacterial (trc) promoter, which does permit selection of plastid transformants in tobacco, and allows low level expression of the reporter gene, gfp, in potato. #

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Section: Pcr and Southern Blot Analyses Of Total Cellular Dnamentioning

confidence: 99%

Generation of homoplasmic plastid transformants of a commercial cultivar of potato (Solanum tuberosum L.)

et al. 2005

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“…For Southern blots, genomic DNA was extracted from leaves according to Frey (1999) with some modifications. Leaf tissue was ground in lysis buffer (20 mM Tris, pH 8.0; 20 mM of EDTA; 2 M NaCl), incubated at 85 °C for 5 min and then placed on ice for 5 min.…”

Section: ⎯⎯⎯⎯mentioning

confidence: 99%

Transformation of potato via Agrobacterium coated microparticle bombardment

Nguyen¹,

Dix²,

Nugent³

2010

Biol. plant.

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The transformation of potato (Solanum tuberosum L. cv. Désirée) was extended by the Agrobacterium-mediated biolistic method. Using this approach transgenic shoots could be obtained at a similar frequency to that achieved through conventional biolistics. Leaves from shoot cultures were bombarded with gold particles coated in Agrobacterium tumefaciens cells harboring a binary plasmid encoding three genes of interest in the T-DNA. Nine shoots were obtained from 20 shots, with selection of transgenic shoots on a series of media containing progressively increasing concentrations of hygromycin from 5 to 20 mg dm -3 .

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“…DNA was extracted from leaves as described by Frey (1998). To detect the ipt gene, two primers were designed to amplify a 395-bp fragment.…”

Section: Polymerase Chain Reactionmentioning

confidence: 99%

“…Genomic DNA was extracted from leaf tissue following the method described by Frey (1998). DNA was digested with HindIII or EcoRI and after electrophoresis was blotted to a nylon filter.…”

Section: Southern Analysismentioning

confidence: 99%

Delayed Senescence in Cauliflower Transformed with an Autoregulated Isopentenyl Transferase Gene

Nguyen

Jordi

Dun

et al. 2008

International Journal of Plant Sciences

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Cauliflower (Brassica oleracea var. botrytis) was transformed, using Agrobacterium tumefaciens, with an autoregulated isopentenyl transferase (ipt) gene under the control of a senescence-associated gene promoter, pSAG 12 , isolated from Arabidopsis thaliana. The effect of introducing this chimeric construct on cytokinin (CK) content, chlorophyll retention, and plant morphology and development were investigated. A range of CK and chlorophyll contents was found among the individual primary transformants. Progeny were studied from one of the primary transformed lines that did not have elevated cytokinin content and was phenotypically similar to the parent line but displayed delayed leaf senescence. The pSAG 12 :ipt gene was inherited in a Mendelian manner, and the effect of this gene on senescence-related parameters was observed in a number of the progeny. While the pSAG 12 :ipt progeny did exhibit delayed leaf senescence, it was accompanied by undesirable agronomic traits, including less synchronous curd initiation, smaller curd size, and greater susceptibility to fungal infection.

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Genetic flexibility of plant chloroplasts

Cited by 33 publications

References 1 publication

Generation of homoplasmic plastid transformants of a commercial cultivar of potato (Solanum tuberosum L.)

Generation of homoplasmic plastid transformants of a commercial cultivar of potato (Solanum tuberosum L.)

Transformation of potato via Agrobacterium coated microparticle bombardment

Delayed Senescence in Cauliflower Transformed with an Autoregulated Isopentenyl Transferase Gene

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