Transformation Systems for Generating Marker–Free Transgenic Plants

Yoder, John I.; Goldsbrough, Andrew P.

doi:10.1038/nbt0394-263

Cited by 193 publications

(85 citation statements)

References 56 publications

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“…The safety of marker genes is another potential problem in plant genetic engineering. These issues were reviewed extensively by Yoder and Goldsbrough (1994). Thus, the demand for marker-free transgenic plants is high.…”

Section: Introductionmentioning

confidence: 99%

Vectors carrying two separate T‐DNAs for co‐transformation of higher plants mediated by Agrobacterium tumefaciens and segregation of transformants free from selection markers

Komari¹,

et al. 1996

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SummaryNovel 'super-binary' vectors that carried two separate T-DNAs were constructed. One T-DNA contained a drugresistance, selection-marker gene and the other contained a gene for 13-glucuronidase (GUS), A large number of tobacco (Nicotiana tabacum L.) and rice (Oryza sativa L.) transformants were produced by Agrobacterium tumefaciens LBA4404 that carried the vectors. Frequency of cotransformation with the two T-DNAs was greater than 47%. GUS-positive, drug-sensitive progeny were obtained from more than half of the co-transformants. Molecular analyses by Southern hybridization and polymerase chain reactions confirmed integration and segregation of the T-DNAs. Thus, the non-selectable T-DNA that was genetically separable from the selection marker was integrated into more than a quarter of the initial, drug-resistant transformants. Since various DNA fragments may be inserted into the non-selectable T-DNA by a simple procedure, these vectors will likely be very useful for the production of marker-free transformants of diverse plant species. Delivery of two T-DNAs to plants from mixtures of A. tumefaciens was also tested, but frequency of cotransformation was relatively low.

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Section: Introductionmentioning

confidence: 99%

Vectors carrying two separate T‐DNAs for co‐transformation of higher plants mediated by Agrobacterium tumefaciens and segregation of transformants free from selection markers

Komari¹,

et al. 1996

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“…Transformation technologies of nuclear genomes have been developed to eliminate antibiotic marker genes, an approach referred to as nuclear genome-clean gene transformation technology (CGTT) (Yoder et al, 1994). Over the past several years, consumer and environmental organizations have expressed ethical and biosafety concerns about the use of antibiotic-and herbicide-resistance genes derived from microorganisms (Miki & McHugh, 2004).…”

Section: Clean Gene Transformation Technology For Chloroplast Engineementioning

confidence: 99%

Application of Mutated Acetolactate Synthase Genes for Herbicide Resistance to Crop Improvement

Shimizu¹,

Kawai²,

Kaku³

et al. 2011

Herbicides, Theory and Applications

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“…Because of the controversy about releasing genetically engineered plants which carry antibiotic resistance marker genes into the environment, initiatives have been taken to generate transgenic plants which do not contain such genes (Yoder and Goldsbrough 1994). One approach was cotransformation using two Agrobacterium stains carrying different T-DNAs with the aim of obtaining uncoupled integration.…”

Section: Agrobacterium Tumefaciens Transformationmentioning

confidence: 99%

“…Various antibiotic resistance genes are used as selectable marker genes in the production of transgenic plants (Yoder and Goldsbrough 1994). In Brassica transformation, the most commonly used is neomycin phosphotransferase (EC 2.7.1.95) gene (nptll) from transposon Tn5 (Bevan et al 1983) which confers resistance towards some aminoglycosides such as kanamycin, neomycin, gentamicin and also paromomycin.…”

Section: Selectable Markersmentioning

confidence: 99%

Genetic transformation of Brassica

Poulsen

1996

Plant Breeding

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Tissue culture regeneration of plants from Brassica species can be achieved from various explant types. Their performance, however, varies with the genotype and age of the explant and is sensitive to medium supplements and Agrobacterium cocultivation. Most Brassica transformations were carried out using Agrobacterium and it was established that nopaline and agropine strains of .4. tumefaciens and A. rhizogenes, respectively, are the most efficient. Application of virulence gene induction is ambiguous. Binary vectors are mostly apphed in spite of indications that cointegrate vectors are more efficient. Cotransformation is successful with two different bacteria or with one bacterium carrying both plasmids, however, it has proved problematic to integrate the two transgenes into separate loci. Transformations with wild type rol genes develop hairy roots, from which transgenic plants can be regenerated. Vectorless transformation is feasible with microprojectile bombardment of microspore derived embryos being the most promising.Antibiotic markers providing resistance to kanamycin, hygromycin, and spectinomycin are applicable as selectable markers, while chloramphenicol is not suitable. Among herbicides, phosphinotricin gives good results while chlorsulfuron is unsuitable for in vitro selection. Major therapeutic antibiotics being successfully applied are carbenicillin, cefotaxime, amoxycillin, and ticarcillin. Standard methods for evaluating transgenic plants also have been applied to Brassica species. Some breeding objectives obtained through genetic transformation are presented.

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Transformation Systems for Generating Marker–Free Transgenic Plants

Cited by 193 publications

References 56 publications

Vectors carrying two separate T‐DNAs for co‐transformation of higher plants mediated by Agrobacterium tumefaciens and segregation of transformants free from selection markers

Vectors carrying two separate T‐DNAs for co‐transformation of higher plants mediated by Agrobacterium tumefaciens and segregation of transformants free from selection markers

Application of Mutated Acetolactate Synthase Genes for Herbicide Resistance to Crop Improvement

Genetic transformation of Brassica

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