Generation of "backbone" free, low transgene copy plants by launching T-DNA from the Agrobacterium chromosome

Oltmanns, Heiko; Frame, Bronwyn; Lee, Lan‐Ying; Johnson, Susan; Li, Bo; Wang, Kan; Gelvin, Stanton B.

doi:10.1038/npre.2009.3497.1

Cited by 19 publications

(43 citation statements)

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“…26 Launching of T-DNA from the Agrobacterium chromosome has also been tested for delivery of transgene. 27 This design, which minimizes copy number in Agrobacterium, results in a high percentage of single copy plant transformants with intact T-DNA, although the overall transformation frequency is lower compared to binary vectors. 27 Delivering one long T-DNA from the Agrobacterium chromosome might be a good alternative to the use of several smaller binary vectors.…”

Section: Current Trait Combination Tools: Breeding Stacks and Moleculmentioning

confidence: 98%

“…27 This design, which minimizes copy number in Agrobacterium, results in a high percentage of single copy plant transformants with intact T-DNA, although the overall transformation frequency is lower compared to binary vectors. 27 Delivering one long T-DNA from the Agrobacterium chromosome might be a good alternative to the use of several smaller binary vectors. However, the additional step of inserting T-DNA into the Agrobacterium chromosome in addition to the reduced transformation efficiency make the method more cumbersome in practice.…”

Section: Current Trait Combination Tools: Breeding Stacks and Moleculmentioning

confidence: 98%

See 1 more Smart Citation

Trait stacking in transgenic crops: Challenges and opportunities

Que¹,

Chilton²,

Fontes³

et al. 2010

GM Crops

177

116

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In recent years, there has been a rapid increase in the planting of transgenic crops with stacked traits. Most of these products have been formed by conventional breeding, i.e. the crossing of transgenic plant (event) containing individual transgenes with other event(s) containing single or double transgenic traits. Many biotech companies are developing stacked trait products with increasing numbers of insect and herbicide tolerance genes for controlling a broad range of insect pests and weeds. There has also been an increase in development of technologies for molecular stacking of multiple traits in a single transgene locus. In this review we look at the status of stacked trait products, crop trait stacking technologies and the technical challenges we are facing. We also review recent progress in developing technology for assembling large transgene arrays in vitro (molecular stacks), their delivery to crop plants and issues they pose for transgene expression.

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Section: Current Trait Combination Tools: Breeding Stacks and Moleculmentioning

confidence: 98%

Section: Current Trait Combination Tools: Breeding Stacks and Moleculmentioning

confidence: 98%

Trait stacking in transgenic crops: Challenges and opportunities

Que¹,

Chilton²,

Fontes³

et al. 2010

GM Crops

177

116

View full text Add to dashboard Cite

show abstract

Section: The Regulatory Environmentmentioning

confidence: 99%

“…The cost of securing regulatory approval is not only associated with the provision of information on selected events, but the associated indirect costs that are incurred whilst managing product development. For example, in the development of the transgenic products it is necessary to select for, or develop strategies for, (Oltmanns et al, 2010;Ye et al, 2011) generation of simple integration events. As well as the cost of dossier preparation other regulatory costs include physical separation requirements imposed by regulators (though these may be impractical or unnecessary), documentation and inspection requirements during trials, waste disposal and staff training.…”

Section: The Cost Of Regulationmentioning

confidence: 99%

Genetic modification; the development of transgenic ornamental plant varieties

Chandler

Sanchez²

2012

Plant Biotechnology Journal

101

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Summary Plant transformation technology (hereafter abbreviated to GM, or genetic modification) has been used to develop many varieties of crop plants, but only a few varieties of ornamental plants. This disparity in the rate and extent of commercialisation, which has been noted for more than a decade, is not because there are no useful traits that can be engineered into ornamentals, is not due to market potential and is not due to a lack of research and development activity. The GM ornamental varieties which have been released commercially have been accepted in the marketplace. In this article, progress in the development of transgenic ornamentals is reviewed and traits useful to both consumers and producers are identified. In considering possible factors limiting the release of genetically modified ornamental products it is concluded that the most significant barrier to market is the difficulty of managing, and the high cost of obtaining, regulatory approval.

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“…A recent study (Oltmanns et al. 2010) demonstrated that starting plant transformation from the A. tumefaciens chromosome could be a way to reduce the portion of transgenic plant lines with vector backbone integrations dramatically, but so far, this procedure to produce Agrobacterium plant transformation strains is laborious and not applicable to the widely used strain A. tumefaciens LBA4404.…”

Section: Confirming the Lack Of Vector Backbonementioning

confidence: 99%

Efficient screening of transgenic plant lines for ecological research

Gase

Weinhold

Bozorov

et al. 2011

Molecular Ecology Resources

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Plants stably transformed to manipulate the expression of genes mediating ecological performance have profoundly altered research in plant ecology. Agrobacterium-mediated transformation remains the most effective method of creating plants harbouring a limited number of transgene integrations of low complexity. For ecological/physiological research, the following requirements must be met: (i) the regenerated plants should have the same ploidy level as the corresponding wild-type plant and (ii) contain a single transgene copy in a homozygous state; (iii) the T-DNA must be completely inserted without vector backbone sequence and all its elements functional; and (iv) the integration should not change the phenotype of the plant by interrupting chromosomal genes or by mutations occurring during the regeneration procedure. The screening process to obtain transformed plants that meet the above criteria is costly and time-consuming, and an optimized screening procedure is presented. We developed a flow chart that optimizes the screening process to efficiently select transformed plants for ecological research. It consists of segregational analyses, which select transgenic T₁ and T₂ generation plants with single T-DNA copies that are homozygous. Indispensable molecular genetic tests (flow cytometry, diagnostic PCRs and Southern blotting) are performed at the earliest and most effective times in the screening process. qPCR to quantify changes in transcript accumulation to confirm gene silencing or overexpression is the last step in the selection process. Because we routinely transform the wild tobacco, Nicotiana attenuata, with constructs that silence or ectopically overexpress ecologically relevant genes, the proposed protocol is supported by examples from this system.

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Generation of "backbone" free, low transgene copy plants by launching T-DNA from the Agrobacterium chromosome

Cited by 19 publications

References 0 publications

Trait stacking in transgenic crops: Challenges and opportunities

Trait stacking in transgenic crops: Challenges and opportunities

Genetic modification; the development of transgenic ornamental plant varieties

Efficient screening of transgenic plant lines for ecological research

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