Enhanced production of single copy backbone-free transgenic plants in multiple crop species using binary vectors with a pRi replication origin in Agrobacterium tumefaciens

Ye, Xudong; Williams, Edward J.; Shen, Junjiang; Johnson, Susan; Lowe, Brenda; Radke, Sharon E.; Strickland, Steve; Esser, James A.; Petersen, Michael W.; Gilbertson, Larry A.

doi:10.1007/s11248-010-9458-6

Cited by 29 publications

(49 citation statements)

References 43 publications

(93 reference statements)

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“…However, pRi ori-based binary vectors are not routinely used due to its low copy numbers and thus require more effort for molecular manipulation. Ye et al (2011) reported that pRi ori binary vectors significantly increased the frequency of single-copy, backbone-free transgenic events relative to RK2 oriV-based binary vectors in multiple crops, including maize.…”

Section: Improvement In Transgenic Event Qualitymentioning

confidence: 99%

Maize transformation technology development for commercial event generation

Que

Sivamani

et al. 2014

Front. Plant Sci.

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Maize is an important food and feed crop in many countries. It is also one of the most important target crops for the application of biotechnology. Currently, there are more biotech traits available on the market in maize than in any other crop. Generation of transgenic events is a crucial step in the development of biotech traits. For commercial applications, a high throughput transformation system producing a large number of high quality events in an elite genetic background is highly desirable. There has been tremendous progress in Agrobacterium-mediated maize transformation since the publication of the Ishida et al. (1996) paper and the technology has been widely adopted for transgenic event production by many labs around the world. We will review general efforts in establishing efficient maize transformation technologies useful for transgenic event production in trait research and development. The review will also discuss transformation systems used for generating commercial maize trait events currently on the market. As the number of traits is increasing steadily and two or more modes of action are used to control key pests, new tools are needed to efficiently transform vectors containing multiple trait genes. We will review general guidelines for assembling binary vectors for commercial transformation. Approaches to increase transformation efficiency and gene expression of large gene stack vectors will be discussed. Finally, recent studies of targeted genome modification and transgene insertion using different site-directed nuclease technologies will be reviewed.

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Section: Improvement In Transgenic Event Qualitymentioning

confidence: 99%

Maize transformation technology development for commercial event generation

Que

Sivamani

et al. 2014

Front. Plant Sci.

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“…Obtaining plants lacking Cas9 becomes more difficult if several copies of the T-DNA are inserted at different chromosomal locations. It is known that binary vectors that replicate at a single copy in Agrobacterium lead to a higher proportion of transformants containing single copy, backbone-free transgenic plants (Ye et al, 2011). The plasmid backbone used in the experiments described above (Figure 1) has a pVS1 origin of replication, which is known to replicate in Agrobacterium at approximately 20 copies (Heeb et al, 2000;Itoh et al, 1984;Zhi et al, 2015).…”

Section: Test Of a Construct With Low Copy Number In Agrobacteriummentioning

confidence: 99%

“…The plasmid backbone used in the experiments described above (Figure 1) has a pVS1 origin of replication, which is known to replicate in Agrobacterium at approximately 20 copies (Heeb et al, 2000;Itoh et al, 1984;Zhi et al, 2015). We therefore tested a binary vector (pAGM37443) that contains the Agrobacterium rhizogenes A4 plasmid origin of replication, which replicates in Agrobacterium at approximately 1 copy per cell (Nishiguchi et al, 1987;Ye et al, 2011). The same constructs as described previously ( Figure 1) were assembled again in vector pAGM37443 and transformed in Arabidopsis by floral dipping.…”

Section: Test Of a Construct With Low Copy Number In Agrobacteriummentioning

confidence: 99%

Addition of Multiple Introns to a Cas9 Gene Results in Dramatic Improvement in Efficiency for Generation of Gene Knockouts in Plants

Grützner

Martin

Horn

et al. 2020

Preprint

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Running title: High efficiency site-targeted mutagenesis with an intronized Cas9 gene AbstractThe recent discovery of the mode of action of the CRISPR/Cas9 system has provided biologists with a useful tool for generating site-specific mutations in genes of interest. In plants, site-targeted mutations are usually obtained by stably transforming a Cas9 expression construct into the plant genome. The efficiency with which mutations are obtained in genes of interest can vary considerably depending on specific features of the constructs, including the source and nature of the promoters and terminators used for expression of the Cas9 gene and the guide RNA, and the sequence of the Cas9 nuclease itself.To optimize the efficiency with which mutations could be obtained in target genes in Arabidopsis thaliana with the Cas9 nuclease, we have investigated several features of its nucleotide and/or amino acid sequence, including the codon usage, the number of nuclear localization signals (NLS) and the presence or absence of introns. We found that the Cas9 gene codon usage had some effect on Cas9 activity and that two NLSs work better than one. However, the most important impact on the efficiency of the constructs was obtained by addition of 13 introns into the Cas9 coding sequence, which dramatically improved editing efficiencies of the constructs; none of the primary transformants obtained with a Cas9 lacking introns displayed a knockout mutant phenotype, whereas between 70% and 100% of primary transformants generated with intronized Cas9 displayed mutant phenotypes. The intronized Cas9 was also found to be effective in other plants such as Nicotiana benthamiana and Catharanthus roseus.

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“…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.…”

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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Enhanced production of single copy backbone-free transgenic plants in multiple crop species using binary vectors with a pRi replication origin in Agrobacterium tumefaciens

Cited by 29 publications

References 43 publications

Maize transformation technology development for commercial event generation

Maize transformation technology development for commercial event generation

Addition of Multiple Introns to a Cas9 Gene Results in Dramatic Improvement in Efficiency for Generation of Gene Knockouts in Plants

Genetic modification; the development of transgenic ornamental plant varieties

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