High-resolution structural analysis of biolistic transgene integration into the genome of wheat

Jackson, Scott A.; Zhang, P.; Chen, W. P.; Phillips, Ronald L.; Friebe, Bernd; Muthukrishnan, S.; Gill, Bikram S.

doi:10.1007/s001220100608

Cited by 59 publications

(23 citation statements)

References 24 publications

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“…The salient finding was that an array of integrated transgenes, genetically segregating as a single unit, could be interrupted with genomic DNA. This was found to be the case in transgenic rice (Kohli et al 1999b) oat (Pawlowski and Somers 1998;Svitashev et al 2000Svitashev et al , 2002Svitashev and Somers 2001;Makarevitch et al 2003) maize (Mehlo et al 2000) wheat (Abranches et al 2000;Jackson et al 2001) and potato (Romano et al 2003b). These studies made it remarkably clear that transgenes can undergo rearrangements before or during integration into the host genome.…”

Section: Particle Bombardment Facilitates Simultaneous Multiple Gene mentioning

confidence: 96%

“…Such studies have been carried out mainly in cereals with large nuclei and chromosomes, such as wheat (Abranches et al 2000;Jackson et al 2001) and oat (Svitashev et al 2000;Svitashev and Somers 2001), although there were also experiments in rice (Dong et al 2001). Following these studies, Kohli et al (2003) proposed a hierarchical model of transgene organization:…”

Section: Particle Bombardment Facilitates Simultaneous Multiple Gene mentioning

confidence: 99%

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Particle bombardment and the genetic enhancement of crops: myths and realities

et al. 2005

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DNA transfer by particle bombardment makes use of physical processes to achieve the transformation of crop plants. There is no dependence on bacteria, so the limitations inherent in organisms such as Agrobacterium tumefaciens do not apply. The absence of biological constraints, at least until DNA has entered the plant cell, means that particle bombardment is a versatile and effective transformation method, not limited by cell type, species or genotype. There are no intrinsic vector requirements so transgenes of any size and arrangement can be introduced, and multiple gene cotransformation is straightforward. The perceived disadvantages of particle bombardment compared to Agrobacterium-mediated transformation, i.e. the tendency to generate large transgene arrays containing rearranged and broken transgene copies, are not borne out by the recent detailed structural analysis of transgene loci produced by each of the methods. There is also little evidence for major differences in the levels of transgene instability and silencing when these transformation methods are compared in agriculturally important cereals and legumes, and other non-model systems. Indeed, a major advantage of particle bombardment is that the delivered DNA can be manipulated to influence the quality and structure of the resultant transgene loci. This has been demonstrated in recently reported strategies that favor the recovery of transgenic plants containing intact, singlecopy integration events, and demonstrating high-level transgene expression. At the current time, particle bombardment is the most efficient way to achieve plastid transformation in plants and is the only method so far used to achieve mitochondrial transformation. In this review, we discuss recent data highlighting the positive impact of particle bombardment on the genetic transformation of plants, focusing on the fate of exogenous DNA, its organization and its expression in the plant cell. We also discuss some of the most important applications of this technology including the deployment of transgenic plants under field conditions.

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Section: Particle Bombardment Facilitates Simultaneous Multiple Gene mentioning

confidence: 96%

Section: Particle Bombardment Facilitates Simultaneous Multiple Gene mentioning

confidence: 99%

Particle bombardment and the genetic enhancement of crops: myths and realities

et al. 2005

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“…Because of the complexities of both the insertions and the wheat genome, only a few such transgene integration sites have been analyzed (Abranches et al, 2000;Jackson et al, 2001;Rooke et al, 2003). These studies reveal that genes originally on separate plasmids are cointegrated into the same sites in multiple tandem and nontandem copies, sometimes interspersed with genomic DNA.…”

Section: Tmentioning

confidence: 99%

Variant high-molecular-weight glutenin subunits arising from biolistic transformation of wheat

Blechl¹,

Vensel²

2013

Journal of Cereal Science

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a b s t r a c tGenetic transformation via the biolistic method has been used to introduce genes encoding natural and novel high-molecular-weight glutenin subunits (HMW-GS) into wheat. The appearance of new seed proteins of sizes not predicted by the transgene coding sequences was noted in some experiments. In this report, the identities of thirteen of these novel proteins were determined by tandem mass spectrometry (MS/MS). Seven different proteins larger than and two proteins smaller than the native protein were shown to contain peptides from 1Dx5. A novel protein found in some progeny of crosses between a transgenic plant and Great Plains winter wheats was larger than but contained several peptides from 1Dy10. In one line, a protein larger than and a protein smaller than HMW-GS each contained peptides from the N-and C-terminus of 1Dx5 and from the repeat region of 1Dy10. In a sixth transgenic line, the native Bx7 gene was apparently replaced by a gene that encodes a larger version of 1Bx7. The variant proteins accumulate in the polymeric protein fraction, indicating that they can form inter-molecular disulfide bonds. These results show that novel proteins found in some transformants are encoded by altered versions of either the transforming or endogenous HMW-GS genes.Published by Elsevier Ltd.

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“…In the allohexaploid genome of bread wheat, biolistic transformation leads to a random distribution of transgene integration sites, and transgene expression is more strongly influenced by the identity of the promoter than by any position effect (Jackson et al 2001). Similarly in barley, both biolistic transformation and Agrobacterium-mediated gene transfer are non-selective in terms of the genomic location of the integration site (SalvoGarrido et al 2004).…”

Section: Transgene Integrationmentioning

confidence: 99%

Genetic transformation technology in the Triticeae

Kumlehn

Hensel

2009

Breed. Sci.

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The Triticeae crops comprise wheat, barley, rye and triticale, which together provide a major portion of the world's food and feed. The growing demand for human nutrition and renewable energy requires an intensification in application-oriented research and the establishment and utilization of current biotechnology in these crops. Genetic transformation provides an important means both to elucidate gene function, and to engineer crop plants in a directed and precise way. This review covers a range of issues surrounding the production of stable transgenic lines within the Triticeae. Some quality aspects of transgenesis of particular relevance to the Triticeae are also discussed.

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High-resolution structural analysis of biolistic transgene integration into the genome of wheat

Cited by 59 publications

References 24 publications

Particle bombardment and the genetic enhancement of crops: myths and realities

Particle bombardment and the genetic enhancement of crops: myths and realities

Variant high-molecular-weight glutenin subunits arising from biolistic transformation of wheat

Genetic transformation technology in the Triticeae

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