Silencing of HMW glutenins in transgenic wheat expressing extra HMW subunits

Alvarez, M. Lucrecia; Guelman, Sebastián; Halford, Nigel G.; Lustig, Shlomo; Reggiardo, M. I.; Ryabushkina, N. A.; Shewry, Peter R.; Stein, Juliana; Vallejos, Rubén H.

doi:10.1007/s001220050042

Cited by 116 publications

(63 citation statements)

References 28 publications

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“…Of these, 3 were sterile and 8 were silenced in later generations. Consequently the final transformation efficiency of transcriptionally active transgenic Alvarez et al (2000), Anand et al (2003), Makandar et al (2006) and Chen et al (2008), who also observed silencing during the early generations of selfing in transgenic plants. Gene silencing is usually associated with integration of transgenes in multiple copies and to sequence homology between a transgene and an endogenous gene.…”

Section: Discussionmentioning

confidence: 95%

Increased tolerance to wheat powdery mildew by heterologous constitutive expression of the Solanum chacoense Snakin-1 gene

Faccio¹,

Rovere²,

Hopp³

et al. 2011

CZECH J GENET PLANT

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Great efforts are currently being devoted to studying the use of transgenes to confer resistance to phytopathogenic fungi. Snakin-1 is a broad-spectrum antimicrobial peptide isolated from Solanum that is active in vitro against bacteria and fungi. Recently, it was reported that overexpression of the snakin-1 (SN1) gene in transgenic potato plants enhanced resistance to Rhizoctonia solani and Erwinia carotovora. In this work wheat transgenic plants that constitutively expressed the S. chacoense SN1 gene were challenged with Blumeria graminis f.sp. tritici. Enhanced resistance to the pathogen was observed in two transgenic lines in which the development of the disease was delayed and reduced compared with the wild type variety ProINTA Federal. An association between high resistance to the pathogen and a high level of snakin-1 transcripts in the plant was observed. This is the first report on SN1 gene expression in Gramineae and its effects on wheat powdery mildew development.

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Section: Discussionmentioning

confidence: 95%

Increased tolerance to wheat powdery mildew by heterologous constitutive expression of the Solanum chacoense Snakin-1 gene

Faccio¹,

Rovere²,

Hopp³

et al. 2011

CZECH J GENET PLANT

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“…And we analyzed the protein expression of Glu-1Dy10 by Western blot with an anti x-type HMW specific antibody. The 11 transgenic plants (2,6,7,9,15,18,20,22,24,26,28) which were shown abnormal morphologies comparing with 'Dongjin' were removed.…”

Section: Resultsmentioning

confidence: 99%

“…We guess that the expression levels of Glu-1Dy10 were too low to detect signal in case of the one-copy inserted lines. In case of Multi-copies inserted lines, this phenomenon may be related to homology-dependent gene silencing in plants [2]. Genetic engineering of plants sometimes results in transgene silencing after integration into the genome, which may relate to a defense mechanism against foreign DNA expression [15,31].…”

Section: Resultsmentioning

confidence: 99%

“…Several HMW-GS genes have been shown to be functional when transformed into Escherichia coli [10], tobacco [23], wheat [1,2,5,6] and tritordeum [24]. The genetic engineering of transgenic plants in most crop species requires the use of selectable marker genes and selective agents, such as herbicides and antibiotics in order to minimize regeneration of non-transformed tissues.…”

Section: Introductionmentioning

confidence: 99%

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Development of Marker-free Transgenic Rice Expressing the Wheat Storage Protein, Glu-1Dy10, for Increasing Quality Processing of Bread and Noodles

Park¹,

Shin²,

Hwang³

et al. 2014

Journal of Life Science

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Rice flour is used in many food products. However, dough made from rice lacks extensibility and elasticity, making it less suitable than wheat for many food products such as bread and noodles. The high-molecular weight glutenin subunits (HMW-GS) of wheat play a crucial role in determining the processing properties of the wheat grain. This paper describes the development of marker-free transgenic rice plants expressing a wheat Glu-Dy10 gene encoding the HMG-GS from the Korean wheat cultivar 'Jokyeong' using Agrobacterium-mediated co-transformation. Two expression cassettes, consisting of separate DNA fragments containing Glu-1Dy10 and hygromycin phosphotransferase II (HPTII) resistance genes, were introduced separately into Agrobacterium tumefaciens EHA105 for co-infection. Each EHA105 strain harboring Glu-1Dy10 or HPTII was infected into rice calli at a 3: 1 ratio of Glu-1Bx7 and HPTII. Among 290 hygromycin-resistant T0 plants, we obtained 29 transgenic lines with both the Glu-1Dy10 and HPTII genes inserted into the rice genome. We reconfirmed the integration of the Glu-1Dy10 gene into the rice genome by Southern blot analysis. Transcripts and proteins of the Glu-1Dy10 in transgenic rice seeds were examined by semi-quantitative RT-PCR and Western blot analysis. The marker-free plants containing only the Glu-1Dy10 gene were successfully screened in the T1 generation.

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“…Cloned HMW glutenin genes have been shown to be functional when introduced into Escherichia coli (Galili 1989), tobacco (Roberts et al 1989), wheat (Altpeter et al 1996;Blechl and Anderson 1996;Barro et al 1997;Alvarez et al 2000) and tritordeum (Rooke et al 1999). Detailed inheritance studies of these transgenes have not been reported, although it has been reported that the transgene is stable through three generations in wheat (Altpeter et al 1996).…”

Section: Introductionmentioning

confidence: 99%

Expression and inheritance of the wheat Glu-1DX5 gene in transgenic maize

et al. 2002

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We have produced transgenic maize plants containing a wheat Glu-1Dx5 gene encoding the high-molecularweight glutenin subunit 1Dx5. Analysis by SDS-PAGE showed that a protein similar in size to the wheat 1Dx5 subunit accumulates in the endosperm of transgenic maize from four independent transformation events. This protein reacts with a monoclonal antibody specific to the wheat 1Dx5 subunit and was not detected in nontransgenic controls or in pollen, anthers, leaves or embryos of plants grown from seeds expressing this protein in endosperm. Genomic Southern-blot analysis is consistent with results from SDS-PAGE and indicates that the transgene integration sites are complex and are different in the four events studied. Using the presence of this protein as a phenotypic marker, we studied the inheritance of this gene through three sexual generations. Reciprocal crosses with nontransgenic plants and self-pollinations were performed, and the resulting kernels were analyzed for the presence of the 1Dx5 subunit. These data, together with PCR analysis for the transgene, suggest that the transgene is inefficiently transmitted through pollen in all four events. Abstract We have produced transgenic maize plants containing a wheat Glu-1Dx5 gene encoding the highmolecular-weight glutenin subunit 1Dx5. Analysis by SDS-PAGE showed that a protein similar in size to the wheat 1Dx5 subunit accumulates in the endosperm of transgenic maize from four independent transformation events. This protein reacts with a monoclonal antibody specific to the wheat 1Dx5 subunit and was not detected in nontransgenic controls or in pollen, anthers, leaves or embryos of plants grown from seeds expressing this protein in endosperm. Genomic Southern-blot analysis is consistent with results from SDS-PAGE and indicates that the transgene integration sites are complex and are different in the four events studied. Using the presence of this protein as a phenotypic marker, we studied the inheritance of this gene through three sexual generations. Reciprocal crosses with nontransgenic plants and selfpollinations were performed, and the resulting kernels were analyzed for the presence of the 1Dx5 subunit. These data, together with PCR analysis for the transgene, suggest that the transgene is inefficiently transmitted through pollen in all four events.

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Silencing of HMW glutenins in transgenic wheat expressing extra HMW subunits

Cited by 116 publications

References 28 publications

Increased tolerance to wheat powdery mildew by heterologous constitutive expression of the Solanum chacoense Snakin-1 gene

Increased tolerance to wheat powdery mildew by heterologous constitutive expression of the Solanum chacoense Snakin-1 gene

Development of Marker-free Transgenic Rice Expressing the Wheat Storage Protein, Glu-1Dy10, for Increasing Quality Processing of Bread and Noodles

Expression and inheritance of the wheat Glu-1DX5 gene in transgenic maize

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