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Przetakiewicz, Anna; Orczyk, Wacław; Nadolska‐Orczyk, Anna

doi:10.1023/a:1023030511800

Cited by 80 publications

(20 citation statements)

References 48 publications

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“…Immature embryos of wheat and triticale were precultured on MSB3 medium containing MS salts (Murashige and Skoog 1962), B5 vitamins (Gamborg et al 1968), and 3 mg l −1 Dicamba, as previously described (Przetakiewicz et al 2003). After 2 to 3 days, the embryos were inoculated with a drop of A. tumefaciens suspension and co-cultured for the next 2–3 days, depending on the growth rate of the bacteria.…”

Section: Methodsmentioning

confidence: 99%

“…The subsequent culture was on MSB6 medium containing 2 mg l −1 Picloram, 1 mg l −1 2,4-D, 500 mg l −1 casein hydrolysate, 150 mg l −1 Timentin, and one of two selection agents, 50 mg l −1 kanamycin or 2 mg l −1 phosphinothricin. After a further 4–5 weeks, the embryonic calli were transferred onto plant regeneration medium R2-MSB and rooted on 0.5 MS medium (Przetakiewicz et al 2003). The media contained the same selection agent.…”

Section: Methodsmentioning

confidence: 99%

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The Agrobacterium-mediated transformation of common wheat (Triticum aestivum L.) and triticale (x Triticosecale Wittmack): role of the binary vector system and selection cassettes

2011

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The influence of two binary vector systems, pGreen and pCAMBIA, on the Agrobacterium-mediated transformation ability of wheat and triticale was studied. Both vectors carried selection cassettes with bar or nptII driven by different promoters. Two cultivars of wheat, Kontesa and Torka, and one cultivar of triticale, Wanad, were tested. The transformation rates for the wheat cultivars ranged from 0.00 to 3.58% and from 0.00 to 6.79% for triticale. The best values for wheat were 3.58% for Kontesa and 3.14% for Torka, and these were obtained after transformation with the pGreen vector carrying the nptII selection gene under the control of 35S promoter. In the case of the bar selection system, the best transformation rates were, respectively, 1.46 and 1.79%. Such rates were obtained when the 35S::bar cassette was carried by the pCAMBIA vector; they were significantly lower with the pGreen vector. The triticale cultivar Wanad had its highest transformation rate after transformation with nptII driven by 35S in pCAMBIA. The bar selection system for the same triticale cultivar was better when the gene was driven by nos and the selection cassette was carried by pGreen. The integration of the transgenes was confirmed with at least three pairs of specific starters amplifying the fragments of nptII, bar, or gus. The expression of selection genes, measured by reverse transcriptase polymerase chain reaction (RT-PCR) in relation to the actin gene, was low, ranging from 0.00 to 0.63 for nptII and from 0.00 to 0.33 for bar. The highest relative transcript accumulation was observed for nptII driven by 35S and expressed in Kontesa that had been transformed with pGreen.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

The Agrobacterium-mediated transformation of common wheat (Triticum aestivum L.) and triticale (x Triticosecale Wittmack): role of the binary vector system and selection cassettes

2011

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“…Agrobacterium -mediated transformation experiments were performed according to our previously described protocols for wheat (Przetakiewicz et al, 2003, 2004) and for triticale (Nadolska-Orczyk et al, 2005). Putative transgenic plants were regenerated and selected on modified MS media containing 20 mg l -1 of hygromycin as a selectable agent.…”

Section: Methodsmentioning

confidence: 99%

Artificial MicroRNA-Based Specific Gene Silencing of Grain Hardness Genes in Polyploid Cereals Appeared to Be Not Stable Over Transgenic Plant Generations

et al. 2017

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Gene silencing by RNA interference is a particularly important tool in the study of gene function in polyploid cereal species for which the collections of natural or induced mutants are very limited. Previously we have been testing small interfering RNA-based approach of gene silencing in wheat and triticale. In this research, artificial microRNAs (amiRs) were studied in the same species and the same target genes to compare effectiveness of both gene silencing pathways. amiR cassettes were designed to silence Puroindoline a (Pina) and Puroindoline b (Pinb) hardness genes in wheat and their orthologues Secaloindoline a (Sina) and Secaloindoline b (Sinb) genes in triticale. Each of the two cassettes contained 21 nt microRNA (miR) precursor derived from conserved regions of Pina/Sina or Pinb/Sinb genes, respectively. Transgenic plants were obtained with high efficiency in two cultivars of wheat and one cultivar of triticale after using the Pinb-derived amiR vector for silencing of Pinb or Sinb, respectively. Lack of transgenic plants in wheat or very low transformation efficiency in triticale was observed using the Pina-derived amiR cassette, despite large numbers of embryos attempted. Silencing of Pinb in wheat and Sinb in triticale was highly efficient in the T1 generation. The transcript level of Pinb in wheat was reduced up to 92% and Sinb in triticale was reduced up to 98%. Moreover, intended silencing of Pinb/Sinb with Pinb-derived amiR cassette was highly correlated with simultaneous silencing of Pina/Sina in the same transgenic plants. High downregulation of Pinb/Pina genes in T1 plants of wheat and Sinb/Sina genes in T1 plants of triticale was associated with strong expression of Pinb-derived amiR. Silencing of the target genes correlated with increased grain hardness in both species. Total protein content in the grains of transgenic wheat was significantly lower. Although, the Pinb-derived amiR cassette was stably inherited in the T2 generation of wheat and triticale the silencing effect including strongly decreased expression of silenced genes as well as strong expression of Pinb-derived amiR was not transmitted. Advantages and disadvantages of posttranscriptional silencing of target genes by means of amiR and siRNA-based approaches in polyploid cereals are discussed.

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“…Plant regeneration from immature inflorescences is also effective with 34 plants per 64 explants with the variety Galan (Havrlentová et al 2001). Research investigating callus induction and plant regeneration from immature embryos from different cultivars of triticale showed that the mean number of plant regeneration coefficient ranged between 9.7 for cultivar Gabo and 15.9 for the cultivar Wanad (Przetakiewicz et al 2003). For microspore and anther cultures of wheat, barley or triticale, both the level of induction and regeneration are significantly higher than in the best forms of rye (Davies and Morton 1998; González and Jouve 2005; Castillo et al 2000; Holme et al 1999).…”

Section: Introductionmentioning

confidence: 99%

The specificity and genetic background of the rye (Secale cereale L.) tissue culture response

et al. 2012

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Rye is one of the most important crops in Eastern and Northern Europe. Despite the numerous beneficial features of rye, its annual production decreases successively which correlates with the lack of progress in its breeding compared with other cereals. Biotechnological methods could effectively improve the breeding of rye. However, their application is highly limited by the absence of an efficient procedure for plant regeneration in vitro, since rye is one of the most recalcitrant cereals with regard to the tissue culture response (TCR), and successful regeneration is highly dependent on genotype. Efforts to understand the genetic mechanisms controlling TCR of rye have elucidated some basic aspects, and several genes and genome regions controlling this trait have been identified. The aim of this review is to summarize the limited current knowledge of this topic.

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Cited by 80 publications

References 48 publications

The Agrobacterium-mediated transformation of common wheat (Triticum aestivum L.) and triticale (x Triticosecale Wittmack): role of the binary vector system and selection cassettes

The Agrobacterium-mediated transformation of common wheat (Triticum aestivum L.) and triticale (x Triticosecale Wittmack): role of the binary vector system and selection cassettes

Artificial MicroRNA-Based Specific Gene Silencing of Grain Hardness Genes in Polyploid Cereals Appeared to Be Not Stable Over Transgenic Plant Generations

The specificity and genetic background of the rye (Secale cereale L.) tissue culture response

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