Genetic transformation technology in the Triticeae

Kumlehn, Jochen; Hensel, Goetz

doi:10.1270/jsbbs.59.553

Cited by 22 publications

(12 citation statements)

References 69 publications

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“…The hpt :: gfp transformation efficiencies achieved in these experiments ranged from 0.7 to 9.6 % (Table 2), rates which are comparable with current protocols based on immature embryo explants of cv. “Golden Promise” (Goedeke et al 2007; Hensel and Kumlehn 2009). Transgenics carrying either the selectable marker only or both transgenes were recovered from each strain/vector combination.…”

Section: Discussionmentioning

confidence: 99%

The elimination of a selectable marker gene in the doubled haploid progeny of co-transformed barley plants

et al. 2012

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Following the production of transgenic plants, the selectable marker gene(s) used in the process are redundant, and their retention may be undesirable. They can be removed by exploiting segregation among the progeny of co-transformants carrying both the selectable marker gene and the effector transgene. Here we show that the doubled haploid technology widely used in conventional barley breeding programmes represents a useful means of fixing a transgene, while simultaneously removing the unwanted selectable marker gene. Primary barley co-transformants involving hpt::gfp (the selectable marker) and gus (a model transgene of interest) were produced via Agrobacterium-mediated gene transfer to immature embryos using two respective T-DNAs. These plants were then subjected to embryogenic pollen culture to separate independently integrated transgenes in doubled haploid progeny. A comparison between 14 combinations, involving two Agrobacterium strains carrying various plasmids, revealed that the highest rate of independent co-transformation was achieved when a single Agrobacterium clone carried two binary vectors. Using this principle along with Agrobacterium strain LBA4404, selectable marker-free, gus homozygous lines were eventually obtained from 1.5 per 100 immature embryos inoculated. Compared to the segregation of uncoupled T-DNAs in conventionally produced progeny, the incorporation of haploid technology improves the time and resource efficiency of producing true-breeding, selectable marker-free transgenic barley.Electronic supplementary materialThe online version of this article (doi:10.1007/s11103-012-9988-9) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

The elimination of a selectable marker gene in the doubled haploid progeny of co-transformed barley plants

et al. 2012

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“…Rice has a small genome (∼430 Mb) with a high degree of chromosomal synteny with other major cereal crops (Bolot et al, 2009). Barley, with its diploid genome, represents another convenient model, particularly for Triticeae crops which share high genomic colinearity, providing a basis for genetic and genomic analyses in polyploid wheats (Bennetzen and Freeling, 1997; Hayes et al, 2003; Kumlehn and Hensel, 2009).…”

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Genetics of Tillering in Rice and Barley

et al. 2014

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Tillering, or the production of lateral branches (i.e., culms), is an important agronomic trait that determines shoot architecture and grain production in grasses. Shoot architecture is based on the actions of the apical and axillary meristems (AXMs). The shoot apical meristem (SAM) produces all aboveground organs, including AXMs, leaves, stems, and inflorescences. In grasses like rice (Oryza sativa L.) and barley (Hordeum vulgare L.), vegetative AXMs form in the leaf axil of lower leaves of the plant and produce tillers (branches). Tiller development is characterized by three stages, including (i) AXM initiation, (ii) bud development, and (iii) outgrowth of the axillary bud into a tiller. Each tiller has the potential to produce a seed-bearing inflorescence and, hence, increase yield. However, a balance between number and vigor of tillers is required, as unproductive tillers consume nutrients and can lead to a decreased grain production. Because of its agronomic and biological importance, tillering has been widely studied, and numerous works demonstrate that the control of AXM initiation, bud development, and tillering in the grasses is via a suite of genes, hormones, and environmental conditions. In this review, we describe the genes and hormones that control tillering in two key cereal crops, rice and barley. In addition, we discuss how the development of new genomics tools and approaches, coupled with the synteny between the rice and barley genomes, are accelerating the isolation of barley genes underlying tillering phenotypes.A t the global level, the most important cereal crops are maize (Zea mays L.), rice, wheat (Triticum aestivum L.) and barley (H. vulgare spp. vulgare), with a total of 2.4 billion tons produced annually at a value of >446 billion

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“…Twenty transformants where chosen for further analysis under green-house conditions. To avoid the chimerism problem associated with the biolistic method used in the present study (Kumlehn and Hensel 2009), 10% of the total T 1 grains from 20 selected T 0 transformants were sown and treated with a 2% Basta™ solution to test for bialaphos resistance. Application of the herbicide three days after the start of vernalization cycle resulted in on average 60% mortality, as wild type wheat plants are known to be highly susceptible to bialaphos at the three to four leaves stage.…”

Section: Resultsmentioning

confidence: 99%

Development of wheat genotypes expressing a glutamine-specific endoprotease from barley and a prolyl endopeptidase from Flavobacterium meningosepticum or Pyrococcus furiosus as a potential remedy to celiac disease

Osorio

Wen

Mejías

et al. 2018

Funct Integr Genomics

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Ubiquitous nature of prolamin proteins dubbed gluten from wheat and allied cereals imposes a major challenge in the treatment of celiac disease, an autoimmune disorder with no known treatment other than abstinence diet. Administration of hydrolytic glutenases as food supplement is an alternative to deliver the therapeutic agents directly to the small intestine, where sensitization of immune system and downstream reactions take place. The aim of the present research was to evaluate the capacity of wheat grain to express and store hydrolytic enzymes capable of gluten detoxification. For this purpose, wheat scutellar calli were biolistically transformed to generate plants expressing a combination of glutenase genes for prolamin detoxification. Digestion of prolamins with barley endoprotease B2 (EP-HvB2) combined with Flavobacterium meningosepticum prolyl endopeptidase (PE-FmPep) or Pyrococcus furiosus prolyl endopeptidase (PE-PfuPep) significantly reduced (up to 67%) the amount of the indigestible gluten peptides of all prolamin families tested. Seven of the 168 generated lines showed inheritance of transgene to the T generation. Reversed phase high-performance liquid chromatography of gluten extracts under simulated gastrointestinal conditions allowed the identification of five T lines that contained significantly reduced amounts of immunogenic, celiac disease-provoking gliadin peptides. These findings were complemented by the R5 ELISA test results where up to 72% reduction was observed in the content of immunogenic peptides. The developed wheat genotypes open new horizons for treating celiac disease by an intraluminal enzyme therapy without compromising their agronomical performance.

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Genetic transformation technology in the Triticeae

Cited by 22 publications

References 69 publications

The elimination of a selectable marker gene in the doubled haploid progeny of co-transformed barley plants

The elimination of a selectable marker gene in the doubled haploid progeny of co-transformed barley plants

Genetics of Tillering in Rice and Barley

Development of wheat genotypes expressing a glutamine-specific endoprotease from barley and a prolyl endopeptidase from Flavobacterium meningosepticum or Pyrococcus furiosus as a potential remedy to celiac disease

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