Genetic Transformation of Triticeae Cereals for Molecular Farming

Hensel, Goetz

doi:10.5772/22430

Cited by 3 publications

(3 citation statements)

References 93 publications

(74 reference statements)

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“…Agrobacterium -mediated transformation of monocotyledonous species has been for a long time technically challenging, especially for cereals [ 48 , 49 ]. It took more than ten years after publishing the first successful Agrobacterium -mediated transformation of tobacco by De Block et al [ 50 ] to produce the transgenic plants in crops such as rice [ 51 ], and maize [ 52 ].…”

Section: Introductionmentioning

confidence: 99%

Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter

et al. 2021

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Background Stipagrostis pennata (Trin.) De Winter is an important species for fixing sand in shifting and semi-fixed sandy lands, for grazing, and potentially as a source of lignocellulose fibres for pulp and paper industry. The seeds have low viability, which limits uses for revegetation. Somatic embryogenesis offers an alternative method for obtaining large numbers of plants from limited seed sources. Results A protocol for plant regeneration from somatic embryos of S. pennata was developed. Somatic embryogenesis was induced on Murashige & Skoog (MS) medium supplemented with 3 mg·L–1 2,4-D subsequently shoots were induced on MS medium and supplemented with 5 mg·L–1 zeatin riboside. The highest shoots induction was obtained when embryogenic callus derived from mature embryos (96%) in combination with MS filter-sterilized medium was used from Khuzestan location. The genetic stability of regenerated plants was analysed using ten simple sequence repeats (SSR) markers from S. pennata which showed no somaclonal variation in regenerated plants from somatic embryos of S. pennata. The regenerated plants of S. pennata showed genetic stability without any somaclonal variation for the four pairs of primers that gave the expected amplicon sizes. This data seems very reliable as three of the PCR products belonged to the coding region of the genome. Furthermore, stable expression of GUS was obtained after Agrobacterium-mediated transformation using a super binary vector carried by a bacterial strain LBA4404. Conclusion To our knowledge, the current work is the first attempt to develop an in vitro protocol for somatic embryogenesis including the SSR marker analyses of regenerated plants, and Agrobacterium-mediated transformation of S. pennata that can be used for its large-scale production for commercial purposes.

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Section: Introductionmentioning

confidence: 99%

Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter

et al. 2021

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“…Another step forward will be to study the relationship between this spatial, compositional information and grain properties, such as biomechanical properties that can affect the milling process. Progress towards establishing plants as a vehicle for the production of plant-made pharmaceuticals is likely to accelerate in the coming years (Hensel, 2011). The achievement of appropriate levels of expression remains somewhat empirical, and to a large extent varies from one recombinant protein to another (Hensel, 2011).…”

Section: Future Perspectives and Conclusionmentioning

confidence: 99%

“…Progress towards establishing plants as a vehicle for the production of plant-made pharmaceuticals is likely to accelerate in the coming years (Hensel, 2011). The achievement of appropriate levels of expression remains somewhat empirical, and to a large extent varies from one recombinant protein to another (Hensel, 2011). Therefore, immunolabeling of recombinant proteins will be necessary in order to understand pathways and biochemical processes yet to be unraveled (Finnie, Sultan, & Grasser, 2011).…”

Section: Future Perspectives and Conclusionmentioning

confidence: 99%

Current potential and limitations of immunolabeling in cereal grain research

Vázquez-Gutiérrez

Langton

2015

Trends in Food Science & Technology

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Immunolabeling techniques have made a valuable contribution to cereal grain research during the past decade in terms of precise localization of specific compounds. While these techniques have several limitations, such as the availability and specificity of the antibodies, immunolabeling has proven especially useful in cereal studies seeking a better understanding of grain development and characterization. According to the literature reviewed in this paper, immunolabeling techniques will continue to be a useful tool in the characterization and localization of cereal grain components.

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Overexpression of Cytokinin Dehydrogenase Genes in Barley (Hordeum vulgare cv. Golden Promise) Fundamentally Affects Morphology and Fertility

et al. 2013

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Barley is one of the most important cereal crops grown worldwide. It has numerous applications, but its utility could potentially be extended by genetically manipulating its hormonal balances. To explore some of this potential we identified gene families of cytokinin dehydrogenases (CKX) and isopentenyl transferases, enzymes that respectively irreversibly degrade and synthesize cytokinin (CK) plant hormones, in the raw sequenced barley genome. We then examined their spatial and temporal expression patterns by immunostaining and qPCR. Two CKX-specific antibodies, anti-HvCKX1 and anti-HvCKX9, predominantly detect proteins in the aleurone layer of maturing grains and leaf vasculature, respectively. In addition, two selected CKX genes were used for stable, Agrobacterium tumefaciens-mediated transformation of the barley cultivar Golden Promise. The results show that constitutive overexpression of CKX causes morphological changes in barley plants and prevents their transition to flowering. In all independent transgenic lines roots proliferated more rapidly and root-to-shoot ratios were higher than in wild-type plants. Only one transgenic line, overexpressing CKX under the control of a promoter from a phosphate transporter gene, which is expressed more strongly in root tissue than in aerial parts, yielded progeny. Analysis of several T1-generation plants indicates that plants tend to compensate for effects of the transgene and restore CK homeostasis later during development. Depleted CK levels during early phases of development are restored by down-regulation of endogenous CKX genes and reinforced de novo biosynthesis of CKs.

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Genetic Transformation of Triticeae Cereals for Molecular Farming

Cited by 3 publications

References 93 publications

Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter

Protocol development for somatic embryogenesis, SSR markers and genetic modification of Stipagrostis pennata (Trin.) De Winter

Current potential and limitations of immunolabeling in cereal grain research

Overexpression of Cytokinin Dehydrogenase Genes in Barley (Hordeum vulgare cv. Golden Promise) Fundamentally Affects Morphology and Fertility

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