Biotechnological advances of electroporation of grapevine and sugar beet cells

Kovalenko, P. G.; Schuman, N.V.; Ponomarenko, S. P.

doi:10.1016/s0302-4598(97)00008-1

Cited by 4 publications

(2 citation statements)

References 11 publications

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“…However, E. coli-based cloning vectors are convenient for direct transformation by protoplast permeation or particle bombardment. In grapevine, Kovalenko et al (1997) observed that higher expression levels could be obtained with a linearized plasmid rather than with a circular vector. Later, the use of minimal gene cassettes, which are linear DNA fragments comprising the gene of interest flanked by regulatory sequences, was shown to be as effective as traditional circular plasmids in V. vinifera Chardonnay (Vidal et al, 2006).…”

Section: Direct Transformation Vectors and Linear Minimal Cassettesmentioning

confidence: 99%

Transient expression assays in grapevine: a step towards genetic improvement

Jelly

Valat

Walter

et al. 2014

Plant Biotechnology Journal

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SummaryIn the past few years, the usefulness of transient expression assays has continuously increased for the characterization of unknown gene function and metabolic pathways. In grapevine (Vitis vinifera L.), one of the most economically important fruit crops in the world, recent systematic sequencing projects produced many gene data sets that require detailed analysis. Due to their rapid nature, transient expression assays are well suited for large-scale genetic studies. Although genes and metabolic pathways of any species can be analysed by transient expression in model plants, a need for homologous systems has emerged to avoid the misinterpretation of results due to a foreign genetic background. Over the last 10 years, various protocols have thus been developed to apply this powerful technology to grapevine. Using cell suspension cultures, somatic embryos, leaves or whole plantlets, transient expression assays enabled the study of the function, regulation and subcellular localization of genes involved in specific metabolic pathways such as the biosynthesis of phenylpropanoids. Disease resistance genes that could be used for marker-assisted selection in conventional breeding or for stable transformation of elite cultivars have also been characterized. Additionally, transient expression assays have proved useful for shaping new tools for grapevine genetic improvement: synthetic promoters, silencing constructs, minimal linear cassettes or viral vectors. This review provides an update on the different tools (DNA constructs, reporter genes, vectors) and methods (Agrobacteriummediated and direct gene transfer methods) available for transient gene expression in grapevine. The most representative results published thus far are then described.

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Section: Direct Transformation Vectors and Linear Minimal Cassettesmentioning

confidence: 99%

Transient expression assays in grapevine: a step towards genetic improvement

Jelly

Valat

Walter

et al. 2014

Plant Biotechnology Journal

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“…As reported in our previous papers [9,15], the plant DNA fragments with transcription promoted activity were selected from the pool of random tobacco nuclear DNA fragments, and one of them (pDNt23) was shown to have the root-specific expression in transgenic plants. The chimeric plasmid pDNt23-35SCaMV-CAT-nos-3 (pDNt23-root specific promo ter, nos-nopaline synthase terminator and had the selectable gene as neomycin phosphotransferase II-NPT-II) was used in our experiments with the electroporation of the licorice suspension protoplasts.…”

Section: Leguminosae Tribe Astragalaceae)mentioning

confidence: 99%

Transient gene expression and total flavonoids production in the electroporated licorice Glycyrrhiza glabra L. suspension protoplasts

Kovalenko

2004

Biopolym. Cell

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The bacterial chloramphenicol acety(transferase (CAT) gene was expressed in licorice G. glabra L. suspension protoplasts by electroporation after introduction of the chimeric plasmid pDNt23-CaMV35Snos-cat (pDNt23-root-specific and CaMV 35S promoters, nopaline synthase nos-terminator, and selectable NPT-II gene). Maximum of CAT activity in the licorice protoplasts was observed in SO h after electroporation Together with recent advances in the cell isolation and electroporation procedures, this system allows to study expression of the root-specific promoter introduced into the licorice cells. The level of total flavonoids production in 4 weeks culture was tested in the electroporated cell lines. Additionally, these electroporated cells were treated by elicitors (yeast extract and fungal bioextract). The total flavonoids production increased by 2-5 times in comparison with the non-electroporated licorice cells.

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