Explants Used for the Generation of Transgenic Plants

Piqueras, Abel; Alburquerque, Nuria; Folta, Kevin M.

doi:10.1007/978-3-642-04809-8_2

Cited by 14 publications

(6 citation statements)

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“…Pre-culture of explants was found crucial in this study, improving explant viability, reducing explant overgrowth from A. tumefaciens and assisting in the identification and removal of bacterial contaminated material prior to A. tumefaciens inoculation. Pre-induction of the regeneration capacity via a pre-culture treatment has been reported to improve the adhesion of A. tumefaciens during co-cultivation [ 29 ]. Pre-culture activates cell division and the phase of the cell-cycle has been shown to influence stable transformation [ 30 ] The presence of actively dividing cells improved cell competence for A. tumefaciens infection in regenerable tissue [ 29 ].…”

Section: Discussionmentioning

confidence: 99%

“…Pre-induction of the regeneration capacity via a pre-culture treatment has been reported to improve the adhesion of A. tumefaciens during co-cultivation [ 29 ]. Pre-culture activates cell division and the phase of the cell-cycle has been shown to influence stable transformation [ 30 ] The presence of actively dividing cells improved cell competence for A. tumefaciens infection in regenerable tissue [ 29 ]. In Brassica rapa , seedling hypocotyls pre-cultured prior to exposure to A. tumefaciens showed greater shoot regeneration and improved transformation efficiency [ 31 ].…”

Section: Discussionmentioning

confidence: 99%

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Agrobacterium-mediated transformation systems of Primula vulgaris

Hayta

Smedley

et al. 2018

Plant Methods

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BackgroundGenetic transformation is a valuable tool and an important procedure in plant functional genomics contributing to gene discovery, allowing powerful insights into gene function and genetically controlled characteristics. Primulaceae species provide one of the best-known examples of heteromorphic flower development, a breeding system which has attracted considerable attention, including that of Charles Darwin. Molecular approaches, including plant transformation give the best opportunity to define and understand the role of genes involved in floral heteromorphy in the common primrose, Primula vulgaris, along with other Primula species.ResultsTwo transformation systems have been developed in P. vulgaris. The first system, Agrobacterium-mediated vacuum infiltration of seedlings, enables the rapid testing of transgenes, transiently in planta. GUS expression was observed in the cotyledons, true leaves, and roots of Primula seedlings. The second system is based on Agrobacterium tumefaciens infection of pedicel explants with an average transformation efficiency of 4.6%. This transformation system, based on regeneration and selection of transformants within in vitro culture, demonstrates stable transgene integration and transmission to the next generation.ConclusionThe two transformation systems reported here will aid fundamental research into important traits in Primula. Although, stable integration of transgenes is the ultimate goal for such analyses, transient gene expression via Agrobacterium-mediated DNA transfer, offers a simple and fast method to analyse transgene functions. The second system describes, for the first time, stable Agrobacterium-mediated transformation of Primula vulgaris, which will be key to characterising the genes responsible for the control of floral heteromorphy.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Agrobacterium-mediated transformation systems of Primula vulgaris

Hayta

Smedley

et al. 2018

Plant Methods

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“…When performing transformation and regeneration experiments, explant selection is a critical parameter because the different explant sources often have varying potential for transformation (Piqueras et al, 2010). In the present study, it can be seen that different explant types resulted in different transformation efficiency (Fig.…”

Section: Effect Of Explant Types On Gene Transformation Efficiencymentioning

confidence: 52%

Agrobacterium-Mediated Transformation of Panax Vietnamensis Ha Et Grushv.

Nhựt¹,

Huy²,

Hương³

et al. 2018

Vietnam J. Biotechnol.

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In recent years, the Agrobacterium-mediated genetic transformation system has become the most useful method widely used for the introduction of foreign genes into plant cells followed by regeneration of genetically improved plants. Panax vietnamensis Ha et Grushv. is a highly valued medicinal plant native to Vietnam with limited area of distribution. This report illustrates the possibilities of biotechnology for genetic transformation aimed at establishing an effective production of secondary metabolites in P. vietnamensis. In the present investigation, 0.5 cm2 leaf blades, 1 cm long leaf petioles and 0.5 cm3 callus clusters were used for the hairy root induction. Results indicated that hairy roots were induced on P. vietnamensis callus clusters co-cultivated with Agrobacterium rhizogenes strain ATCC15834 at OD600 of 0.5 with an infection time of 20 min and a supplementation of 100 mM acetosyringone. PCR amplification of the DNA isolated from the resulting hairy roots was used to confirm the presence of rol genes. Compared to in vitro rhizome cultures, hairy root cultures appear to be potential for continuous production of valuable secondary metabolites with similar saponin profiles. The protocol described in this study is simple and rapid and therefore, can be used for large-scale experiments for the rapid production of valuable compounds.

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“…Application of genomic techniques provides great tools for the improvement of Rhododendron traits (Ueno et al 2000; Madej et al 2006). The leaf explant culture is the most promising platform for genetic improvement through Agrobacterium-mediated transformation, isolation and transfection of protoplasts or microprojectile bombardment (Piqueras et al 2010). For obtaining transgenic genotypes, it is necessary to transfer new genes to the leaf cells being able to de novo morphogenesis (San Jose et al 2014).…”

Section: Introductionmentioning

confidence: 99%

Direct regeneration and morpho-histological study of de novo shoot development from leaf explants of Rhododendron mucronulatum Turcz.

Zaytseva

2023

Preprint

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Rhododendron mucronulatum Turcz., a rare medicinal and ornamental deciduous multi-branched shrub native to northern parts of East Asia, is a valuable genetic resource for breeding and biotechnological producing bioactive metabolites. To create propagation system of R. mucronulatum, an efficient method of direct shoot regeneration from leaf explants based on thidiazuron (TDZ) and dark treatment was developed, and detailed histological analysis for revealing localization of cell competent to morphogenesis was carried out for the first time. The highest regeneration rate (75%) and number of shoots per explant (an average of 51.7) were achieved under 0.5 µM TDZ and dark treatment. The dark treatment was found to increase the regeneration rate more than two-fold compared to culturing under light conditions. The histological analysis showed that tested TDZ concentrations induced direct organogenesis pathway. The competent to morphogenesis cells were found to locate in epidermal tissue of leaf explants on adaxial side of petiole and lamina base next to main vein. The formation of embryo-like structures was observed on the late morphogenesis stages. It was discovered that parenchymal and epidermal tissues of embryo-like structures were well-separated from explant tissue but its vascular bundle was connected with vascular bundle of explant and had a joining vascular system, and led to adventitious shoots development. The result obtained can promote further establishment of efficient and stable systems for genetic transformation and biomass production of R. mucronulatum.

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Explants Used for the Generation of Transgenic Plants

Cited by 14 publications

References 159 publications

Agrobacterium-mediated transformation systems of Primula vulgaris

Agrobacterium-mediated transformation systems of Primula vulgaris

Agrobacterium-Mediated Transformation of Panax Vietnamensis Ha Et Grushv.

Direct regeneration and morpho-histological study of de novo shoot development from leaf explants of Rhododendron mucronulatum Turcz.

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