Transposable elements can be used to study cell lineages in transgenic plants.

Finnegan, E. Jean; Taylor, Brian H.; Craig, Stuart; Dennis, Elizabeth S.

doi:10.1105/tpc.1.8.757

Cited by 53 publications

(18 citation statements)

References 23 publications

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“…This last observation is in agreement with the results of earlier studies on both Ac-excision from the rolC gene [15] and Spm-excision from the GUS gene [23] in tobacco. Our results diverge from those obtained by Finnegan et al [14] who studied excision of Ac from the GUS gene in tobacco. They found extensive or complete staining of certain tissues in leaves of 2 out of 3 analyzed plants.…”

Section: Resultscontrasting

confidence: 99%

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A transposon tagging strategy with Ac on plant cell level in heterologous plant species

Rommens¹,

Kneppers²,

Haring³

et al. 1991

Plant Science

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Disclaimer/Complaints regulationsIf you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library: http://uba.uva.nl/en/contact, or a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible. The maize transposable element Ac can have a 'late' excision time during leaf development in certain transgenic tobacco plants. This was visualized with an assay based on Ac-excision restoring GUS-expression. Leaves of the described plants contain over 103 small blue spots, each of these spots representing an independent excision event. Leaves showing this 'late' excision phenomenon may be used for transposon tagging experiments at plant cell level. Plants which display 'late' Ac-excision do not detectably express GUS during the preceding callus phase, thus allowing transformants to be preselected for a 'late' timing of excision. To examine the applicability of this phenomenon a phenotypic selection assay for excision of Ac was used. Transformed calli containing Ac within the hygromycin resistance gene were regenerated and protoplasts isolated from leaves of regenerated plants were selected on hygromycin. Up to 0.8% of these protoplasts displayed hygromycin resistance. The hygromycin resistant derivatives analyzed were shown to represent independent transposition events. Ac-insertions which can be generated in this way may be used for transposon tagging experiments at cell level.

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Section: Resultscontrasting

confidence: 99%

“…The timing of Ac-excision can be studied with visual assays. These assays are based on Ac-excision restoring activity of/3-glucuronidase (GUS) [14], rolC [15] or streptomycin resistance [16].…”

Section: )mentioning

confidence: 99%

A transposon tagging strategy with Ac on plant cell level in heterologous plant species

Rommens¹,

Kneppers²,

Haring³

et al. 1991

Plant Science

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“…This relatively low transformation frequency precludes the use of DNA gel blot hybridization to monitor Ds excision and probably prevents monitoring of low-level replication dependence for Ds excision in our transient assay system. While assays for Ac-mediated Ds excision from a Gus reporter gene have been described, these have relied on the use of protoplasts (Houba-Herin et al, 1990), Agrobacterium-infected plants (Shen and Hohn, 1992) or transgenic plants (Finnegan et al, 1989). The simple assay described here allowed rapid evaluation of the components of a proposed gene tagging strategy prior to the timeconsuming process of stable transformation of barley.…”

Section: Discussionmentioning

confidence: 99%

Development of a simple transient assay for Ac/Ds activity in cells of intact barley tissue

et al. 1997

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The development of a barley (Hordeurn vulgare L.) transformation system made it possible to consider the use of maize Activator/Dissociation (Ac/Ds) transposable elements for gene tagging in transgenic barley plants. However, barley transformation is time‐consuming, and therefore a simple transient assay for Ac/Ds activity in intact barley tissues was developed to test the components of a proposed gene tagging system, prior to their stable introduction into plants. In this assay, barley scutellar tissue is co‐transformed with constructs containing the maize Ac transposase gene and an Escherichia coli uidA reporter gene (Gus), the expression of which is interrupted by a maize Ds element. In transformed barley scutellar cells, Ac transposase‐mediated excision of the Ds element generates a functional Gus gene, leading to histochemically detectable GUS activity. Characterization of the excision products showed that they had a pattern of nucleotide deletions and/or transversions similar to that found in maize and other heterologous plant systems. In addition, although contrary to the situation observed in heterologous dicot systems, efficient Ds excision in barley, a heterologous monocot system, appears to be inversely associated with Ac copy number, a finding similar to the Ac dosage effects observed in maize. The transient assay was used to demonstrate functional transposase activity in barley callus lines stably transformed with an Ac transposase gene.

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“…A special feature of the phenotypic assays employing a cell autonomous marker gene [36,54] is the fact that the timing of Ac excision can be defined, while even plant cell lineages can be followed [ 19] when marker gene re-activation takes place in specific cells. Other phenomena caused by Ac or Ds transposition are deletions and inversion occurring in the maize DNA [49].…”

Section: Activity Of Autonomous Plant Transposable Elements In New Gementioning

confidence: 99%

“…screened will be chimaeric for the mutated phenotype the trait that is examined should preferably be expressed in a cell-autonomous way (compare the results with the phenotypic assays employing cell autonomous marker genes [19,36,54]). …”

Section: Prospects Of the Application Ofheterologous Mobile Dna Elemementioning

confidence: 99%

The use of transgenic plants to understand transposition mechanisms and to develop transposon tagging strategies

et al. 1991

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The use of transgenic plants to understand transposition mechanisms and to develop transposon tagging strategies Haring, M.A.; Rommens, C.M.T.; Nijkamp, H.J.J.; Hille, J. Disclaimer/Complaints regulationsIf you believe that digital publication of certain material infringes any of your rights or (privacy) interests, please let the Library know, stating your reasons. In case of a legitimate complaint, the Library will make the material inaccessible and/or remove it from the website. Please Ask the Library: http://uba.uva.nl/en/contact, or a letter to: Library of the University of Amsterdam, Secretariat, Singel 425, 1012 WP Amsterdam, The Netherlands. You will be contacted as soon as possible. AbstractThis review compares the activity of the plant transposable elements Ac, Tam3, En/Spm and Mu in heterologous plant species and in their original host. Mutational analysis of the autonomous transposable elements and two-element systems have supplied data that revealed some fundamental properties of the transposition mechanism. Functional parts of Ac and En/Spm were detected by in vitro binding studies of purified transposase protein and have been tested for their importance in the function of these transposable elements in heterologous plant species. Experiments that have been carried out to regulate the activity of the Ac transposable element are in progress and preliminary results have been compiled. Perspectives for manipulated transposable elements in transposon tagging strategies within heterologous plant species are discussed.

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Transposable elements can be used to study cell lineages in transgenic plants.

Cited by 53 publications

References 23 publications

A transposon tagging strategy with Ac on plant cell level in heterologous plant species

A transposon tagging strategy with Ac on plant cell level in heterologous plant species

Development of a simple transient assay for Ac/Ds activity in cells of intact barley tissue

The use of transgenic plants to understand transposition mechanisms and to develop transposon tagging strategies

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