RNA-mediated transposition of the tobacco retrotransposon Tnt1 in Arabidopsis thaliana.

Lucas, Hélène; Feuerbach, Frank; Kunert, K.; Grandbastien, Marie–Angèle; Caboche, Michel

doi:10.1002/j.1460-2075.1995.tb07231.x

Cited by 82 publications

(92 citation statements)

References 68 publications

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“…Furthermore, several different single-copy insert plants were easily obtained after only three backcrosses and one self-pollination, starting with a plant containing 28 copies of Tnt1, demonstrating the feasibility of separating a given Tnt1 insertion from the others even from a genotype containing large numbers of transposed copies. No new transposition events were observed in any of the progenies studied, as already observed in Arabidopsis and Medicago (Lucas et al, 1995;d'Erfurth et al, 2003). Thus, the insertions of Tnt1 are stable and the element does not transpose at a detectable frequency in the germ cells of lettuce under standard growth conditions in the greenhouse.…”

Section: Discussionsupporting

confidence: 67%

“…To investigate whether Tnt1 transposes in lettuce, leaf fragments from two different varieties of butterhead lettuce, 'Mariska' and 'Jessy,' were cocultivated with an Agrobacterium strain carrying the tnk23 plasmid (Fig. 1A) containing in its T-DNA an autonomous copy of Tnt1 (Lucas et al, 1995). After 2 d of cocultivation, leaf explants were transferred to an in vitro selective regeneration medium until a number of transformed bushes appeared, as described by Dinant et al (1997).…”

Section: Transposition Of Tnt1 During In Vitro Transformation Of Lettmentioning

confidence: 99%

“…1A) is a derivative of the pBin19 vector carrying the autonomous Tnt1 retroelement (Lucas et al, 1995). The pHLV55501 vector is also derived from pBin19 and carries a translational LTR-GUS-intron fusion (Courtial et al, 2001).…”

Section: T-dna Vectors and Bacterial Strainmentioning

confidence: 99%

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Successful Gene Tagging in Lettuce Using the Tnt1 Retrotransposon from Tobacco

Mazier¹,

Botton²,

Flamain³

et al. 2007

Plant Physiol.

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The tobacco (Nicotiana tabacum) element Tnt1 is one of the few identified active retrotransposons in plants. These elements possess unique properties that make them ideal genetic tools for gene tagging. Here, we demonstrate the feasibility of gene tagging using the retrotransposon Tnt1 in lettuce (Lactuca sativa), which is the largest genome tested for retrotransposon mutagenesis so far. Of 10 different transgenic bushes carrying a complete Tnt1 containing T-DNA, eight contained multiple transposed copies of Tnt1. The number of transposed copies of the element per plant was particularly high, the smallest number being 28. Tnt1 transposition in lettuce can be induced by a very simple in vitro culture protocol. Tnt1 insertions were stable in the progeny of the primary transformants and could be segregated genetically. Characterization of the sequences flanking some insertion sites revealed that Tnt1 often inserted into genes. The progeny of some primary transformants showed phenotypic alterations due to recessive mutations. One of these mutations was due to Tnt1 insertion in the gibberellin 3b-hydroxylase gene. Taken together, these results indicate that Tnt1 is a powerful tool for insertion mutagenesis especially in plants with a large genome.

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Section: Discussionsupporting

confidence: 67%

Section: Transposition Of Tnt1 During In Vitro Transformation Of Lettmentioning

confidence: 99%

See 1 more Smart Citation

Successful Gene Tagging in Lettuce Using the Tnt1 Retrotransposon from Tobacco

Mazier¹,

Botton²,

Flamain³

et al. 2007

Plant Physiol.

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“…In plants, direct evidence for retrotransposition was demonstrated for only a few elements, such as Tos17 of rice (Hirochika et al, 1996) and Tto1 , Tto2 , and Tnt1 of tobacco (Hirochika, 1993;Lucas et al, 1995). This finding may reflect technical problems, because special techniques, such as transposon display, are required to monitor the mobility of highly repetitive elements (Melayah et al, 2001).…”

Section: Introductionmentioning

confidence: 99%

Two-Step Regulation and Continuous Retrotransposition of the Rice LINE-Type Retrotransposon Karma

2003

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Here, we report the identification of Karma , a LINE-type retrotransposon of plants for which continuous retrotransposition was observed in consecutive generations. The transcription of Karma is activated in cultured cells of rice upon DNA hypomethylation. However, transcription is insufficient for retrotransposition, because no increase in the copy number was observed in cultured cells or in the first generation of plants regenerated from them. Despite that finding, copy number increase was detected in the next generation of regenerated plants as well as in later generations, suggesting that the posttranscriptional regulation of Karma retrotransposition is development dependent. Our results indicate that two different mechanisms, one transcriptional and the other developmental, control the mobilization of Karma . In addition, unlike other known active plant retrotransposons, Karma is not subject to de novo methylation, and retrotransposition persists through several generations.

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“…The Tto1 element, from tobacco (Nicotiana tabacum), has also been used for mutagenesis in Arabidopsis (Okamoto and Hirochika, 2000) and rice (Hirochika et al, 1996). Tnt1, originally isolated from tobacco, has been successfully used in several heterologous hosts, including Medicago truncatula (d'Erfurth et al, 2003;Tadege et al, 2005Tadege et al, , 2008Iantcheva et al, 2009), Arabidopsis (Lucas et al, 1995;Courtial et al, 2001), and lettuce (Lactuca sativa; Mazier et al, 2007). Collectively, these studies demonstrate that retrotransposons transpose preferentially into gene-rich regions, thus making them highly mutagenic.…”

mentioning

confidence: 99%

Tnt1 Retrotransposon Mutagenesis: A Tool for Soybean Functional Genomics

et al. 2012

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Insertional mutagenesis is a powerful tool for determining gene function in both model and crop plant species. Tnt1, the transposable element of tobacco (Nicotiana tabacum) cell type 1, is a retrotransposon that replicates via an RNA copy that is reverse transcribed and integrated elsewhere in the plant genome. Based on studies in a variety of plants, Tnt1 appears to be inactive in normal plant tissue but can be reactivated by tissue culture. Our goal was to evaluate the utility of the Tnt1 retrotransposon as a mutagenesis strategy in soybean (Glycine max). Experiments showed that the Tnt1 element was stably transformed into soybean plants by Agrobacterium tumefaciens-mediated transformation. Twenty-seven independent transgenic lines carrying Tnt1 insertions were generated. Southernblot analysis revealed that the copy number of transposed Tnt1 elements ranged from four to 19 insertions, with an average of approximately eight copies per line. These insertions showed Mendelian segregation and did not transpose under normal growth conditions. Analysis of 99 Tnt1 flanking sequences revealed insertions into 62 (62%) annotated genes, indicating that the element preferentially inserts into protein-coding regions. Tnt1 insertions were found in all 20 soybean chromosomes, indicating that Tnt1 transposed throughout the soybean genome. Furthermore, fluorescence in situ hybridization experiments validated that Tnt1 inserted into multiple chromosomes. Passage of transgenic lines through two different tissue culture treatments resulted in Tnt1 transposition, significantly increasing the number of insertions per line. Thus, our data demonstrate the Tnt1 retrotransposon to be a powerful system that can be used for effective large-scale insertional mutagenesis in soybean.

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RNA-mediated transposition of the tobacco retrotransposon Tnt1 in Arabidopsis thaliana.

Cited by 82 publications

References 68 publications

Successful Gene Tagging in Lettuce Using the Tnt1 Retrotransposon from Tobacco

Successful Gene Tagging in Lettuce Using the Tnt1 Retrotransposon from Tobacco

Two-Step Regulation and Continuous Retrotransposition of the Rice LINE-Type Retrotransposon Karma

Tnt1 Retrotransposon Mutagenesis: A Tool for Soybean Functional Genomics

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