Transcription and somatic transposition of the maize En / Spm transposon system in rice View Full Text


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Article Info

DATE

2003-11-14

AUTHORS

R. Greco, P. B. F. Ouwerkerk, A. J. C. Taal, C. Sallaud, E. Guiderdoni, A. H. Meijer, J. H. C. Hoge, A. Pereira

ABSTRACT

Transposition of the maize En/Spm system in rice was investigated using a two-component construct consisting of an immobilised transposase source driven by the CaMV 35S-promoter, and a modified I/dSpm transposon. Mobilization of I/dSpm in somatic sectors was demonstrated by sequencing of excision products and isolation of flanking genomic sequences in T0 and T1 progeny plants. Since the transposition efficiency appeared to be considerably lower than that observed in maize or in other heterologous systems like Arabidopsis, we examined En/Spm transcription and splicing in the transgenic rice plants. Northern analysis revealed the presence of transcripts encoding the active TnpA and TnpD transposases, with the latter predominating; this is the reverse of what is seen in maize and Arabidopsis. RT-PCR analysis confirmed the occurrence of correct splicing and the formation of the two other alternatively spliced transcripts (TnpB and TnpC), as previously described for maize. Two alternative splice donor sites at the end of exon 1 were identified in maize at positions 578 and 704. We observe that rice is similar to maize in that TnpA is preferentially spliced at position 578. We also show that in Arabidopsis splicing occurs preferentially at position 704, as in other dicots like tobacco. These observations indicate differences in the splicing of transcripts of the maize En/Spm element between dicot and monocot hosts. Nevertheless, the ratio in which the transcripts for the active transposases are produced seems to determine the efficiency of transposition, irrespective of the host considered. A limiting amount of TnpA might therefore be responsible for the lower transposition activity of En/Spm in rice. Alternatively, reduced mobility of the modified I/dSpm element used may have resulted from the absence of critical sequences necessary for transposition. The influence of endogenous, autonomous, En/Spm -related elements present in the rice genome on the transposition behaviour of the exogenous maize element is also considered. More... »

PAGES

514-523

References to SciGraph publications

  • 2001-10. Molecular analysis of the Doppia transposable element of maize in PLANT MOLECULAR BIOLOGY
  • 1989-05. The maize En-1/Spm element transposes in potato in MOLECULAR GENETICS AND GENOMICS
  • 2000-08. A transgenic perspective on plant functional genomics in TRANSGENIC RESEARCH
  • 1993-10. Definition and characterization of an artificial En/Spm-based transposon tagging system in transgenic tobacco in PLANT MOLECULAR BIOLOGY
  • 1998-02. Excision of Ds1 from the genome of maize streak virus in response to different transposase-encoding genes in PLANT MOLECULAR BIOLOGY
  • 1998-08. The behaviour of the autonomous maize transposable element En/Spm in Arabidopsis thaliana allows efficient mutagenesis in PLANT MOLECULAR BIOLOGY
  • 1993-02. Genetic and molecular analysis of a three-component transposable-element system in maize in MOLECULAR GENETICS AND GENOMICS
  • 2002-11. The genome sequence and structure of rice chromosome 1 in NATURE
  • 1997-09. Transposon tagging in rice in PLANT MOLECULAR BIOLOGY
  • 2001-05. Early and multiple Ac transpositions in rice suitable for efficient insertional mutagenesis in PLANT MOLECULAR BIOLOGY
  • 2002-11. Sequence and analysis of rice chromosome 4 in NATURE
  • 1993-01. A 168 bp derivative of Suppressor-mutator/Enhancer is responsible for the maize o2–23 mutation in PLANT MOLECULAR BIOLOGY
  • 1996-05. Ubiquitin promoter-based vectors for high-level expression of selectable and/or screenable marker genes in monocotyledonous plants in TRANSGENIC RESEARCH
  • 1995-09. A two-element Enhancer-Inhibitor transposon system in Arabidopsis thaliana in MOLECULAR GENETICS AND GENOMICS
  • 2001-09. Tagged Transcriptome Display (TTD) in indica rice using Ac transposition in MOLECULAR GENETICS AND GENOMICS
  • 1970-01. The En mutable system in maize in THEORETICAL AND APPLIED GENETICS
  • 2003-01. An active DNA transposon family in rice in NATURE
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1007/s00438-003-0942-z

    DOI

    http://dx.doi.org/10.1007/s00438-003-0942-z

    DIMENSIONS

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    PUBMED

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    32 schema:description Transposition of the maize En/Spm system in rice was investigated using a two-component construct consisting of an immobilised transposase source driven by the CaMV 35S-promoter, and a modified I/dSpm transposon. Mobilization of I/dSpm in somatic sectors was demonstrated by sequencing of excision products and isolation of flanking genomic sequences in T0 and T1 progeny plants. Since the transposition efficiency appeared to be considerably lower than that observed in maize or in other heterologous systems like Arabidopsis, we examined En/Spm transcription and splicing in the transgenic rice plants. Northern analysis revealed the presence of transcripts encoding the active TnpA and TnpD transposases, with the latter predominating; this is the reverse of what is seen in maize and Arabidopsis. RT-PCR analysis confirmed the occurrence of correct splicing and the formation of the two other alternatively spliced transcripts (TnpB and TnpC), as previously described for maize. Two alternative splice donor sites at the end of exon 1 were identified in maize at positions 578 and 704. We observe that rice is similar to maize in that TnpA is preferentially spliced at position 578. We also show that in Arabidopsis splicing occurs preferentially at position 704, as in other dicots like tobacco. These observations indicate differences in the splicing of transcripts of the maize En/Spm element between dicot and monocot hosts. Nevertheless, the ratio in which the transcripts for the active transposases are produced seems to determine the efficiency of transposition, irrespective of the host considered. A limiting amount of TnpA might therefore be responsible for the lower transposition activity of En/Spm in rice. Alternatively, reduced mobility of the modified I/dSpm element used may have resulted from the absence of critical sequences necessary for transposition. The influence of endogenous, autonomous, En/Spm -related elements present in the rice genome on the transposition behaviour of the exogenous maize element is also considered.
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    41 CaMV
    42 DSPM
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