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Efficient insertional mutagenesis in rice using the maize En/Spm elements.

Chellian Santhosh Kumar1, Rod A Wing, Venkatesan Sundaresan

  • 1Department of Plant Sciences, Life Sciences Addition 1002, University of California, Davis, 95616, USA.

The Plant Journal : for Cell and Molecular Biology
|November 22, 2005
PubMed
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Researchers created a new system for insertional mutagenesis in rice using the maize enhancer/suppressor mutator (En/Spm) element. This novel method, utilizing fluorescent markers, efficiently generates stable dSpm insertions in rice genes.

Area of Science:

  • Plant genetics
  • Molecular biology
  • Genomics

Background:

  • Insertional mutagenesis is crucial for understanding gene function in plants.
  • Existing systems like Activator-Dissociation (Ac-Ds) have limitations in genomic distribution and transposition patterns.
  • The maize enhancer/suppressor mutator (En/Spm) element offers potential for novel insertional mutagenesis strategies.

Purpose of the Study:

  • To develop and validate a novel insertional mutagenesis system in rice (Oryza sativa) utilizing the maize (Zea mays) En/Spm element.
  • To assess the efficiency and distribution patterns of dSpm insertions in the rice genome.
  • To evaluate the utility of a fluorescence-based selection system for identifying stable transpositions.

Main Methods:

  • Developed a single T-DNA construct containing Spm-transposase and the non-autonomous defective suppressor mutator (dSpm) element.

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  • Incorporated green fluorescent protein (GFP) and Discosoma sp. Red Fluorescence Protein (DsRed) as fluorescent markers for selection.
  • Generated and analyzed 353 stable dSpm insertion lines in rice.

    • Main Results:

    • Achieved high frequencies of unlinked germinal transposition of dSpm in rice.
    • dSpm insertions were widely distributed across rice chromosomes, with a significant preference (70%) for genic regions.
    • dSpm insertions exhibited a different genomic distribution compared to Ac-Ds elements, with a higher proportion of unlinked transpositions.

    Conclusions:

    • The maize En/Spm element is an effective tool for functional genomics in rice, complementing existing insertional mutagens.
    • The developed fluorescence-based selection system is efficient and shows promise for application in other crop species.
    • This system provides a valuable resource for rice functional genomics and crop improvement efforts.