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Plant Cell Reports

, Volume 23, Issue 12, pp 819–833 | Cite as

Isolation and characterization of a Ds-tagged rice (Oryza sativa L.) GA-responsive dwarf mutant defective in an early step of the gibberellin biosynthesis pathway

  • Marcia Margis-Pinheiro
  • Xue-Rong Zhou
  • Qian-Hao Zhu
  • Elizabeth S. Dennis
  • Narayana M. UpadhyayaEmail author
Genetics and Genomics

Abstract

We have isolated a severe dwarf transposon (Ds) insertion mutant in rice (Oryza sativa L.), which could be differentiated early in the seedling stage by reduced shoot growth and dark green leaves, and later by severe dwarfism and failure to initiate flowering. These mutants, however, showed normal seed germination and root growth. One of the sequences flanking Ds, rescued from the mutant, was of a chromosome 4-located putative ent-kaurene synthase (KS) gene, encoding the enzyme catalyzing the second step of the gibberellin (GA) biosynthesis pathway. Dwarf mutants were always homozygous for this Ds insertion and no normal plants homozygous for this mutation were recovered in the segregating progeny, indicating that the Ds insertion mutation is recessive. As mutations in three recently reported rice GA-responsive dwarf mutant alleles and the dwarf mutation identified in this study mapped to the same locus, we designate the corresponding gene OsKS1. The osks1 mutant seedlings were responsive to exogenous gibberellin (GA3). OsKS1 transcripts of about 2.3 kb were detected in leaves and stem of wild-type plants, but not in germinating seeds or roots, suggesting that OsKS1 is not involved in germination or root growth. There are at least five OsKS1-like genes in the rice genome, four of which are also represented in rice expressed sequence tag (EST) databases. All OsKS1-like genes are transcribed with different expression patterns. ESTs corresponding to all six OsKS genes are represented in other cereal databases including barley, wheat and maize, suggesting that they are biologically active.

Keywords

iAc/Ds insertional mutagenesis GA-responsive dwarf mutants Gibberellin biosynthesis pathway ent- Kaurene synthase Oryza sativa 

Abbreviations

CDP

ent-Copalyl diphosphate

CPS

CDP synthase

Ds

Dissociation

DTC

Diterpene cyclase

EST

Expressed sequence tag

GA

Gibberellic acid

GFP

Green fluorescent protein

GGDP

trans-Geranylgeranyl diphosphate

hpRNA

Hairpin RNA

iAc

Immobile activator

KS

ent-Kaurene synthase

Os

Oryza sativa L.

PCR

Polymerase chain reaction

TAIL-PCR

Thermal asymmetric interlaced-PCR

Notes

Acknowledgements

The authors wish to thank Kerrie Ramm, Ramani Shivakkumar and Shamsul Hoque for their excellent technical support. Thanks to Rogerio Margis, Andrew Eamens, Ming-Bo Wang, Chris Helliwell and Peter Chandler for their valuable comments, suggestions and critical reading of the manuscript. The authors are grateful to their previous collaborators Chellian Santhoshkumar and Kottaram K. Narayanan, for providing pSK200 and pSK300 constructs. M. Margis-Pinheiro’s visit to CSIRO Plant Industry was supported by the Brazilian “Conselho Nacional de Desenvolvimento Científico e Tecnológico—CNPq”. Rice insertional mutagenesis work of CSIRO Plant Industry’s “Rice Functional Genomics” group (http://www.pi.csiro.au/fgrttpub/home.htm) is supported by GrainGene (Australia), Rural Industries Research and Development Corporation (RIRDC), Australia and the NSW Agricultural Genomics Centre.

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Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • Marcia Margis-Pinheiro
    • 1
  • Xue-Rong Zhou
    • 1
  • Qian-Hao Zhu
    • 1
  • Elizabeth S. Dennis
    • 1
  • Narayana M. Upadhyaya
    • 1
    Email author
  1. 1. Plant IndustryCommonwealth Scientific and Industrial Research Organization (CSIRO)CanberraAustralia

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