Skip to main content
SpringerLink
Log in

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

  • Genetics and Genomics
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2a,b
Fig. 3a–c
Fig. 4
Fig. 5
Fig. 6
Fig. 7a–d

Similar content being viewed by others

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

References

  • Ait-Ali T, Swain SM, Reid JB, Sun TP, Kamiya Y (1997) The LS locus of pea encodes the gibberellin biosynthesis enzyme ent-kaurene synthase A. Plant J 11:443–454

    Article  Google Scholar 

  • Ashikari M, Matsuoka M (2002) Application of rice genomics to plant biology and breeding. Bot Bull Acad Sin 43:1–11

    Google Scholar 

  • Bensen RJ, Johal GS, Crane VC, Tossberg JT, Schbable PS, Meeley RB, Briggs SP (1995) Cloning and characterization of the maize An1 gene. Plant Cell 7:75–84

    Article  Google Scholar 

  • Börner A, Korzun V, Malyshev S, Ivandic V, Graner A (1999) Molecular mapping of two dwarfing genes differing in their GA response on chromosome 2H of barley. Theor Appl Genet 99:670–675

    Article  Google Scholar 

  • Chiang HH, Hwang I, Goodman HM (1995) Isolation of the Arabidopsis GA4 locus. Plant Cell 7:195–201

    Article  Google Scholar 

  • Cho E-M, Okada A, Kenmoku H, Otomo K, Toyomasu T, Mitsuhashi W, Sassa T, Yajima A, Yabuta G, Mori K et al (2004) Molecular cloning and characterization of a cDNA encoding ent-cassa-12,15-diene synthase, a putative diterpenoid phytoalexin biosynthetic enzyme, from suspension-cultured rice cells treated with a chitin elicitor. Plant J 37:1–8

    Google Scholar 

  • Davidson SE, Elliott RC, Helliwell CA, Poole AT, Reid JB (2003) The pea gene NA encodes ent-kaurenoic acid oxidase. Plant Physiol 131:335–344

    Article  Google Scholar 

  • Davis GL, McMullen MD, Baysdorfer C, Musket T, Grant D, Staebell M, Xu G, Polacco M, Koster L, Melia-Hancock S, Houchins K, Chao S, Coe EH Jr (1999) A maize map standard with sequenced core markers, grass genome reference points and 932 expressed sequence tagged sites (ESTs) in a 1736-locus map. Genetics 152:1137–1172

    CAS  PubMed  Google Scholar 

  • Debeaujon I, Koornneef M (2000) Gibberellin requirement for Arabidopsis seed germination is determined both by testa characteristics and embryonic abscisic acid. Plant Physiol 122:415–424

    Article  Google Scholar 

  • Devereux J, Haeberli P, Smithies O (1984) A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res 12:387–396

    CAS  PubMed  Google Scholar 

  • Futsuhara Y, Kikuchi F (1993) In: Matsuo T, Kumazawa K, Ishihara R, Hirata H (eds) Science of rice plant, vol 3. Food and Agriculture Policy Research Center, Tokyo, pp 300–308

    Google Scholar 

  • Gale M, Morre G, Devos K (2001) Rice: a central genome for the genetics of all cereals. In: Khush GS, Brar DS, Hardy B (eds) Rice genetics IV. Proceedings of the fourth international rice genetics symposium. International Rice Research Institute, Los Banos, Philippines, pp 79–88

  • Gallardo K, Job C, Groot SP, Puype M, Demol H, Vandekerckhove J, Job D (2002) Proteomics of Arabidopsis seed germination. A comparative study of wild-type and gibberellin-deficient seeds. Plant Physiol 129:823–837

    Article  CAS  PubMed  Google Scholar 

  • Hedden P, Phillips AL (2000) Gibberellin metabolism: new insights revealed by the genes. Trends Plant Sci 5:523–530

    Article  CAS  PubMed  Google Scholar 

  • Helliwell CA, Sheldon CC, Olive MR, Walker AR, Zeevaart JAD, Peacock WJ, Dennis ES (1998) Cloning of the Arabidopsis ent-kaurene oxidase gene GA3. Proc Natl Acad Sci USA 95:9019–9024 http://www.pnas.org/cgi/content/abstract/95/15/9019?ijkey=f4ae86d716d53fa27c536edfa91bdf99b03f14e7&keytype2=tf_ipsecsha

    Google Scholar 

  • Itoh H, Ueguchi-Tanaka M, Sentoku N, Kitano H, Matsuoka M, Kobayashi M (2001) Cloning and functional analysis of two gibberellin 3 beta-hydroxylase genes that are differently expressed during the growth of rice. Proc Natl Acad Sci USA 98:8909–8914

    Article  Google Scholar 

  • Kaneko M, Itoh H, Ueguchi-Tanaka M, Ashikari M, Matsuoka M (2002) The alpha-amylase induction in endosperm during rice seed germination is caused by gibberellin synthesized in epithelium. Plant Physiol 128:1264–1270

    Article  Google Scholar 

  • Kawaide H, Sassa T, Kamiya Y (2000) Functional analysis of the two interacting cyclase domains in ent-kaurene synthase from the fungus Phaeosphaeria sp. L487 and a comparison with cyclases from higher plants. J Biol Chem 275:2276–2280

    Article  Google Scholar 

  • Khandjian EW (1987) Optimized hybridization of DNA blotted and fixed to nitrocellulose and nylon membranes. Biotechnology 5:165–167

    Google Scholar 

  • King RW, Evans LT (2003) Gibberellins and flowering of grasses and cereals: prizing open the lid of the “Florigen” black box. Annu Rev Plant Biol 54:307–328

    Article  Google Scholar 

  • Koornneef M, van der Veen J (1980) Induction and analysis of gibberellin sensitive mutants in Arabidopsis thaliana (L.) Heynh. Theor Appl Genet 58:257–263

    Article  Google Scholar 

  • Kumar S, Tamura K, Jacobsen I, Nei M (2000) MEGA2: molecular evolutionary genetics analysis, version 2.0. Pennsylvania State Universities, University Park, Pa. (http://www.megasoftware.net/)

    Google Scholar 

  • Liu YG, Mitsukawa N, Ooosumi T, Whittier RF (1995) Efficient isolation and mapping of Arabidopsis thaliana T-DNA insert junctions by thermal asymmetric interlaced PCR. Plant J 8:457–463

    Article  CAS  PubMed  Google Scholar 

  • Murakani Y (1995) In: Matsuo T, Kumazawa K, Ishihara R, Hirata H (eds) Science of rice plant, vol 2. Food and Agriculture Policy Research Center, Tokyo, pp 182–189

    Google Scholar 

  • Ogawa S, Toyomasu T, Yamane H, Murofushi N, Ikeda R, Morimoto Y, Nishimura Y, Omori T (1996) A step in the biosynthesis of gibberellins that is controlled by the mutation in the semi-dwarf rice cultivar Tan-ginbozu. Plant Cell Physiol 37:363–368 http://go5.isiknowledge.com/portal.cgi?DestApp=WOS&DestParams=%3F%26CustomersID%3DHighwire%26CustomersIP%3D152.83.165.114%26Func%3DLinks%26PointOfEntry%3DFullRecord%26PublisherID%3DHighwire%26ServiceName%3DTransferToWos%26ServiceUser%3DLinks%26UT%3DA1996UJ28900017&DestFail=http%3A%2F%2Fcel.isiknowledge.com%2FCIW.cgi%3F%26CustomersID%3DHighwire%26Func%3DLinks%26PointOfEntry%3DFullRecord%26PublisherID%3DHighwire%26ServiceName%3DTransferToWos%26ServiceUser%3DLinks%26UT%3DA1996UJ28900017%26e%3D__MkYKVqXGQ.ZZZH2c3IqOmOjsMoVd2fnw2e0fUi2Rzwqu59u4tjan7JHyolPz6U&SrcApp=Highwire&SrcAuth=Highwire

    Google Scholar 

  • Olszewski N, Sun TP, Gubler F (2002) Gibberellin signalling: biosynthesis, catabolism, and response pathways. Plant Cell 14 [Suppl]:61–80 http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=151248

    Google Scholar 

  • Peng J, Harberd NP (2002) The role of GA-mediated signalling in the control of seed germination. Curr Opin Plant Biol 5:376–381

    Article  Google Scholar 

  • Phinney BO (1984) Gibberellin A1, dwarfism, and the control of shoot elongation in higher plants. In: Crozier A, Hillman FR (eds) The biosynthesis and metabolism of plant hormones. Cambridge University Press, Cambridge, pp 17–41

    Google Scholar 

  • Sakamoto T, Miura K, Itoh H, Tatsumi T, Ueguchi-Tanaka M, Ishiyama K, Kobayashi M, Agrawal GK, Takeda S, Abe K et al (2004) An overview of gibberellin metabolism enzyme genes and their related mutants in rice. Plant Physiol 134:1642–1653

    Article  Google Scholar 

  • Sasaki A, Ashikari M, Ueguchi-Tanaka M, Itoh H, Nishimura A, Swapan D, Ishiyama K, Saito T, Kobayashi M, Khush GS, Kitano H, Matsuoka M (2002) Green revolution: a mutant gibberellin-synthesis gene in rice. Nature 416:701–702

    Article  CAS  PubMed  Google Scholar 

  • Silverstone AL, Chang CW, Krol E, Sun TP (1997) Developmental regulation of the gibberellin biosynthetic gene GA1 in Arabidopsis thaliana. Plant J 12:9–19

    Article  Google Scholar 

  • Singh DP, Jermakow AM, Swain SM (2002) Gibberellins are required for seed development and pollen tube growth in Arabidopsis. Plant Cell 14:3133–3147

    Article  Google Scholar 

  • Sitnikova T, Rzhetsky A, Nei M (1995) Interior-branch and bootstrap tests of phylogenetic trees. Mol Biol Evol 12:319–333 http://mbe.oupjournals.org/cgi/content/abstract/12/2/319

    Google Scholar 

  • Spielmeyer W, Ellis MH, Chandler PM (2002) Semidwarf (sd-1), “green revolution” rice, contains a defective gibberellin 20-oxidase gene. Proc Natl Acad Sci USA 99:9043–9048

    Article  Google Scholar 

  • Sponsel VM, Schmidt FM, Porter SG, Nakayama M, Kohlstruk S, Estelle M (1997) Characterization of new gibberellin-responsive semidwarf mutants of Arabidopsis. Plant Physiol 115:1009–1020

    Article  Google Scholar 

  • Sun TP, Goodman HM, Ausubel FM (1992) Cloning the Arabidopsis GA1 locus by genomic subtraction. Plant Cell 4:119–128

    Article  Google Scholar 

  • Swain S, Reid JB, Kamiya Y (1997) Gibberellins are required for embryo growth and seed development in pea. Plant J 12:1329–1338

    Article  Google Scholar 

  • Upadhyaya NM, Surin B, Ramm K, Gaudron J, Schunmann PHD, Taylor W, Waterhouse PM, Wang MB (2000) Agrobacterium-mediated transformation of Australian rice cultivars Jarrah and Amaroo using modified promoters and selectable markers. Aust J Plant Physiol 27:201–210 http://www.publish.csiro.au/nid/102/paper/PP01205.htm

    Google Scholar 

  • Upadhyaya NM, Zhou XR, Zhu QH, Ramm K, Wu LM, Eamens A, Sivakumar R, Kato T, Yun DW, Santhoshkumar C, Narayanan KK, Peacock JW, Dennis ES (2002) An iAc/Ds gene and enhancer trapping system for insertional mutagenesis in rice. Funct Plant Biol 29:547–559 http://www.publish.csiro.au/nid/102/paper/PP99078.htm

    Google Scholar 

  • Wang M-B, Waterhouse PM (1997) A rapid and simple method of assaying plants transformed with hygromycin or PPT resistance genes. Plant Mol Biol Rep 15:209–215

    Article  Google Scholar 

  • Wang M-B, Abbott DC, Waterhouse PM (2000) A single copy of a virus-derived transgene encoding hairpin RNA gives immunity to barley yellow dwarf virus. Mol Plant Pathol 1:347–356

    Article  Google Scholar 

  • Wang M-B, Boulter D, Gatehouse JA (1992) A complete sequence of the rice sucrose synthase-1 (RSs1) gene. Plant Mol Biol 19:881–885

    Article  Google Scholar 

  • Wang M-B, Li Z, Mathews PR, Upadhyaya NM, Waterhouse PM (1998) Improved vectors for Agrobacterium tumefaciens-mediated transformation of monocot plants. Acta Hortic 461:401–407

    Google Scholar 

  • Xu YL, Li L, Wu KQ, Peeters AJM, Gage DA, Zeevaart JAD (1995) The GA5 locus of Arabidopsis thaliana encodes a multifuctional gibberellin 20-oxidase—molecular cloning and functional expression. Proc Natl Acad Sci USA 92:6640–6644

    CAS  PubMed  Google Scholar 

  • Yamaguchi S, Saito T, Abe H, Yamane H, Murofushi N, Kamiya Y (1996) Molecular cloning and characterization of a cDNA encoding the gibberellin biosynthetic enzyme ent-kaurene synthase B from pumpkin (Cucurbita maxima L.). Plant J 10:201–213

    Article  Google Scholar 

  • Yamaguchi S, Sun TP, Kawaide H, Kamiya Y (1998) The GA2 locus of Arabidopsis thaliana encodes ent-kaurene synthase of gibberellin biosynthesis. Plant Physiol 116:1271–1278

    Article  Google Scholar 

  • Yaxley JR, Ross JJ, Sherriff LJ, Reid JB (2001) Gibberellin biosynthesis mutations and root development in pea. Plant Physiol 125:627–633

    Article  Google Scholar 

Download references

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.

Author information

Author notes

  1. Marcia Margis-Pinheiro

    Present address: Laboratório de Genética Molecular Vegetal (LGMV) – Depto de Genética, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil

Authors and Affiliations

  1. Plant Industry, Commonwealth Scientific and Industrial Research Organization (CSIRO), GPO Box 1600, Canberra, ACT 2601, Australia

    Marcia Margis-Pinheiro, Xue-Rong Zhou, Qian-Hao Zhu, Elizabeth S. Dennis & Narayana M. Upadhyaya

Authors
  1. Marcia Margis-Pinheiro
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xue-Rong Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Qian-Hao Zhu
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Elizabeth S. Dennis
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Narayana M. Upadhyaya
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Narayana M. Upadhyaya.

Additional information

Communicated by P. Lakshmanan

Rights and permissions

Reprints and permissions

About this article

Cite this article

Margis-Pinheiro, M., Zhou, XR., Zhu, QH. et al. 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. Plant Cell Rep 23, 819–833 (2005). https://doi.org/10.1007/s00299-004-0896-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-004-0896-6

Keywords

Search

Navigation