Abstract
The gibberellin (GA) 2-oxidase (PcGA2ox1) from bean catalyses the 2β-hydroxylation of some precursor and bioactive GAs resulting in their inactivation. We have expressed PcGA2ox1 under the control of the estrogen receptor-based chemical-inducible system, XVE, to modify plant architecture and assess whether transgene expression is localised. Applications of estradiol to the shoot apical region of inducible PcGA2ox1 overexpressors exhibited delays in both bolting (maximum of 46 days) and times to anthesis (maximum of 62 days) compared to wildtype (36 and 41 days, respectively), without altering leaf area. Individual treated leaves showed signs of epinasty and became dark green; such estradiol-treated regions maintained these ‘green-islands’ well beyond the onset of leaf senescence. Northern blots revealed that the PcGA2ox1 transcript could be detected within 1 h of treatment. The level of PcGA2ox1 transcript appeared to peak 3–5 h after estradiol application in both high and semi expressors. Quantitative Reverse Transcription (QRT)-PCR data showed that GA down-regulated genes AtGA3ox1, AtGA20ox1 and SCARECROW-LIKE3 (SCL3) were up-regulated and the GA up-regulated genes AtGA2ox1 and AtExp1 were down-regulated in estradiol-treated leaves of inducible PcGA2ox1 overexpressors; neighbouring non-treated leaves showing no significant changes. Further molecular analyses revealed that expression of the transgene was confined to estradiol-treated leaves only. Expression profiles of GA down- and up-regulated genes in inducer-treated overexpressors appeared to be synchronised with changes in leaf phenotype. These observations suggest that PcGA2ox1 under the control of the XVE system can be used effectively to alter plant architecture in Arabidopsis by localised 2β-hydroxylation of GAs at estradiol-treated sites.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- CaMV:
-
Cauliflower mosaic virus
- GA:
-
Gibberellin
- GUS:
-
β-Glucuronidase
- DIG:
-
Digoxigenin
- PcGA2ox1:
-
Phaseolus coccineus L. GA 2-oxidase1
- QRT-PCR:
-
Quantitative reverse transcription-PCR
- SCL3 :
-
SCARECROW-LIKE3
References
Bishop ON (1971) Statistics for biology, 2nd edn. Longman Group Ltd, London
Caddick MX, Greenland AJ, Jepson I, Krause K-P, Qu N, Riddell KV, Salter MG, Schuch W, Sonnewald U, Tomsett AB (1998) An ethanol inducible gene switch for plants used to manipulate carbon metabolism. Nat Biotechnol 16:177–180
Chiang HH, Hwang I, Goodman HM (1995) Isolation of the Arabidopsis GA4 locus. Plant Cell 7:195–201
Chomczynski P, Mackey K (1995) Modification of the TRI ReagentTM procedure for isolation of RNA from polysaccharide—and proteoglycan—rich sources. Biotechniq 19:942–945
Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743
Coles JP, Phillips AL, Croker SJ, Garcia-Lepe R, Lewis MJ, Hedden P (1999) Modification of gibberellin production and plant development in Arabidopsis by sense and antisense expression of gibberellin 20-oxidase genes. Plant J 17:547–556
Curtis IS, Power JB, Hedden P, Phillips A, Lowe KC, Ward DA, Davey MR (2000a) Transformation and characterization of transgenic plants of Solanum dulcamara L—incidence of transgene silencing. Ann Bot 86:63–71
Curtis IS, Ward DA, Thomas SG, Phillips AL, Davey MR, Power JB, Lowe KC, Croker SJ, Lewis MJ, Magness SL, Hedden P (2000b) Induction of dwarfism in transgenic Solanum dulcamara by over-expression of a gibberellin 20-oxidase cDNA from pumpkin. Plant J 23:329–338
Curtis IS, Nam HG, Yun JY, Seo KH (2002) Expression of an antisense GIGANTEA (GI) gene fragment in transgenic radish causes delayed bolting and flowering. Trans Res 11:249–256
Fire A (1999) RNA-triggered gene silencing. Trends Genet 15:358–363
Gatz C, Lenk I (1998) Promoters that respond to chemical inducers. Trends Plant Sci 3:352–358
Gawronska H, Yang Y-Y, Furukawa K, Kendrick RE, Takahashi N, Kamiya Y (1995) Effects of low irradiance stress on gibberellin levels in pea seedlings. Plant Cell Physiol 36:1361–1367
Hedden P, Phillips AL (2000) Gibberellin metabolism: new insights revealed by the genes. Trends Plant Sci 5:523–530
Lazo GR, Stein PA, Ludwig RA (1991) A DNA transformation-competent Arabidopsis genomic library in Agrobacterium. Bio/Technol 9:963–967
Lester DR, Ross JJ, Smith JJ, Elliot RC, Reid JB (1999) Gibberellin 2-oxidation and the SLN gene of Pisum sativum. Plant J 19:65–73
Martin DN, Proebsting WM, Hedden P (1997) Mendel’s dwarfing gene: cDNAs from the Le alleles and function of the expressed proteins. Proc Natl Acad Sci USA 94:8907–8911
Martin DN, Proebsting WM, Hedden P (1999) The SLENDER gene of pea encodes a gibberellin 2-oxidase. Plant Physiol 121:775–781
Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
Niki T, Nishijima T, Nakayama M, Hisamatsu T, Oyama-Okubo N, Yamazaki H, Hedden P, Lange T, Mander LN, Koshioka M (2001) Production of dwarf lettuce by overexpressing a pumpkin gibberellin 20-oxidase gene. Plant Physiol 126:965–972
Ogawa M, Hanada A, Yamauchi Y, Kuwahara A, Kamiya Y, Yamaguchi S (2003) Gibberellin biosynthesis and response during Arabidopsis seed germination. Plant Cell 15:1591–1604
Phillips AL, Ward DA, Uknes S, Appleford NEJ, Lange T, Huttly AK, Gaskin P, Graebe JE, Hedden P (1995) Isolation and expression of three gibberellin 20-oxidase cDNA clones from Arabidopsis. Plant Physiol 108:1049–1057
Sakamoto T, Kobayashi M, Itoh H, Tagiri A, Kayano T, Tanaka H, Iwahori S, Matsuoka M (2001) Expression of a gibberellin 2-oxidase gene around the shoot apex is related to phase transition in rice. Plant Physiol 125:1508–1516
Sakamoto T, Morinaka Y, Ishiyama K, Kobayashi M, Itoh H, Kayano T, Iwahori S, Matsuoka M, Tanaka H (2003) Genetic manipulation of gibberellin metabolism in transgenic rice. Nat Biotechnol 21:909–913
Sauter M, Kende H (1992) Gibberellin-induced growth and regulation of the cell division cycle in deepwater rice. Planta 188:362–368
Schomburg FM, Bizzell CM, Lee DJ, Zeevaart JAD, Amasino RM (2003) Overexpression of a novel class of gibberellin 2-oxidases decreases gibberellin levels and creates dwarf plants. Plant Cell 15:151–163
Singh DP, Jermakow AM, Swain SM (2002) Gibberellins are required for seed development and pollen tube growth in Arabidopsis. Plant Cell 14:3133–3147
Thomas SG, Phillips AL, Hedden P (1999) Molecular cloning and functional expression of gibberellin 2-oxidases, multifunctional enzymes involved in gibberellin deactivation. Proc Natl Acad Sci USA 96:4698–4703
Yamauchi Y, Ogawa M, Kuwahara A, Hanada A, Kamiya Y, Yamaguchi S (2004) Activation of gibberellin biosynthesis and response pathways by low temperature during imbibition of Arabidopsis thaliana seeds. Plant Cell 16:367–378
Zuo J, Niu Q-W, Chua N-H (2000) An estrogen receptor-based transactivator XVE mediates highly inducible gene expression in transgenic plants. Plant J 24:265–273
Acknowledgements
We would like to thank N.-H. Chua and A.W. Mellon (The Rockefeller University, New York, USA) for providing pER8 and P. Hedden (Rothamsted Research, Harpenden, Herts, UK) for cDNA of PcGA2ox1. This work was supported by a RIKEN Fellow Award (to I.S.C.).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Curtis, I.S., Hanada, A., Yamaguchi, S. et al. Modification of plant architecture through the expression of GA 2-oxidase under the control of an estrogen inducible promoter in Arabidopsis thaliana L. Planta 222, 957–967 (2005). https://doi.org/10.1007/s00425-005-0037-7
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00425-005-0037-7