Expression of vacuolar H+-pyrophosphatase (OVP3) is under control of an anoxia-inducible promoter in rice
- 386 Downloads
Vacuolar H+-pyrophosphatase (V-PPase) expression increases in a number of abiotic stresses and is thought to play a role in adaptation to abiotic stresses. This paper reports on the regulation of six V-PPase genes in rice (Oryza sativa L.) coleoptiles under anoxia, using flood tolerant and intolerant cultivars to test our hypothesis. Quantitative PCR analysis showed that one vacuolar H+-pyrophosphatase (OVP3) was induced by anoxia, particularly in flood-tolerant rice. Regulation of OVP3 expression under anoxia was investigated by analysing putative OVP promoters. The putative OVP3 promoter contained more previously identified anoxia-inducible motifs than the putative promoters of the other five OVP genes. GUS activity in transgenic rice plants containing the OVP3 promoter region linked to the GUS reporter gene was induced only by anoxia. Salt and cold treatments had little effect on OVP3 promoter-driven GUS expression when compared to the anoxic treatment.
KeywordsAnoxia Gene expression Promoter Rice Vacuolar H+-pyrophosphatase
We thank Olivier Cotsaftis for expert assistance with rice transformation, Andrew Harvey for sequence analysis, Ezaz Mamun for vacuole isolation and Gwenda Mayo for technical support involving microscopy. We are also grateful to Masayoshi Maeshima (Nagoya University, Japan) for the gift of the anti-V-PPase and anti-V-ATPase antibodies. This research was made possible through the generosity of the Australian Centre for Plant Functional Genomics, School of Agriculture, Food and Wine, University of Adelaide, Australia.
- Atwell BJ, Rees T (1986) Distribution of protein synthesized by seedlings of Oryza sativa grown in anoxia. J Plant Physiol 123:401–408Google Scholar
- Howell KA, Narsai R, Carroll A, Ivanova A, Lohse M, Usadel B, Millar AH, Whelan J (2009) Mapping metabolic and transcript temporal switches during germination in rice highlights specific transcription factors and the role of RNA instability in the germination process. Plant Physiol 149:961–980CrossRefPubMedGoogle Scholar
- Lehr A, Kirsch M, Viereck R, Schiemann J, Rausch T (1999) cDNA and genomic cloning of sugar beet V-type H+-ATPase subunit A and c isoforms: evidence for coordinate expression during plant development and coordinate induction in response to high salinity. Plant Mol Biol 39:463–475CrossRefPubMedGoogle Scholar
- Peter Verrijzer C, Tjian R (1996) TAFs mediate transcriptional activation and promoter selectivity. Trends Biochem Sci 21:338–342Google Scholar
- Rea PA, Poole RJ (1993) Vacuolar H+ -translocating pyrophosphatase. Annu Rev Plant Physiol Plant Mol Biol 44:157–180Google Scholar
- Sallaud C, Meynard D, van Boxtel J, Gay C, Bes M, Brizard JP, Larmande P, Ortega D, Raynal M, Portefaix M, Ouwerkerk PB, Rueb S, Delseny M, Guiderdoni E (2003) Highly efficient production and characterization of T-DNA plants for rice (Oryza sativa L.) functional genomics. Theor Appl Genet 106:1396–1408PubMedGoogle Scholar
- Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. A laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, pp 121–131Google Scholar
- Stitt M (1998) Pyrophosphate as an energy donor in the cytosol of plant cells: an enigmatic alternative to ATP. Bot Acta 111:167–175Google Scholar
- Stomp A (1992) Histochemical localization of β-glucuronidase. In: Gallagher SR (ed) GUS protocols: using the GUS gene as a reporter of gene expression. Academic Press, San Diego, pp 103–113Google Scholar
- Xia J-H, Saglio P, Roberts J (1985) Nucleotide levels do not critically determine survival of maize root tips acclimated to a low-oxygen environment. Plant Physiol 1995:589–595Google Scholar