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BMC Proceedings

Open Access

Investigating the expression pattern of the OsAPx1 gene promoter in rice

  • Julio Garighan1,
  • Carolina Ribeiro1 and
  • Márcia Margis-Pinheiro1
BMC Proceedings20148(Suppl 4):P92

https://doi.org/10.1186/1753-6561-8-S4-P92

Published: 1 October 2014

Background

Ascorbate peroxidase (APx) is a key enzyme of the antioxidant metabolism, catalyzing the decomposition of hydrogen peroxide (H2O2) in water, using ascorbate as an electron donor. The H2O2 is a reactive oxygen species (ROS) produced constantly by aerobic metabolism. Under biotic and abiotic stress the level of H2O2 increases and, in large quantities, can cause cellular damage. In rice, there are eight APx genes that encode products target to different subcellular compartments: cytosol, peroxisoma, mitochondria and chloroplast. OsAPx1 gene encodes a cytosolic isoform of APx. The study of promoters is an important tool that allows to analyze the overall expression pattern of genes in plants.

Methods

A sequence of approximately 2kb preceding the translation initiation site of the OsAPx1 gene was isolated, cloned into pENTR vector and recombined in pHGWFS7 vector, whichallows the fusion of the promoter sequence with two report genes, Gfp and Gus, and confers resistence to hygromycin. The construction was named pPROM1. The transformation of rice calli, originated from nipponbare cultivar seeds, was performed via Agrobacterium tumefaciens. The transformed calli were grown in selection medium with hygromycin, regenerated into plants, acclimatized in a greenhouse and the confirmation of transgene was verified by PCR using specific primers for the Hpt and Gus genes. For visualization of expression pattern of the promoter, by GUS histochemical assay, samples of plants were collected and analyzed by X-gluc histochemical assays. The segments were incubated in 1 mMX-gluc solution at 37°C for 16h. After reaction, green tissues were incubated in 70% ethanol for chlorophyll discoloration. In the in silico analysis of cis-elements in the promoter region of OsAPx1 was used the following databases available online:-PlantPan (http://plantpan.mbc.nctu.edu.tw/) and PlantCare (http://bioinformatics.psb.ugent.be/webtools/plantcare/html/)

Results and conclusions

Nine lines of transgenic plants expressing Gus under the control of the OsAPx1 promoter were obtained. The GUSexpression was observed in leaf (especially in leaf mesophyll), ligule and in wounded regions. These results show that OsAPx1 gene seems to be expressed in green tissues and to respond to damage. Apparently, there is no change in the expression pattern during different development stages. The in silico analysis demonstrates the presence cis-elements responsive to hormones, drought and light.

Declarations

Acknowledgements

CNPq, FAPERGS, CAPES and ICGEB.

Authors’ Affiliations

(1)
Universidade Federal do Rio Grande do Sul

References

  1. Jefferson RA, Kavanagh TA, Bevan MW: GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 1987, 6 (13): 3901-3907.PubMed CentralPubMedGoogle Scholar
  2. Upadhyaya NM, Surin B, Ramm K, Gaudron J, Schünmann PHD, Taylor W, Waterhouse PM, Wang MB: Agrobacterium-mediated transformation of Australian rice cultivars Jarrah and Amaroo using modified promoters and selectable markers. Australian Journal Plant Physiology. 2000, 27 (3): 201-210.Google Scholar
  3. Teixeira FK, Menezes-Benavente L, Margis R, Margis-Pinheiro M: Analysis of the molecular evolutionary history of the ascorbate peroxidase gene family: inferences from the rice genome. J Mol Evol. 2004, 59 (6): 761-770.View ArticlePubMedGoogle Scholar

Copyright

© Garighan et al.; licensee BioMed Central Ltd. 2014

This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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