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Published on January 28, 2009; 10.1104/pp.108.133371

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Received November 28, 2008
Accepted January 26, 2009

Alternative Splicing Studies of the ROS Gene Network in Populus Reveal Two Isoforms of High Iso-electric Point Superoxide Dismutase

Vaibhav Srivastava , Manoj Kumar Srivastava , Kamel Chibani , Robert Nilsson , Nicolas Rouhier , Michael Melzer , and Gunnar Wingsle *

Umea Plant Science Centre, Department of Forest Genetics and Plant Physiology, Swedish University of Agricultural Sciences, 901 83 Umea, Sweden; Department of Molecular Cell Biology, Institute of Plant Genetics and Crop Plant Research, 06466 Gatersleben, Germany; Unite Mixte de recherche INRA-UHP 1136, Interactions Arbres/Micro-organismes, Nancy Universite, IFR110 GEEF, BP 239, 54506 Vandoeuvre Cedex France

* Corresponding author; email: Gunnar.Wingsle{at}genfys.slu.se.

Recent evidence has shown that alternative splicing (AS) is widely involved in the regulation of gene expression, substantially extending the diversity of numerous proteins. In this study, a subset of ESTs representing members of the ROS gene network was selected from the PopulusDB to investigate AS mechanisms in Populus. Examples of all known types of AS were detected, but intron retention (IntronR) was the most common. Interestingly, the closest Arabidopsis (Arabidopsis thaliana) homologues of half of the AS genes identified in Populus are not reportedly alternatively spliced. Two genes encoding the protein of most interest in our study (hipI-SOD) have been found in Populus trichocarpa, designated PthipI-SODC1 and PthipI-SODC2. Analysis of the EST libraries has indicated the presence of two transcripts of PthipI-SODC1 (hipI-SODC1b and hipI-SODC1s). Alignment of these sequences with the PthipI-SODC1 gene showed that hipI- SODC1b was 69 bp longer than hipI-SODC1s due to an AS event involving the use of an alternative donor splice site in the 6th intron. Transcript analysis showed that the splice variant hipI-SODC1b was differentially expressed, being clearly expressed in cambial and xylem, but not phloem, regions. In addition, immunolocalisation and mass spectrometric data confirmed the presence of hipI-SOD proteins in vascular tissue. The functionalities of the spliced gene products were assessed by expressing recombinant hipI-SOD proteins and in vitro SOD activity assays.






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