Molecular Plant Breeding 2016, Vol.7, No.17, 1-7
6
spectrophotometer at 460 nm as o-dianizidine oxidation, with 3 replications. Enzyme activity was expressed as
µmole of o-dianizidine oxidation. FM/ming of fresh matter (FM).
The aerial (shoot and leaves) and root tissues peroxidase pattern were resolved by native in 10% separating
polyacrylamide gelelectrophoresis on vertical slab gel (Hoffer, USA) using the procedure of Laemmli (1970).
Equal amounts of proteins (15 µl) were loaded on to each lane. The gel electrophoresis experiment was repeated
tree times. To determine the pattern of peroxidase isoforms gels were staining and visualized by immersing the
gels in 100 ml 0.2 M acetate buffer pH 4.6 added with 1% o-dianizidine (Sigma), dissolved in 2 ml 95% ethanol,
and 200 µl 3% H
2
O
2
at room temperature until the brown color appeared. Scanned Isozymes profiles gels were
analyzed by Software GELANALYZER (Istvan Lazar Hungary Copyright 2010) to determine Rfs bands.
Acknowledgments
This work was supported by Research PNR projectentitled ″Biotechnologie des Rhizobiums et Amélioration des Plantes″ (code
Project 61) affiliated to the Algerian Ministry of Higher Education and Scientific Research. We are very grateful two anonymous
referees and editor of this paper for their helpful and constructive comments.
Authors' contributions
YN conceived of the study, design it has and performed the statistical analysis and analysis results. BA and CC carried out the
research as part of their MSc project. F-LFZ have been involved in the revising it critically for intellectual content. All authors have
participated sufficiently in the work and all authors read and approved the final manuscript.
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