Ismaeel alsaleh*(1), Abu-ghorrah Mahmoud(1) and Aroub Almasri(2)
(1). Department of plant protection -faculty of agriculture, Damascus University, Damascus, Syria.
(2). National commission for biotechnology( NCBT), Ministry of higher education and scientific research, Damascus, Syria.
(*Corresponding author: Ismaeel Alsale, E-Mail: ismaeel.alsaleh@damascusuniversity.edu.sy).
Received: 1/10/2023 Accepted: 5/11/2023
Abstract:
This study was conducted in Plant Bacterial Diseases lab at Damascus university – faculty of Agriculture, and Molecular Biology lab at National commission for Biotecnology , We evaluated the efficiency of spraying wheat seedlings with the recombinant HarpinEa protein in increasing the activity of the peroxidase enzyme (POD), we used for that spectrophotometry and polyacrylamide gel electrophoresis (native-PAGE) assays, the results of the measurement enzyme activity by spectrophotometer showed increasing in POD activity at wheat seedlings which treated with Harbin compared to seedlings treated with water (control). the highest activity of the enzyme was at 72 hours post-treatment with Harpin compared to control (it reached 1.828 times concentration of the enzyme in the control), while it decreased slightly at 96 hours after spraying. Detection of activity of POD enzyme at wheat seedlings by polyacrylamide electrophoresis (Native-PAGE) confirmed last result, electrophoresis showed that the density and intensity of the peroxidase bands gradually increased in samples treated with harbin and reached the highest effectiveness (the highest density and intensity of the resulting band) at 72 hours post-treatment with Harpin, then the band density decreased slightly at 72 hours post-treatment with Harpin. Also at this study, we conducted gene expression of GPX (one of the genes responsible for peroxidase enzyme), and gene expression of Taβ-actin gene (housekeeping gene as internal control) on agarose gel. The results showed stability in gene expression by the time for both genes, whether in wheat seedlings treated with water or seedlings treated with Harbin, i.e. there was’nt any effect of treating wheat with harbin protein on the GPX gene expression, which indicates that the effectiveness of Harbin protein in stimulating activity of peroxidase enzyme didn’t match with gene expression of GPX, maybe there are other genes play a role in increasing its activity. This study showed the possibility of of treating wheat with Harbin protein which produced locally to increase its immunity and resistance to various pests.
Key words: Wheat, harbin protein, peroxidase enzyme.
Full Paper in Arabic: pdf