Shahid Bahonar University of Kerman and Iranian Biotechnology Society Agricultural Biotechnology Journal 2228-6705 18 4 2026 12 22 Effects of sodium azide (NaN3) treatment on in vitro growth and genetic variation in the Garnem rootstock Effects of sodium azide (NaN3) treatment on in vitro growth and genetic variation in the Garnem rootstock 615 628 5686 10.22103/jab.2026.27271.1922 FA Sarah Ali Muhammad Al- Hayany Horticulture Department, College of Agriculture, University of Diyala, Iraq 0000-0002-7295-9574 Mohammed D. Abdulhadi Horticulture Department, College of Agriculture, University of Diyala, Iraq 0000-0002-9186-1564 Ayad Assi Obaid Horticulture Department, College of Agriculture, University of Diyala, Iraq 0000-0001-5853-9664 Journal Article 2026 05 23 ObjectiveSodium azide is a mutagenic chemical used to create genetic diversity in plants. The aim of this study was to investigate the effect of different concentrations of sodium azide on the in vitro growth of Garnem rootstock and to identify possible changes in the sequence of the chloroplast rbcL gene after treatment.Materials and methodsThis study was conducted on Garnem rootstock under in vitro conditions from August 2021 to June 2022. Homogenized explants were treated with sodium azide at concentrations of 13 and 26 mg/L for 60 minutes, and untreated samples were considered as controls. Each treatment consisted of 10 independent replicates. After treatment, the explants were cultured in proliferation medium containing 6 mg/L kinetin (Kin) and 1 mg/L 6-benzylaminopurine (BA). Mortality percentage, number of branches, branch length and number of leaves were evaluated. Also, a fragment of the rbcL gene was amplified by PCR and sequenced in the control and treated samples. Sequence similarity was examined using BLAST. ResultsSodium azide affected the survival and growth of branches. The highest mortality of explants (70%) was observed at a concentration of 26 mg/L, while no mortality was recorded in the control treatment. The highest average number of branches and leaves was obtained in the control. The length of branches also differed between the treatments; the highest value was observed at a concentration of 26 and the lowest value at a concentration of 13 mg/L. Sequence comparison showed that in the studied region of the rbcL gene of the treated samples, there were possible changes in some nucleotides. The studied sequences had a high similarity with the reference sequence of Prunus persica rbcL with accession number JN114836.1 in GenBank.ConclusionSodium azide affected the viability and in vitro growth of Garnemo basal explants in a concentration-dependent manner. Also, sequence changes in the studied region of the chloroplast rbcL gene were observed after treatment. However, to confirm the mutational nature, heritability, and extent of these changes, further biological replicates and sequencing of additional genomic regions are necessary. ObjectiveSodium azide is a mutagenic chemical used to create genetic diversity in plants. The aim of this study was to investigate the effect of different concentrations of sodium azide on the in vitro growth of Garnem rootstock and to identify possible changes in the sequence of the chloroplast rbcL gene after treatment.Materials and methodsThis study was conducted on Garnem rootstock under in vitro conditions from August 2021 to June 2022. Homogenized explants were treated with sodium azide at concentrations of 13 and 26 mg/L for 60 minutes, and untreated samples were considered as controls. Each treatment consisted of 10 independent replicates. After treatment, the explants were cultured in proliferation medium containing 6 mg/L kinetin (Kin) and 1 mg/L 6-benzylaminopurine (BA). Mortality percentage, number of branches, branch length and number of leaves were evaluated. Also, a fragment of the rbcL gene was amplified by PCR and sequenced in the control and treated samples. Sequence similarity was examined using BLAST. ResultsSodium azide affected the survival and growth of branches. The highest mortality of explants (70%) was observed at a concentration of 26 mg/L, while no mortality was recorded in the control treatment. The highest average number of branches and leaves was obtained in the control. The length of branches also differed between the treatments; the highest value was observed at a concentration of 26 and the lowest value at a concentration of 13 mg/L. Sequence comparison showed that in the studied region of the rbcL gene of the treated samples, there were possible changes in some nucleotides. The studied sequences had a high similarity with the reference sequence of Prunus persica rbcL with accession number JN114836.1 in GenBank.ConclusionSodium azide affected the viability and in vitro growth of Garnemo basal explants in a concentration-dependent manner. Also, sequence changes in the studied region of the chloroplast rbcL gene were observed after treatment. However, to confirm the mutational nature, heritability, and extent of these changes, further biological replicates and sequencing of additional genomic regions are necessary. https://jab.uk.ac.ir/article_5686_07bb5fdef1ee99d35eaccce14f8b5540.pdf