IMPACT OF DIFFERENT PLOIDY LEVELS CHANGE ON DIANTHALEXIN CONTENT IN DIANTHUS CARYOPHYLLUSL. BUSHRA M.J. ALWASHA, SATTAR A. SHLAHIB AND SUMAYA F. HAMADC
The research was conducted to study the effect of elicitor (Fusarium oxysporum) and precursor (anthranilic acid) on accumulation of Dianthalexin in callus culture initiated from leaf of Dianthus caryophyllus. Callus was induced in Murashige and Skoog (MS) media in the presence of 1.0 mg/L Dichlorophenoxy acetic acid (2,4-D) and 0.5 mg/L Benzyl Adenine (BA), supplemented with F. oxysporum (0.0, 2.0,4.0 or 6.0) mL/L and anthranilic acid (0.0, 1.0, 5.0 or 10.0) mg/L. Quantitative and qualitative assessment of Dianthalexin was performed by using high performance liquid chromatography (HPLC) after three and six days culture. Cytological study was performed on callus treated with F. oxysporum and anthranilic acid by using Paradichlorobenzen (PDB) for chromosomes pretreatment and dimeric cyanine nucleic acid stain (YOYO-1) for staining. Anthranilic acid at 10.0 mg/L was the most effective for stimulated Dianthalexin in callus after three days gave 102.46 μg/mL followed by F. oxysporum at concentration 4.0 mL/L after three days too, gave 73.74 μg/ml comparison with Dianthalexin in leaf of intact plant 6.33 μg/mL and in untreated callus 28.15 μg/mL. Addition of F. oxysporum and anthranilic acid lead to change in chromosome numbers. Dianthalexin accumulation in treated callus coupled with chromosomal variation especially in callus treated with 4.0 mL/L F. oxysporum and 10.0 mg/l anthranilic acid which have tetraploid cells.
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