OPTIMIZATION OF AGROBACTERIUM-MEDIATED GENE TRANSFORMATION IN AJOWAN MEDICINAL PLANT (TRACHYSPERMUM AMMI L.) USING FLD GENE

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NIAZIAN M.; SADAT NOORI S.A.; TOHIDFAR M.; GALUSZKA P.; MORTAZAVIAN S.M.M.

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Journal: Journal of Molecular and Cellular Researches Year:2017 | Volume:30 | Issue:3 Page(s): 452-463

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Title

OPTIMIZATION OF AGROBACTERIUM-MEDIATED GENE TRANSFORMATION IN AJOWAN MEDICINAL PLANT (TRACHYSPERMUM AMMI L.) USING FLD GENE

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Keywords

AGROBACTRIUMQ2

AJOWANQ1

FLAVODOXINQ1

HYGROMAYCINQ1

TRANSFORMATION FACTORSQ1

Abstract

To optimize gene transformation condition in AJOWAN medicinal plant effect of individual parameters such as optical density ofAgrobacterium cell culture, Agrobacterumstrain, type of antibiotic to elimination of Agrobacterium, acetosyringone concentration and inoculation duration was investigated using p6U-Ubi-FVT1 plasmid contain HPT gene as selectable marker andfld gene.20 mg/L of HYGROMAYCIN was identified as minimum concentration of this selectable agent to prevent of callus induction in non-transgenic cells of AJOWAN. Interaction effect of optical density of Agrobacterium, Agrobacterium strain and type of Agrobacterium killing antibiotic on number of regenerated plant was significant at 1% probability level and less inhibitory effect on regenerated plant was related to OD600=0.6-0.8´GV3101 Agrobacterium starin´160 mg/L of timentin antibiotic. More regenerated plant resistance to HYGROMAYCIN was achieved by 254.8 mm/L of acetosyringone. More regenerated plant was achieved from 30 minutes inoculation duration than 1 and 20 minutes inoculation duration, nonetheless there was no statistically significant difference between means of regenerated plant from 20 and 30 minutes inoculation duration. The presence of the target gene was evaluated using PCR analysis on regenerated plants. Results of PCR analysis confirm entity of at least one copy of the target gene on plants genome. The superiority of transgenic plants against non-transformed plants for tolerance to -0.3 Mpa osmotic stress was proved through osmotic stress tolerance bioassay.

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APA: Copy

NIAZIAN, M., SADAT NOORI, S.A., TOHIDFAR, M., GALUSZKA, P., & MORTAZAVIAN, S.M.M.. (2017). OPTIMIZATION OF AGROBACTERIUM-MEDIATED GENE TRANSFORMATION IN AJOWAN MEDICINAL PLANT (TRACHYSPERMUM AMMI L.) USING FLD GENE. JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), 30(3 ), 452-463. SID. https://sid.ir/paper/248539/en

Vancouver: Copy

NIAZIAN M., SADAT NOORI S.A., TOHIDFAR M., GALUSZKA P., MORTAZAVIAN S.M.M.. OPTIMIZATION OF AGROBACTERIUM-MEDIATED GENE TRANSFORMATION IN AJOWAN MEDICINAL PLANT (TRACHYSPERMUM AMMI L.) USING FLD GENE. JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY)[Internet]. 2017;30(3 ):452-463. Available from: https://sid.ir/paper/248539/en

IEEE: Copy

M. NIAZIAN, S.A. SADAT NOORI, M. TOHIDFAR, P. GALUSZKA, and S.M.M. MORTAZAVIAN, “OPTIMIZATION OF AGROBACTERIUM-MEDIATED GENE TRANSFORMATION IN AJOWAN MEDICINAL PLANT (TRACHYSPERMUM AMMI L.) USING FLD GENE,” JOURNAL OF MOLECULAR AND CELLULAR RESEARCH (IRANIAN JOURNAL OF BIOLOGY), vol. 30, no. 3 , pp. 452–463, 2017, [Online]. Available: https://sid.ir/paper/248539/en

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