Author/Authors :
Sharafi Ali نويسنده National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran , Danafar Hossein نويسنده Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran. , Azadi Pejman نويسنده Agricultural Biotechnology Research Institute of Iran (ABRII), Mahdasht Road, Karaj, Iran , Jaberi Mehdi نويسنده Novin Giti Gene Biotech Company, Biotechnology Incubator Center of National Institute of Genetic Engineering and Biotechnology, Tehran, Iran , Sharafi Ata Allah نويسنده Novin Giti Gene Biotech Company, Biotechnology Incubator Center of National Institute of Genetic Engineering and Biotechnology, Tehran, Iran , Kheiri-Manjili Hamidreza نويسنده Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran , Ahmadnia Alireza نويسنده Zanjan Pharmaceutical Biotechnology Research Center, Zanjan University of Medical Sciences, Zanjan, Iran
Abstract :
[Background]Cosmos bipinnatus is an important medicinal plant with antioxidative, antigenotoxic, anti-inflammatory, and anti-proliferative effects on several cancer cell lines. It has great potential for development as a promising cancer chemo-preventive agent. Hairy root-based culture technology is a new sustainable production platform for producing specific pharmaceutical secondary metabolites.[Objectives]The current study developed and introduced a reliable transformation system for C. bipinnatus by optimization of aspects important in transformation frequency using Agrobacterium rhizogenes.[Methods]Five bacterial strains, including ATCC 15834, ATCC 31798, A7, MAFF-02-10266, and MSU440, 2 explant types (leaf and stem), and 2 co-cultivation media (full MS and ½ MS) were examined. Genomic DNA was extracted using a modified CTAB protocol from putative transgenic root lines and the control root. Transgenic hairy root lines were approved by means of Polymerase Chain Reaction (PCR) using specific rolB gene primers.[Results]The highest ratio of genetically transformed root induction was found from leaf explants using A. rhisogenes strains ATCC15834 and MSU440 (72% to 73%). When ½ MS medium was used as a co-cultivation medium, a significant increase in transformation frequency (84%) was observed.[Conclusions]The MSU440 Agrobacterium strain and ½ MS co-cultivation medium could significantly improve genetic transformation efficiency for establishment of hairy root-based cultures for C. bipinnatus.
