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Hairy root induction and plant regeneration of medicinal plant Dracocephalum kotschyi
Authors:Ali Sharafi  Haleh Hashemi Sohi  Pejman Azadi  Ata Allah Sharafi
Institution:1. Pharmacognosy and Pharmaceutical Biotechnology Department, Faculty of Pharmacy, Zanjan University of Medical Sciences, P.O.Box 45195-1338, Zanjan, Iran
2. National Institute of Genetic Engineering and Biotechnology (NIGEB), Shahrak-e Pajoohesh, km 15, Tehran - Karaj Highway, Tehran, Iran
3. Tissue Culture and Genetic Engineering Department, Agricultural Biotechnology Research Institute of Iran (ABRII), P. O. Box 31535-1897, Karaj, Iran
4. Novin Giti Gene Biotechnology, Co., Biotechnology Incubator Center of National Institute of Genetic Engineering and Biotechnology (NIGEB), P. O. Box:1417863171, Tehran, Iran
Abstract:An efficient hairy root induction system for an important endangered medicinal plant, Dracocephalum kotschyi, was developed through Agrobacterium rhizogenes-mediated transformation by modifying the co-cultivation medium using five bacterial strains, A4, ATCC15834, LBA9402, MSU440, and A13 (MAFF-02-10266). A drastic increase in transformation frequency was observed when a Murashige and Skoog medium lacking NH4NO3 KH2PO4, KNO3 and CaCl2 was used, resulting in hairy root induction frequencies of 52.3 %, 69.6 %, 48.6 %, 89.0 %, and 80.0 % by A4, A13, LBA9402, MSU440, and ATCC15834 strains, respectively. For shoot induction, hairy roots and unorganized tumors induced by strain ATCC15834 were placed on an MS media supplemented with 0.1, 0.25, 0.5, and 1 mg/l BA plus 0.1 mg/l NAA. The high frequency of shoot regeneration and number of shoot were obtained in the medium containing 0.25 mg/l BA and 0.1 mg/l NAA. Root induction occurred from the base of regenerated shoots on the MS medium supplemented with 0.5 mg/l IBA after 10 days.
Keywords:Agrobacterium strains  Co-cultivation medium  Dracocephalum kotschyi  Hairy root  Regeneration
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