title: A reliable and efficient protocol for inducing genetically transformed roots in medicinal plant Nepeta pogonosperma.
abstract: Nepeta pogonosperma is an important medicinal plant with anti-inflammatory effects. An efficient and reliable transformation system for this plant was developed through optimization of several factors which affected the rate of Agrobacterium rhizogenes mediated transformation. Five bacterial strains, A4, ATCC15834, LBA9402, MSU440 and A13, two explant types, leaves and stems, and several co-cultivation media were examined. The maximum rate of hairy root induction was obtained from stem explants using MSU440 and ATCC15834 bacterial strains. A drastic increase in the frequency of transformation (91 %) was observed when MS medium lacking NH4NO3, KH2PO4, KNO3 and CaCl2. Hairy root lines were confirmed by polymerase chain reaction (PCR) using primers of the rolB gene. According to Southern blot analysis, one T-DNA copy was inserted into each of the hairy root lines. In the present study, transgenic hairy roots have been obtained trough genetic transformation by A. rhizogenes harbouring two plasmids, the Ri plasmid and pBI121 binary vector harbouring gus reporter gene. Expression of the gus gene in transgenic hairy root was confirmed by histochemical GUS assay.
authors: National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
authors: Sepideh
authors: S
authors: Valimehr
authors: National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
authors: Forough
authors: F
authors: Sanjarian
authors: National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
authors: Haleh Hashemi
authors: HH
authors: Sohi
authors: Pharmaceutical Biotechnology Department, School of Pharmacy, Zanjan University of Medical Sciences, P.O.Box 451951338, Zanjan, Iran ; Novin Giti Gene Biotech. Co., Biotechnology Incubator Center of NIGEB, P.O.Box:1417863171, Tehran, Iran ; Synthetic Biology Department, Biofuel Research Team (BRTeam), Karaj, Iran.
authors: Ali
authors: A
authors: Sharafi
authors: National Institute of Genetic Engineering and Biotechnology (NIGEB), Tehran, Iran.
authors: Farzaneh
authors: F
authors: Sabouni
keywords: Agrobacterium rhizogenes; Hairy root; Nepeta pogonosperma