Effects of different tissue culture conditions in Hairy roots induction in Hypericumperforatum L.
Journal Title: International Research Journal of Applied and Basic Sciences - Year 2014, Vol 8, Issue 5
Abstract
Hypericum perforatum L. has been used as a traditional medicinal plant. It contains a lot of constituents with documented biological activity including phenolics, abroad range of flavonoids, naphthodianthrones and phloroglucinols. An efficient and reliable transformation system for a very important medicinal plant Hypericum perforatum L. was developed through optimization of several factors that affect the rate of effective A.rhizogenes- mediated transformation and growth rate of hairy root. Three bacterial strains, A4, ATCC15834 and 11325and three explants types, leaves, shoots and seedlings were examined. The highest frequency of transformation was achieved using ATCC15834 strain in the excised seedling. Several inoculation and co-cultivation media and different concentration of sucrose (0, 20, 30, and 40 gr/l), activated carbon (0, 1, 2, and 3 gr/l). ATCC15834 strain and the excised seedling as explant produced the highest number of hairy roots. In the study maximum number and percent of hairy roots were obtained by treatment of 45 min inoculation and 72 h co-cultivation,also we observed hairy roots proliferationincreased around the explants in media 1/2MS and B5 during co-cultivation compared to the MS full strength and 1/4MS medium. Maximum induction in 1/2MS medium was obtained with 1 gr/lactivated charcoal.The maximum induction and production of hairy root was registered at 20 gr/l sucrose. Transgenic hairy root lines were confirmed by polymerase chain reaction (PCR).
Authors and Affiliations
VahidehBivadi *| M.Sc.student of plant biotechnology ,Mohaghegh e Ardabili University, Ardebil, Iran. email: vbivadi@yahoo.com, RasoolAsghariZakaria| Associate professor of Department of Agronomy and Plant Breeding, Faculty of Agriculture, University of Mohaghegh e Ardabili, Ardabil, Iran., Nasser Zare| Assistant professor of Department of Agronomy and Plant Breeding, Faculty of Agriculture, University of Mohaghegh e Ardabili, Ardabil, Iran., BeniaminYazdani| Ph.D. student of molecular genetics and genetic engineering, Mohaghegh e Ardabili University, Ardabil, Iran.
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