Induction of transgenic hairy roots in Trigonella foenum-graceum co-cultivated with Agrobacterium rhizogenes harboring a
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J. Crop Sci. Biotech. 2013 (December) 16 (4) : 263 ~ 268 DOI No. 10.1007/s12892-013-0082-x RESEARCH ARTICLE
Induction of Transgenic Hairy Roots in Trigonella foenumgraceum Co-cultivated with Agrobacterium rhizogenes Harboring a GFP Gene Zeynab Shahabzadeh1, Bahram Heidari1,*, Reza Faramarzi Hafez2 Department of Crop Production and Plant Breeding, College of Agriculture, 7144165186, Shiraz University, Shiraz, Iran Department of Biotechnology, College of Agriculture, 5715944931, Urmia University, Urmia, Iran
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Received: June 11, 2013 / Revised: September 12, 2013 / Accepted: November 25, 2013 Ⓒ Korean Society of Crop Science and Springer 2013
Abstract An important feature of A. rhizogenes-induced hairy roots is their unique ability for investigation of gene function and production of secondary metabolites such as diosgenin in fenugreek. In order to evaluate the transformation frequency and the efficiency of transgenic hairy root induction, leaf and stem explants from two fenugreek ecotypes, Karaj and Bushehr, were infected with three concentrations of OD600= 0.8, 1.2, and 1.6 of A. rhizogenes strain K599 harboring a GFP gene. Regardless of ecotype, the ability of stem explants for the induction of hairy roots (8.09) and the transformation frequency (81.3%) was higher compared with leaf explants with the values of 5.97 and 71.88%, respectively. The number of transgenic GFP-positive hairy roots ranged from 4.2 to 13.5 in the Karaj ecotype and 3.8 to 9.9 in Bushehr. The highest transgenic hairy root (8.76), the transformation frequency (79.76%), and the growth rate of transgenic roots (0.77 d-1) were obtained from infection with K599 at OD600= 1.2, while the lowest belonged to the bacterial concentration of OD600=1.6. Although the ecotype Bushehr had lower total roots (7.53) and transgenic hairy roots (6.08), it showed higher transformation frequency (79.56%) than Karaj (73.63%). Therefore, the results indicate the importance of genotype, type of explant and bacterial concentration in breeding for induction of transgenic hairy roots and consequently, production of secondary metabolites in fenugreek. Key words: fenugreek, GFP, hairy root, transformation
Introduction Fenugreek (Trigonella foenum-graecum L.) is an annual plant belonged to the family Papilionaceae. It is extensively cultivated in India and northern Africa. Its seeds and leaves have been used not only as food but also as traditional medicines (Petropoulos 2002). Fenugreek is known to have antidiabetic (Anitha Devi et al. 2003; Vijayakumar and Bhat 2008), antifungal (Abbasoglu and Turkoz 1995), antibacterial (Bhatti et al. 1996; Sauvaire et al. 1996), immune-modulator (Ghanadi 2005), anti-inflammatory, and antipyretic effects (Max 1992). The two biologically active alkaloid metabolites, galactomannan and diosgenin, can be found in the seed of fenuBahram Heidari (
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The Korean Society of Crop Science
greek plants. Diosgenin is a naturally occurring steroidal saponin produced by
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