Regenerating salt tolerant saffron ( Crocus sativus ) using tissue culture with increased pharmaceutical ingredients
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J. Crop Sci. Biotech. 2013 (Sep) 16 (3) :209~217 DOI No. 10.1007/s12892-013-0031-8 RESEARCH ARTICLE
Regenerating Salt Tolerant Saffron (Crocus sativus) Using Tissue Culture with Increased Pharmaceutical Ingredients Zeynab Shahabzadeh, Bahram Heidari*, Ali Dadkhodaie Department of Crop Production and Plant Breeding, College of Agriculture, Shiraz University, Shiraz, 7144165186, Iran Received: February 28, 2013 / Revised: May 20, 2013 / Accepted: July 23, 2013 Ⓒ Korean Society of Crop Science and Springer 2013
Abstract Due to its vegetative reproduction, saffron has a narrow genetic base and induced in vitro variations provide opportunities for expanding new cultivars. The objectives of this study were to evaluate sodium azide-induced variations in saffron’s corm culture in order to increase salt tolerance and pharmaceutical ingredients. Corm explants from the well-known ecotypes, Estahban and Kashmar, were subjected to various concentrations of sodium azide (NaN3) (0.09, 0.12, and 0.22 mg L-1) and NaCl (1.5, 2.5, and 4.0 dS equivalent to 0.07, 0.12, and 0.20 g NaCl in 100 mL water) in Murashige and Skoog medium supplemented with 1 mg L-1 2-4-D, 1 mg L-1 BAP, and 30 g L-1 sucrose and in a second pot culture experiment. The active pharmaceutical ingredients (crocin, picrocrocin, and safranal) were measured by high-performance liquid chromatography (HPLC). Variations in sodium azide-treated plants were more broadened for callus fresh weight (0.57 - 7.57 g), embryo weight (1.24 - 10.29 g), and regenerated seedlings (3.0 - 21.25) compared with those (0.12 - 3.77 g, 0.56 - 4.56 g, and 0.25 - 11.50, respectively) that were not treated with sodium azide. Under 0.20% salt, flowering failed in some of plants developed from sodium azide-untreated corms. HPLC analysis indicated wider ranges for crocin (11.92 - 18.03 mg g-1), picrocrocin (8.99 - 14.76 mg g-1), and safranal (2.13 - 7.36 mg g-1) in sodium azide-treated plants compared to the ranges (0.0 - 16.1, 0.0 - 12.5, and 0.0 - 6.66 mg g-1, respectively) in untreated plants. From a breeding perspective, induced variations found in this study would be useful to improve saffron's quality and salt tolerance. Key words: in vitro culture, pharmaceuticals, saffron, salinity, sodium azide
Introduction Saffron is a term used for the branched and red stigmas of Crocus sativus which belongs to the Iridaceae family (Ghaffari et al. 2009). The plants within this family are herbs with rhizomes, corms, or bulbs. The saffron plant, unknown in the wild, likely descended from Crocus cartwrightianus, C. thomasii, and C. pallasii (Grigg 1974; Hayes 2001; Hill 2004). It has been assumed that it originated from Iran, Asia Minor, or Greece and subsequently became widespread in India, China, the Mediterranean Basin, and Eastern Europe (Grilli Caiola et al. 2004; Nair et al. 1991; Negbi 1999; Tammaro 1987). Iran and Spain are known as the main saffron producers although above 90% of the world production belongs to Iranian saffron (Fernandez 2004). Saffron possesses unique characteristics such as
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