كميت‌ سنجي مقدار ناخالصي نمونه‌هاي زعفران با روش Real Time PCR و ارزيابي تنوع ژنتيكي برخي نمونه‌هاي زعفران

چكيده انگليسي :

59Quantification of Impurities in Adulterated Samples Using Real Time PCR and Evaluation of Genetic Variation in Saffron Fatemeh Asgari Dehaghani Fatemehasgari5331@gmail com Department of Agricultural Biotechnology Isfahan University of Technology Isfahan 84156 83111 IranDegree M Sc Language FarsiMajid Talebi mtalebi@cc iut ac irAbstractSaffron Crocus sativus is a crop with high economical and medicinal values This plant isconsidered as the most expensive agricultural and medicine products in the world and it hasspecial position among industrial and exportial products of Iran Iran now is the largestproducer and exporter of saffron in all over the world Because of the importance of saffronand its high price it is supply in market often mixed with other ingredients such as safflower maize stigma Calendula and daylily Separating impurities is done using decompositionreactions such as coloured reactions high performance liquid chromatography HPLC nearinfrared spectroscopy NIR and proton nuclear magnetic resonance NMR which are highlycostly and time consuming DNA molecules have been recently adapted to detect impuritiessuch as safflower yet they are not widely used since the method does not reveal the exactquantity of impurity Real Time PCR is nowadays employed to precisely detect and quantifychemical substances added to the main material or compound To this reason the presentstudy used DNA molecule quantification to estimate purity of fake saffron compared to thegenuine product To reach this goal several chloroplastic genes such as rbcl mat K trn L trnF and ITS region were received from NCBI for several species of saffron and safflower Theprimers designed based on these regions were analyzed and the results showed that the ITSprimers designed could specifically amplify the region in saffron and safflower Afteroptimizing conditions performed Real Time PCR reaction with SYBR Green method Thesuitable dilutions for the standard curve was optimized and used in next experiments withmixed sample of safflower saffron and commercial sample In this study observed that theconcentration of tested commercial sample is similar to concentration of DNA of pure saffron Also the limit of saffron detection was almost equal to 0 0064 Therefore the proposedmethod is recommended as a precise and trustful procedure to quantify impurities in theavailable saffron products In this study genetic diversity among 28 genotypes of saffronwhich were collected from different regions of Iran was also assessed using the SCoTmolecular marker Using 13 SCoT primers 188 polymorphic bands were amplified withaverage of 14 46 bands and the average PIC was equal to 0 228 Drawing phylogenetic treebased on the data from SCoT and UPGMA algorithm revealed that most of the genotypes inthe study were categorized into a same class Samples of wild saffron collected formGolestankooh Khansar and Tarq Natanz were however categorized separately from othergenotypes The average gene diversity and Shannon index of genotypes was 0 226 and 0 363 respectively Entirely concluded that the genetic diversity among genotypes and geneticexchange among populations and genotypes was not so sustainable Key words Absolute Quantitation Real Time PCR Saffron ITS SCoT

عسگري دهاقاني، فاطمه

كميت‌ سنجي مقدار ناخالصي نمونه‌هاي زعفران با روش Real Time PCR و ارزيابي تنوع ژنتيكي برخي نمونه‌هاي زعفران

كميت سنجي مطلق , ITS , SCoT