7383

6889

كارشناسي ارشد

بيوتكنولوژي

اصفهان: دانشگاه صنعتي اصفهان، دانشكده كشاورزي

1391

سيزده، 74ص.: مصور، جدول، نمودار

ص.ع. به فارسي و انگليسي

مجيد طالبي، مسعود بهار

حسين زينلي

21/9/91

بدرالدين ابراهيم طباطبائي، آقا فخر ميرلوحي

1396/09/21

كتابنامه

كشاورزي

مهندسي كشاورزي

ID6889

به فارسي و انگليسي: قابل رويت در نسخه ديجيتالي

Evaluation of Genetic Variation and Detection of Impurities in Saffron Using PCR based Methods Saeid Babaei gharah ghani sdbabaei@gmail com s babaei@ag iut ac ir Date of Submission September 10 2012 Department of Agricultural Biotechnology Isfahan University of Technology Isfahan 84156 83111 Iran Degree M Sc Langaueg Farsi 1 M Talebi Assist professor Supervisor m talebi@cc iut ac ir 2 M Bahar Associate professor Suppervisor m bahar@cc iut ac irAbstractSaffron as the most expensive spice in the world is obtained from dried stigmas of Crocussativus L flowers and used mainly as food coloring and flavoring in food industry and alsothe use of its effective components in medicine In this study genetic diversity among 28saffron genotypes collected from different regions of Iran was evaluated using sequencerelated amplified polymorphism SRAP marker Nineteen SRAP primer combinationsamplified a total of 147 polymorphic fragments with an average of 7 74 fragments per eachprimer combination and average of polymorphic information content PIC of 0 15 Clusteranalysis using NJ Neighbor Joining method was divided the genotypes into four groupsthat most of them clustered in a major group Principal component analysis PCA showedthat cluster analysis is more appropriate for revealing genetic relationship of saffrongenotypes The close relationships among saffron genotypes revealed in this study can bedue to vegetative propagation human selection of superior genotypes and existence ofnarrow genetic base of saffron The results confirmed that the SRAP markers don t haveability for discrimination of genetic diversity among saffron genotypes In order to designspecific primers for amplification of saffron DNA we used RAPD markers Twenty fiverandom primers were tested among them 6 primers produced bands with different size andappropriate amplification Specific bands were cloned for sequencing and designed specificprimers After optimization of PCR conditions for designed specific primers they weretested with mixed of saffron safflower and corn DNAs as misused saffron The results ofthese experiments confirmed the specific amplification of saffron Specific primer was alsodesigned and tested for safflower using ITS sequences and the results revealed specificamplification of safflower under mixed saffron and safflower Since the results of PCR isexpressed as qualitative presence or absence bands for quantification of saffron inmisused samples we used Real Time PCR reaction using designed saffron specific primersbased on DNA fragments of RAPD marker and using sybergreen kit Because of unknownreign of fragment amplified with RAPD primers and high sensitive Real Time PCRreaction to nonspecific amplification in low values and also low nonspecific amplificationin sybergreen method the results in Real Time PCR reaction were not reliable There for designing of specific primers from coding region of saffron genome for specificamplification in Real Time PCR and quantification of saffron in misused samples wererecommended Key words saffron SRAP specific primer RAPD Real Time PCR

مجيد طالبي، مسعود بهار

حسين زينلي

بدرالدين ابراهيم طباطبائي، آقا فخر ميرلوحي