5444

5104

كارشناسي ارشد

بيو تكنولوژي

اصفهان: دانشگاه صنعتي اصفهان، دانشكده كشاورزي

1389

يازده،134ص.: مصور،جدول،نمودار

ص.ع.به فارسي و انگليسي

مسعود بهار، بدرالدين ابراهيم سيد طباطبائي

ابوالقاسم محمدي

6/7/89

آقا فخر مير لوحي، مجيد طالبي

مهندسي كشاورزي

ID5104

به فارسي و انگليسي: قابل رويت در نسخه ديجيتالي

Identification and Isolation of Microsatellite Markers for Assessing Genetic Diversity among Genotypes of Carthamus tinctorius Niloofar Mokhtari niloomokhtari@ yahoo com Date of submission March 23 2010 Department of Agricultural Biotechnology Isfahan University of Technology Isfahan 84156 83111 Iran Degree M Sc Language Farsi 1 M Bahar Ph D Assoc Professor Supervisor mbahar@cc iut ac ir 2 B E Seyed Tabatabaei Professor Supervisor Sayedt @cc iut ac ir Abstract Safflower Carthamus tinctorius L belongs to Asteraceae family Safflower is known as one of the major oilseed crops in the world with commercial uses Iran have been considered as the center of origins of this plant so the assessment of genetic diversity is a prerequistic for evoloutionary studing in this plant Precise genetic relationships among cultivars using molecular DNA markers is necessary to make better breeding programs The objectives of this study were 1 to isolate and identify microsatellites from the safflower genome and 2 to detect the polymorphisms among safflower genotypes using microsatellites In this study Hamilton et al 1999 procedure was used to develop new safflower SSR primers Genomic DNA extracted from Koose 1 cultivar The extracted DNA was digested with NheI RsaI AluI HaeIII and XmnI Enrichment was done by hybridizing a microsatellite containting fragments with biotin labeled probes such as AG 15 or ATG 10 These probes were captured using magnetic beads coated with streptavidin Biotinylated oligoucleotides had extremely high affinity with streptavidin The purified fragments were ligated to pBluescript plasmid vector and transferred in to E coli MC1061 Colonies containing microsatellite motifs were selected using Colony PCR procedure Thirty five colonies were screened out of which 24 68 5 had SSR motifs Among these colonies twenty were found to be suitable for primer designing After optimization of primers they were used to amplify the genomic DNA extracted from 22 genotypes These genotypes included 13 cultivated safflower and nine wild accessions Twenty one primers were designed in which only 16 primer pairs produced the expected amplification product in safflower cultivars Of 21 SSR loci 15 showed polymorphic patterns The scored bands were analysed using Powermarker V3 25 and Mega 4 softwares The average number of polymorphic alleles and PIC value were 2 85 and 0 3665 respectively Results showed that cultivated and wild genotypes were clustered in two major groups The dendrogram separated Iranian cultivars from

مسعود بهار، بدرالدين ابراهيم سيد طباطبائي

ابوالقاسم محمدي

آقا فخر مير لوحي، مجيد طالبي