تشخيص چند شكلي ژنتيكي بين برخي از ارقام انار براساس آناليز متوالي ITS ژن هاي ريبوزومي هسته اي و نشانگر SRAP

چكيده انگليسي :

Assessment of Genetic Diversity within Some of Pomegranate Cultivars Based on Sequences Analysis of Internal Transcribed Spacers ITSs of Nuclear Ribosomal DNA and SRAP Marker Mohammad Hossein Soleimani Najafabadi mh soleimani@ag iut ac ir Date of submission 2012 1 31 Department of Agricultural Biotechnology Isfahan University of Technology Isfahan 84156 83111 Iran Degree M Sc Language Farsi 1 B E Sayed Tabatabaei Professor Supervisor Sayedt@cc iut ac ir 2 M Talebi Assist Professor Supervisor mtalebi@cc iut ac irAbstractThe pomegranate Punica granatum L is one of the oldest known fruit species This plant is native to Iran and perhaps somesurrounding areas Pomegranate cultivars distinction is mainly based on morphological characteristics As morphologicalcharacteristic like husk and aril color are affected by agro ecosystems for this ancient and widespread fruit it is likely that thesame genotypes have different names in different regions synonymy and or homonymy A precise method for determinationand discrimination of the genotypes is requested for breeding and commercialization of Iranian pomegranate cultivars which itcan be achieved using molecular markers Nowadays different molecular markers have been used to determine the geneticdiversity among some pomegranate genotypes Notwithstanding high morphological diversity among the genotypes low level ofpolymorphism is indicated in these methods In this research sequence related amplified polymorphism SRAP markers whichtarget ORFs as functional regions of pomegranate genome and nuclear ribosomal DNA sequences encoding the 5 8S RNA andthe flanking internal transcribed spacers ITS1 and ITS2 were used to detect the genetic diversity within the genotypes andphylogenetic relationships of pomegranate SRAPs were used to assess genetic diversity of 63 cultivated wild and ornamentalpomegranate genotypes from five different geographical regions of Iran A total of 250 fragments were amplified using 13 primercombinations PCs among which 133 bands 53 were polymorphic Average PIC value was 0 28 over all PCs The geneticdistance among genotypes ranged from 0 10 to 0 37 with an average of 0 24 Cluster analysis using the Neighbor Joining NJ method suggested that there are close relationships between ornamental and some wild genotypes Although the AMOVA resultshad shown the significant differences in the genetic diversity among the regions P value 0 0048 the genetic variation wasmainly caused by the variation of intra regions The results showed low genetic differentiation Fst 0 025 and high gene flow Nm 2 28 among regions ITS region was also used to evaluate the genetic diversity and phylogenetic relationships of 20pomegranate genotypes These data exhibited the presence of polymorphism among pomegranate genotypes Variation amonggenotypes demonstrated both in the length and sequence content of ITS region The size of the spacer s sequences ranged from246 to 250 bases for ITS2 and 206 bases for ITS1 ITS2 seems to be relatively more informative than ITS1 Cluster analysisusing the maximum parsimony method grouped the genotypes into three clusters Also predicted conformation of the secondarystructure of ITS2 transcripts and thermodynamic stability subjected to detailed inspection in comparative analysis amonggenotypes Five distinct conformations constructed from ITS2 region of pomegranate genotypes This method was able to setapart some genotypes as supplement of cluster analysis method Cluster analysis of ITS2 sequences confirmed previous studieswhich suggested that Punica genus is actually derived members of the Lythraceae family In this family Punica genus standbeside Capuronia Galphinia and Laofensia which is in agreement with previous studies Keywords Pomegranate Genetic diversity SRAP Phylogenetic relationships ITS sequence

سليماني نجف آبادي، محمدحسين

تشخيص چند شكلي ژنتيكي بين برخي از ارقام انار براساس آناليز متوالي ITS ژن هاي ريبوزومي هسته اي و نشانگر SRAP

تنوع ژنتيكي , روابط فيلوژنتيكي