10511

9694

كارشناسي ارشد

بيوتكنولوژي كشاورزي

اصفهان: دانشگاه صنعتي اصفهان، دانشكده كشاورزي

1394

پانزده، 80ص.: مصور، جدول، نمودار

مجيد طالبي

بدرالدين ابراهيم سيد طباطبايي، مريم تاتاري

1394/07/19

بهرام شريف نبي، سعيد انصاري

1396/10/04

كتابنامه

كشاورزي

مهندسي كشاورزي

ID9694

17Introduction of Reference Genes for qPCR Data Normalization inDifferent Tissues of Punica granatum Fatemeh Fazli F Fazli@ag iut ac ir 19 August 2015 Department of Agricultural Biotechnology Isfahan University of Technology Isfahan 84156 83111 IranDegree M Sc Language PersianSupervisor M Talebi mtalebi@cc iut ac irAbstractGene expression analysis is one of the important stages of modern molecular biology and increase a betterunderstanding of complex biological processes Real time PCR is the most precise method to evaluate thepattern of gene expression Use of reference genes is the best method to normalize the data obtained fromthe mRNA molecule Beta carotene pathway from the geranyl geranyl pp which includes four differentenzymes has not been identified in pomegranate On the other hand a proper reference gene as anormalizer in real time PCR has not been introduced for this plant For this purpose the RNA wasextracted using a CTAB based method from the different tissues of the pomegranate leaf flower stem seed and peel and cDNA was synthetized The stability of eight reference genes including rps9 18s actin ubq pal cyp cpr and ef1 was evaluated in the qPCR reaction and finally the beta carotene pathwaygenes were partially sequenced and their expression was analyzed using the introduced reference gene The results of the electrophoresis and the spectrophotometry demonstrated that the RNAs from thedifferent tissues were extracted with high quality and quantity Analysis of the eight reference genes inqPCR by the Norm Finder software revealed that the least 0 0246 and the most 0 2472 standarddeviation SD is related to the cyp and 18s respectively In leaf and seed ubq flower cpr stem cyp andpeel rps9 showed more expression stability Mean comparison showed that the expression of lyc psy andzdc genes was significant in the seed tissue zdc psy lyc In the peel tissue it showed that theexpression of psy and lyc zdc and lyc was not significant while the expression of psy and zdc wassignificant psy lyc zdc The mean comparison also showed that the expression of three genes in theShirin Poost Siyah Yazd genotype was not significant while their expression in Qomi Poost Qermez andTorsh Poost Sefid Ryjab were significant lyc psy zdc Cyp was introduced as a stable internalreference gene in pomegranate Lycopene pigment involved in the production of color therefore peelscolor is probably associated with lyc gene expression The expression of lyc at the end of the pathway ismore in the peel tissue therefore it can be concluded that the beta carotene synthesis in the peel tissue ismore than seed Keywords pomegranate RNA extraction qPCR Carotenoid gene expression

مجيد طالبي

بدرالدين ابراهيم سيد طباطبايي، مريم تاتاري

بهرام شريف نبي، سعيد انصاري