3565

3368

كارشناسي ارشد

بيوتكنولوژي

اصفهان: دانشگاه صنعتي اصفهان، دانشكده كشاورزي

1385

شانزده، 145، [II]ص.: مصور، جدول، نمودار

ص. ع. به فارسي و انگليسي

مسعود بهار

علي تاج آبادي پور

18/4/86

آقا فخر مير لوحي، بهرام باني نسب

مهندسي كشاورزي

ID3368

به فارسي و انگليسي: قابل رويت در نسخه ديجيتال

Abstract In order to understand the evolutionary trend of the genus Pistacia and geneticvariation among Iranian pistachio genotypes two DNA libraries enriched fordinucleotide AG n and trinucleotide ATG n microsatellite motifs were developedfrom Pistacia khinjuk genome Total genomic DNA was extracted from Khinjukleaves and digested with a mixture of four restriction enzymes NheI RsaI AluI andHaeIII Overhanging ends of the obtained DNA fragments were removed using mungbean nuclease dephosphorylated and ligated with phosphorylated SNX linkers Thelinker ligated fragments were selected through selective hybridization to biotinylatedoligonucleotides AG 15 ATG 10 and capturing with streptavidin coated magneticbeads The selected fragments containing AG and ATG repeats were inserted inpBluescript vector transferred into Escherichia coli MC1061 strain and thenenriched libraries were screened by colony PCR technique The plasmids of 44positive clones were extracted and sequenced For two clones the sequences were notclear enough however the repeat sequences were obvious in 35 clones 83 3 among the 42 clones showed high quality sequences The comparison of enrichmentand sequencing results showed that AG microsatellite motifs are more frequentcomparing to ATG motifs and they have more repeated number For clones containedthe repeats 27 pair of primers were designed from flanking regions of the repeatsusing Primer3 software After optimization of their PCR amplifications the primersets were used to assay the polymorphism of Pistachio species and Iranian genotypesof P vera The amplification patterns of the fragments on polyacrylamid gel wereanalyzed using PowerMarker V3 25 and MEGA3 softwares and the dendrogram werethen represented These microsatellites were found to be polymorphic in differentrelated species and the dendrogram showed that all wild species of pistachio werederived from P vera The dendogram also revealed an interesting evolutionary trendfrom P vera to P khinjuk followed by P integerrima P palaestina and then toP atlantica The genotypes of domestic pistachio were classified into three differentgroups including group I sabz peste noogh ravar2 moosa abadi group II momtaztajabadi momtaz gholam rezai hasani hasan zade nish kalaghi kale ghoochi ahmad aghai group III badami zarand ohadi fandoghi riz akbari khanjaridamghan badami ravar seifadini and three independent genotypes of Sarakhs Ghazvini zoodras and Italiai zoodras According to the dendroram it is postulated thatthe domesticated genotypes of P vera are evolved from wild Sarakhs and then thiswild genotypes likely develops to Ghazvini zoodras Italiai zoodras and otherdomesticated pistachios Hence it seems that the wild Sarakhs pistachio plays animportant role in evolutionary trend of the genus Pistacia The results in this studyindicated that microsatellites developed in Pistacia khinjuk are distributed in thegenome of other pistachio species including P vera genotypes and they are highlyuseful in characterization of pistachio species and genotypes

مسعود بهار

علي تاج آبادي پور

آقا فخر مير لوحي، بهرام باني نسب