پديد آورنده :
مهاجر، مهرداد
عنوان :
تشخصيص همزمان ويروس هاي مهم سيب زميني با استفاده روش multiplex RT-PCR
مقطع تحصيلي :
كارشناسي ارشد
گرايش تحصيلي :
بيماري شناسي گياهي
محل تحصيل :
اصفهان: دانشگاه صنعتي اصفهان، دانشكده كشاورزي
صفحه شمار :
[سيزده]، 84ص.: مصور، جدول
يادداشت :
ص.ع. به فارسي و انگليسي
توصيفگر ها :
ويروس واي سيب زميني , ويروس اس سيب زميني , ويروس پيچيدگي برگ سيب زميني و ويروس موزاييك يونجه
تاريخ نمايه سازي :
94/2/5
استاد داور :
بهرام شريف نبي، بدرالدين ابراهيم طباطبايي
تاريخ ورود اطلاعات :
1396/09/26
چكيده انگليسي :
85Simultaneous Detection of Potato Viruses by Multiplex RT PCR Mehrdad Mohajer Mehrdad hotmail68@yahoo com November 2 2014 Department of Plant Protection Isfahan University of Technology Isfahan 84156 83111 IranDegree M Sc Language FarsiSupervisor A Massah amassah@cc iut ac irAbstractPotato Solanum tuberosum L is economically and nutritionally the fifth most important crop after wheat rice maize and barley Virus diseases are the most important destructive agents in potato worldwide Potatovirus Y PVY Potato leaf roll virus PLRV Potato virus S PVS and Alfalfa mosaic virus AMV causegreat yield losses in potato Isfahan and Chaharmahal o bakhtiari provinces are the major areas in potatoproduction in Iran and mentioned viruses are widely spread in these areas So far different methods have beendeveloped for detection of potato viruses but polymerase chain reaction PCR as a sensitive rapid and easymethod is increasingly used more than other methods in detection of potato viruses The unique feature of thismethod is the simultaneous detection of several viruses multiplex RT PCR which makes reduce detectioncosts and time Regarding importance of virus infection in potato fields in Isfahan and Chaharmahal o bakhtiari provinces and requiring a rapid and sensitive detection method for producing virus free potato seedtubers designing and optimizing of a multiplex RT PCR method for simultaneous detection of important potatoviruses seems to be an essential need of research plant pathology laboratories and plant disease clinics For thispurpose samples showing viral symptoms including mosaic leaf rolling and bright yellow spots calico werecollected from Isfahan Chadegan Daran Damaneh Frieden in Isfahan province and Broujen and Faradonbehin Chaharmahal o bakhtiari province In addition several samples available in greenhouse of IsfahanUniversity of technology were used Total RNA was extracted from leaf samples according to lithium chloridemethod cDNA synthesis and PCR were performed for detection of PVY PVS PLRV AMV using specificprimer pairs PVYCPcEcoRII PVYCPvBamHI PLRV fkhR PLRV fkhF PVS R PVS F AMV R AMV F respectively In addition primer pair 18s rRNA R 18s rRNA F was used as internal control gene The PCR wasdone using primer pairs individually binary combination of each primer pairs and primer pairs of all fourviruses Duplex and multiplex RT PCR were optimized with changes in primer concentration magnesiumchloride concentration and primers annealing temperature Simultaneous detection of PVY PVS PLRV andAMV was performed as duplex RT PCR with binary combination of each primer pairs and then multiplex RT PCR with all four primer pairs In PCR duplex and multiplex RT PCR primer pairs PVYCPcEcoRII PVYCPvBamHI PVS R PVS F AM R AMV f PLRV fkhR PLRV fkhR and 18s rRNA R 18s rRNA Famplified 801bp 684bp 415bp 336bp and 255bp segments respectively It seems that the optimized multiplexRT PCR in this study as a sensitive accurate and rapid method could be used in simultaneous detection ofimportant potato viruses Keywords multiplex RT PCR Potato virus Y Potato leaf roll virus Potato virus S and Alfalfa mosaic virus 18s rRNA gene
استاد داور :
بهرام شريف نبي، بدرالدين ابراهيم طباطبايي
