10369

9568

كارشناسي ارشد

بيماري شناسي گياهي

اصفهان: دانشگاه صنعتي اصفهان، دانشكده كشاورزي

1394

چهارده، 65ص.: مصور، جدول، نمودار

امير مساح

بهرام شريف نبي

1394/05/27

آذر شاه پيري، نفيسه پورجواد

1396/10/04

كتابنامه

كشاورزي

مهندسي كشاورزي

ID9568

به فارسي و انگليسي

66 Simultaneous Detection of Important Apple Viruses by Multiplex RT PCR Hamzeh Abkhesht Abkhesht@gmail com Date of submission Department of Plant Protection Isfahan University of Technology Isfahan 84156 83111 Iran Degree M Sc Language FarsiA Massah Ph D Assist professor Supervisor amassah@cc iut ac irAbstractApple Malus domestica Borkh in the Rosaceae family is a fruit tree compatible with cool climate Apple stemgrooving virus ASGV Apple stem pitting virus ASPV and Apple chlorotic leaf spot virus ACLSV typemembers of Capilovirus Foveavirus and Trichovirus respectively in the family Betafexiviridae are the majorcausal agents of viral diseases of apple trees These viruses are usually symptomless on trees and cause over 60percent reduction of apple fruit production particularly in mixed infections Development of rapid accurateand sensitive detection methods is essential to make correct management decisions for control of viral diseases The aim of this study was to optimize a multiplex RT PCR for simultaneous detection of ASGV ASPV andACLSV in apple trees For this purpose sampling was done in the different seasons of 2013 and 2014 fromIsfahan Lorestan Chaharmahal va Bakhtiari and Markazi provinces Total RNA was extracted from leaves andblossoms of apple trees and subjected to RT PCR method using specific primer pairs including ASGV F ASGV R for ASGV 713 bp ASPA ASPC for ASPV 264 bp and A52 A53 for ACLSV 358bp Theexpected size bands were obtained in RT PCR assays The amplicons were sequenced and BLAST searchconfirmed the infection of the samples with ASGV ASPV and ACLSV Simultaneous detection of ASGV ASPV and ACLSV was performed using duplex RT PCR assay with binary combinations of specific primerpairs and then using multiplex RT PCR with three primer pairs ASGV F ASGV R ASPA ASPC andA52 A53 The results indicated the multiplex RT PCR method is an efficient method for simultaneousdetection of ASGV ASPV and ACLSV One hundred and ten collected samples were tested with multiplexRT PCR method The results showed 70 53 and 35 samples were infected with ASGV ASPV and ACLSV respectively In addition the mixed infection of viruses was identified 37 24 19 and 16 forASGV ASPV ACLSV ASGV ACLSV ASPV and ACLSV ASGV ASPV respectively To ourknowledge this is the first use of multiplex RT PCR method for simultaneous detection of viral infections ofapple trees in Iran According to the results multiplex RT PCR is a sensitive accurate affordable and reliablemethod for detection of apple tree viruses Keywords Apple stem grooving virus Apple stem pitting virus Apple chlorotic leaf spot virus MultiplexRT PCR Mixed Infection

امير مساح

بهرام شريف نبي

آذر شاه پيري، نفيسه پورجواد