رديابي و تعيين برخي خصوصيات مولكولي ويروس برگ قاشقي گيلاس ﴿Cherry leaf roll virus﴾ در استان چهارمحال بختياري

چكيده انگليسي :

Detection and Molecular Characterization of Cherry leaf roll virus in Charmahal va Bakhtiari Provinces Saba Gharaati S gharaati@ag iut ac ir Date of submission May 8 2011 Department of Plant Protection Isfahan University of Technology Isfahan 84156 83111 IranDegree M Sc Language FarsiAmir Massah amassah@ cc iut ac ir AbstractCherry leaf roll virus CLRV belongs to the genus Nepovirus in the familly Comoviridae occurs worldwide Thevirus is widespread in North America Germany Chile New Zealand Australia China Hungary Yugoslavia Spain Portugal Japan and Turkey Cherry leaf roll virus naturally infects a wide range of woody and herbaceousplants that the most of them belong to the genera Betula Fagus Juglans and Prunus The virus infects elm blackberry raspberry walnut blueberry stone fruits and other trees and causes severe damage It has been alsodetected in many herbaceous plants such as Rheum Cherry leaf roll virus causes symptoms such as chlorotic spots leaf curling enation dieback skin cracking and gum in sour cherry and sweet cherry In cherry trees flowering andleaves opening could be delayed The fruits of infected trees show early blight and swell before maturation CLRVhas been previously reported from olive trees in Zanjan province but there are no available molecular data for theIranian isolates of CLRV During the years 2009 and 2010 for detection and determination of certain molecularproperties of CLRV leaf samples showing CLRV symptoms were collected from stone fruits orchards in Charmahalva bakhtiari and Isfahan provinces The large number of the collected samples was positive for CLRV in doubleantibody sandwich enzyme linked immunosorbent assay DAS ELISA using a commercial polyclonal antiserumagainst CLRV The infection of some ELISA positive samples was confirmed by the amplification of a DNAfragment of the expected size 286 bp in nested reverse transcription polymerase chain reaction nested RT PCR and immunocapture nested RT PCR IC nested RT PCR with CLRVF CLRVR primer pair The amplicons in fourisolates including S1 Zayanderoud border almond S2 Saman almond S3 Natanz apricot S15 Cham chang apricot were cloned and sequenced The BLAST search revealed high similarity between the Iranian isolates andother CLRV isolates available in GenBank and the amplified segment is a part of 3 non coding region NCR ofRNA2 Multiple alignment of the 3 NCR of RNA2 nucleotide sequences of the Iranian isolates and those of otherCLRV isolates available in GenBank showed high similarity ranging from 93 5 97 5 between Iranian isolates The highest and lowest similarity were found between Cham chang apricot isolate and a German isolate fromwalnut with accession number AJ877146 98 2 and Saman almond isolate and a German isolate from Sambuscusnigra with accession number AJ877138 86 9 respectively Phylogenetic analysis indicated that CLRV isolateswere divided into six different phyogenetic groups including A B C E D1 and D2 The S2 isolate was placed inthe phylogenetic group D2 and S15 and S1 isolates were placed in phylogenetic group D1with some isolates fromwalnut No correlation was found between host origin and geographical regions in phylogenetic analysis Considering the economic importance and high incidence of CLRV on stone fruits in Charmahal va Bakhtiariprovince more research for identifying CLRV on other hosts and regions of Iran should be conducted Key words Cherry leaf roll virus 3 non coding region Phylogenetic analysis IC nested RT PCR DAS ELISA

قرائتي، صبا

رديابي و تعيين برخي خصوصيات مولكولي ويروس برگ قاشقي گيلاس ﴿Cherry leaf roll virus﴾ در استان چهارمحال بختياري

Das-ELISa , درخت تبارزايي , Ic-nested-RT-PCR