رديابي و تعيين خصوصيات مولكولي ويروس لكه حلقوي بافت مرده هسته دارها Prunus necrotic ringspot virus در استان هاي اصفهان و چهارمحال بختياري

چكيده انگليسي :

Detection and molecular characterization of Prunus necrotic ring spot virus in Isfahan and Charmahal va Bakhtiari provinces Foroogh Asakereh F asakereh@ag iut ac ir Date of submission March 14 2011 Department of Plant Protection Isfahan University of Technology Isfahan 84156 83111 IranDegree M Sc Language FarsiAmir Massah amassah@cc iut ac ir AbstractStone fruit trees are affected by a large number of viruses which cause significant economic losses Prunus necrotic ringspot virus PNRSV a member of the genus Ilarvirus in the family Bromoviridae occurs worldwide and infects mainly stone fruits The virus is widely distributed in temperate zones onstone fruit trees including sweet cherry sour cherry almond peach apricot plum and many ornamentaland wild varieties of sweet cherry peach and plum and some ornamental species such as rose The virusalso infects apple quince hawthorn and walnut Some isolates of PNRSV are symptomless but most ofthem cause symptoms such as chlorotic and necrotic spot shot hole and leaf deformation PNRSV hasbeen previously reported from Iran but there are no molecular data available for the Iranian isolates ofPNRSV This study was carried out to detect PNRSV in Charmahal va Bakhtiari and Isfahan provincesand determine certain molecular properties of PNRSV Iranian isolates For this purpose leaf samplesshowing the symptoms of PNRSV were collected from stone fruit orchards in Charmahal va Bakhtiariand Isfahan provinces during 2009 and 2010 The collected samples were tested by double antibodysandwich enzyme linked immunosorbent assay DAS ELISA and indirect ELISA using a commercialpolyclonal antiserum against PNRSV or immunocapture reverse transcription polymerase chainreaction IC RT PCR with Ilar1 Ilar2 primers The infection of large numbers of the samples wasconfirmed by the use of ELISA and by the amplification of a DNA fragment of the expected size 206bp The virus coat protein gene CP was amplified by IC RT PCR using PNRSV PNRSV primersin some isolates for sequencing and molecular comparisons Considering the host species and geographicregions the PCR product of the expected size 612 bp in 6 isolates including FA1 Emamieh almond FA2 Lordegan almond FA3 Saman almond FA4 Natanz peach FA5 Cham chang sour cherry and FA6 Farsan sweet cherry were cloned and sequenced The BLAST search revealed high similaritybetween the Iranian isolates and other PNRSV isolates available in GenBank Multiple alignment of theCP amino acid sequences showed high similarity ranging from 96 5 100 between Iranian isolates Cham chang sour cherry isolate showed the lowest similarity 69 5 with Emamieh almond isolate Two isolates of Farsan sweet cherry and Natanz peach showed 100 similarity Multiple alignment ofthe CP amino acid sequence of the Iranian isolates and those of other PNRSV isolates available inGenBank revealed the highest and lowest similarity between Lordegan almond isolate and an Italianpeach isolate 99 5 with accession number AJ133205 and an American sweet cherry isolate 86 7 with accession number AF465235 respectively Phylogenetic analysis indicated that PNRSV isolateswere divided into four different phyogenetic groups including I II III and CH30 The Iranian isolateswere placed in the phylogenetic group II No association was found between host origin and geographicregion in phylogenetic analysis Considering the economic importance of PNRSV and high incidence ofthis virus in Isfahan and Charmahal va Bakhtiari provinces conducting more research on PNRSV and theuse of different techniques for making virus free seedlings seems necessary Key words Prunus necrotic ringspot virus ELISA Coat protein gene Phylogeneticanalysis 79

عساكره، فروغ

رديابي و تعيين خصوصيات مولكولي ويروس لكه حلقوي بافت مرده هسته دارها Prunus necrotic ringspot virus در استان هاي اصفهان و چهارمحال بختياري

توالي ژن پروتئين پوششي , درخت تبارزايي