رديابي و تعيين خصوصيات مولكولي ويروس زردي نكروتيك باقلا (Faba bean necrotic yellows virus) در استان هاي مركزي و غربي ايران

چكيده انگليسي :

Detection and Determination of Certain Molecular Properties of Faba bean necrotic yellows virus in Central and Western Provinces Mahsa Mansourpour Mahsa mp86@yahoo com Date of submission April 7 2010 Department of Plant Protection Isfahan University of Technology Isfahan 84156 83111 Iran Degree M Sc Language Farsi A Massah Assist Prof Supervisor amassah@cc iut ac ir A Ahoonmanesh Prof Supervisor aliahoon@cc iut ac irAbstract Faba bean necrotic yellows virus FBNYV causes severe yield losses and crop failure in fabaceousplants in western Asia and northern Africa This virus belongs to Nanoviridae with a multipartite genomeconsisting of ten circular ssDNA components FBNYV has been detected by TBIA method in chickpeaand lentil in East Azerbaijan and West Azerbaijan Qazvin Kermanshah and Kordestan provinces Themain purpose of this research was detection and determination of certain molecular properties of FBNYVin central and western provinces of Iran During a survey from 2008 to 2009 legume samples chickpea lentil faba bean and common bean showing yellowing dwarfing and stem distortion were collected fromIsfahan Markazi Chaharmahal O Bakhtiyari Kermanshah Kordestan and Lorestan provinces Thesamples collected from Kermanshah Kordestan and Lorestan provinces were tested by indirect ELISAusing a polyclonal antiserum In indirect ELISA seven out of 53 faba bean samples from Lorestan six outof 56 chickpea samples from Kermanshah three out of 18 chickpea samples from Kordestan and two outof six lentil samples from Kermanshah were tested positive Total DNA was extracted from ELISA positive samples and samples collected from Isfahan Markazi and Chaharmahal O Bakhtiyari provinces Nested PCR was carried out using primers derived from the DNA 1 of Egyptian isolate of FBNYV Novisible band was detected in the first step of Nested PCR reaction with DNA1F2 DNA1R2 primer pair The expected PCR product 387 bp was amplified in second step with DNA1F6 DNA1R4 primer pair in17 out of 48 chickpea samples from Isfahan four out of 22 common bean samples from Markazi andseven out of 12 common bean samples from Chaharmahal O Bakhtiyari Eleven isolates representingdifferent hosts and geographical regions including A5 and A13 faba bean Lorestan D13 chickpea Kermanshah M28 lentil Kermanshah G10 chickpea Kordestan SA4 SN9 and LN4 chickpea Isfahan LG3 and LO4 common bean Markazi and SH5 common bean Chaharmahal O Bakhtiyari were sequenced The blast comparison revealed high similarity between nucleotide sequence of Iranianisolates and DNA1of Egyptian FBNYV isolate available at GenBank Pairwise nucleotide sequencealignment of A5 A13 D13 G10 M28 SA4 SH5 and SN9 isolates with Egyptian and Syrian FBNYVisolates revealed 98 68 98 55 98 55 97 39 97 10 96 23 98 55 97 39 and 74 68 75 65 75 65 74 78 74 49 75 43 75 65 74 78 identity respectively These results show higher similarity between Iranianisolates and Egyptian isolate Phylogenetic analysis confirmed closer relationship between Iranian isolatesand Egyptian isolate No correlation was found between hosts and geographical regions in phylogeneticanalysis Multiple amino acid sequence alignment revealed the presence of GXGKS G GE GKS motifin Iranian Egyptian and Syrian isolates This is the first report of FBNYV in Isfahan Markazi andChaharmahal O Bakhtiyari provinces and natural infection of common bean by FBNYV in Iran Keywords Faba bean necrotic yellows virus Indirect ELISA Nested PCR

منصورپور، مهسا

رديابي و تعيين خصوصيات مولكولي ويروس زردي نكروتيك باقلا (Faba bean necrotic yellows virus) در استان هاي مركزي و غربي ايران

الايزاي غير مستقيم , Nested-PCR