شناسايي و تعيين برخي خصوصيات مولكولي ويروييدهاي مو در استان هاي اصفهان، مركزي، چهارمحال بختياري و يزد

چكيده انگليسي :

Identification and Partial Molecular Characterization of Grapevine Viroids in Isfahan Markazi Chahar mahal va Bakhtyari and Yazd Provinces Abdolrahman Khezerpour kamrankhezerpour@yahoo com Date of submission September 18 2013 Department of Plant Protection Isfahan University of Technology Isfahan 84156 83111 Iran Degree M Sc Language Farsi Supervisor A Massah Assist Prof amassah@cc iut ac ir Abstract Viroids are the smallest plant pathogens consisting a single stranded RNA molecule and cause destructive diseases in plants So far five viroids species including Grapevine yellow speckle viroid 1 GYSVd 1 Grapevine yellow speckle viroid 2 GYSVd 2 Hop stunt viroid HSVd Australian grapevine viroid AGVd and Citrus exocortis viroid CEVd have been identified in grapevine There is no available information on distribution and molecular properties of grapevine viroids in central provinces of Iran In 2012 grapevine leaves showing vein banding and yellow speckle symptoms and symptomless samples were collected from Isfahan Chaharmahal va Bakhtiari Yazd and Markazi provinces Total RNA was extracted from grapevine leaves and subjected to RT PCR using the following specific primer pairs including GYSVd1 mf GYSVd1 mr GYSVd1 H GYSVd1 c4 GYSVd1 H GYSVd1 C GYSVd1c4 GYSVd2 h1 GYSVd2 c1 HSVd h2 HSVd c3 HSVd 87P HSVd 83M AGVd h2 AGVd c2 and AGVd h1 AGVd c1 The expected PCR products confirm infection of samples to GYSVd1 GYSVd2 HSVd and AGVd in RT PCR Forty 33 46 and 120 out of 120 collected samples were infected to GYSVd2 AGVd GYSVd2 and HSVd but none of them amplified expected PCR products of CEVd For simultaneous detection of grapevine viroids multiplex RT PCR was carried out using the primer pairs mix containing GYSVd1 mf GYSVd1 mr GYSVd2 h1 GYSVd2 c1 HSVd 87P HSVd 83M and AGVd h2 AGVd c2 The expected size of PCR products 130bp for AGVd 250 bp for GYSVd2 300 bp for HSVd and 370 bp for GYSVd1 were amplified in multiplex RT PCR The PCR products or cloned of full length sequences of 23 isolates of different viroids representing different geographical regions were sequenced Multiple alignments revealed high homology more than 90 between Iranian viroid isolates and other viroid isolates available at Genbank The Meyme and Khatam isolates of AGVd showed the highest 99 7 and the lowest 97 3 homology with the Maragheh isolate JQ686701 and Iranian isolate FJ940923 respectively The highest 100 and lowest 92 3 homology were found between Saman Mahallat and Shahreza isolates of HSVd and china isolate DQ371459 and between Khatam isolate and Iranian isolate JX430795 The Meyme Mahallat and Shahreza isolates of GYSVd1 showed the highest homology 97 8 with Italian isolate AF462167 and the lowest homology 95 1 was found between Gahro isolate of GYSVd1and China GU170805 Iran JN008866 and Italian AF462167 isolates The Gahro isolate of GYSVd2 showed the highest homology 99 4 with china DQ377131 and Meymeh and Tiran isolates of GYSVd2 showed the lowest homology 98 3 with Iranian isolate JN008867 Phylogenetic analyses based on Neighbor Joining method revealed no separation based on geographical regions These results showed high incidence of grapevine viroids in central provinces of Iran and high nucleotide homology between isolates of central provinces of Iran and other isolates available at GenBank Key Words Grapevine yellow speckle viroid 1 Grapevine yellow speckle viroid 2 Hop stunt viroid Australian grapevine viroid Molecular properties

خضرپور، عبدالرحمان

شناسايي و تعيين برخي خصوصيات مولكولي ويروييدهاي مو در استان هاي اصفهان، مركزي، چهارمحال بختياري و يزد

ويروييد لكه زرد 1مو , ويروييد لكه زرد2مو , ويروييد كوتولگي رازك , ويروييد استراليايي مو , خصوصيات مولكولي