5446

5106

كارشناسي ارشد

بيماري شناسي گياهي

اصفهان: دانشگاه صنعتي اصفهان، دانشكده كشاورزي

1389

نه،108ص.: مصور،جدول،نمودار

ص.ع.به فارسي و انگليسي

مسعود بهار، علي آهون منش

جواد كريم زاده اصفهاني

6/7/89

امير مساح، آقا فخر مير لوحي

مهندسي كشاورزي

ID5106

به فارسي و انگليسي: قابل رويت در نسخه ديجيتالي

Molecular Identification and Classification of Phytoplasmas Associated with Forage Crops in Central Region of Iran Somayeh Hoseini Somaieh hoseini@yahoo com Date of submission May 12 2011 Department of Plant Protection Isfahan University of Technology Isfahan 84156 83111 Iran Degree M Sc Language Farsi M Bahar Ph D Assoc Professor Supervisor mbahar@cc iut ac ir A Ahoonmanesh Ph D Professor Supervisor Aliahoon@cc iut ac ir Abstract Phytoplasmas are known as a serious threat for cultivation of important plants and cause 700 diseases in fruit trees forage crops ornamental vegetables and wild plants In central part of Iran phytoplasma diseases are the causative agent of yellowing whitches broom stunting and malformation in many important crops Recently several symptoms of Phytoplasmal diseases have been found associated with forage crops in different areas of Isfahan province Affected plants exhibit chlorosis leaf stunting growth reduction rosette and witches broom symptoms and the damages are significantly increasing in alfalfa growing regions Since different phytoplasmas are prevalent in crop growing area of Isfahan and seems that phytoplasmas have expanded their host range in the region to include legume crops therefore the present investigation was aimed at identification and classifying the phytoplasmas that associated with main forage crops in central part of Iran including Isfahan province During 2008 2009 alfalfa Medicago sativa clover Trifolium sp and sainfoin Onobrychis vicifolia plants showing yellowing little leaf appearance and stunting symptoms were collected from different regions of Isfahan Chahar mahal o bakhtiari and Yazd provinces Total DNA was extracted from midribs and phloem tissues of the samples and subjected for PCR amplification using universal and specific primers for detection of phytoplasmas The PCR reaction was carried out employing the universal phytoplasma primer pair P1 P7 followed by R16F2n R16R2 as nested primer pair Total DNA from healthy alfalfa clover and sainfoin plants were used as negative controls In the most cases due to low concentration of phytoplasma in plant materials no band was observed in first round PCR but in nested PCR using primer pair P1 P7 followed by primer pair R16F2n R16R2 an expected 16S rDNA fragment 1 2kb was amplified from 57 infected plants Detection of Phytoplasmal DNA fragment in alfalfa clover and sainfoin showing phytoplasma symptoms indicated that phytoplasmas are involved to cause disease in reviewed fields No PCR amplification was observed in healthy plants To classify the phytoplasmas detected in the samples collected from different region PCR RFLP analysis were performed using different restriction enzymes For this purpose 1239 bp fragment amplified in nested PCR were digested with CfoI MseI KpnI RsaI TaqI AluI HaeIII Sau96I enzymes Based on comparison of PCR RFLP profiles of the resultant fragments with reference strains the samples were separated into three groups I II III The PCR RFLP patterns of samples in group I were correlated to the profile of Candidatus Phytoplasma solani group 1 while the members of group II were identified as Ca Phytoplasma aurantifolia The samples yielded indistinguishablePDF created with pdfFactory Pro trial version www pdffactory com

مسعود بهار، علي آهون منش

جواد كريم زاده اصفهاني

امير مساح، آقا فخر مير لوحي