Investigation on the genetic structure of Gnomonia leptostyla populations by PCR-RFLP in Azarbayejan-e-Sharqi, Iran

Subject Areas : Agroecology Journal

Siyamak Salahi ¹ , Mohammad Javan Nikkhah ² , Soleiman Jamshidi ³

1 - M.Sc. of plant pathology, Islamic Azad University, Miyaneh Branch
2 - Scientific Board of Plant Protection Department, Agriculture faculty, Tehran University
3 - Scientific Board of Islamic Azad University, Miyaneh Branch

Received: 2007-01-04 Accepted : 2007-05-16 Published : 2007-03-21

Keywords: Genetic variation, Walnut Anthracnose, Gnomonia leptostyla, Ribosomal DNA, ITS,

Abstract :

    Gnomonia leptostyla (Fr.) Ces. et de Not., the teleomorph of Marssonina juglandis (Lib.) Magn., is the causal agent of the walnut anthracnose, a wide spread disease in almost all the walnut growing areas and causes severe damages particularly in the north of iran such as Kermanshah, Azarbayejan, Khorasan and Karaj regions. Sampling was conducted from several areas of Azarbayejan-e-Sharqi province during 2005-2006. 60 Fungus isolates were isolated from samples and cultured as streaked single spore on OMA. Molecular techniques based on PCR-RFLP applied to investigate the genetic variability of G. leptostyla.Total DNA was isolated from mycelial powder by rapid mini-preparation method. A region of 60 isolates coding for the small-subunit ribosomal RNA (SrDNA-18s) and the internal transcribed spacer (ITS) were amplified and analyzed by restriction enzyme digestion. The results of PCR-RFLP showed no polymorphism either in length or in pattern among all the isolates tested.

References:

1. بهداد، ا. 1369. بیماری های درختان میوه ایران، چاپ دوم. انتشارات نشاط اصفهان، 293 ص.       

2. صارمی، ح. و ر. رزاز هاشمی. 1379. پراکنش و اپیدمی لکه سیاه گردو در غرب کشور. چهاردهمین کنگره گیاه پزشکی ایران. دانشگاه صنعتی اصفهان. صفحه332.      

3. جوان نیکخواه، م. 1380. تحقیق روی تنوع ژنتیکی جمعیت قارچ Magnaporthe grisea (Hebert) عامل بیماری بلاست برنج، با استفاده از خصوصیات مولکولی، بیماریزایی و سازگار رویشی در استان گیلان. رساله دکتری در رشته بیماری شناسی گیاهی. دانشگاه تهران، کرج، ایران. 170 ص.

4. شهریاری، ف. و ع. امام جمعه. 1381. واکنش زنجیره ای پلیمراز PCR ، مقدمه‌ای بر تکنیکهای مولکولی (ترجمه). دانشگاه امام رضا (ع). 228 ص.

5. شیخ الاسلامی، م. 1384. بررسی برخی خصوصیات بیولوژی و تنوع ژنتیکیErysiphe betae عامل بیماری سفیدک پودری چغندرقند. رساله دکتری بیماری شناسی. گروه گیاهپزشکی. دانشکده کشاورزی.  دانشگاه تهران. 134 ص.  

1. Aanen, D.K., T.W. Kuiper and R.F. Hoekstra. 2001. A widely distributed DNA polymorphism within a biological species of the ectomycorrhizal fungus  Hebeloma velutipes. Mycol. Res. 105 (3): 284-290.


2. Belisario, A., M. Hubbes. 1992. Genetic Variability in Gnomonia leptostyla.  ISHS Acta Horticulturae. 385-387.    


3. Black, W.M., and D. Neely. 1974. Effect of select environmental. American PhytopathologicalSociety, 3: 284. USA.      


4. Bruns, T.D., T.J. White, and J.W. Taylor. 1992. Evolutionary relationship within the fungi: analyses of small subunit rRNA sequences. Mol. Phylogen. Evol. 1: 231-243.     


5. Buchko, J., and G.R. Klassen. 1990. Detection of length heterogeneity in the ribosomal DNA of Pythium ultimumby PCR amplification of the intergenic region. Curr. Genet. 18: 203-205.       


6. Gardes, M. and T.D. Bruns. 1993. ITS primers with enhanced specificityfor Basidiomycetes application to the identification of mycorrhizae and rusts. Mol. Ecol. 2: 113-118.


7. Gargas, A., P.T. Depriest. 1996. A nomenclature for fungal PCR primers with example from intron-containing SSU rDNA. Mycologia 88: 745-748.


8. Harrington, T.C., J. Steimel, F. Workneh and X.B. Yang. 2000. Molecular identification of fungi associated with vascular discoloration of soybean in the north central United States. Plant dis. 84: 83-89.


9. Liu, D., S. Coloe, R. Baird, and J. Pedersen. 2000. Rapid Mini-preparation of Fungal DNA for PCR. Journal of Clinical Microbiology 38: 471.     


10. White, T.J., T. Bruns,  and J. Taylor. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In PCR protocols, A guide to methods and amplification. Academic press San Diego, 315-322.            


11. Weising, K., H. Nybom, K. Wolff and W. Meyer. 1995. DNA fingerprinting   in plant and fungi. CRC Press. PP. 322.