بررسی تنوع ژنتیکی قارچ عامل بیماری آنتراکنوز گردو Ophiognomonia leptostyla (Fr.) Sogonov در منطقه ITS و IGS با استفاده از نشانگر CAPS در شمال غرب ایران

نوع مقاله: پژوهشی

نویسندگان

1 کارشناس ارشد بیو تکنولوژی دانشگاه آزاد اسلامی واحد میانه

2 عضو هیأت علمی گروه بیوتکنولوژی کشاورزی دانشگاه آزاد اسلامی تهران. واحد علوم تحقیقات.

3 عضو هیأت علمی گروه گیاهپزشکی دانشگاه آزاد اسلامی واحد میانه.

چکیده

به منظور ارزیابی تنوع ژنتیکی قارچ عامل بیماری آنتراکنوز گردو Ophiognomonia leptostyla در شمال­غرب ایران تعداد 25 جدایه از مناطق مختلف جمع­آوری و با استفاده از روش تک اسپورسازی خالص شدند. استخراج DNA از میسلیوم قارچ با استفاده از روش لیو و همکاران انجام شد. تکثیر منطقه ITS-rDNA توسط جفت آغازگر اختصاصی، ITS1 و ITS4 انجام گردید. هم‎چنین منطقه IGS-rDNA با استفاده از جفت آغازگر NS1R و  LR13R تکثیر شد. نوارهایی به طول 600 جفت­باز و 2320-2300 جفت باز از تکثیر منطقه ITS و IGS به­ دست آمد. قطعات تکثیر شده توسط پنج آنزیم برش­دهنده EcoRHindIII، TagHinfI و BamHI مورد هضم قرار گرفتند. از مجموع­ آنزیم­های برشی مورد استفاده EcorTagI و BamHI فاقد محل برش بر محصولات تکثیری منطقه ITS و HinfI و BamHI نیز فاقد محل برش بر محصولات تکثیری منطقه IGS بودند. گروه­بندی جدایه­های مورد مطالعه بر اساس الگوهای نواری هر دو منطقه در سطح شباهت 75 درصد، آن‎ها را در چهار گروه طبقه­بندی نمود.

کلیدواژه‌ها


عنوان مقاله [English]

Genetic diversity in walnut anthracnose casual agent fungi, Ophiognomonia leptostyla (Fr.) Sogonov by ITS and IGS CAPS in northwest of Iran

نویسندگان [English]

  • Mahdi Mianaji 1
  • hamid abdollahi 2
  • solaiman jamshidi 3
1 M.Sc. in Biotechnology, Islamic Azad University, Miyaneh Branch, Miyaneh, Iran.
2 Scientific Board of Islamic Azad University, Science and Research Branch, Tehran, Iran .
3 Scientific Board of Islamic Azad University, Miyaneh Branch, Miyaneh, Iran.
چکیده [English]

Genetic diversity of Ophiognomonia leptostyla, walnut anthracnose casual agent in northwest of Iran was studied. 30 isolates were collected  from different regions and purified as single spore cultures. DNA extraction from fungi mycelium was carried out. The ITS-rDNA region amplified  with ITS1 and ITS4 primers. The IGS-rDNA region was amplified by NS1R and LR13R primers. Bands with 600 and 2300-2320 bp length were obtained in ITS and IGS amplification, respectively. Amplified fragments were digested by EcorI, HindIII, TagI, HinfI and BamHI restriction enzymes. EcorI, TagI and BamHI had no restriction site on amplified ITS region and HinfI and BamHI had no restriction site on IGS region. All isolates were grouped in four clusters based on ITS and IGS CAPS with 75% similarities.

کلیدواژه‌ها [English]

  • Anthracnose
  • Walnut black spot
  • PCR-RFLP
  • Marssoniella juglandis
Ashena S, Zamanizade HR, Mahmodi SB (2008) Pathogenic variability and genetic diversity of Fusarium solani isolates and its association with sugerbeet root rot. Suger Beet Journal 24(1): 77-95. [In Persian with English Abstract].

 Baghaee Ravari S, Falahati Rastegar M, Jafarpour B, Shokoohifar F (2007) The study of probable variation in ITS-DNA region of Fusarium solani in potato and its correlation with pathogenicity and geographical origin in Razavi and Northen Khorasan provenance. Journal of Agricultural Science and Natural Resources, 14(3): 32-41. [In Persian with English Abstract].

Behdad E (1979) Disease of fruits crops in Iran. Neshat publication, Inc, second edition. Esfahan. [In Persian with English Abstract].

Belisario A (2002) Anthracnose. pp. 77-78. In: Teviotdale BL, Michailides TJ and Pscheidt JW (eds.), Compendium of nut crop diseases in temperate zones. USA: APS Press.

 Belisario A, Scotton M, Santori A, Onofri S (2008) Variability in the Italian population of Gnomonia leptostyla, homothallism and resistance of Juglans species to anthracnose. Forest Pathology 38: 129–145.

Burns TD, White TJ, Tylor JW (1991) Fungal molecular systematics. Annual Review of Ecological System 22: 525-564

 Cesati V, De Notaris G. (1863). Schema di classificazione degle sferiacei italici aschigeri piu' o meno appartenenti al genere Sphaeria nell'antico significato attribuitoglide Persono. Commentario della Società Crittogamologica Italiana 1(4): 205. 

Groom MJ, Meffe GK, Carroll CR (2006) Principles of conservation biology (3rd ed). Sunderland, MA: Sinauer Associates. http://www.sinauer.com/groom.

 Konstantinova P, Yli-Mattila T (2004) IGS–RFLP analysis and development of molecular markers for identification of Fusarium poae, Fusarium langsethiae, Fusarium sporotrichioides and Fusarium kyushuense. International Journal of Food Microbiology 95: 321– 331.

 Liu D, Sue C, Baird R, Pedersen J (2000) Rapid mini-preparation of fungal DNA for PCR. Journal of Clinical Microbiology 38 (1): 471-471.

 Luciana B, Lourenço L, Paulo C, Garcia A, Shirlei M, Pimentel R (2003) Restriction fragment analysis of the ribosomal DNA of Paratelmatobius and Scythrophrys species (Anura, Leptodactylidae). Genetics and Molecular Biology 26 (2): 139-143.

 Naghavi MR, Ghariiazi B, Hosseini Salekdeh G (2006) Molecular markers. Tehran University, Tehran. ISBN 964-03-5107-5. [In Persian with English Abstract].

 Salahi, S (2007) Study of genetic diversity in Gnomonia leptostyla casual agent of walnut anthracnose by PCR and evaluation of its distribution and survival in east azarbaijan, Iran. M.Sc. Thesis, University of Tehran, Tehran, Iran. [In Persian with English Abstract].

 Santos SM, Martinez-Bermejo D, Rodri guez-Molina MC, Diez JJ (2007) Cultural characteristics, pathogenicity and genetic diversity of Fusarium oxysporum isolates from tobacco fields in Spain. Physiological and  Molecular Plant Pathology 71: 26 – 32.

 Saremi H, Hashemi SR, Okhovvat SM (2006) Severity of walnut anthracnose and its relatively resistant in Iran. Communication in Agricultural and Applied Biological Science 71: 1267-74. [In Persian with English Abstract].

 Sogonov MV, Castlebury LA, Rossman AY, Mejia LC, White JF (2008) Leaf-inhabiting genera of the Gnomoniaceae, Diaporthales. Mycology 62: 1-77.

 Tabatabaei M, Dehlavi A, Ahmadi AR (1998) Walnut, hicory and pecan. Jihad-e-Daneshgahi press. Tehran. [In Persian with English Abstract].

 Val Verde M, Vandemark G, Martinez O, Paredes- Lope Z (1999) Genetic diversity of Ustilago maydis strains. World Journal of Microbiology and Biotechnology 16: 49-55.

White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. pp. 315-322. In: Innis, M A. Gelfand, D H Sninsky, J J, White T J.(eds), PCR protocols: a guide to methods and applications. MA Innis Academic Press, San Diego. CA. USA.