با همکاری انجمن‏‌ بیماری شناسی گیاهی ایران

نوع مقاله : بیماری‌شناسی گیاهی

نویسندگان

1 دانشیار، بخش تحقیقات گیاه‌پزشکی، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان زنجان، سازمان تحقیقات، آموزش و ترویج کشاورزی،

2 محقق، بخش تحقیقات گیاه‌پزشکی، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان زنجان، سازمان تحقیقات، آموزش و ترویج کشاورزی، زنجان،

3 بخش تحقیقات خاک و آب، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان زنجان، سازمان تحقیقات، آموزش و ترویج کشاورزی، زنجان، ایران

4 پژوهشکده میوه‌های معتدله و سردسیری، موسسه تحقیقات علوم باغبانی، سازمان تحقیقات، آموزش و ترویج کشاورزی، کرج، ایران.

5 محقق، بخش تحقیقات گیاه‌پزشکی، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان زنجان، سازمان تحقیقات، آموزش و ترویج کشاورزی، زنجان، ایران.

چکیده

بیماری پژمردگی ورتیسیلیومی در اثر قارچ Verticillium dahliae یکی از مخربترین عوامل بیماری­زای زیتون دردنیا ومناطق مختلف زیتون‌کاری ایران محسوب می‌شود. در این تحقیق عکس‌العمل 15 رقم امید­بخش و 15 ژنوتیپ بومی زیتون نسبت به قارچ عامل بیماری بررسی شد. نهال‌های نه ماهه زیتون برای ارزیابی کمی قارچ عامل بیماری و تعیین میزان حساسیت یا مقاومت نسبت به بیماری با ‌روش فرو بردن ریشه در زاد مایه انجام شد. میزان افزایش کمی DNA قارچ موجود در نهال­ها با استفاده از تکنیک Real time PCR و سیستم SYBR Green همراه با آغازگرهای اختصاصی برای ژن بتا توبولین2 در فواصل زمانی صفر،2، 15، 45 و80 روز پس از مایه زنی، ارزیابی شد. ژنوتیپ­های QG-11، DD2-1، PS-5 و رقم مانزانیلا به‌ترتیب بالاترین حساسیت را نسبت به سایر ارقام و ژنوتیپ­ها در برابر بیماری داشتنند. ژنوتیپ‌های KH-13، TTS-1، کد 5 لرستان و DS-5 به‌ترتیب کم‌ترین غلظت DNA قارچ در DNA کل ریشه را به خود اختصاص دادند. علاوه بر این، در این تحقیق مشخص گردید نتایج روش متعارف ارزیابی مقاومت ارقام زیتون مبتنیبر ثبت علایم ظاهری بیماری با نتایج حاصل از کمیت سنجی قارچ با استفاده از Real time PCR دارای هم‌پوشانی نسبتاً بالایی است به‌طوری که ارقامی که در روش ارزیابی متعارف  جزء ارقام مقاوم یا بسیار حساس طبقه‌بندی شدند، درروش ارزیابی ملکولی نیز در گروه‌های مشابهی قرار گرفتند. با توجه به‌دقت وحساسیت بالای روش ارزیابی ملکولی می‌توان از روش اخیر برای غربالگری منابع مختلف زیتون برای مقاومت به بیماری پژمردگی ورتیسیلیومی استفاده کرد.
 

کلیدواژه‌ها

عنوان مقاله [English]

Screening of promising olive cultivars and local genotypes for resistance to Verticillium dahliae in Iranian collection

نویسندگان [English]

  • Hossein Jafary 1
  • Neda Zand 2
  • Mahdi Taheri 3
  • Aliasghar Zeinanloo 4
  • Shamsollah Najafi 5

1 Associate Professor of Plant Protection Research Department, Zanjan Agricultural and Natural Resources Research and Education Center, AREEO, Zanjan, Iran.

2 Researcher, Plant Protection Research Department, Zanjan Agricultural and Natural Resources Research and Education Center, AREEO, Zanjan, Iran.

3 Associate Professor, Soil and Water Research Department, , Zanjan Agricultural and Natural Resources Research and Education Center, AREEO, Zanjan, Iran.

4 Horticultural Sciences Research Institute, AREEO, Karaj, Iran.

5 Researcher, Plant Protection Research Department, Zanjan Agricultural and Natural Resources Research and Education Center, AREEO, Zanjan, Iran.

چکیده [English]

Verticilliumwilt of olive (VWO) caused by fungus Verticillium dahliae, is one of the most important diseases of olive worldwide. The response of 15 promising olive cultivars and 15 local varieties was evaluated to the fungus. The olive saplings of each genotype were inoculated with suspension of the fungus using root dip method. The degree of susceptibility/resistance of each genotype was evaluated based on both morphological feature and quantification of fungus in the root system of different genotypes. Total DNA was extracted from root of saplings at different sampling intervals: 0, 2, 12, 45 and 80 days post inoculation. The gradual increase for DNA of fungus in saplings was quantified by Real-time PCR technique and utilizing of SYBR Green system with specific primer pairs for ß-tubulin2 gene. Based on both morphological and quantitative Real-time PCR assays: QG-11, DD2-1, treeNo2, PS-5 and Manzanilla were highly susceptible and KH-13, TTS-1, DS-5 code 5 Lorestan and DS-5 were evaluated as resistant genotypes to the VWO. Moreover, we found parallel results for conventional method of screening (based on disease symptoms) and quantification of DNA in root system of olive by Real-time PCR technique. Olive genotypes evaluated as highly susceptible or resistant to the disease in conventional method showed the similar reaction to the fungus in Q-PCR method. This may suggest that conventional method of evaluation for resistance could be replaced by Q-PCR molecular method in high thropout screening of olive germplasm.

کلیدواژه‌ها [English]

  • Olive cultivars
  • Quantitative assessment
  • Real time PCR
  • Resistance
Al-Ahmad, MA. and Mosli, MN. 1993. Verticillium wilt of olive in Syria. EPPO Bulletin 23: 521–529.
Blanco-Lopez, MA. and Jimenez-Dıaz, RM. 1995. Una propuesta de lucha integrada contra la verticilosis del olivo. Fruticultura Profesional. Especial Produccion Integrada 70: 52–58.
Blanco-Lopez, MA. Jimenez-Dıaz, RM. and Caballero, JM. 1984. Symptomatology, incidence and distribution of Verticillium wilt of olive trees in Andalucıa. Phytopathologia Mediterranea, 23: 1–8.
Cirulli, M. and Montemurro, G .1976. A comparation of pathogenic isolates of Verticillium dahliae and sources of resistance in olive. Agriculturae Conspectus Scientificus, 39: 469–476.
Colella, C. Miacola, C. Amenduni, M. DAmico, M. Bubici, G. and Cirulli, M. 2008. Sources of Verticillium wilt resistance in wild olive germplasm from the Mediterranean region. Plant Pathology. 57:533-539.
El, Said. S, Hegazi Ayman, A. Hegazi, and Abdou, M. Abd Allatif, 2012. Resistance of Some Olive Cultivars to Verticillium Wilt. Journal of Applied Sciences Research, 8(5): 2758-2765.
Fradin, E. F. and Thomma, B. P. H. J. 2006. Physiology and molecular aspects of Verticillium wilt diseases caused by of Verticillium dahliae and Verticillium albo-atrum. Molecular Plant Pathology 7(2): 71-86.
Gao, X. Jackson, T. A. Lambert, K. N. Li, S. 2004. Detection and Quantification of Fusarium solani f. sp. glycines in Soybean Roots with Real-Time Quantitative Polymerase Chain Reaction. Plant Disease, 88:  372- 380.
 Gayoso, C. and pomar, F. 2007. Assessment of real-time PCR as a method for determining the presence of Verticillium dahliae in different Solanaceae cultivares. European Journal of Plant pathology 118: 199-209.
Gramaje, D. Pérez-Serrano, V. Montes-Borrego, M. Navas-Cortés, J. A. Jiménez-Díaz, R. M. and Landa, B. B. 2013. A Comparison of Real-Time PCR Protocols for the Quantitative Monitoring of Asymptomatic Olive Infections by Verticillium dahliae Pathotypes. Phytopathology. 103(10):1058-68.
Heale, JB .1988. Verticillium spp., the cause of vascular wilt diseases in many species. Advances in Plant Pathology, 6: 291–312.
Hiemstra, J. and Harris, D. .1998. Compendium of Verticillium Wilt in Tree Species, Ponsen and Looijen, Wageningen, the Netherlands.
Hussain, T. Singh and B. P. Anwar, F. 2013. A Quantative Real-Time PCR of Phytophthora infestans in different Indian potato cultivars. Journal of Agriculture and Veterinary Science, 4:17- 26.
Jafary, H. Khanmohammadi, S. and Mehri, Nastaran, 2014. Detection of pathotypes of Verticillium dahliae, the causal agent of olive Verticillium wilt in olive orchards of Tarom using Nested-PCR technique. Applied Researches in Plant Protection. 2(2): 47-58.
Jimenez-Diaz, R. M., Bubici, G. Jimenez-Gasco, M. M. Antoniou, P. P. and Tjamos, E. C., 2012. Verticillium wilt, a major threat to olive production: Current status and future prospects for its management. Plant Dis. 96:304-329.
Kim, K. Y. Xiao, C. L. and Rogers, D. J. 2005. Influence of culture media and environmental factors on mycelial growth and pycnidial production of Sphaeropsis pyriputrescens. Mycologia 97: 25-32.
Lopez-Escudero, F. J. and Blanco-Lopez, M.A. 2001. Effect of a single or double soil solarization to control Verticillium wilt in established olive orchards in Spain. Plant Disease, 85: 489–496
Lopez-Escudero, L. F. J. and. Blanco-Lopez, M. A. 2005. Recovery of young olive trees from Verticillium dahliae. European Journal of Plant Pathology, 113: 367–375.
Lopez-Escudero, L. F. J. del Rio, C. Caballero, J. M. and Blanco-Lopez, M. A. 2004. Evaluation of olive cultivars for resistance to Verticillium dahliae. European Journal of Plant Pathology, 110: 79–85.
Markakis, E. A. Tjamos, S. E. Antoniou, P. P. Paplomatas, E. J. and Tjamos, E. C. 2009. Symptom development, pathogen isolation and Real-Time QPCR quantification as factors for evaluating the resistance of olive cultivars to Verticillium pathotypes. European Journal of Plant Pathology, 82: 47-51.
Mercado-Blanco, J. Collado-Romero, M. Parrilla-Araujo, S. Rodrigues-Jurado, D. and Jimenes-Diaz, D. 2003. Quantitative monitoring of colonization of olive genotypes by Verticillium dahliae pathotypes with real-time polymerase chain reaction. Physiological and Molecular Plant Pathology, 63: 91-105.
Mercado-Blanco, J. Rodriguez-Jurado, D. Parrilla-Araujo, S. and Jimenez-Diaz, R. M., 2003. Simultaneous detection of the defoliating and nondefoliating Verticillium infected olive plants by duplex, nested polymerase chain reaction. Plant Disease, 87: 1487-1494.
Moretti, C. Quaglia, M. Cerri, M. Nicosia, D. E. and Buonaurio, R. 2015. A real-time PCR assay for detection and quantification of Botrytis cinerea in Pelargonium x hortorum plants, and its use for evaluation of plant resistance. European Journal of Plant pathology, 143: 159.
Muwarfaq, R. K. 2006. Seed transmission of Verticillium dahliae in olive as detected by a highly sensitive Nested-PCR based assay. European Journal of Phytopathology. Mediterr, 45: 15-23.
Novaes, R. M. L. Rodrigues, J. G. and Lovato, M. B. 2009. An efficient protocol for tissue sampling and DNA isolation from the stem bark of Leguminosae trees. Genetics and Molecular Research, 8: 86-96.
Perez-Artes, E. Garcıa-Pedrajas, M. D. Bejarano-Alcazar, J. and Jimenez-Dıaz, R. M. 2000. Differentiation of cotton-defoliating and nondefoliating pathotypes of Verticillium dahliae by RAPD and specific PCR analyses.European Journal of Plant Pathology,106: 507–517.
Rahnama, K. Razavi, S. R. Latifi, N. and Zarei, H. 1998. Investigation the occurrence of decay of branches and olive trees in Golestan province. The 13th Iranian Plant Protection Congress. P222. ( In Persian with English summary).
Rodriguez, E. Garcia-garrido, J. García, M. and Campos, M. 2008. Agricultural factors affecting Verticillium wilt in olive orchards in Spain. European Journal of Plant Pathology 22: 287–295.
Rodrıguez-Jurado, D. 1993. Interacciones huesped-parasito en la marchitez del olivo (Olea europaea L.) inducida por Verticillium dahliae Kleb. Ph.D. Thesis, Universidad de Cordoba, Spain
Sanei, S. J. Okhovat, S. M. Hejarod, GH. A. Saremi, H. and Nikkhah, M. J. 2003.  The role of therapeutic therapy of olive cuttings in reducing contamination with Verticillium wilt in Iran. The 16th Iranian Plant Protection Congress. P432. (In Persian with English summary).
Sanei, S.J. and S.E. Razavi, 2011. Reaction of some olive cultivars to verticillium dahliae isolates agent of vascular wilt: A comparative study. American Journal of  Experimental  Agriculture, 1(4): 320-330.
Schena, L. and Ippolito, A. 2003. Rapid and sensitive detection of Rosellinia necatrix in roots and soils by real time Scorpion- PCR. Journal of Plant Pathology 85: 15–25.
Schena, L.  Nigro, F. Ippolito, A. and Gallitelli, A. 2004. Real-time quantitative PCR: a new technology to detect and study phytopathogenic and antagonistic fungi. European Journal of Plant Pathology 110: 893–908.
Serrhini, M.N. and Zeroual, A. 1995. La Verticilosis del olivo en Marruecos. Olivae, 58: 58–61.
Sinclair, W.A. Lyon, H.H. and Johnson, W.T. 1987. Diseases of Trees and Shrubs, Comstock Publishing Associates, Cornell University Press, Ithaca and London.
Song, P. Cai, C. Q. Skokut, M. Kosegi, B. D. and Petoline, J. F. 2002. Quantitative real- time PCR as a screeng tool for estiminating transgen copy number in maze. PCR Plant Cellule Reproduction 20: 948-954.
Sousa, M. V. Machado, J. C. Simmons and H. E. Munkvold, G. P. 2014. Real-time quantitative PCR assays for the rapid detection and quantification of Fusarium oxysporum f. sp. phaseoli in Phaseolus vulgaris (common bean) seeds. Plant Pathology, 64: 478-488.
Sun Y, Skinner DZ and Liang GH, Hulbert SH. 1994. Phylogenetic analysis of sorghum and related taxa using internal transcribed spacers of nuclear ribosomal DNA. Theoretical and Applied Genetics 89:26–32.
Thanassoulopoulos, C.C. Biris, D.A. and Tjamos, E.C. 1979. Survey of Verticillium wilt of olive trees in Greece. Plant Disease Reporter, 63: 936–940
Tjamos, E.C. and Jimenez-Dıaz, R.M. 1998. Management of disease. In: Hiemstra J and Harris D (eds) Compendium of VerticilliumWilt in Tree Species (pp 55–57) Ponsen&Looijen, Wageningen, the Netherlands
Vargas-Machuca, R. Martin, C. and Galindez, W. 1987. Recovery of Verticillium dahliae from weed plants in farmers fields in Peru. Plant Disease, 71: 756–758
Wilhelm, S .1955. Longevity of the Verticillium wilt fungus in the laboratory and field. Phytopathology, 45: 180–181
Wilhelm, S. and Taylor, J.B. 1965. Control of Verticillium wilt in olive trough natural recovery and resistance. Phytopathology, 55: 310–316