تنوع ژنتیکی جدایه هایAlternaria alternata عامل لکه موجی گوجه فرنگی در استان خوزستان با کمک نشانگرهای ریزماهواره

پژوهش های حفاظت گیاهان ایران

مقاله 6، دوره 30، شماره 4 - شماره پیاپی 34، دی 1395، صفحه 573-586   

نوع مقاله: مقالات پژوهشی

شناسه دیجیتال (DOI): 10.22067/jpp.v30i4.42644

نویسندگان

خشنود نوراللهی^* ؛ محسن حسنی

دانشگاه ایلام

چکیده

بیماری لکه موجی با عامل Alternaria alternata یکی از بیماری‌های مهم گوجه فرنگی در استان خوزستان می‌باشد. به منظور ارزیابی تنوع ژنتیکی جدایههای A. alternata نمونهبرداری از مزارع گوجه فرنگی در مناطق مختلف انجام گرفت. تعداد 64 جدایه جداسازی گردید. پس از جداسازی، خالص‌سازی و شناسایی جدایه‌ها، آزمون مولکولی با استفاده از پنج جفت آغازگر ریزماهواره انجام گردید. در نهایت 21 آلل در همه جمعیت‌ها شناسایی شد. میانگین تعداد آلل در هر لوکوس برابر با 2/4 بود. بیشترین و کمترین تعداد آلل به ترتیب مربوط به لوکوس AEM13 با هشت آلل و لوکوس‌های AEM6 وAEM9 با دو آلل بود. نتایج تجزیه واریانس مولکولی نشان داد که 85 درصد از تنوع ژنتیکی در بین کلیه جدایه‌ها و 14 درصد، به مناطق مختلف جغرافیایی اختصاص دارد. بین جدایه‌ها از مناطق مختلف شباهت ژنتیکی بالایی وجود داشت. شباهت ژنتیکی بالا را می‌توان به مهاجرت ژن یا ژنوتیپ در اثر عوامل مختلف نسبت داد. بیشترین و کمترین تنوع درون جمعیت به ترتیب مربوط به جمعیت شوشتر و جمعیت امیدیه بود. پس از تجزیه دندروگرام داده ها بر اساس UPGMA و ضریب تشابه دایس در سطح 62 درصد، جدایه‌ها در هشت گروه قرار گرفتند. وجود تنوع بالا به علت جهش، نوترکیبی و تولید مثل جنسی می‌باشد. نتایج حاصل از این مطالعه برای اصلاح ارقام مقاوم و گسترش روش‌های کنترل بیماری لکه موجی مفید خواهد بود.

کلیدواژه‌ها

شباهت ژنتیکی؛ لکه موجی؛ واریانس مولکولی؛ SSR

مراجع

1. Aradhya M.K., Chan H.M., and Parfitt D.E. 2001. Genetic variability in the pistachio late blight fungus, Alternaria alternate. Mycological Research, 105: 300–306.

2. Baymani M. 2012. Determination of dominant causal agent of early blight of tomato and loss estimation and its chemical control in Khuzestan province. M.Sc thesis, Shahid Chamran University, Ahvaz, Iran.

3. Benichou S.A., Dongo D.E., Henni P., and Simoneau P. 2009. Isolation and characterization of microsatellite markers from the phytopathogenic fungus Alternaria dauci. Molecular Ecology Resources, 9: 390–392.

4. Bock C.H., Thrall P.H., Brubaker C.L., and Burdon J.J. 2002. Detection of genetic variation in Alternaria brassicicola using AFLP fingerprinting. Mycological Research, 106: 428-434.

5. Burdon J.J., and Silk J. 1997. Sources and patterns of diversity in plant-pathogenic fungi. Phytopathology, 87: 664-669.

6. Chowdhury A.K., Yonemoto Y., Kato H., and Macha M.M. 2005. Cultivar identification by morphometric descriptors and RAPD markers among some Acerola (Malpighia glabra Linn.) cultivars. Japanese Journal Of Tropical Agriculture, 49: 41-42.

7. Dini-Andreote F., Cristina Pietrobon V., Andreote F.D., Romao A.S, BellatoSposito M., and Araujo W.L. 2009. Genetic variability of Brazilian Isolates of Alternaria alternata detected by AFLP and RAPD techniques. Brazilian Journal of Microbiology, 40: 670-677.

8. Doyle J.J., and Doyle J.L. 1990. A rapid total DNA preparation procedure for fresh plant tissue. Focus, 12: 13-15.

9. Eatebarian H. 2006. Vegetable disease and their control. Third edition. Tehran University Press. (In Persian).

10. Esphandiari A. 1947. Field crop and fruit tree disease in north subtropical region of Iran. Issues No. 5, Institute Of Plant Pests And Diseases. (In Persian).

11. Geisler D.M., Pitt J.I., and Taylor J.W. 1998. Cryptic speciation and recombination in the aflatoxin-producing fungus Aspergillus flavus. Proceedings of National Academy of Sciences, USA, 95: 388-393.

12. Ghosta Y. 2004. A taxonomic study on the genus Alternaria from Iran, Ph.D Thesis, Tarbiat Modarres University, Tehran, Iran.

13. Hajianfar R., and Zarbakhsh A. 2006. Identification of causal organism of early blight and stem canker diseases on tomato in major production regions of country. Iranian Plant Protection Journal, 17: 182. (In Persian with English abstract)

14. Hajipour Jarchelo Z., Ghosta Y., and Rezaee S. 2012. Study of pathogenic species of Alternaria on tomato and potato in West Azerbaijan province. Journal of Plant Protection Science, 43: 155-163. (In Persian with English abstract).

15. Izadpanah K., Ashkan M., Banihashemi Z., Rahimian H., and Minasian V. 2011. Plant pathology. Volume II, Fifth Edition (translation), Tehran Aeezh publication. (Translated in Persian).

16. Jaafarnia S., and Homae M. 2008. Comprehensive Illustrated Guide greenhouse cucumber and tomato. Cultivaton, Sokhan Gostare Publishing, Mashhad. (in Persian with English abstract)

17. Jaliani N. 1992. Tomato early blight disease and its chemical control in Jiroft and Bam. 10th plant protection congress Kerman. Iran 118p (in Persian).

18. Kakvan N., Zamanizadeh H., Morid B., Taheri H., and Hajmansor S. 2012. Study on pathogenic and genetic diversity of Alternaria alternata isolated from citrus hybrids of Iran, based on RAPD-PCR technique. European Journal of Experimental Biology, 2 (3): 570-576.

19. Kale M.S., Pardeshi V.C., Gurjar G.S., Gupta V.S., Gohokar, R.S., Ghoropade P.B., and Kadoo N.Y. 2012. Inter simple sequence repeat markers reveal high genetic diversity among A. alternata isolates of Indian origin. Journal of Mycology and Plant Pathology, 42(2): 194-200.

20. Kolliker R., Jones E.S., Drayton M.C., Dupal M. P., and Forster J. W. 2001. Development and characterization of simple sequence repeat (SSR) marker for white clover (Trifolium repens L.). Theoretical and Applied Genetics, 102: 416-424.

21. Kumar M., Mishra G. P., Singh R., Kumar J., Naik P.K., and Singh S. B. 2009. Correspondence of ISSR and RAPD markers for comparative analysis of genetic diversity among different apricot genotypes from cold arid deserts of trans-Himalayas. Physiology and Molecular Biology of Plants, 15: 225-236.

22. Leiminger J., Bahnweg G., and Hausladen H. 2010. Population genetics consequences on early blight disease. Twelfth EuroBlight workshop.

23. Liang-Dong G., and Li X. 2004. Genetic variation of alternaria alternata, an endophytic fungus isolated from Pinus tabulaeformis as determined by random amplified microsatellites (RAMS). Fungal Diversity, 16: 53-65.

24. Litt M., and Luty J.A. 1989. A hyper variable microsatellite revealed by in vitro amplification of a dinunleotide repeat within the cardiac muscle actin gene. American Journal of Human Genetics. 44: 397-401.

25. Lourenzo V., Rodriguez T., Campos A., Braganza C., Scheuermann K., Reis A., Brommonschenkel S., Maffia L., and Mizubuti S. 2011. Genetic structure of the population of Alternaria solani in Brazil. Journal of Phytopathology, 159: 233-240.

26. Martinezi S.P., Snowdon R., and Kuhnemann J.P. 2004. Variability of Cuban and international populations of Alternaria solani from different hosts and localities AFLP genetic analysis. European Journal of Plant Pathology 110: 399–409.

27. McDonald B.A., Zhan J., and Burdon J.J. 1999. Genetic structure of Rhynchosporium secalis in Australia. Phytopathology, 89: 639-645.

28. Mohammadi S.A., and Prasanna B.M. 2003. Analysis of genetic diversity in crop plant: Salien statistical tools and considerations. Crop Science, 43: 1248-1235.

29. Montazeri M., Greaves M.P., Pei M.H., and Ruiz C. 2005. An analysis of genetic diversity in hyphal tip isolates of promising mycoherbicide Alternaria alternata for control of Amaranthus retroflexus. Iranian Journal of Weed Science, 1: 51-65.

30. Morris P.F., Connolly M.S., and St-Clair D.A. 2000. Genetic diversity of Alternaria alternata isolated from tomato in California assessed using RAPDs. Mycological Research Journal, 104: 286-292.

31. Nasim G.H., Khan S., and Khokhar I. 2012. Molecular polymorphism and phylogenetic relationship of some Alternaria alternata isolates. Pakistan Journal of Botany, 44: 1267-1270.

32. Nourollahi K., Haghi Z., and Mehrabi Oladi A. 2014. Study of genetic diversity of Fusarium verticillioides isolates the causal agent of crown and root rot in rice in ilam province using SSR marker. Iranian Journal of plant protection science, 45: 29-37. (In Persian with English abstract)

33. Peakall R., and Smouse P.E. 2012. GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research-an update. Bioinformatics, 28: 2537-2539.

34. Perrier X., and Jacquemoud-Collet J.P. 2006. DARwin software, http://darwin.cirad.fr/darwin

35. Roder M.S., Korzun V., Wendehake K., Plaschke J., Tixier M.H., Leroy P., and Ganal M.W. 1998. A microsatellite map of wheat, Genetics, 149:20-23.

36. Rohlf F.J. 1998. NTSYS-pc. Numerical Taxonomy and Multivariate Analysis System, version 2.02, Exeter Software, Setauket, NY.

37. Rotem J. 1994. The genus Alternaria: biology, epidemiology, and pathogenicity. St Paul, Minnesota, USA: American Phytopathological Society, 326 pp.

38. Shahryari D., and Karimi Rozbahani A. 1998. Stem canker of tomato in Varamin. Journal of Plant Pests and Diseases, 65: 12-19.

39. Sharma T.R.T., and Tewari J.P. 1998. RAPD analysis of three Alternaria species pathogenic to crucifers. Mycological Research, 102: 807–814.

40. Slatkin M. 1987. Gene flow and the geographic structure of natural populations. Science, 236: 787-792.

41. Tautz D. 1989. Hyper variability of simple sequences as a general source for polymorphic DNA markers. Nucleic Acids Research, 17: 6463-6470.

42. Timmer L.W., Peever T.L., Solel Z., and Akimitsu K. 2003. Alternaria diseases of citrus – novel pathosystems. Phytopathologia Mediterranea, 42: 99-112.

43. Tran-Dinh N., and Hocking A. 2006. Isolation and characterization of polymorphic microsatellite markers for Alternaria alternata. Molecular Ecology, 6: 405–407.

44. Vander Waals J. E., Korsten L., and Slippers B. 2004. Genetic diversity among Alternaria solani isolates from potatoes in South Africa. Plant Disease, 88: 959-964.

45. Vicent A., Armengol J., and Garcıa Jimenez J. 2007. Rain fastness and persistence of fungicides for control of Alternaria brown spot of citrus. Plant Disease, 91: 393–399.

46. Weber J. L., and May P.E. 1989. Abundant class of human DNA polymorphisms which can be typed using the polymerase chain reaction. American Journal of Human Genetics, 44: 338-396.

47. Weir T.L., Huff D. R., Christ B.J., and Romaine C.P. 1998. RAPD-PCR analysis of genetic variation among isolates of Alternaria solani and A. alternata from potato and tomato. Mycologia, 90: 813-821.

48. Yeh F.C., Yang R.C., and Boyle T. 1999. Microsoft Window-based freeware for population genetic analysis (POPGENE), ver.1.31, ftp://ftp.microsoft.com/softlib/MSLFILES/HPGL.EXE.

49. Youssuf A.M., and Gherbawy H. 2005. Genetic variation among isolates of Alternaria spp. from select Egyptian crops. Phytopathology and Plant Protection, 38: 77- 89.

50. Zane L., Bargelloni L., and Patarnello T. 2002. Strategies for microsatellite isolation. A review, Molecular Ecology, 11:1-16.