Türkiye’de Antepfıstığından Elde Edilen Alternaria spp.’nin PCR-RFLP ile Karakterizasyonu ve Klonal Soy Hattına İlişkin Bulgular

Öz

Alternaria spp., antep fıstığında (Pistacia vera) ve diğer birçok üründe önemli verim kayıplarına neden olmaktadır. Bu çalışmada, Türkiye’de yerel antep fıstığı çeşitlerinden elde edilen Alternaria izolatları, ITS, Actin ve Tef1-α bölgelerine dayalı PCR-RFLP yöntemi kullanılarak karakterize edilmiştir. PCR amplifikasyonu sonucunda sırasıyla yaklaşık 600, 250 ve 280 bp büyüklüğünde ürünler elde edilmiştir. Restriksiyon analizi, izolatlar arasında herhangi bir polimorfizm bulunmadığını göstermiş ve bu izolatların tek bir klonal soya ait olabileceğini düşündürmüştür.

Anahtar Kelimeler

• Antepfıstığı
• Alternaria
• PCR-RFLP

Destekleyen Kurum

Proje çalışmaları Gaziantep Üniversitesi tarafından desteklenmiştir.

Etik Beyan

Etik beyan gerekli değildir.

Teşekkür

Safa J. Tuama Ali ve Ali Alayedi Irak hükümeti tarafından, Selçuk Başbuğa TÜBİTAK tarafından desteklenmiştir.

PCR-RFLP Characterization of Alternaria spp. from Pistachio in Türkiye and Evidence for a Putative Clonal Lineage

Öz

Alternaria spp. cause significant yield losses in pistachio (Pistacia vera) and other crops. In this study, Alternaria isolates collected from local pistachio cultivars in Türkiye were characterized using PCR-RFLP based on ITS, Actin, and Tef1-α regions. PCR amplification generated fragments of approximately 600, 250, and 280 bp, respectively. Restriction analysis revealed no polymorphism among the isolates, suggesting that they may belong to a single clonal lineage.

Anahtar Kelimeler

• Pistachio
• Alternaria
• PCR-RFLP

Destekleyen Kurum

This research was supported by Gaziantep University

Etik Beyan

Not applicable to this article.

Teşekkür

Safa J. Tuama Ali and Ali Alayedi were supported by scholarships provided by the Government of Iraq. Selçuk Başbuğa was supported by a scholarship from TÜBİTAK.

Kaynakça

1. 1. Al-Saghir M, Porter D. Taxonomic revision of the genus Pistacia L. (Anacardiaceae). Am J Plant Sci. 2012;3:12–32.
2. 2. FAOSTAT. Food and Agriculture Organization of the United Nations [Internet]. 2024 [cited 2026 March 6]. Available from: http://faostat.fao.org/site/339/default.aspx
3. 3. Sarpkaya K. Antepfıstığında karazenk hastalığına neden olan Pseudocercospora pistacina’nın biyolojisi, epidemiyolojisi ve mücadelesine yönelik çalışmalar [doctoral dissertation]. Adana: Çukurova Üniversitesi; 2014. 4. Eskalen A, Küsek M, Danisti L, Karadaş S. Fungal diseases in pistachio trees in East-Mediterranean and Southeast Anatolian regions. In: Ak BE, editor. XI GREMPA Seminar on Pistachios and Almonds. Zaragoza: CIHEAM; 2001. Cah Options Mediterr. 56.
4. 5. Konukoğlu F, Küsek M, Sarpkaya K, Karadağ S, Can C. Discoloration of pistachio nuts grown in the Gaziantep Province of Türkiye. Acta Hortic. 2011;912:761–763.
5. 6. Pryor BM, Michailides TJ. Morphological, pathogenic, and molecular characterization of Alternaria isolates associated with Alternaria late blight of pistachio. Phytopathology. 2002;92:406–416. 7. Michailides TJ. Above ground fungal diseases. In: Ferguson L, editor. Pistachio Production Manual. California (CA): 2005.
6. 8. Özkılınç H, Kurt S. Screening fungicide resistance of Alternaria pathogens causing Alternaria blight of pistachio in Türkiye. YYU J Agric Sci. 2017;27(4):543–549.
7. 9. Özkılınç H, Sarpkaya K, Kurt S, Can C, Polatbilek H, Yaşar A, et al. Pathogenicity, morpho-species, and mating types of Alternaria spp. causing Alternaria blight in Pistacia spp. in Türkiye. Phytoparasitica.2017;45:645–658.
8. 10. Özkılınç H, Sevinç U. Molecular phylogenetic species in Alternaria pathogens infecting pistachio and wild relatives. 3 Biotech. 2018;8:250.

9. 11. Michailides TJ, Morgan DP, Goldhammer DA. Surface drip irrigation reduces Alternaria late blight of pistachio caused by Alternaria alternata. In: California Pistachio Industry Annual Report. Crop year 1995–1996. Fresno (CA): California Pistachio Commission; 1996. p. 129–132.
10. 12. Gusella G, López-Moral A, Antón-Domínguez BI, Trapero C, Polizzi G, Michailides TJ, et al. Current status of pistachio diseases in countries of the Mediterranean Basin. Plant Pathol. 2024;73:2005–2029.
11. 13. Rhouma A, Rhouma R, Hajji-Hedfi L. The threat of Alternaria late blight of pistachio and its integrated management. J Oasis Agric Sustain Dev. 2024;30:2024.
12. 14. Can C, Konukoğlu F, Özgün E, Küsek M, Özkılınç H, Aktan A, et al. Alternaria blight infections of pistachio fruits in the Southeastern Anatolia region of Türkiye. Acta Hortic. 2014;1028:171–177.
13. 15. Rotem J. The genus Alternaria: biology, epidemiology and pathogenicity. St. Paul (MN): American Phytopathological Society Press; 1994.
14. 16. Simmons EG. Alternaria: an identification manual. Utrecht: CBS Fungal Biodiversity Centre; 2007.
15. 17. Peever TL, Su G, Carpenter-Boggs L, Timmer LW. Molecular systematics of citrus-associated Alternaria species. Mycologia. 2004;96:119–134.
16. 18. Andrew M, Peever TL, Pryor BM. An expanded multilocus phylogeny does not resolve morphological species within the small-spored Alternaria species complex. Mycologia. 2009;101:95–109.
17. 19. Stewart JE, Özkılınç H, Timmer LM, Pryor BM, Peever TL. Evolutionary history and species boundaries of the citrus brown spot fungus. Phytopathology. 2011;101:171.
18. 20. Crous PW, Quaedvlieg W, Sarpkaya K, Can C, Erkılıç A. Septoria-like pathogens causing leaf and fruit spot of pistachio. IMA Fungus. 2013;4(2):187–199.
19. 21. White TJ, Bruns TD, Lee SB, Taylor JW. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sninsky JJ, White TJ, editors. PCR Protocols: A Guide to Methods and Applications. New York: Academic Press; 1990. p. 315–322.
20. 22. Samson RA, Hong S, Peterson SW, Frisvad JC, Varga J. Polyphasic taxonomy of Aspergillus section Fumigati and its teleomorph Neosartorya. Stud Mycol. 2007;59:147–203.
21. 23. Balajee SA, Houbraken J, Verweij PE, Hong SB, Yaguchi T, Varga J, et al. Aspergillus species identification in the clinical setting. Stud Mycol. 2007;59:39–46.
22. 24. Milgroom MG, Peever TL. Population biology of plant pathogens. Plant Dis. 2003;87:608–617.
23. 25. Adachi Y, Watanabe H, Tanabe K, Doke N, Nishimura S, Tsuge T. Nuclear ribosomal DNA as a probe for genetic variability in the Japanese pear pathotype of Alternaria alternata. Appl Environ Microbiol. 1993;59:3197–3205.
24. 26. Weir TL, Huff DR, Christ BJ, Romaine CP. RAPD-PCR analysis of genetic variation among isolates of Alternaria solani and Alternaria alternata from potato and tomato. Mycologia. 1998;90:813–821.
25. 27. Mirhendi SH, Diba K, Kordbacheh P, et al. Identification of pathogenic Aspergillus species by a PCR-restriction enzyme method. J Med Microbiol. 2007;56:1568–1570.
26. 28. Mohankumar M, Vijayasamundeeswari A, Karthikeyan M, Mathiyazhagan S, Paranidharan V, Velazhahan R. Analysis of molecular variability among isolates of Aspergillus flavus by PCR-RFLP of the ITS regions of rDNA. J Plant Prot Res. 2010;50:446–451.
27. 29. Özgün E. Antep fıstığında (Pistacia vera L.) aflatoksin oluşumuna neden olan Aspergillus türlerinin belirlenmesi [master’s thesis]. Gaziantep: Gaziantep Üniversitesi; 2013.
28. 30. Peever TL, Canihos Y, Olsen L, Ibanez A, Liu YC, Timmer LW. Population genetic structure and host specificity of Alternaria spp. causing brown spot of Minneola tangelo and rough lemon in Florida. Phytopathology. 1999;89:851–860.
29. 31. Berbee ML, Payne BP, Zhang G, Roberts RG, Turgeon BG. Shared ITS DNA polymorphisms in isolates of opposite mating type reveal a recombining history for three presumed asexual species in the filamentous ascomycete genus Alternaria. Mycol Res. 2003;107:169–182.
30. 32. Slavov S, Mayama S, Atanassov A. Some aspects of epidemiology of Alternaria alternata tobacco pathotype. Biotechnol Biotechnol Equip. 2004;18:85–89.
31. 33. Lawrence DP, Park MS, Pryor BM. Nimbya and Embellisia revisited, with new combinations for Alternaria celosiae and A. perpunctulata. Mycol Prog. 2012;11:799–815.
32. 34. Guo-yin T, Zhi-ling Y, Zhi-lin Y, Shou-an Z. Morphological, molecular and pathogenic characterization of Alternaria longipes, the fungal pathogen causing leaf spot on Atractylodes macrocephala. Afr J Microbiol Res. 2013;7:2589–2595.
33. 35. Gardes M, White TJ, Fortin JA, Bruns TD, Taylor JW. Identification of indigenous and introduced symbiotic fungi in ectomycorrhizae by amplification of nuclear and mitochondrial ribosomal DNA. Can J Bot. 1991;69:180–190.
34. 36. Anderson JB, Stasovski E. Molecular phylogeny of northern hemisphere species of Armillaria. Mycologia. 1992;84:505–516.
35. 37. Kang JC, Crous PW, Mchau GRA, Serdani M, Song SM. Phylogenetic analysis of Alternaria spp. associated with apple core rot and citrus black rot in South Africa. Mycol Res. 2002;106:1151–1162.
36. 38. Achilonu CC, Gryzenhout M, Marais GJ, Ghosh S. Differential detection of Alternaria alternata haplotypes isolated from Carya illinoinensis using PCR-RFLP analysis of Alt a1 gene region. Genes. 2023;14:1115. 39. Habib W, Masiello M, El Ghorayeb R, Gerges E, Susca A, Meca G, et al. Mycotoxin profile and phylogeny of pathogenic Alternaria species isolated from symptomatic tomato plants in Lebanon. Toxins (Basel). 2021;13:513.
37. 40. Woudenberg JHC, Groenewald JZ, Binder M, Crous PW. Alternaria redefined. Stud Mycol. 2013;75:171–212.
38. 41. Chou HH, Wu WS. Phylogenetic analysis of internal transcribed spacer regions of the genus Alternaria, and the significance of filament-beaked conidia. Mycol Res. 2002;106:164–169.
39. 42. Salotti I, Giorni P, Battilani P. Biology, ecology, and epidemiology of Alternaria species affecting tomato: Ground information for the development of a predictive model. Front Plant Sci. 2024;15:1430965.
40. 43. Tymon LS, Peever TL, Johnson DA. Identification and enumeration of small-spored Alternaria species associated with potato in the U.S. Northwest. Plant Dis. 2016;100:465–472.
41. 44. Kusaba M, Tsuge T. Nuclear ribosomal DNA variation and pathogenic specialization in Alternaria fungi known to produce host-specific toxins. Appl Environ Microbiol. 1994;60:3055–3062.
42. 45. Aradhya MK, Chan HM, Parfitt DE. Genetic variability in the pistachio late blight fungus, Alternaria alternata. Mycol Res. 2001;105:300–306.
43. 46. Kusaba M, Tsuge T. Phylogeny of Alternaria fungi known to produce host-specific toxins on the basis of variation in internal transcribed spacers of ribosomal DNA. Curr Genet. 1995;28:491–498.
44. 47. Roberts RG, Reymond ST, Andersen B. RAPD fragment pattern analysis and morphological segregation of small-spored Alternaria species and species groups. Mycol Res. 2000;104:151–160.
45. 48. Yessimseitova A, Abdrakhmanova A, Tokbergenova Z, Abdullaeva B, Muranets A, Nurtaza A, et al. Identification and characterization of Alternaria species causing early blight on tomato in Kazakhstan. Agronomy. 2025;15:1251.
46. 49. Kılınç B, Güldür M, Dikilitaş M. Şanlıurfa ilinde Antepfıstığı (Pistacia vera L.) ağaçlarında Neoscytalidium novaehollandiae’nın bulaşıklık oranının belirlenmesi, morfolojik ve genetik karakterizasyonu. Harran Tarım ve Gıda Bilimleri Dergisi. 2022; 26(1), 25-39.