Haşhaş (Papaver somniferum) Kök Boğazı ve Kapsül Yanıklığı Etmeni Alternaria papavericola’nın Moleküler Karakterizasyonu

Öz

Hem dünyada hem de ülkemizde önemli tıbbi ve aromatik bitkiler arasında yer alan haşhaş (Papaver somniferum L.) bitkisinin tohumlarından ve yağından da gıda olarak yararlanılmaktadır. Bu çalışmada Isparta ilinden toplanan haşhaş tohumlarından izole edilen, haşhaşta kök boğazı ve kapsül yanıklığına neden olan Alternaria papavericola Woudenb. & Crous’nın moleküler karakterizasyonunun yapılması amaçlanmıştır. Bu amaçla rDNA’nın ITS (internal transcribed spacer) gen bölgesi ITS1 ve ITS4 evrensel primerleri kullanılarak çoğaltılmış ve dizilenmiştir. DNA nükleotid dizisi, NCBI GenBank veri tabanında BLAST işlemine sokulmuş ve veri tabanındaki diğer ilgili diziler ile karşılaştırılmıştır. Microsoft Office Word programı yardımı ile FASTA formatına çevrilen A.papavericola’nın DNA dizileri, MEGA 6.0 programı kullanılarak Tamura-Nei modellemesiyle Maximum Likelihood (ML) metodu kullanılarak filogenetik ağaç oluşturulmuştur.

Anahtar Kelimeler

• Alternaria papavericola
• haşhaş
• moleküler karakterizasyon

Molecular Characterization of Alternaria papavericola, the Causal Agent of Crown and Capsule Blight of Poppy (Papaver somniferum)

Abstract

Both globally and in our country, the poppy (Papaver somniferum L.) is a significant medicinal and aromatic plant, with its seeds and oil serving as food. This work aimed to molecularly characterize Alternaria papavericola Woudenb. & Crous, the fungus that causes crown and capsule blight in poppies. The fungus was isolated from poppy seeds that were collected from the province of Isparta. For this purpose, the ITS (internal transcribed spacer) gene region of rDNA was amplified and sequenced using ITS1 and ITS4 universal primers. The DNA nucleotide sequence was BLASTed in the NCBI GenBank database and compared with other related sequences in the database. The DNA sequences of A.papavericola were converted to FASTA format with the help of the Microsoft Office Word program, and a phylogenetic tree was constructed using the Maximum Likelihood (ML) method with Tamura-Nei modeling using the MEGA 6.0 program.

Keywords

• Alternaria papavericola
• poppy
• molecular characterization

Kaynakça

1. İşler, N. 2024. Haşhaş Tarımı. https://www.mku.edu.tr/files/898-595293aa-9378-4c9f-8650-e3066e00eaab.pdf.
2. Scott, J., Quarrell, S. 2016. Potential biosecurity risks associated with poppy straw and pellet importation. Tasmanian Institute of Agriculture, University of Tasmania, pp:31.
3. Woudenberg, J.H.C., Groenewald, J.Z., Binder, M., Crous, P.W. 2013. Alternaria redefined. Studies in Mycology 75:171-212.
4. Mraz, F. 1960. Influence of the harmfull effect of the fungus Helminthosporium papaveris on poppy yields in the region Karlovy Vary in the years 1957 and 1958. Sbornik Cesk Akad Zemedelske 33:1083-1094.
5. Ellis, M.B. 1971. Dematiaceous hyphomycetes. Commonwealth Mycological Institute, Kew, Surrey, England.
6. Karahan, O., Maden, S. 1978. Haşhaşta kök boğazı yanıklığı hastalığı (Dendryphıon papaveris (Saw.) Sawada), tohumla taşınma durumu ve tohum ilaçlarının bu etmene etkisi üzerinde çalışmalar. Bitki Koruma Bülteni 18:1-9.
7. Inderbitzin, P., Shoemaker, R.A., O’Neill, N.R., Turgeon, B.G., Berbee, M.L. 2006. Systematics and mating systems of two fungal pathogens of opium poppy: the heterothallic Crivellia papaveracea with a Brachycladium penicillatum asexual state and a homothallic species with a Brachycladium papaveris asexual state. Canadian Journal of Botany 84:1304‐1326.
8. Gülcan, C., 2021. Uşak ilinde haşhaş (Papaver somniferum L.) üretiminde kapsül yanıklığına neden olan fungal etmenlerin belirlenmesi (Yüksek Lisans Tezi). Uşak Üniversitesi Lisansüstü Eğitim Enstitüsü Tarım Bilimleri Anabilim Dalı, Uşak, 97s.

9. Sneh, B., Burpee, L., Ogoshi, A. 1991. Identification of Rhizoctonia species. APS Press, Minnesota, USA, pp:133.
10. Taylor, J.W., Jacobson, D.J., Kroken, S., Kasuga, T., Geiser, D.M., Hibbett, D.S., Fisher, M.C. 2000. Phylogenetic species recognition and species concepts in fungi. Fungal Genetic and Biology 31:21-32.
11. Kaunitz, J.D. 2015. The discovery of PCR: ProCuRement of divine power. Digestive Diseases and Sciences 60(8):2230-2231.
12. White, T.J., Bruns, T., Lee, S., Taylor, J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: M.A. Innis, D.H. Gelfand, J.J. Sninsky, T.J. White (Eds.): PCR protocols: A guide to methods and applications. San Diego: Academic Press. pp:315-322.
13. Bruns, T.D., Vilgays, T.J., Taylor, J.W. 1991. Fungal molecular systematics. Annual Review of Ecology and Systematics 22:525-564.
14. Eken, C., Hasenekoğlu, İ., Genç, T., Eryücel, D. 2014. Erzurum ilinde fiğ (Vicia sativa L.) tohumlarından izole edilen Aspergillus ve Penicillium türleri. Uluslararası Katılımlı Türkiye 5. Tohumculuk Kongresi ve Sektörel İş Forumu, 19-23 Ekim 2014, Diyarbakır, s:657-660.
15. Hall, T.A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Window 95/98/NT. Nucleic Acids Symposium Series 41:95-98.
16. Tamura, K., Stecher, G., Peterson, D., Filipski, A., Kumar, S. 2013. MEGA6: molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution 30(12):2725-2729.
17. Felsenstein J. 1985. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783-791.
18. Kılıçoğlu, M.Ç., Özkoç İ. 2008. Fungal sistematikteki moleküler gelişmeler. OMÜ Ziraat Fakültesi Dergisi 23(1):65-72.
19. Schoch, C.L., Seifert, K.A., Huhndorf, S., Robert, V., Spouge, J.L., André Levesque, C., Chen, W. 2012. Fungal Barcoding Consortiuma. Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. Proceedings of the National Academy of Sciences 109(16):6241-6246.
20. Carling, D.E., Baird, R.E., Gitaitis, R.D., Brainard, K., Kuninaga, S. 2002. Characterization of AG-13, a newly reported anastomosis group of Rhizoctonia solani. Phytopathology 92(8):893-899.
21. Hyakumachi, M., Priyatmojo, A., Kubota, M., Fukui, H. 2005. New anastomosis groups, AG-T and AG-U of binucleate Rhizoctonia spp. causing root and stem rot of cut- flower and miniature roses. Phytopathology 95:784-792.
22. García, V.G., Onco, M.P., Susan, V.R. 2006. Biology and systematics of the form genus Rhizoctonia. Spanish Journal of Agricultural Research 4(1):55-79.
23. Sharon, M., Kuninaga, S., Hyakumachi, M., Naito, S., Sneh, B. 2008. Classification of Rhizoctonia spp. using rDNA-ITS sequence analysis supports the genetic basis of the classical anastomosis grouping. Mycoscience 49(2):93-114.
24. Kılıçoğlu, M.Ç., Özkoç, İ. 2013. Phylogenetic analysis of Rhizoctonia solani AG-4 isolates from common beans in Black Sea coastal region, Turkey, based on ITS-5.8 S Rdna. Turkish Journal of Biology 37(1):18-24.
25. Aiello, D., Guarnaccia, V., Formica, P.T., Hyakumachi, M., Polizzi, G. 2017. Occurrence and characterization of Rhizoctonia species causing diseases of ornamental plants in Italy. European Journal of Plant Pathology 148(4):967-982.
26. Eken, C., Demir, D., Uysal, G., Çalışkan, S., Sevindik E., Çağlayan, K. 2024. Isolation, identification, and pathogenicity of Rhizoctonia spp. recovered from soil samples of the greenhouse tomato growing area of the West Mediterranean Region, Türkiye. Journal of Phytopathology 172:e13297.
27. Eken, C., Demir, D., Uysal, G., Çalışkan, S., Sevindik E., Çağlayan, K. 2024. First report of the nematode-trapping fungus Arthrobotrys thaumasia in Türkiye and its biocontrol potential against Meloidogyne incognita. Journal of Phytopathology 172:e13354.
28. Keeling, P.J., Luker, M.A., Palmer, J.D. 2000. Evidence from beta-tubulin phylogeny that microsporidia evolved from within the fungi. Molecular Biology and Evolution 17(1):23-31.
29. Mohammed, A.S., Kadar, N.H., Kihal, M., Henni, J.E., Sanchez, J., Gallego, E., Garrido-Cardenas, J.A. 2016. Characterization of Fusarium oxysporum isolates from tomato plants in Algeria. African Journal of Microbiology Research 10(30):1156-1163.
30. Tuney, E., Asan, A., Şen, B. 2018. Saf kültür olarak stoklanmış bazı mikrofungusların ITS, β-tubulin ve Aktin gen dizilerine göre moleküler tanısı. Mantar Dergisi 9(1):1-17.
31. Huang, S.K., Jeewon, R., Hyde, K.D., Bhat, D.J., Chomnunti, P., Wen, T.C. 2018. Beta-tubulin and Actin gene phylogeny supports Phaeoacremonium ovale as a new species from freshwater habitats in China. MycoKeys 41:1-15.
32. Pothiraj, G., Hussain, Z., Singh, A.K., Solanke, A.U., Aggarwal, R., Ramesh, R. Shanmugam, V. 2021. Characterization of Fusarium spp. inciting vascular wilt of tomato and its management by a chaetomium-based biocontrol consortium. Frontiers in Plant Science 12:748013.