Alternaria burnsii’nin gelişimi üzerine farklı kültürel koşulların etkisi ve rDNA-ITS sekansına dayanan filogenetik analizi

Yıl 2017, Cilt: 32 Sayı: 3, 284 - 290, 15.10.2017

Harun Bayraktar Olgac Yılmaz Göksel Özer

https://doi.org/10.7161/omuanajas.343719

Öz

Alternaria burnsii’nin neden olduğu kimyon yanıklığı birçok ülkede
kimyon üretimini sınırlandıran önemli bir fungal hastalıktır. Patojen ülkemizde
de hemen hemen tüm kimyon ekim alanlarında yaygın olup önemli ekonomik
kayıplara neden olmaktadır. Bununla birlikte, Alternaria burnsii’ nin morfolojik, fenotipik ve filogenetik
özellikleri hakkında çok fazla bilgi bulunmamaktadır.  Bu çalışmada Alternaria burnsii’nin miseliyal gelişimi ve sporulasyonu üzerine
farklı besi ortamları ve çevresel faktörlerin etkisi araştırılmıştır. Ayrıca
patojen izolatları arasındaki morfolojik farklılıklar değerlendirilmiştir. Bu
amaçla patojen izolatlar, yedi farklı besi ortamı (Patates Dekstroz Agar ,
Patates Havuç Agar, Mısır Unu Agar, Czapek Dox Agar, Sukroz Agar, Su Agarı,
Domates Suyu Agarı, V88) üzerinde iki farklı inkubasyon koşulu altında
geliştirilmiştir. Değerlendirilen kültür ortamları arasında en yüksek
sporulasyon değişken sıcaklık ve ışık koşularına maruz bırakılan V88 ortamı
üzerinde elde edilmiştir. Ayrıca kültür ortamı ve gelişme koşullarına bağlı
olarak konidi uzunluğu, genişliği ve septa sayısı bakımından istatiksel olarak önemli
farklılıklar gözlemlenmiştir. Sıcaklık patojen gelişimini etkileyen önemli bir
sınırlayıcı faktör olup en iyi gelişme oranı 25 °C’ de gözlemlenmiştir. Ayrıca,
UPGMA metodu ile gerçekleştirilen ITS sekansının filogenetik analizi ise test
edilen tüm izolatların aynı grup içerisinde yer aldığını ve bu izolatların
GenBankasından elde edilen ve farklı seksiyonları temsil eden Alternaria tür gruplarına ait
izolatlardan filogenetik olarak farklı olduğunu göstermiştir. Bu sonuçların
gelecekteki araştırmalar için patojen inokulumunun hazırlanmasının yanısıra
patojen tespit ve tanılanması için farklı metotların geliştirilmesinde faydalı
bilgiler sağlayacağı düşünülmektedir. 

Anahtar Kelimeler

Kimyon, Fungus, Besi ortamı, Patojen gelişimi, rDNA-ITS

Effect of different cultural conditions on the growth of Alternaria burnsii and its phylogenetic analysis based on rDNA-ITS sequences

Öz

Cumin blight caused by Alternaria burnsii is an important fungal
disease restricting cumin production in many countries. The pathogen is widespread in almost all cumin growing
areas in Turkey and
causes major economic losses. However, little information is known about morphologic,
phenotypic and phylogenetic characteristics of Alternaria burnsii. In this study, the effect of different nutrient
media and environmental factors on mycelial growth and sporulation of Alternaria burnsii was studied, and
morphological differences among the pathogen isolates were evaluated. For this
purpose, pathogen isolates were grown on seven different nutrient media (Potato
Dekstrose Agar, Potato Carrot Agar, Corn Meal Agar, Czapek Dox Agar, Sucrose
Agar, Water Agar, Tomato Juice Agar, V88) under two different incubation conditions.
Among the culture media evaluated, the highest sporulation was obtained on V88
media exposed to the alternating temperature and light regimes. Also, the
length, width and septa number of the conidia showed statistically differences
depending on culture media and growing conditions. The temperature was a
significant limiting factor affecting the pathogen growth. The best growth was
observed at 25 °C. Phylogenetic analysis of the rDNA-ITS sequences performed by
UPGMA method indicated that all tested isolates clustered in the same
group and these isolates were phylogenetically distinct from the isolates of Alternaria-species groups, representing
different sections retrieved from GenBank. These results could provide useful
information in the pathogen inoculum production for further studies as well as
the development of different methods for the identification and determination
of the pathogen.

Anahtar Kelimeler

Cumin, Fungus, Growth medium, Pathogen growth, rDNA-ITS

Kaynakça

• Alhussaen, K.M. 2012. Altemaria solani isolated from Tomato in Jordan Valley. Research Journal of Biological Sciences, 7(8): 316-319.
• Bayraktar, H., Özer, G., Oksal, E. 2013. Phylogenetic analysis of Alternaria spp. and related fungi associated with cumin blight based upon analysis of nuclear ITS and Alt a1 gene sequences. 1st Mediterranean Symposium on Medicinal and Aromatic Plants (MESMAP-2013) April 17-20, The Northern Cyprus.
• Benlioğlu, S., Delen, N. 1996. Studies on the Sporulation of the Early Blight Agent [Alternaria solani (Ell. and Mart.) Jones and Grout] of Tomatoes. Journal of Turkish Phytopathology, 25: 23-28.
• Castro, M.E.A., Zambolim, L., Chaves, G.M., Cruz, C.D., Matsuoka, K. 2000. Pathogenic variability of Alternaria solani, the causal agent of tomato early blight. Summa Phytopathologica, 26:24-28.
• Douglas, D.R. 1972. The effect of light and temperature on the sporulation of different isolates of Alternaria solani. Canadian Journal of Botany, 50: 629-634.
• Ellis, M.B. 1971. Dematiaceous Hyphomycetes. CMI, Kew, Surrey, England, pp. 1-608.
• Ellis, M.B. 1976. More Dematiaceous Hyphomycetes. CMI, Kew. Surrey, England, pp. 1-507.
• Gemawat, P.D., Prasad, N. 1971. Alternaria blight of Cuminum cyminum L. Proceedings of the Indian National Science Academy: Biological Sciences, 38: 38-43.
• Holiday, P. 1980. Fungus Diseases of Tropical Crops. Cambridge University Press.
• Hong, S.G., Cramer, R.A., Lawrence, C.B., Pryor, B.M. 2005. Alt a1 allergen homologs from Alternaria and related taxa: analysis of phylogenetic content and secondary structure. Fungal Genetics and Biology, 42:119-129.
• Jadhav, B.M., Perane, R.R., Kale, A.A., Pawar, N.B. 2011. Morphological, pathological and molecular variability among Alternaria macrospora isolates causing leaf blight of cotton. Indian Phytopathology, 64(3): 54-257.
• Kocatürk, S. 1988. The important cumin diseases in Cen¬tral Anatolia. Journal of Turkish Phytopathology 17:121.
• Lawrence, D.P., Gannibal, P.B., Peever, T.L., Pryor, B.M. 2013. The sections of Alternaria: formalizing species-group concepts. Mycologia, 105: 530-546.
• Lukens, R.J. 1960. Conidial production from filter paper culture of Helminthosporium vagans and Alternaria solani. Phytopathology, 50: 867-868.
• Naik, M. K., Prasad, Y., Bhat, K. V., Rani, G. D. 2010. Morphological, physiological, pathogenic and molecular variability among isolates of Alternaria solani from tomato. Indian Phytopathology, 63(2): 168-173.
• Özer, G., Bayraktar, H. 2015. Determination of fungal pathogens associated with Cuminum cyminum in Turkey. Plant Protection Science, 51:74-79.
• Prasad, M.S., Sujatha, L.M., Rao, S.C. 2009. Analysis of cultural and genetic diversity in Alternaria helianthi and determination of pathogenic variability using wild helianthus species. Journal of Phytopathology, 157:609-617.
• Pryor, B.M., Bigelow, D.M. 2003. Molecular characterization of Embellisia and Nimbya species and their relationship to Alternaria, Ulocladium and Stemphylium. Mycologia, 95(6):1141-1154.
• Ramjegathesh, R., Ebenezar, E.G. 2012. Morphological and physiological characters of Alternaria alternata causing leaf blight disease of onion. International Journal of Plant Pathology, 3(2):34-44.
• Reader, U., Broda, P. 1985. Rapid preparation of DNA from filamentous fungi. Lett. Appl. Microbiol., 1: 17-20.
• Rodrigues, T.T., Maffia, L.A., Dhingra, O.D., Mizubuti, E.S. 2010. In vitro production of conidia of Alternaria solani. Tropical Plant Pathology, 35: 203-212.
• Rotem, J. 1994. The genus Alternaria: biology, epidemiology and pathogenicity. The American Phytopathological Society, St. Paul, Minnesota.
• Sambrook, J., Fritsch, E.F., Maniatis, T. 1989. Molecular cloning: A Laboratory manual, 2nd edn. New York, Cold Spring Harbor Laboratory Press, pp. 1659.
• Shahzad, A. 2003. Studies on Alternaria leaf blotch of apple in Kashmir (Doctoral dissertation, Ph. D.(Agric.) Thesis submitted to Post Graduate Faculty, Skuastk, Shalimar, Kashmir.
• Simmons, E.G. 1992. Alternaria taxonomy: current status, viewpoint, challenge. In: Alternaria Biology, Plant Diseases and Metabolites. J.
• Chelkowski and A. Visconti, eds. Amsterdam, Netherlands:Elsevier Science Publishers, 1-35.
• Simmons, E.G. 2007. Alternaria: an Identification Manual. Utrecht, CBS Fungal Diversity Centre.
• Tamura, K., Peterson, D., Peterson, N., Stecher, G., Nei, M., Kumar, S. 2011. MEGA5: molecular evolutionary ge-netics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Molecular Biology and Evolution, 28: 2731–2739.
• Udayashankar, A.C., Nayaka, S.C., Archana, B., Anjana, G., Niranjana, S.R., Mortensen, C.N., Lund, O.S., Prakash, H.S. 2012. Specific PCR-based detection of Alternaria helianthi: the cause of blight and leaf spot in sunflower. Archives of Microbiology, 194(11): 923-932.
• Waggoner, P.E., Horsfall, J.G. 1969. EPIDEM. A simulator of plant disease written for a computer. Bulletin of the Connecticut Agricultural Experiment Station, New Haven., 698:1-80. White, T.J., Bruns, T.D., Lee, S., Taylor, J. 1990. Amplification and direct sequencing of fungal ribosomal RNA for phylogentics. In: Innis MA, Gelfland DH, Sninsky JJ, White TJ, eds. PCR Protocols: A guide to methods and applications. San Diego, CA, USA: Academic Press, pp. 315-22.