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Investigation of the Effects of Eugenol on Fusarium culmorum

Yıl 2018, Cilt: 8 Sayı: 2, 215 - 221, 30.06.2018
https://doi.org/10.21597/jist.428382

Öz

Fusarium culmorum is one of the causal agents lead to economic loses in small grain cereals. In

this study, alterations in spore production, and linear growth rates in F. culmorum 15 isolate subjected to increased

concentrations of eugenol (0, 200, 400, 800 µg mL-1) were examined. Additionally, the and expression of FcMgv1

and FcStuA genes which are essential in asexual stage and cell wall structure formation were also examined tested

in F. culmorum eugenol treated and non-treated fungal cultures. F15 isolate subjected to increased concentrations of

eugenol (0, 200, 400, 800 µg mL-1). Minimum inhibitory concentration was determined as 400 µg mL-1 eugenol.

In comparison of control and experiment sets, there were significantly different (p<0.01) the decrease in spore

production and linear growth rate (LGR) were significantly different (p<0.01). In real time polymerase chain

reaction (qPCR) analysis, β-tubulin was used as endogenous control and the expression of FcMgv1 and FcStuA

genes were determined by using cDNAs converted from total RNAs of control and experiment sets were used

in Eva-Green fluorophore dyebased real time PCR to examine the FcMgv1 and FcStuA expression. Significant

differences (p<0.05) were also determined in fold changes in gene expression. Normalization results showed that

fold changes in FcMgv1 and FcStuA genes were as +4.35±0.25 and +2.04±0.13, respectively. qPCR results were

also confirmed via reverse transcription PCR (RT-PCR) analysis. The late apoptosis was detected in the cells by

using acridine orange/ethidium bromide staining. Findings obtained from this study showed that eugenol have the

potential antifungal effects and it could be used in struggling with head blight disease caused by F. culmorum via its

in vitro antifungal effects. This study presents the original data on about is first report to evaluatinge the phenotypic

and transcriptomic effects of pure eugenol compound on F. culmorum.

Kaynakça

  • Anand A, Zhou T, Trick HN, Gill BS, Bockus WW, Muthukrishnanl S, 2003. Greenhouse and Field Testing of Transgenic Wheat Plants Stably Expressing Genes for Thaumatin-Like Protein, Chitinase and Glucanase Against Fusarium graminearum. Journal of Experimental Biology, 54 (384): 1101-1111.
  • Arif T, Bhosale JD, Kumar N, et al., 2003. Natural products- Antifungal agents derived from plants. Journal of Asian Natural Products Research, 11 (7): 621-638.
  • Arslan Ü, Baykal N, 2002. Kök ve kökboğazı fungal patojenlerine karşı bazı buğday çeşitlerinin reaksiyonları ve tohum koruyucu fungusitlerin Fusarium culmorum (w.g.sm.) sacc.’a etkisi. Uludağ Üniversitesi Ziraat Fakültesi Dergisi, 16: 69-76.
  • Bai G, Shaner G, 2004. Management and resistance in wheat and barley to Fusarium head blight. Annual Review of Phytopathology, 42: 135-161.
  • Bernardo A, Bai G, Guo P, Xiao K, Guenzi AC, Ayoubı P, 2007. Fusarium graminearum-induced changes in gene expression between Fusarium head blight-resistant and susceptible wheat cultivar. Functional and Integrative Genomics, 7: 69-77.
  • Braga PC, Dal Sasso M, Culici M, Alfieri M, 2007. Eugenol and thymol, alone or in combination, induce morphological alterations in the envelope of Candida albicans. Fitoterapia, 78(6) : 396-400.
  • Campaniello D, Corbo MR, Sinigaglia M, 2010. Antifungal activity of eugenol against Penicillium, Aspergillus, and Fusarium species. Journal of Food Protection, 73 (6): 1124-1128.
  • Castillo F, Hernández D, Gallegos G, Rodriguez R, Aguilar CN, 2012. Antifungal Properties of Bioactive Compounds from Plants, Fungicides for Plant and Animal Diseases. 1st Edition, InTechOpen. 81-106 pp.
  • Chung WH, Ishii H, Nishimura K, Ohshima M, Iwama T, Yoshimatsu H, 2008. Genetic analysis and PCR-based identification of major Fusarium species causing head blight on wheat in Japan. Journal of General Plant Pathology, 10: 110-118.
  • Karababa M, Coste AL, Rognon B, Bille J, Sanglard D, 2004. Comparison of gene expression profiles of Candida albicans azole-resistant clinical isolates and laboratory strains exposed to drug inducing multidrug transporters. Antimicrobial Agents and Chemotherapy, 48 (8): 3064-3079.
  • Livak JK, Schmittgen TD, 2001. Analysis of relative gene expression data using real time quantitative PCR and the 2-ΔΔCT method. Methods, 25: 402-408.
  • McGahon AJ, Martin SJ, Bissonnette RP, Mahboubi A, et al.,1995. The end of the (cell) line: methods for the study of apoptosis in vitro. Methods Cell Biol. 46: 153-185.
  • Miedaner T, Cumagun CJR, Chakraborty S, 2008. Population genetics of three important head blight pathogens Fusarium graminearum, F. pseudograminearum and F. culmorum. Journal of Phytopathology, 156: 129-139.
  • Obanor F, Erginbas-Orakci G, Tunali B, Nicol JM, Chakraborty S, 2010. Fusarium culmorum is a single phylogenetic species based on multilocus sequence analysis. Fungal Biology, 114: 753-765.
  • Pasquali M, Migheli Q, 2014. Genetic approaches to chemotype determination in type B-trichothecene producing Fusaria. International Journal of Food Microbiology, 189: 164–182.
  • Pereira FO, Mendes JM, Lima ED, 2013. Investigation on mechanism of antifungal activity of eugenol against Trichophyton rubrum. Medical Mycology, 51(5) : 507–513.
  • Sanglard D, Kuchler K, Ischer F, Pagani JL, Monod M, Bille J, 1995. Mechanisms of resistance to azole antifungal agents in Candida albicans isolates from AIDS patients involve specific multidrug transporters. Antimicrobial Agents and Chemotherapy, 39(11): 2378-2386.
  • Scherm B, Orru M. Balmas V, Spanu F, Azara E, Delogu G, Hammond TM, Keller NP, Migheli Q, 2011. Altered trichothecene biosynthesis in tri6-silenced transformants of Fusarium culmorum influences the severity of crown and foot rot on durum wheat seedlings. Molecular Plant Pathology, 12(8): 759-771.
  • Sefer Ö, Yörük E, Develi ES, Sezer AS, Konukcu Z, 2017. Investigation of the Effects of 2, 4-Dimethylpyrrole on Fusarium culmorum Causal Agent of Head Blight Disease. International Journal of Secondary Metabolite, 4(2) : 103-113.
  • Trajano VN, Lima EDO, Souza FSD, 2012. Antifungal activity of the essential oil of Cinnamomum zeylanicum Blume and eugenol on Aspergillus flavus. Journal of Essential Oil Bearing Plants, 15(5) : 785-793.
  • Yörük E, Tunali B, Kansu B, Ölmez F, Uz G, Zümrüt IM, Sarıkaya A, Meyva G, 2016. Characterization of high-level deoxynivalenol producer Fusarium graminearum and F. culmorum isolates caused head blight and crown rot diseases in Turkey. Journal of Plant Diseases and Protection, 123: 177–186.

Eugenol’ün Fusarium culmorum Üzerindeki Etkilerinin İncelenmesi

Yıl 2018, Cilt: 8 Sayı: 2, 215 - 221, 30.06.2018
https://doi.org/10.21597/jist.428382

Öz

Fusarium culmorum küçük daneli tahıllarda ekonomik kayıplara yol açan hastalık etmenlerinden birisidir.
Bu çalışmada, arttırılmış eugenol konsantrasyonlarına (0, 200, 400, 800 µg mL-1) maruz bırakılmış F. culmorum
F15 izolatında spor üretimi ve doğrusal büyüme oranı belirlendi. İlaveten, eugenol uygulanmış ve uygulanmamış
fungal kültürlerde aseksüel üreme ve hücre çeperi yapısının oluşumunda elzem olan FcMgv1 ve FcStuA genlerinin
anlatımı da test edildi. Üremeyi engelleyen minimum konsantrasyon 400 µg mL-1 eugenol olarak belirlendi.
Kontrol ve deney setleri karşılaştırıldığında, spor üretimi ve doğrusal büyüme oranında bilimsel olarak anlamlı
azalma belirlendi (p<0.01). Gerçek zamanlı polimeraz zincir reaksiyonu (qPCR) analizlerinde, β-tubulin endojen
kontrol olarak kullanıldı ve FcMgv1 ve FcStuA genlerinin anlatımı Eva-Green florofor boyası kullanılarak saptandı.
Gen anlatımındaki oransal değişimlerde bilimsel olarak anlamlı farklılıklar (p<0.01) saptandı. Normalizasyon
bulguları FcMgv1 ve FcStuA genlerindeki oransal değişimlerin sırasıyla +4.35±0.25 and +2.04±0.13 olduğunu
gösterdi. qPCR bulguları ayrıca ters transkripsiyon PCR (RT-PCR) analizleri ile doğrulandı. Akridin orange/
Etidyum bromür dual boyamasında hücrelerde geç apoptoza rastlanmıştır. Çalışmadan elde edilen bulgular
eugenolün potansiyel antifungal etkilere sahip olduğunu ve F. culmorum’un neden olduğu başak yanıklığı ile
mücadelede kullanılabileceğini göstermiştir. Bu çalışma saf eugenol bileşiğinin F. culmorum üzerindeki fenotipik
ve transkriptomik etkilerinin değerlendirilmesini açısından özgün bulgular sunmaktadır.

Kaynakça

  • Anand A, Zhou T, Trick HN, Gill BS, Bockus WW, Muthukrishnanl S, 2003. Greenhouse and Field Testing of Transgenic Wheat Plants Stably Expressing Genes for Thaumatin-Like Protein, Chitinase and Glucanase Against Fusarium graminearum. Journal of Experimental Biology, 54 (384): 1101-1111.
  • Arif T, Bhosale JD, Kumar N, et al., 2003. Natural products- Antifungal agents derived from plants. Journal of Asian Natural Products Research, 11 (7): 621-638.
  • Arslan Ü, Baykal N, 2002. Kök ve kökboğazı fungal patojenlerine karşı bazı buğday çeşitlerinin reaksiyonları ve tohum koruyucu fungusitlerin Fusarium culmorum (w.g.sm.) sacc.’a etkisi. Uludağ Üniversitesi Ziraat Fakültesi Dergisi, 16: 69-76.
  • Bai G, Shaner G, 2004. Management and resistance in wheat and barley to Fusarium head blight. Annual Review of Phytopathology, 42: 135-161.
  • Bernardo A, Bai G, Guo P, Xiao K, Guenzi AC, Ayoubı P, 2007. Fusarium graminearum-induced changes in gene expression between Fusarium head blight-resistant and susceptible wheat cultivar. Functional and Integrative Genomics, 7: 69-77.
  • Braga PC, Dal Sasso M, Culici M, Alfieri M, 2007. Eugenol and thymol, alone or in combination, induce morphological alterations in the envelope of Candida albicans. Fitoterapia, 78(6) : 396-400.
  • Campaniello D, Corbo MR, Sinigaglia M, 2010. Antifungal activity of eugenol against Penicillium, Aspergillus, and Fusarium species. Journal of Food Protection, 73 (6): 1124-1128.
  • Castillo F, Hernández D, Gallegos G, Rodriguez R, Aguilar CN, 2012. Antifungal Properties of Bioactive Compounds from Plants, Fungicides for Plant and Animal Diseases. 1st Edition, InTechOpen. 81-106 pp.
  • Chung WH, Ishii H, Nishimura K, Ohshima M, Iwama T, Yoshimatsu H, 2008. Genetic analysis and PCR-based identification of major Fusarium species causing head blight on wheat in Japan. Journal of General Plant Pathology, 10: 110-118.
  • Karababa M, Coste AL, Rognon B, Bille J, Sanglard D, 2004. Comparison of gene expression profiles of Candida albicans azole-resistant clinical isolates and laboratory strains exposed to drug inducing multidrug transporters. Antimicrobial Agents and Chemotherapy, 48 (8): 3064-3079.
  • Livak JK, Schmittgen TD, 2001. Analysis of relative gene expression data using real time quantitative PCR and the 2-ΔΔCT method. Methods, 25: 402-408.
  • McGahon AJ, Martin SJ, Bissonnette RP, Mahboubi A, et al.,1995. The end of the (cell) line: methods for the study of apoptosis in vitro. Methods Cell Biol. 46: 153-185.
  • Miedaner T, Cumagun CJR, Chakraborty S, 2008. Population genetics of three important head blight pathogens Fusarium graminearum, F. pseudograminearum and F. culmorum. Journal of Phytopathology, 156: 129-139.
  • Obanor F, Erginbas-Orakci G, Tunali B, Nicol JM, Chakraborty S, 2010. Fusarium culmorum is a single phylogenetic species based on multilocus sequence analysis. Fungal Biology, 114: 753-765.
  • Pasquali M, Migheli Q, 2014. Genetic approaches to chemotype determination in type B-trichothecene producing Fusaria. International Journal of Food Microbiology, 189: 164–182.
  • Pereira FO, Mendes JM, Lima ED, 2013. Investigation on mechanism of antifungal activity of eugenol against Trichophyton rubrum. Medical Mycology, 51(5) : 507–513.
  • Sanglard D, Kuchler K, Ischer F, Pagani JL, Monod M, Bille J, 1995. Mechanisms of resistance to azole antifungal agents in Candida albicans isolates from AIDS patients involve specific multidrug transporters. Antimicrobial Agents and Chemotherapy, 39(11): 2378-2386.
  • Scherm B, Orru M. Balmas V, Spanu F, Azara E, Delogu G, Hammond TM, Keller NP, Migheli Q, 2011. Altered trichothecene biosynthesis in tri6-silenced transformants of Fusarium culmorum influences the severity of crown and foot rot on durum wheat seedlings. Molecular Plant Pathology, 12(8): 759-771.
  • Sefer Ö, Yörük E, Develi ES, Sezer AS, Konukcu Z, 2017. Investigation of the Effects of 2, 4-Dimethylpyrrole on Fusarium culmorum Causal Agent of Head Blight Disease. International Journal of Secondary Metabolite, 4(2) : 103-113.
  • Trajano VN, Lima EDO, Souza FSD, 2012. Antifungal activity of the essential oil of Cinnamomum zeylanicum Blume and eugenol on Aspergillus flavus. Journal of Essential Oil Bearing Plants, 15(5) : 785-793.
  • Yörük E, Tunali B, Kansu B, Ölmez F, Uz G, Zümrüt IM, Sarıkaya A, Meyva G, 2016. Characterization of high-level deoxynivalenol producer Fusarium graminearum and F. culmorum isolates caused head blight and crown rot diseases in Turkey. Journal of Plant Diseases and Protection, 123: 177–186.
Toplam 21 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Yapısal Biyoloji
Bölüm Moleküler Biyoloji ve Genetik / Moleculer Biology and Genetic
Yazarlar

Emre Yörük 0000-0003-2770-0157

Özlem Sefer Bu kişi benim 0000-0002-2711-5938

Ayşe Server Sezer Bu kişi benim 0000-0001-6869-8757

Zeynep Konukcu Bu kişi benim 0000-0002-7854-3131

Elif Sedef Develi Bu kişi benim 0000-0001-6696-7001

Yayımlanma Tarihi 30 Haziran 2018
Gönderilme Tarihi 15 Aralık 2017
Kabul Tarihi 25 Ocak 2018
Yayımlandığı Sayı Yıl 2018 Cilt: 8 Sayı: 2

Kaynak Göster

APA Yörük, E., Sefer, Ö., Sezer, A. S., Konukcu, Z., vd. (2018). Eugenol’ün Fusarium culmorum Üzerindeki Etkilerinin İncelenmesi. Journal of the Institute of Science and Technology, 8(2), 215-221. https://doi.org/10.21597/jist.428382
AMA Yörük E, Sefer Ö, Sezer AS, Konukcu Z, Develi ES. Eugenol’ün Fusarium culmorum Üzerindeki Etkilerinin İncelenmesi. Iğdır Üniv. Fen Bil Enst. Der. Haziran 2018;8(2):215-221. doi:10.21597/jist.428382
Chicago Yörük, Emre, Özlem Sefer, Ayşe Server Sezer, Zeynep Konukcu, ve Elif Sedef Develi. “Eugenol’ün Fusarium Culmorum Üzerindeki Etkilerinin İncelenmesi”. Journal of the Institute of Science and Technology 8, sy. 2 (Haziran 2018): 215-21. https://doi.org/10.21597/jist.428382.
EndNote Yörük E, Sefer Ö, Sezer AS, Konukcu Z, Develi ES (01 Haziran 2018) Eugenol’ün Fusarium culmorum Üzerindeki Etkilerinin İncelenmesi. Journal of the Institute of Science and Technology 8 2 215–221.
IEEE E. Yörük, Ö. Sefer, A. S. Sezer, Z. Konukcu, ve E. S. Develi, “Eugenol’ün Fusarium culmorum Üzerindeki Etkilerinin İncelenmesi”, Iğdır Üniv. Fen Bil Enst. Der., c. 8, sy. 2, ss. 215–221, 2018, doi: 10.21597/jist.428382.
ISNAD Yörük, Emre vd. “Eugenol’ün Fusarium Culmorum Üzerindeki Etkilerinin İncelenmesi”. Journal of the Institute of Science and Technology 8/2 (Haziran 2018), 215-221. https://doi.org/10.21597/jist.428382.
JAMA Yörük E, Sefer Ö, Sezer AS, Konukcu Z, Develi ES. Eugenol’ün Fusarium culmorum Üzerindeki Etkilerinin İncelenmesi. Iğdır Üniv. Fen Bil Enst. Der. 2018;8:215–221.
MLA Yörük, Emre vd. “Eugenol’ün Fusarium Culmorum Üzerindeki Etkilerinin İncelenmesi”. Journal of the Institute of Science and Technology, c. 8, sy. 2, 2018, ss. 215-21, doi:10.21597/jist.428382.
Vancouver Yörük E, Sefer Ö, Sezer AS, Konukcu Z, Develi ES. Eugenol’ün Fusarium culmorum Üzerindeki Etkilerinin İncelenmesi. Iğdır Üniv. Fen Bil Enst. Der. 2018;8(2):215-21.

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