Investigation of the Effects of Eugenol on Fusarium culmorum

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.

Anahtar Kelimeler

• Antifungal,FcStua,FcMgv1,Linear growth rate,qPCR

Eugenol’ün Fusarium culmorum Üzerindeki Etkilerinin İncelenmesi

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.

Anahtar Kelimeler

• Antifungal,Doğrusal büyüme oranı,FcStua,FcMgv1,qPCR

Kaynakça

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