Effects of Growth Conditions on tri4 Gene Expression in Fusarium culmorum

Year 2017, , 91 - 97, 09.09.2017

Emre Yörük Aylin Gazdağlı Gülruh Albayrak

https://doi.org/10.13002/jafag1079

Abstract

Expression of tri4, found in the tri5 gene cluster, is essential for DON production. In this study, effects of different growth conditions on tri4 expression, as well indirectly on DON production, were investigated in F15 isolate of Fusarium culmorum via qPCR (real time polymerase chain reaction). Control group was grown on potato dextrose agar (PDA) at 25°C (pH 5.6). The effects of pH 3.0 and -7.0 were examined on cultures grown at 25°C. Moreover, 0.5 mM hydrogen peroxide (H2O2) was concurrently added to medium. High quality (A260/280= 1.9-2.0) and quantity (2-3µg/µL) of total RNAs were isolated from all groups. β-tubulin expression was used as internal control and relative quantification values were recorded. tri4 expression was detected in all experiments except F15 grown on pH 3.0. Cp values were calculated as 22.26±1.14-26.84±4.79. tri4 expression levels in experiments were lower than control. Their ΔΔCT and 2-ΔΔCT values were 0-5.54 and 0-0.582, respectively. While maximum tri4 expression was recorded in control, minimum expression was detected in the conditions consisting of pH 5.6 and at 15°C. Findings showed that different pH and temperature values and supplementation of H2O2 resulted in decreasing of tri4 expression. Also, it was detected that acidic pH was a potential repressor for DON production. Findings support the importance of kit development requirement for mycotoxin detection based on gene expression analysis in the field or harvested crops.

Keywords

Fusarium culmorum, qPCR, tri4 gene expression

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