CLASS B TRICHOTHECENE CHEMOTYPING IN Fusarium SPECIES BY PCR ASSAY

Fusarium isolates are divided into three chemotypes according to produce of class B trichothecenes; 3-acetyldeoxynivalenol (3ADON) 15-acetyldeoxynivalenol (15ADON) and nivalenol (NIV) and 4-acetyldeoxynivalenol (NIV) chemotypes. In this study, chemotyping of seventeen isolates from Turkey and Iran belonging to F. graminearum, F. culmorum, F. poae and F. pseudograminearum species were carried out by polymerase chain reaction (PCR) analysis. While all F. culmorum and F. poae isolates determined as 3ADON, remaining F. graminearum and F. pseudograminearum isolates were either 3ADON or 15ADON chemotypes. A common band of 583 bp long DNA fragment was amplified in all of F. culmorum and F. poae, one F. pseudograminearum (21F) and four F. graminearum (14F, sh14, sh15, sh7) isolates with 3ADON chemotype. However, remaining two F. pseudograminearum and four F. graminearum isolates with 15ADON chemotype, yielded amplicons that of 863 bp. It was shown that 3ADON was more predominant chemotype from other class B trichothecenes. This is the first report on chemotyping of F. poae and F. pseudograminearum isolates and also to show presence of 3ADON chemotype in F. graminearum isolate from Turkey

GENETIC DIVERSITY AMONG FUSARIUM GRAMINEARUM AND F. CULMORUM ISOLATES BASED ON ISSR MARKERS

To characterize the isolates of F. graminearum and F. culmorum fungi from Turkey and Iran, we performed ISSR analysis with 18 non-anchored and 23 anchored (including ten novel) primers. Amplification product sizes were 0.2-3.5 kb. In total, 405 bands were scored, 24 of which (5.92%) were polymorphic. The similarities among F. graminearum isolates were calculated as 62.3-99%, and among F. culmorum as 65.7-94.3%. Moderate genetic variation at intra-and inter-specific levels was determined, and the average intraspecific genetic diversity values were 80.65% for F. graminearum, and 80% for F. culmorum. Cluster analysis separated the isolates into two main clades. Group I consisted of F. culmorum isolates from Turkey that produced DON mycotoxin. Group II contained all F. graminearum isolates that were deoxynivalenol (DON) and nivalenol (NIV) chemotypes from Turkey and Iran. Both groups I and II were divided into two subgroups including their divisions. Phenons in group II included isolates distributed in the same geographic region. ISSR markers clustered isolates within a definite order according to their species. Isolates from the same agro-ecological locations were also kept together in subdivisions. The novel ISSR markers developed for the first time in this study contribute to differentiating between Fusarium isolates according to their species and geographic regions

Expression Analysis of PKS13, FG08079.1 and PKS10 Genes in Fusarium graminearum and Fusarium culmorum

Background: Identification and quantification of mycotoxins produced by Fusarium species are important in controlling fungal diseases. Objectives: Potential of zearalenone, butenolide and fusarin C production was investigated in five Fusarium graminearum and five F. culmorum isolates at molecular level

The effects of polycyclic aromatic hydrocarbon compounds on antioxidant enzyme activities in Mytilus galloprovincialis Lamarck, 1819 from Istanbul Strait

This study aimed to determine the levels of various polycyclic aromatic hydrocarbon compounds in Mytilus galloprovincialis, seasonally obtained from nine stations in the Istanbul Strait, and their effects on antioxidant enzyme activities. Samples were sorted into two groups based on length as 3-5 cm and 5-7 cm, and all analyses were carried out separately for both groups. Salinity, dissolved oxygen, temperature and pH of sea-water at sampling stations were measured. Very high levels of PAHs were detected, especially in Spring and Summer, hence, the risk of consuming mussels in those seasons was indicated. Mean total PAH was highest in Summer (617,86 ng/g), followed by Spring, Winter and Autumn (536,77 ng/g, 293,34 ng/g, 203,70 ng/g, respectively). The level of Benzo(a)pyrene, potentially the most carcinogenic compound among PAHs, was four times higher than the allowed limits of the European Union. Also, the total level of four PAH compounds (Benzo(a)pyrene, Chrysene, Benz(a)anthracene, Benzo(b)fluoranthene), considered as an indicator of carcinogenic risk, was three times higher than the allowed limits. Catalase, peroxidase and superoxide dismutase enzyme activities were calculated. Mean values of catalase and peroxidase activities were highest in Autumn, followed by Summer, while superoxide dismutase activity was highest in Summer, followed by Autumn. The lowest level of enzyme activity for all three enzymes was in Spring. Positive correlations between enzyme activities and PAH compounds were observed only in Winter and Spring. Catalase was positively correlated with six different PAH compounds, peroxidase with five different and superoxide dismutase with two different PAH compounds. The highest number of negative correlations between enzyme activities and PAH compounds was detected for superoxide dismutase, which was negatively correlated with total PAH and seven different PAH compounds, most of which were observed in Autumn

Investigation of Camphor Effects on Fusarium graminearum and F. culmorum at Different Molecular Levels

Fusarium graminearum and F. culmorum are phytopathogens, which cause destructive diseases in cereals. Epidemics of these phytopathogens are caused by mycotoxin contamination and the reduction of crop quality. In this study, the alteration due to in vitro camphor treatment on F. culmorum 9F and F. graminearum H11 isolates was investigated in terms of epigenetic, cellular, and transcription levels. Camphor with different concentrations (0.2, 0.4, 0.8, 1, 2, and 4 mu g/mu L) was applied to potato dextrose agar (PDA) growth media. The minimum inhibitory concentration (MIC) and the half maximal inhibitory concentration (IC50) were calculated as 2 and 1 mu g/mu L respectively. hog1, mst20, CAT, POD, mgv1, stuA, and tri5 genes, which are related to various cellular processes and pathogenesis, were examined by qPCR assay. qPCR analysis showed that camphor treatment leads to the downregulation of tri5 expression but the upregulation of the remaining genes. Apoptosis and oxidative stress were confirmed via acridine orange/ethidium bromide (AO/EB) and dichlorofluorescin diacetate (DCF-DA) staining, respectively. Moreover, coupled restriction enzyme digestion-random amplification (CRED-RA) assay, used for DNA methylation analysis, was carried out to evaluate epigenetic alterations. The decrease in genomic template stability (GTS) values, which resulted due to the alterations in random amplified polymorphic DNA (RAPD) profiles caused by camphor treatment, were detected as 97.60% in F. culmorum 9F and 66.27% in F. graminearum H-11. The outer and inner methylated cytosine profiles are determined by CRED-RA assay as type I-IV epigenetic alterations. The outcomes indicated that camphor could lead to alterations at several molecular levels of F. graminearum and F. culmorum