Germination of Fusarium graminearum Ascospores and Wheat Infection are Affected by Dry Periods and by Temperature and Humidity During Dry Periods.

The effects of temperature and relative humidity (RH) on germination of Fusarium graminearum ascospores, and of dry periods (DP) of different lengths and of temperature and RH during DP on ascospore survival were studied both in vitro and in planta. Optimal temperatures for ascospore germination at 100% RH were 20 and 25°C; germination was ≤5% when ascospores were incubated at 20°C and RH ≤ 93.5%. Viable ascospores were found at all tested combinations of DP duration (0 to 48 h) × temperature (5 to 40°C) or RH (32.5 to 100% RH). Germination declined as DP duration and temperature increased. Germination was lower for ascospores kept at 65.5% RH during the DP than at 76.0, 32.5, or 93.5% RH. Equations were developed describing the relationships between ascospore germination, DP duration and temperature or RH during DP. Durum wheat spikes were inoculated with ascospores and kept dry for 0 to 48 h at approximately 15°C and 65% RH; plants were then kept in saturated atmosphere for 48 h to favor infection. Fungal biomass, measured as F. graminearum DNA by quantitative polymerase chain reaction, declined as DP increased to 24 and 48 h at 3 and 9 days postinfection but not in spikes at maturity.

Influence of temperature on infection, growth, and mycotoxin production by Fusarium langsethiae and F. sporotrichioides in durum wheat.

Information concerning the temperature requirements of the species causing Fusarium head blight of small grains is essential for understanding which species cause the disease in different areas and years, for developing weather-driven disease models, and for predicting mycotoxin type and quantity in kernels. The optimal temperature range for growth was 20-25 °C for Fusarium langsethiae and 25-30 °C for F. sporotrichioides, and the optimum for production of both T-2 and HT-2 toxins was 15 °C for F. langsethiae and 10-15 °C for F. sporotrichioides. Floret infection occurred from 10 to 40 °C for F. sporotrichioides (69.8% average incidence of infected florets) and from 10 to 35 °C for F. langsethiae (17.6% of infected florets). The optimal temperature for spike colonisation was 25 °C for F. langsethiae and 30 °C for F. sporotrichioides, and the optimal temperature range for mycotoxin production was 15-35 °C for F. langsethiae and 20-25 °C for F. sporotrichioides. The quantity of fungal DNA in inoculated spikes was 5.5-times greater for F. sporotrichioides than for F. langsethiae; F. langsethiae DNA was first detected 2 days post-inoculation (dpi), and F. sporotrichioides DNA was first detected 4 dpi. Toxins were first detected 4 and 2 dpi for F. langsethiae and F. sporotrichioides, respectively.

In vitro antifungal activity of terpinen-4-ol, eugenol, carvone, 1,8-cineole (eucalyptol) and thymol against mycotoxigenic plant pathogens.

The aim of this study was to examine the effect of five naturally occurring compounds from essential oils on 10 different species of mycotoxigenic fungi involved in several plant diseases. The antifungal activities of terpinen-4-ol, eugenol, carvone, 1,8-cineole (eucalyptol) and thymol were observed in vitro on Fusarium subglutinans, Fusarium cerealis, Fusarium verticillioides, Fusarium proliferatum, Fusarium oxysporum, Fusarium sporotrichioides, Aspergillus tubingensis, Aspergillus carbonarius, Alternaria alternata and Penicillium sp. The naturally occurring compounds tested showed toxic effects on in vitro mycelium growth of all fungal species but with different level of potency. The results are encouraging for further investigations of in planta antifungal activities of these essential oils components.

Identification and validation of reference genes for gene expression studies in water buffalo.

In gene expression analysis, a key step to obtain informative data from reverse transcription quantitative PCR (RT qPCR) assay is normalization, that is usually achieved by ratio to correct the abundance of the gene of interest against that of an endogenous reference gene. The finding of such reference genes, ideally expressed in a stable way in multiple tissue samples and in different experimental conditions, is a non-trivial problem. In this work, a set of genes potentially useful as reference for gene expression studies in water buffalo has been identified and evaluated. In the first step, a publicly available Bos taurus expressed sequence tags database has been downloaded from the TIGR Gene Index and mined by some simple frequency algorithms to find out which tentative consensuses are present in a remarkable number of different cDNA libraries and, consequently, are more suitable to be included in a starter set of candidate reference genes. To validate the potential of such candidates for their use as normalizers in buffalo gene expression analysis, an RT qPCR analysis has been carried out, in which the expression stability of these genes has been evaluated on a panel of buffalo tissues and organs. Our results indicate that ribosomal proteins L4 and L5 and Gek protein encoding genes can be useful as normalizers to compare gene expression levels across tissues and organs in buffalo.

Assessment of Fusarium infection in wheat heads using a quantitative polymerase chain reaction (qPCR) assay.

The accuracy of a quantitative polymerase chain reaction assay in quantifying the DNA of trichothecene-producing F. culmorum and F. graminearum within harvested wheat grains and head tissue was evaluated in comparison with incidences of infected kernels and deoxynivalenol levels. In a first experiment, six durum and bread wheat varieties were grown in randomized plots for a 2-year period, and inoculated with Fusarium macroconidia at six growth stages between heading and dough ripening, to obtain a wide range of Fusarium head blight incidences. There was a close relationship between fungal DNA and the amount of deoxynivalenol, and this relationship was consistent over Fusarium species, wheat species and varieties, and over a wide range of Fusarium head blight infection. In a second experiment potted wheat plants were grown under environmentally controlled conditions and inoculated with the two Fusarium species at full flowering; head samples were collected before inoculation and after 6 h to 12 days, and processed by the quantitative polymerase chain reaction assay. This assay made it possible to detect the dynamic of fungal invasion in planta after infection had occurred, and to single out the presence of infection before the onset of the disease symptoms: A robust detection of the infection occurred within 18-24 h for F. culmorum, and within 2-9 days for F. graminearum.

In vitro antifungal activity of the tea tree (Melaleuca alternifolia) essential oil and its major components against plant pathogens.

AIMS: The aim of this study was to examine the effect of Melaleuca alternifolia essential oil (TTO) and its principal components on four cereal-pathogenic fungi. METHODS AND RESULTS: The antimycotic properties of TTO and of terpinen-4-ol, gamma-terpinen and 1,8-cineole (eucalyptol) were evaluated in vitro on Fusarium graminearum, Fusarium culmorum and Pyrenophora graminea. Moreover, barley leaves infected with Blumeria graminis were treated with whole TTO. All the tested fungi were susceptible to TTO and its components. CONCLUSIONS: TTO exerted a wide spectrum of antimycotic activity. Single TTO purified components were more active than the whole oil in reducing in vitro growth of fungal mycelium and, among the tested compounds, terpinen-4-ol was the most effective. SIGNIFICANCE AND IMPACT OF THE STUDY: TTO and its components can be considered potential alternative natural fungicides.

From single genes to co-expression networks: extracting knowledge from barley functional genomics.

The paper reports an 'in silico' approach to gene expression analysis based on a barley gene co-expression network resulting from the study of several publicly available cDNA libraries. The work is an application of Systems Biology to plant science: at the end of the computational step we identified groups of potentially related genes. The communities of co-expressed genes constructed from the network are remarkably characterized from the functional point of view, as shown by the statistical analysis of the Gene Ontology annotations of their members. Experimental, lab-based testing has been carried out to check the relationship between network and biological properties and to identify and suggest effective strategies of information extraction from the network-derived data.