Kinga Treder

Development of an FgMito assay: A highly sensitive mitochondrial based qPCR assay for quantification of Fusarium graminearum sensu stricto

An ascomycete fungus, Fusarium graminearum sensu stricto (s.s.), is the major cause of Fusarium head blight (FHB), a devastating disease of cereals worldwide. The fungus contaminates crops with mycotoxins, which pose a serious threat to food and feed safety. In this study, we developed a highly sensitive mitochondrial based qPCR assay (FgMito qPCR) for quantification of F. graminearum s.s. To ensure high sensitivity of the assay, primers and a Minor-groove binding (MGB) probe were designed based on multi-copy mitochondrial DNA. The FgMito assay was successfully validated against a range of geographically diverse F. graminearum s.s. strains to ensure uniformity of the assay at an intraspecific level, as well as with other fungal species to ensure specificity. The assay was further evaluated in terms of efficiency and sensitivity against a test panel of different F. graminearum s.s. strains with various levels of pure fungal DNA and in the presence of wheat background DNA. The results showed a high efficiency of the assay developed, ranging from 93% to 101% with r(2)-values of >0.99. We further showed that three low concentrations of fungal template 2 pg, 0.6 pg and 0.2 pg could be reliably quantified in the presence of wheat background DNA. The FgMito assay was used to quantify F. graminearum s.s. DNA on 65 field samples from a range of hosts with defined levels of trichothecenes. We revealed a significant positive correlation between fungal DNA quantity and the sum of trichothecenes. Lastly, we showed a higher sensitivity of the FgMito assay than the nuclear based qPCR assay for F. graminearum s.s. by comparing Ct-values from both assays.

Development of a Highly Sensitive FcMito qPCR Assay for the Quantification of the Toxigenic Fungal Plant Pathogen Fusarium culmorum

Fusarium culmorum is a ubiquitous, soil-borne fungus (ascomycete) causing foot and root rot and Fusarium head blight on cereals. It is responsible for yield and quality losses as well as grain contamination with mycotoxins, which are a potential health hazard. An extremely sensitive mitochondrial-based qPCR assay (FcMito qPCR) for quantification of F. culmorum was developed in this study. To provide specificity, the FcMito assay was successfully validated against 85 F. culmorum strains and 53 isolates of 30 other fungal species. The assay efficiency and sensitivity were evaluated against different F. culmorum strains with various amounts of pure fungal DNA and in the presence of background wheat DNA. The results demonstrated the high efficiency of the assay (97.2–106.0%, R2-values > 0.99). It was also shown that, in the presence of background DNA, 0.01 pg of fungal template could be reliably quantified. The FcMito assay was used to quantify F. culmorum DNA using 108 grain samples with different trichothecene levels. A significant positive correlation was found between fungal DNA quantity and the total trichothecene content. The obtained results showed that the sensitivity of the FcMito assay was much higher than the nuclear-based qPCR assay for F. culmorum.

Ocena właściwości użytkowych zawiesinowego bionawozu fosforowego z popiołu. Cz. 2, Wpływ na akumulację biomasy i cechy fizjologiczne pszenicy jarej

Badania byly finansowane ze środkow grantu PBS 2/A1/11/2013, pt.: Odnawialne źrodla fosforu – bazą surowcową nowej generacji nawozow, przyznanego przez Narodowe Centrum Badan i Rozwoju.

Ocena właściwości użytkowych bionawozów fosforowych z popiołu i kości. Cz. 1, Wpływ na wybrane cechy morfologiczne oraz zdrowotność pszenicy

Badania byly finansowane ze środkow grantu PBS 2/A1/11/2013 „Odnawialne źrodla fosforu – bazą surowcową nowej generacji nawozow”, przyznanego przez Narodowe Centrum Badan i Rozwoju.

Evaluation of functional properties of ash and bone-based phosphorus biofertilizers. Part 1. Impact on selected morphological attributes and the health of spring wheat

Badania byly finansowane ze środkow grantu PBS 2/A1/11/2013 „Odnawialne źrodla fosforu – bazą surowcową nowej generacji nawozow”, przyznanego przez Narodowe Centrum Badan i Rozwoju.

Effect of a sowing regime and water conditions on nitrogen content and accumulation in the aerial biomass of spring barley (Hordeum vulgare L.) and Italian ryegrass (Lolium multiflorum Lam.)

Competitive interactions are common in plant communities, but the underlying mechanisms and effects of competition have not been fully elucidated to date. A pot experiment (3 series in 20092011) was conducted to evaluate the effect of a sowing method of spring barley and Italian ryegrass, and different water condition on the nitrogen (N) content and accumulation in the aerial biomass of plants at different stages. The experimental factors were as follows: 1) sowing regime – pure sowing and mixed sowing – spring barley undersown with Italian ryegrass, 2) water supply – plants supplied with water to meet their full requirements and 50% water supply reduction. The N content of the aerial biomass of plants was determined at five phenological development stages of spring barley grown in a pure stand under optimal soil moisture conditions: leaf development, tillering, stem elongation, heading and ripening. Nitrogen uptake was determined based on the N content and dry matter accumulation in plants. Nitrogen concentrations in the aerial biomass of barley and ryegrass varied throughout the growing season. Nitrogen content was particularly high in leaves, and it decreased steadily during plant development. Neither the sowing regime nor the water supply exerted significant effects on the N content of the aerial biomass of spring barley. In Italian ryegrass, under optimal water supply, mixed sowing with spring barley decreased N concentrations in shoots only during tillering. In spring barley, mixed sowing reduced N uptake by biomass, and the observed decrease was exacerbated under water deficit. In Italian ryegrass, reduced water supply decreased N uptake, but the effect of competitive interactions between the analyzed species was usually stronger than the influence of water deficit. Differences in N accumulation in aerial biomass were determined by the dr hab. inż. Marta K. Kostrzewska, Chair of Agroecosystems, University of Warmia and Mazury in Olsztyn, 10-718 Olsztyn, Pl. Łódzki 3, e-mail: marta.kostrzewska@uwm.edu.pl * The study was supported financially by the Ministry of Science and Higher Education in 2009-2012 (grant No. N N310 082836).