Comparative Analysis of Native <Emphasis Type="Italic">Crocus</Emphasis> Taxa as a Great Source of Flavonoids with High Antioxidant Activity

Native Crocus taxa are abundant in regions with moderate and dry climate; however, their potential in the food industry has thus far been neglected. The objective of this study was to compare the quantitative and qualitative flavonoid content and antioxidant and cytotoxic activity in organs of several populations of Croatian native taxa and assess their potential for application in the food industry. The dominant flavonoids in the native Crocus taxa were kaempferol and quercetin. Tepals contained similar or higher concentrations of flavonoid-glycosides than other organs. Tepals from Cvv1 and Cvv2 populations contained more flavonoid-glycosides than the commonly used spice saffron (C. sativus stigmas). The FRAP antioxidant activity of Cvv1 and Cvv5 tepals was similar to that of standard Trolox. DPPH inhibition of Cvv1 and Cvv3 tepals was within the range of that recorded for saffron. Cvv1 tepals significantly reduced reactive oxygen species in the broadest concentration range (50–1000 μg/ml), and showed considerable antioxidant activity in the ABTS assay, equal to 82% of standard Trolox antioxidant activity. A significantly higher concentration of kaempferol-rutinoside was recorded in this than in other taxa. The flavonoid showed a very strong or strong correlation with antioxidant assays results, and a negative correlation with cellular reactive oxygen species concentration. We therefore presumed that kaempferol-rutinoside is one of the main antioxidant phenolics in Crocus tepals. None of the tested extracts showed cytotoxicity toward Caco-2 cells. The results revealed that Cvv tepals have potential as a food supplement and are a promising material for further food safety tests.     

Differences in the crude protein fractions of lucerne leaves and stems under different stand structures

Protein degradability in forage legumes is of global importance because utilization efficiency of forage has economic and environmental consequences. However, there are no published studies on the effect of legume stand structure on differences in crude protein (CP) fractions. The main objective of the present research was therefore to investigate differences in CP fractions in leaves and stems of lucerne (Medicago sativa L.) during the growing season. Stand traits were measured over 2 years, and forage was sampled at the early bud and early flower stages in the first, second and third cuts. Stems had significantly higher concentrations (in g kg^?1 CP) of non‐protein (fraction A: 430 g kg^?1 CP) and indigestible nitrogen (fraction C: 92 g kg^?1 CP) than leaves and had lower relative content of true protein (fraction B: 478 g kg^?1 CP). In the total forage (stems and leaves combined), about 80% of the variation in CP fractions was explained by year, cut and maturity. Year was the most important factor, particularly for the B fractions. Cut was the second‐most important factor; its main effect was that the relative abundance of fraction A declined from 394 g kg^?1 CP in the first cut to 293 in g kg^?1 CP the third cut. Maturity increased the amounts of indigestible fraction C and protein fractions B₁ and B₃. This was associated with the leaf weight ratio, which had an inverse relationship with maximal stem length and dry matter yield. Variation partitioning showed that 75% of CP fraction variability associated with cut, maturity and year could be explained by the evaluated stand traits. This research has highlighted the need to consider plant morphological traits when legume CP fractions are evaluated.     

Soil chemical properties,plant species composition,herbage quality,production and nutrient uptake of an alluvial meadow after 45 years of N,P and K application

Little is known about the long-term effects of mineral N, P and K application on the nutritional status of mown alluvial grasslands. We asked how long-term fertilizer application affected soil chemical properties, plant species composition, herbage production, nutrient concentrations in soils and plants and balance of nutrients. Six treatments (control, PK, N50PK, N100PK, N150PK and N200PK) were investigated at the Černíkovice Experiment (Czech Republic) established in 1966 on an Alopecurus pratensis meadow, using annual application rates of 50, 100, 150 and 200 kg N, 40 kg P and 100 kg K ha⁻¹. Data were collected and analysed for 2007, 2008 and 2009. Although fertilizers had been applied over 45 years, differences in soil chemical properties between fertilization treatments were small. The legumes Lathyrus pratensis and Trifolium repens responded highly positively to PK application, and tall grasses, A. pratensis in particular, to NPK application. Herbage quality was high in terms of content of major nutrients, and its chemical properties varied considerably between treatments, cuts and years. Mean annual herbage yield ranged from 6·1 in the control to 9·7 t ha⁻¹ in the N200PK treatment. Herbage production was N-limited in 2007 and 2009, but not in 2008. Seasonal N agronomical efficiency ranged from 4·2 to 22·9 kg of DM herbage per kg of applied N. The herbage N:P and N:K ratios did not reflect the actual response of herbage production to N application. A negative balance between N applied and N removed in harvested herbage was recorded in all treatments. We concluded that in highly productive alluvial grasslands, mineral-rich soils can respond weakly to N, P and K application, fertilizer application modifies plant species composition and herbage production is not N-limited in all years. Nutrient ratios must be interpreted with caution for the estimation of nutrient limitation.     

Yearly variation in maize silage fermentation and nutritive quality

A significant effect of year on maize nutritive value has been reported in many studies; however, the direct effect of year on both nutritive values and fermentation traits of maize silage has not been directly investigated. During a 7‐year period (2006–2012), we investigated the following variables of 597 maize silages commercially produced in bunker silos on farms in the Czech Republic: dry‐matter content (DM), crude protein (CP), starch, amylase‐treated neutral detergent fibre (aNDF), acid detergent fibre (ADF) and net energy lactation (NEL), as well as fermentation traits such as pH, contents of lactic acid (LA), acetic acid (AA), butyric acid (BA) and the ratio between LA and volatile fatty acids. The effect of year and DM explained 16 and 30% of the variability in nutritive and fermentation traits respectively. Although a common silage DM content was used for each year, the climatic effect of year significantly changed almost all tested variables, but particularly aNDF, ADF, starch, AA and BA. The year contribution to trait variability effect was as much as two times higher for fermentation traits (12·6%) as for nutritive value (6·1%). Temperature–precipitation relationships associated with achieved maize yield were considered as factors explaining the year effect. Temperature and precipitation averaged over the full vegetation period had low relevance to nutritive and fermentation traits. This study reveals the importance of year‐to‐year variation in silage nutritive value and fermentation, which could be useful for understanding the contribution of year to these changes in both field experiments and national assessments.     

LAMP assay and rapid sample preparation method for on‐site detection of flavescence dorée phytoplasma in grapevine

In Europe the most devastating phytoplasma associated with grapevine yellows (GY) diseases is a quarantine pest, flavescence dorée (FDp), from the 16SrV taxonomic group. The on‐site detection of FDp with an affordable device would contribute to faster and more efficient decisions on the control measures for FDp. Therefore, a real‐time isothermal LAMP assay for detection of FDp was validated according to the EPPO standards and MIQE guidelines. The LAMP assay was shown to be specific and extremely sensitive, because it detected FDp in all leaf samples that were determined to be FDp infected using quantitative real‐time PCR. The whole procedure of sample preparation and testing was designed and optimized for on‐site detection and can be completed in one hour. The homogenization procedure of the grapevine samples (leaf vein, flower or berry) was optimized to allow direct testing of crude homogenates with the LAMP assay, without the need for DNA extraction, and was shown to be extremely sensitive.     

Characterization of Potato spindle tuber viroid (PSTVd) incidence and new variants from ornamentals

We analyzed the spreading and persistence of PSTVd variants in several ornamentals in the territory of the Czech Republic. The pool of PSTVd variants detected in Solanum jasminoides, S. muricatum, Datura sp. and Brugmansia sp. was biolistically transferred to Matricaria chamomilla, Argyranthemum frutescens and Diascia sp., species which we found as sensitive hosts for PSTVd from ornamentals. The PSTVd pool showed sequence changes and increased variation after its transfer to potato, suggesting a wide adaptation potential of PSTVd in this crop. Potato exhibited genotype-dependent leaf and spindle tuber symptoms, when inoculated with the sap from S. jasminoides infected with the predominant and sequence-stable PSTVd-S1.     

Phytosanitary status of grapevine in Montenegro

During 2006 and 2007, a survey on the incidence and distribution of fourteen grapevine viruses was carried out in the Skadar Lake basin, one of the two main grapevine‐growing areas of Montenegro. In total 165 samples were collected from four red (‘Vranac’, ‘Krato?ija’, ‘Merlot’ and ‘Cardinal’), two white (‘Chardonnay’ and ‘Rkaciteli’) and a few unknown grapevine varieties in the vicinity of Podgorica and Bar. The phytosanitary status of the collected samples was analysed by DAS‐ELISA and the presence of Grapevine fanleaf virus (GFLV), Grapevine leafroll‐associated virus 1 (GLRaV‐1), Grapevine leafroll‐associated virus 2 (GLRaV‐2) and Grapevine leafroll‐associated virus 3 (GLRaV‐3) was confirmed in some of them. The most frequently found virus in assayed samples was GLRaV‐3 (54.5%), followed by GFLV (23%), GLRaV‐1 (20%) and GLRaV‐2 (0.6%). These serological analyses showed the absence of Grapevine leafroll‐associated virus 6 (GLRaV‐6), Grapevine leafroll‐associated virus 7 (GLRaV‐7), Raspberry bushy dwarf virus (RBDV), Strawberry latent ringspot virus (SLRSV), Tomato ringspot virus (ToRSV), Raspberry ringspot virus (RpRSV), Arabis mosaic virus (ArMV), Tobacco ringspot virus (TRSV), Tomato black ring virus (TBRV) and Cherry leaf roll virus (CLRV) from all tested samples.