Genome sequencing and phylogenetic analyses of common and dwarf bunt of wheat provide insights into their genomic diversity and species boundaries, and enable the development of a detection assay for Tilletia controversa

The fungal genus Tilletia is generally recognized by the production of darkly pigmented teliospores, which replace mostly the host ovary. Currently, nearly 200 species of Tilletia species infecting Poaceae are included in this smut genus. Three species, namely T. caries, T. controversa, and T. laevis, cause economically important diseases of wheat in central Europe. Common bunt is caused by T. caries and T. laevis, whereas dwarf bunt is caused by T. controversa. The three species are described based on the morphology of their teliospores. However, they could not be reliably distinguished by using molecular phylogenetic analyses. To obtain deeper insights into the inter- and intraspecies genetic variation and to compare gene contents of these three species we sequenced and functionally and structurally annotated the whole-genomes of four strains of T. caries, five of T. controversa, and two of T. laevis. The obtained data was analysed together with five publicly available genomes (one T. caries, two T. controversa, and two T. laevis strains). In general, our findings demonstrated that the three species were highly similar with regards to genome size and predicted gene content. There was no evidence for expansion or decrease of transposable elements in any of the species. The nine predicted secondary metabolites gene clusters, 84% of the total carbohydrate active enzymes, 72% of secreted proteins, and 50% effector-like proteins were conserved across 16 studied strains. The species-specific proteins made only 0.1% of all predicted proteins, and their function were mainly unknown. In non-repetitive regions, the number of single nucleotide polymorphisms (SNPs) and small insertions or deletions (indels) were lowest within T. laevis (max. 0.52 SNPs/kb and 1.09 bp indels/kb), while they were highest within T controversa (max. 1.47 SNPs/kb and 2.48 indels bp/kb). We also observed extensive sequence identity between the two species of T. caries and T. laevis (0.51 SNPs/kb and 1.04 indels bp/kb on average). At the same time both species showed comparable distances to T. controversa. Accordingly, phylogenomic analysis of 241 protein coding genes revealed two groupings where isolates of T. caries and T. laevis were intermingled in a monophyletic group together, but separated from those of T. controversa, which formed another monophyletic group. Taken together these results suggest that T. caries and T. laevis have either diverged very recently or could be conspecific. These observations also correlate well with the fact that the two species are causing identical disease symptoms, need the same germination conditions, and have similar infection biology. These characteristics are different from those of T. controversa. Dwarf bunt is a quarantine pathogen in several countries. Consequently, its accurate identification is of high priority for plant health as well as to wheat exporters. The current international diagnostic protocol for detection of dwarf bunt in wheat seeds is based on a filtration method and the teliospores morphology. The method is however laborious and requires expert knowledge. To facilitate identification of T. controversa, a loop-mediated isothermal amplification (LAMP) assay was developed. To do this, the generated genomic data were extended further with publicly available genomic data from ten Tilletia isolates in order to identify DNA segments that were conserved in and unique to T. controversa. The developed assay was based on one of these genome regions. The assay specificity was validated against 223 fungal phytopathogens including 11 Tilletia species. The end-point colorimetric based detection LAMP assay had 5 pg limit of detection and showed 100% sensitivity and 97.7% specificity in an interlaboratory test performance study. The majority of smuts are reported to have relatively narrow host ranges including Tilletia species. Tilletia caries and T. controversa however are reported to infect different host species representing several grass genera. To clarify whether they are indeed generalist species with a broad host range or rather represent complexes of cryptic species with narrow host ranges phylogenetic relationships of those species and close relatives were inferred using sequences of three gene regions (ITS rDNA, EF1α, and RPBII). In total 70 specimens were analysed of which 20 specimens were newly produced for this study. In general, the multi locus phylogenetic analysis resolved various species with narrow host ranges parasitizing wild grasses as distinct lineages (i.e., the basal lineages from T. fusca up to T. olida representing 12 species and the crown lineage with T. bromi and T. puccinelliae, respectively). Several small clusters of T. controversa from wild grasses (Thinopyrum intermedium, Bromus marginatus, Agropyron cristatum) and rye (Secale cereale), respectively, clustered as subgroups in a polytomous manner between different clusters of T. caries, T. controversa and T. laevis on wheat. Interestingly, one group of T. controversa sequences obtained from Elymus repens, Th. intermedium, and Agropyron sp. clustered with high support values clearly separate from this polytomous group and together with T. brevifaciens isolates also obtained from Th. intermedium. These representatives of T. controversa with high likelihood represent at least one cryptic species restricted to these wild grasses as hosts and might potentially be conspecific with T. brevifaciens. In the latter case, T. brevifaciens would represent a species with board host range.2021-11-3

Selenized yeast in production of selenium enriched Pleurotus ostreatus mushroom with good flavour

The aim of this study was to investigate the potential influence of selenized yeast (Sel Plex, Alltech Inc., Lexington, USA) on chemical composition and flavour of solid state grown mushroom Pleurotus ostreatus. Amino acid composition that influences the flavour of selenium-enriched P.ostreatus P80 (137.84 ppm of selenium in d.w.) and non enriched cultivated strains with particular emphasis on selenomethionine was determined by HPLC method, after complete hydrolysis. Volatile flavour compounds of mushroom cultures were analyzed by GC-MS using Headspace sampler. In mushrooms with high selenium content, selenium in the form of L-selenomethionine was present. High selenium concentration in fruit body did not significantly change the amino acid composition of mushrooms. The major amino acids of fruit body sample were glutamic acid, alanine, aspartic acid and tryptophan. In the fruit body of P.ostreatus P80 control, 30 volatile compounds were detected, and in selenium enriched sample number of detected compounds was 25. Compounds that were detected in control sample, but not in eriched fruit body are 2-amino-N-ethylpropanamide, 2-methylpropyl pentan-2-yl sulfite, 2,3-pentanedione, heptan-2-one, pentan-1-ol, 2-methyldihydrofuran-3(2H)-one,1-hydroxyacetone, 3-hydroxy-2-butanon, 2,5-dimethylpyrazine, 2,6-dimethylpyrazine and benzaldehyde. Compounds that were detected in enriched P.ostreatus P80, but not in control are acetone, ethylacetate, 2-propanol, acetonitrile, 2-methylbenzaldehyde and 2-hydroxyethylmethacrylate. Selenized yeast presents good source of selenium for selenium enriched mushroom production with good flavour, which is probably safety for consumption

Water productivity indices of the soybean grown on silty clay soil under sprinkler irrigation

The objective of this research was to compare the effects of different irrigation treatments on soybean [Glycine max (L.) Merr.] productivity and water use efficiency on experimental fields of the Maize Research Institute of Zemun Polje(Serbia), in 2007 and 2008. Four irrigation levels were investigated: full irrigation (I100), 65% and 40% of I100 (I65 and I40) and a rain-fed (I0) system. The crop water use efficiency (CWUE, also known as crop water productivity –CWP), irrigation water use efficiency (IWUE) and evapotranspiration water use efficiency (ETWUE) were used to assess the water productivity of each studied treatment. The efficiency of the same treatment differed between the years as it depended on seasonal water availability, weather conditions and their impact on seed yields. Maximum and minimum yields were obtained in the I65 and I0 treatments, averaging 3.41 t ha–1 and 2.26 t ha–1, respectively. Water use efficiency values were influenced by the irrigation levels. In general, CWUE values increased with the increased level of irrigation. In both growing seasons, IWUE and ETWUE decreased with increasing the seasonal water consumption and irrigation depth. On average, treatments I40 and I65 resulted in similar or higher CWUE and ETWUE than I100, in both growing seasons. I65 resulted in the highest IWUE, averaged over the two seasons, while I100 had the lowest IWUE. I65 could be proper for the soybean irrigated in Vojvodina when there is no water shortage and I45 could be used as a good basis for reduced sprinkler irrigation strategy development under water shortage

Effects of Irrigation Rate and Planting Density on Maize Yield and Water Use Efficiency in the Temperate Climate of Serbia

Scarce water resources severely limit maize (Zea mays L.) cultivation in the temperate regions of northern Serbia. A two-year field experiment was conducted to investigate the effects of irrigation and planting density on yield and water use efficiency in temperate climate under sprinkler irrigation. The experiment included five irrigation treatments (full irrigated treatment – FIT; 80% FIT, 60% FIT, 40% FIT, and rainfed) and three planting densities (PD1: 54,900 plants ha–1 ; PD2: 64,900 plants ha–1; PD3: 75,200 plants ha–1). There was increase in yield with the irrigation (1.05–80.00%) as compared to the rainfed crop. Results showed that decreasing irrigation rates resulted in a decrease in yield, crop water use efficiency (WUE), and irrigation water use efficiency (IWUE). Planting density had significant effects on yield, WUE, and IWUE which differed in both years. Increasing planting density gradually increased yield, WUE, and IWUE. For the pooled data, irrigation rate, planting density and their interaction was significant (P < 0.05). The highest two-year average yield, WUE, and IWUE were found for FIT-PD3 (14,612 kg ha–1), rainfed-PD2 (2.764 kg m–3), and 60% FITPD3 (2.356 kg m–3), respectively. The results revealed that irrigation is necessary for maize cultivation because rainfall is insufficient to meet the crop water needs. In addition, if water becomes a limiting factor, 80% FIT-PD3 with average yield loss of 15% would be the best agronomic practices for growing maize with a sprinkler irrigation system in a temperate climate of Serbia

The frequency of falls in children judo training

Purpose: Falling techniques are inseparable part of youth judo training. Falling techniques are related to avoiding injuries exercises (Nauta et al., 2013). There is not good evidence about the ratio of falling during the training in children. Methods: 26 children (age 8.88±1.88) were video recorded on ten training sessions for further indirect observation and performance analysis. Results: Research protocol consisted from recording falls and falling techniques (Reguli et al., 2015) in warming up, combat games, falling techniques, throwing techniques and free fighting (randori) part of the training session. While children were taught almost exclusively forward slapping roll, backward slapping roll and sideward direct slapping fall, in other parts of training also other types of falling, as forward fall on knees, naturally occurred. Conclusions: Judo coaches should stress also on teaching unorthodox falls adding to standard judo curriculum (Koshida et al., 2014). Various falling games to teach children safe falling in different conditions should be incorporated into judo training. Further research to gain more data from groups of different age in various combat and non-combat sports is needed

Fear of crime and victimization among the elderly participating in the self-defence course

Purpose. Self-defence training could enhance seniors´ defensive skills and fitness. There is lack of evidence about fear and concerns of seniors participating in the self-defence course. Methods. 18 elderly persons (16 female, 1 male; age 66.2, SD=5.86) participated in the self-defence course lasting 8 training units (each unit 60 minutes). Standardized tool for fear of crime and victimization analysis previously used in Euro-Justis project in the Czech Republic (2011) was used in pretest and posttest. Results. We explored the highest fear of crime by participants in their residence area after dark (mean=2,77; median=3; SD=0,80), lower fear at the night in their homes (mean=2,29; median=2; SD=0,75) and in their residence area at the daytime (mean=2,00; median=2; SD=0,77) at the beginning of the course. We noticed certain decrease of fear of crime after the intervention. Participant were less afraid of crime in their residence area after dark (mean=2,38; median=2; SD=0,77), they felt lower fear of crime at the night in their homes (mean=2,00; median=2; SD=0,48) and in their residence area at the daytime (mean=1,82; median=2; SD=0,63). Conclusions. The approach to self-defence teaching for elderly should be focused not just on the motor development, but also on their emotional state, fear of crime, perception of dangerousness of diverse situations and total wellbeing. Fear of crime analysis can contribute to create tailor made structure of the self-defence course for specific groups of citizens

Optimization of Operation Sequencing in CAPP Using Hybrid Genetic Algorithm and Simulated Annealing Approach

In any CAPP system, one of the most important process planning functions is selection of the operations and corresponding machines in order to generate the optimal operation sequence. In this paper, the hybrid GA-SA algorithm is used to solve this combinatorial optimization NP (Non-deterministic Polynomial) problem. The network representation is adopted to describe operation and sequencing flexibility in process planning and the mathematical model for process planning is described with the objective of minimizing the production time. Experimental results show effectiveness of the hybrid algorithm that, in comparison with the GA and SA standalone algorithms, gives optimal operation sequence with lesser computational time and lesser number of iterations