Exploitation of epigenetic variation of crop wild relatives for crop improvement and agrobiodiversity preservation

Crop wild relatives (CWRs) are recognized as the best potential source of traits for crop improvement. However, successful crop improvement using CWR relies on identifying variation in genes controlling desired traits in plant germplasms and subsequently incorporating them into cultivars. Epigenetic diversity may provide an additional layer of variation within CWR and can contribute novel epialleles for key traits for crop improvement. There is emerging evidence that epigenetic variants of functional and/or agronomic importance exist in CWR gene pools. This provides a rationale for the conservation of epigenotypes of interest, thus contributing to agrobiodiversity preservation through conservation and (epi)genetic monitoring. Concepts and techniques of classical and modern breeding should consider integrating recent progress in epigenetics, initially by identifying their association with phenotypic variations and then by assessing their heritability and stability in subsequent generations. New tools available for epigenomic analysis ofer the opportunity to capture epigenetic variation and integrate it into advanced (epi)breeding programmes. Advances in -omics have provided new insights into the sources and inheritance of epigenetic variation and enabled the efcient introduction of epi-traits from CWR into crops using epigenetic molecular markers, such as epiQTLs

Determination of Population Structure of Wheat Core Collection for Association Mapping

The microsatellites, as one of the most robust markers for identification of wheat varieties, were used for assessment of genetic diversity and population structure to promote effective use of genetic resources. In this study, the set of 284 wheat varieties were genotyped using 30 microsatellite markers. The chosen SSR markers were located among almost all linkage groups and covered all three genomes. The genotypes used originate from 24 different breeding centers worldwide and are included in an extensive core collection of the Institute of Field and Vegetable Crops in Novi Sad, Serbia. The total number of detected alleles was 349 at all analyzed loci. The average number of detected allelic variant per locus was 11.5. The mean value of polymorphic information content was 0.68. According to the probability of data obtained by program Structure, the results have shown presence of 6 subpopulations within the studied set of genotypes. The population structure positively correlated to some extent with geographic origin. The available pedigree data were included for additional explanation of population structure. The results of this study should provide valuable information for future association studies using the diverse wheat breeding material

Targeted plant improvement through genome editing: from laboratory to field

This review illustrates how far we have come since the emergence of GE technologies and how they could be applied to obtain superior and sustainable crop production. The main challenges of today's agriculture are maintaining and raising productivity, reducing its negative impact on the environment, and adapting to climate change. Efficient plant breeding can generate elite varieties that will rapidly replace obsolete ones and address ongoing challenges in an efficient and sustainable manner. Site-specific genome editing in plants is a rapidly evolving field with tangible results. The technology is equipped with a powerful toolbox of molecular scissors to cut DNA at a pre-determined site with different efficiencies for designing an approach that best suits the objectives of each plant breeding strategy. Genome editing (GE) not only revolutionizes plant biology, but provides the means to solve challenges related to plant architecture, food security, nutrient content, adaptation to the environment, resistance to diseases and production of plant-based materials. This review illustrates how far we have come since the emergence of these technologies and how these technologies could be applied to obtain superior, safe and sustainable crop production. Synergies of genome editing with other technological platforms that are gaining significance in plants lead to an exciting new, post-genomic era for plant research and production. In previous months, we have seen what global changes might arise from one new virus, reminding us of what drastic effects such events could have on food production. This demonstrates how important science, technology, and tools are to meet the current time and the future. Plant GE can make a real difference to future sustainable food production to the benefit of both mankind and our environment.European Cooperation in Science and Technology (COST) CA18111info:eu-repo/semantics/publishedVersio

High-performance liquid chromatography determination of tocopherols in wheat bran

Natural vitamin E is significant phytochemical compound with antioxidant activity and potential benefit for human health. The physiological role of vitamin E is in its ability to react with free radicals in cell membranes and preventing lipid peroxidation. Vitamin E is composed of eight different structurally related compounds, vitamers (a-, B-, y- and 65-tocopherols and a-, B-, y- and otocotrienols) which are known to occur in nature. Among cereals, wheat grain and wheat bran represent rich sources of tocopherols. The aim of this study was to investigate tocophérols composition (a-, B-, y- tocopherols) in selected 20 wheat genotypes. The samples consisted the bran portion of the outer layer of wheat grain that were the by-product of milling on the MLU 202 laboratory flour mill. Tocopherols were determined in the wheat bran samples by using the simple analytical method which includes extraction with hexane and normalphase high-performance liquid chromatography with fluorescence detection. The mobile phase consisted a mixture of hexane:ethyl acetate (70:30, viv). Total tocopherols content in wheat bran extracts ranged from 28.08 in cv. Partizanka to 107.49 mg/kg in cv. Skopljanka The average contents of a-, B- and ytocopherols were in the range from 3.16 (a-tocopherol in cv. Partizanka) to 81.04 mg/kg (y- tocopherol in cv. Skopljanka). Major vitamers for those tested genotypes were a- and y- tocopherols. The results have shown that there were significant differences in tocopherols composition and content among the genotypes, indicating the great genetic potential for improvement. ords: Sulfor. finociception

Laboratory simulated dissipation of rimsulfuron, prosulfuron and oxasulfuron in soil

Rimsulfuron, prosulfuron and oxasulfuron are a group of sulfonylurea herbicides that have been widely used for controlling weeds in a variety of vegetables and other crops. Because of the high herbicidal activity of this sulfonylureas, they are beneficial at low application rates which are approximately 1000 times less than other of convencional herbicides. Hence the using of this herbicides is increasing constantly worldwide. The fate of rimsulfuron, prosulfuron and oxasulfuron in soil type Calcic Chernozem Clayic. Pachic (according to the FAO-WRB classification) were investigated using ultra-performance liquid chromatography with diode array detection for herbicide detection. Their dissipation behaviour in soil were evaluated under laboratory incubated condition at 70% water holding capacity of soil and two temperature (25°C and 30°C) were maintained in termostat. In order to investigate dissipation selected sulfonylureas were used as aqueous solution of commercial formulation herbicides. The recommended dosage of rimsulfuron, prosulfuron and oxasulfuron were 50, 20 and 80 g active ingredient (a.i.) per hectare, respectively. Representative soil samples were collected after 2 hours, and then after 2, 6, 15, 30 and 50 days after the application of herbicides. Initial residues of all investigated herbicides were higher at temperature 25°C than 30°C. The residues of rimsulfuron on both analysed temperature for tested dose were less than LOQ (3.04 ug kg!) after 2 days and 6 days for prosulfuron (2.97 yg kg!) and oxasulfuron (3.40 pg kg). The dissipation of this three sulfonylureas in studied soil was described by using the Mittag-Leffler function ¢ E, (-bt). Model coefficients a, b, c were received from the experimental data by using a fitting procedure. Rimsulfuron and prosulfuron showed faster degradation in tested soil with half lives of 0.49 and 0.03 days at temperature 30°C, respectively. The degradation rate of oxasulfuron appeared something slower with half lives of 0.64 day at temperature 25°C. These results indicate how herbicides belonging to the identical chemical class can behave in various way concerning dissipation in soil

Ekstrakcija nikosulfurona iz zemljišta

Zemijište predstavlja glavni recipijent u kome se adsorbuju i razgrađuju pesticidi, koji u njega dospevaju direktnim putem, kao posledica kontrolisane primene pri suzbijanju korova. Nikosulfuron je selektivni sistematični herbicid, apsorbuje se listom i korenom, relativno brzo se prenosi do meristemskih tkiva. Ovaj herbicid inhibira sintezu acetolaktat sintetazu (ALS), ključni enizim koji učestvuje u biosintezi amino-kiselina sa razgranatim alifatičnim nizom (valin, leucin i izoleucin) u biljkama i zaustavlja deobu ćelija i rast biljke. Nikosulfuron se primenjuje nakon nicanja kukuruza za suzbijanje jednogodišnjih i višegodišnjih uskolisnih i širokolisnih travnih korova. U poređenju sa tradicionalnim herbicidima koji se primenjuju u količini >1kg/ha, nikosulfuron je efikasan u malim količinama <100 g/ha. Postojanost nikosulfurona u zemljištu zavisi od pH, temperature, vlažnosti zemljišta i sadržaja organske materije u zemljištu. S porastom temperature ubrzava se razlaganje nikosulfurona, kako pod uticajem hemijskih faktora (hidroliza, oksidacija) tako i pod uticajem mikroorganizama

Disappearance of nicosulfuron residues in soil under laboratory conditions

Sulfunyluares (SUs) present a new generation of herbicides which selectively control broad- and narrow-leaved weeds. Nonethel iiened utilization of SUs leads to increased concern about herbicides residues as the « msequence of their prolonged persistence in some soils. Nicosulfuron belongs to the group of SUs herbicide. A laboratory experiment at termostat (temperature of 25°C) was carried out to evaluate the effect of temn< ‘ture on the dissipation dynamics and final residues of nicosulfuron in soil type Calcie Chernozem Clayic, Pachic. The dissipation of nicosulfuron in soi! was described by using the Mittag-Leffler function ¢ E, (-hr), ere obtained from the experimental data. The residues of nicosulfuron iphy with diode array detection in the cted herbicide was applied as aqueous solution of commercial formulation nicosulfuron (K elvin", 40 ¢ L'). The starting coneentrations of nicosulfuron (measured in the samples soil taken on the ( th day, ! houirs afier the treatment) Sulfunyluares (SUs) pre 1 new Model coefficients a, h. « v were determined by uitra-performance liquid chromatog ultraviolet region. In order to invest gate dissipation rye cs on 25°C were 389.84 ug ke ‘9.18 = : | ee E ¢ coe . pg kg, 649.02 ug kg 'and 797.69 ug kg! after the treatment with 40 grams active ingredient (a.i.) per hectare of nicosulfuron ig ai. ha'), 50 gal. ha”, 80.g ai. ha” and 100 gai. ha’. respectively. Atier two days, residues of nicosulfuron were lower more than 86% compared with the initial concentra ‘on. Pifty aays aficr the application of all doses the content of residues were below the LOQ for nicosulfuron (3.16 Hg kg'). The obtained half-time degradation (DT:») values of nicosulfuron vere in range of 0.34 d (80 g a.i. ha’ ') to 0.68 d (40 £att Significant reater Cissipation of nicosulfuron observed in this laboratory testing was influenced, most probably, by abiouc and biotic processes. Nicosulfuron poses low risks to the . imilar type soil because of their rapid dissipation

Validation of the method for the determination of dithiocarbamates in fruits

Residues of active substances, which were frequently found in fruits and vegetables of market producers belonged to the dithiocarbamate group pesticide. According to their use dithiocarbamates belong to fungicides in terms dithiocarbamic acid. They are a group of compounds that exist as strong complexes with various metal ions, often in a polymeric form. This makes them difficult to analyse directly because of their limited solubility in most organic solvents. It is important to have a reliable method which enables the quick and simple detection of this group for official monitoring. This work reports a simple, rapid and sensitive method for the assessment residues dithiocarbamates in fruits. Dithiocarbamates were determined indirectly by measuring the amount of carbon disulfide (CS>) that is liberated by the chemical reaction. The fruit samples in undisturbed condition were heated with a solution of stannous (II) chloride and hydrochloric acid yielding carbon disulphide. Incurred carbon disulfide (CS) are determined "head-space" technique of gas-chromatography with mass detector. Linearity was verified by using the solutions of carbon disulphide in acetone. A linear dynamic range was obtained over a range of concentrations from 0.02 to 0.12 mg/kg for carbon disulfide with correlation coefficient r > 0.995. The accuracy of the method was acceptable since the average recoveries measured at four fortification levels were in the range of 83-103% (n = 4). The precision of the developed procedure expressed as the relative standard deviations (RSDs) were lower than 3.7% in all cases. Quantification was based on external standard calibration curves made with spiked blankmatrices

Relationships among oil content, protein content and grain yield in wheat (Triticum aestivum, L.)

Assessment of health-beneficial chemicals in wheat grain and oran, such as proteins and oil is important for the breeding due to increased consume: cernands ior healthier food. Also, it is very important to know their relationships with the grein yield and to what extent they could be modified by growing conditions. The objective of this study was to investigate oil and protein contents, as well as grain yield of 25 winter wheat (Triticum aestivum, L.) varieties and to establish correlations among these traits in different environments. The field studies were performed on three different experimental sites in Serbia during two growing seasons (2009/10 and 2010/11). The investigated locations representing Serbia’s most important wheat-growing areas: Rimski Sancevi (45°20’N, 19°51'F 37 m altitude), Sremska Mitrovica (46°06'N, 19°33'E, 83 m altitude) and Pancevo (44°50 N, 20°40°E, 76 m altitude). The oil was extracted from wheat bran obtained by laboratory mi! MLU 202. Classical Rushkovsky method was used to determine oil content, while protein content was determined by the ICC 105/2 method. Significant variability was found among the genotypes for all analysed traits. In the six environments, the oil content varied from 2 “> 5.6%. with the average value of 3.8% and coefficient of variation (CV) 15.7%. The protein content ranged from 11.2% to 17.8% with the CV of 5.4%, while the grain yield varied fram 2.3 to 9.8 t/ha, with the CV of 17.8%. The average genotypic values for all environments have shown negative correlation between protein content and grain yield (r=-0.739**). This correlation was highly significant in three (E3, E4 and £6) out of six environments, while significant positive correlation between oil content and grain yield (r=0.441*) was found only in Ee. The established correlations between the traits are highly environmentally dependant and co. » modified by variable growing conditions. It will be very valuable for breeding process to identufy climatic variables and growing conditions that modify unwanted correlations, 2s well as to identify genotypes in which these correlations are not expressed !n orce iow successful breeding for improvement of both traits at the same time. For positive correlations it is also useful to know the favourable conditions in which they can be maximiz

Dissipation of nicosulfuron and oxasulfuron in soil under field condition

Sulfonylureas represent a major advancement in global crop protection technology and they have revolutionized weed control by introducing a unique mode of action. These herbicides have low toxicity to mammals while they are highly toxic to plants. The fate of sulfonylurea herbicides is directly related to their chemical structure and mainly to the ionisation of the sulfonylurea bridge. The dissipation behavior of nicosulfuron and oxasulfuron in soil were investigated. Soil was cored on experimental site Rimski Sanéevi in Serbia four times (0-30 days after the spray). The residue levels and dissipation rates of nicosulfuron and oxasulfuron in soil were determined by liquid chromatography with diode array detection. The pesticide formulations were applied in two different rates for nicosulfuron (50 g ai ha-1 and 100 g ai ha Nicogan 40SC) and oxasulfuron (80 g ai ha’' and 160 g ai ha’' Dynox). The fortified recoveries were in the following range for nicosulfuron from 85.90% to 98.71% with relative standard deviations (RSDs) of 0.16—4.80% and for oxasulfuron from 88.09% to 99.01% with relative standard deviations (RSDs) of 1.6-5.66%. The limit of detections (LODs) for nicosulfuron and oxasulfuron in soil were 0.002 mg/kg and 0.003 mg/kg, respectively. The dissipation of nicosulfuron and oxasulfuron residues over the time in soil was described by the Mittag-Leffler function a*Ea,B (- bt). Coefficients a, b, a, 8 were obtained from the experimental data, by using fitting procedure. We got a=0.8, B=3.71, a=159.11, b=12.79 for nicosulfuron and a=0.8, B=4.11, a=82.53, b=7.0 for oxasulfuron. The field dissipation half-life time for nicosulfuron at the topsoil (0-30cm soil depth) was 5.2 days while for oxasulfuron was 9.67days after application. These results could be utilized for the environmental risks assessment and minimizing risk for contamination of natural water resources and damage to following crops