Crop-Specific Responses to Cold Stress and Priming: Insights from Chlorophyll Fluorescence and Spectral Reflectance Analysis in Maize and Soybean.

This study aimed to investigate the impact of cold stress and priming on photosynthesis in the early development of maize and soybean, crops with diverse photosynthetic pathways. The main objectives were to determine the effect of cold stress on chlorophyll a fluorescence parameters and spectral reflectance indices, to determine the effect of cold stress priming and possible stress memory and to determine the relationship between different parameters used in determining the stress response. Fourteen maize inbred lines and twelve soybean cultivars were subjected to control, cold stress, and priming followed by cold stress in a walk-in growth chamber. Measurements were conducted using a portable fluorometer and a handheld reflectance instrument. Cold stress induced an overall downregulation of PSII-related specific energy fluxes and efficiencies, the inactivation of RCs resulting in higher energy dissipation, and electron transport chain impairment in both crops. Spectral reflectance indices suggested cold stress resulted in pigment differences between crops. The effect of priming was more pronounced in maize than in soybean with mostly a cumulatively negative effect. However, priming stabilized the electron trapping efficiency and upregulated the electron transfer system in maize, indicating an adaptive response. Overall, this comprehensive analysis provides insights into the complex physiological responses of maize and soybean to cold stress, emphasizing the need for further genotype-specific cold stress response and priming effect research.

Genetic diversity and selection signatures in a gene bank panel of maize inbred lines from Southeast Europe compared with two West European panels.

Southeast Europe (SEE) is a very important maize-growing region, comparable to the Corn belt region of the United States, with similar dent germplasm (dent by dent hybrids). Historically, this region has undergone several genetic material swaps, following the trends in the US, with one of the most significant swaps related to US aid programs after WWII. The imported accessions used to make double-cross hybrids were also mixed with previously adapted germplasm originating from several more distant OPVs, supporting the transition to single cross-breeding. Many of these materials were deposited at the Maize Gene Bank of the Maize Research Institute Zemun Polje (MRIZP) between the 1960s and 1980s. A part of this Gene Bank (572 inbreds) was genotyped with Affymetrix Axiom Maize Genotyping Array with 616,201 polymorphic variants. Data were merged with two other genotyping datasets with mostly European flint (TUM dataset) and dent (DROPS dataset) germplasm. The final pan-European dataset consisted of 974 inbreds and 460,243 markers. Admixture analysis showed seven ancestral populations representing European flint, B73/B14, Lancaster, B37, Wf9/Oh07, A374, and Iodent pools. Subpanel of inbreds with SEE origin showed a lack of Iodent germplasm, marking its historical context. Several signatures of selection were identified at chromosomes 1, 3, 6, 7, 8, 9, and 10. The regions under selection were mined for protein-coding genes and were used for gene ontology (GO) analysis, showing a highly significant overrepresentation of genes involved in response to stress. Our results suggest the accumulation of favorable allelic diversity, especially in the context of changing climate in the genetic resources of SEE.

Contrasting Water Withholding Responses of Young Maize Plants Reveal Link Between Lipid Peroxidation and Osmotic Regulation Corroborated by Genetic Analysis.

Linking biochemistry and genetics of tolerance to osmotic stress is of interest for understanding plant adaptations to unfavorable conditions. The aims of this study were to investigate the variability in responses of panel of elite maize inbred lines to water withholding for stress-related traits through association study and to identify pathways linked to detected associations for better understanding of maize stress responses. Densely genotyped public and expired Plant Variety Protection Certificate (ex-PVP) inbred lines were planted in controlled conditions (16-h/8-h day/night, 25°C, 50% RH) in control (CO) and exposed to 10-day water withholding (WW). Traits analyzed were guaiacol peroxidase activity (GPOD), total protein content (PROT), lipid peroxidation (TBARS), hydrogen peroxide accumulation (H2O2), proline accumulation (proline), and current water content (CWC). Proline accumulation was found to be influenced by H2O2 and TBARS signaling pathways acting as an accumulation-switching mechanism. Most of the associations detected were for proline (29.4%) and TBARS (44.1%). Gene ontology (GO) enrichment analysis showed significant enrichment in regulation of integral membrane parts and peroxisomes along with regulation of transcription and polysaccharide catabolism. Dynamic studies involving inbreds with extreme phenotypes are needed to elucidate the role of this signaling mechanism in regulation of response to water deficit.

Mycotoxins Biocontrol Methods for Healthier Crops and Stored Products.

Contamination of crops with phytopathogenic genera such as Fusarium, Aspergillus, Alternaria, and Penicillium usually results in mycotoxins in the stored crops or the final products (bread, beer, etc.). To reduce the damage and suppress the fungal growth, it is common to add antifungal substances during growth in the field or storage. Many of these antifungal substances are also harmful to human health and the reduction of their concentration would be of immense importance to food safety. Many eminent researchers are seeking a way to reduce the use of synthetic antifungal compounds and to implement more eco-friendly and healthier bioweapons against fungal proliferation and mycotoxin synthesis. This paper aims to address the recent advances in the effectiveness of biological antifungal compounds application against the aforementioned fungal genera and their species to enhance the protection of ecological and environmental systems involved in crop growing (water, soil, air) and to reduce fungicide contamination of food derived from these commodities.

Comparative Study of Drought Stress Effects on Traditional and Modern Apple Cultivars.

Genotype-dependent responses of apples to drought stress were evaluated between commercial and traditional apple cultivars. The results indicate different mechanisms of tolerance to investigated drought stress conditions. Chlorophyll fluorescence induction (OJIP) parameters, chlorophyll and carotenoid content, malondialdehyde (MDA), hydrogen peroxide (H2O2), proline, phenols and leaf water content (WC) were measured. The traditional cultivar "Crvenka" confirmed the best tolerance to a drought stress condition, presenting higher photosynthetic efficiency, higher leaf water content, higher levels of chlorophyll content and lower lipid peroxidation with greater membrane stability. The commercial cultivar "Golden Delicious Reinders" showed decreased water content in leaves, increased lipid peroxidation levels and photoinhibition. Considering all results, the commercial cultivar "Golden Delicious Reinders" was adversely affected by drought, while traditional cultivars exhibited better tolerance to drought stress.

Phenolic Acid Profiles and Antioxidant Activity of Major Cereal Crops.

Phenolic acids (PAs) are a dominant group of phenolic compounds in cereals, existing mostly bound to compounds of cell wall. In this study, a total of 25 cereal grain samples, including wheat, winter and spring barley, corn, and popcorn, were evaluated for bound PAs and antioxidant activity in a two-year field trial. The PA contents, determined by HPLC, were significantly affected by cereal type. The mean total PA content was highest in popcorn and corn (3298 and 2213 µg/gdm, respectively), followed by winter and spring barley (991 and 908 µg/gdm, respectively) and wheat (604 µg/gdm). Ferulic acid was the most abundant, accounting from 62% to 83% of total PAs (in popcorn and winter and spring barley, respectively). Across cereals, p-coumaric (35-259 µg/gdm) and p-hidroxybenzoic (45-79 µg/gdm) were also dominant, while in corn and popcorn o-coumaric (71 and 89 µg/gdm, respectively) also occurred in higher content. The mean total phenol content ranged from 853 µg GAE/gdm (wheat) to 1403 µg GAE/gdm (winter barley) with DPPH scavenging activity from 14% to 67%, respectively. A significant influence of crop years on the ferulic acid and total PA content was found, while the variability of other PAs was dependent on the cereal type. The results indicated a high health benefit potential of selected cereals.

Seed Weight as a Covariate in Association and Prediction Studies for Biomass Traits in Maize Seedlings.

BACKGROUND: The seedling stage has received little attention in maize breeding to identify genotypes tolerant to water deficit. The aim of this study is to evaluate incorporation of seed weight (expressed as hundred kernel weight, HKW) as a covariate into genomic association and prediction studies for three biomass traits in a panel of elite inbred lines challenged by water withholding at seedling stage. METHODS: 109 genotyped-by-sequencing (GBS) elite maize inbreds were phenotyped for HKW and planted in controlled conditions (16/8 day/night, 25 °C, 50% RH, 200 µMol/m2/s) in trays filled with soil. Plants in control (C) were watered every two days, while watering was stopped for 10 days in water withholding (WW). Fresh weight (FW), dry weight (DW), and dry matter content (DMC) were measured. RESULTS: Adding HKW as a covariate increased the power of detection of associations in FW and DW by 44% and increased genomic prediction accuracy in C and decreased in WW. CONCLUSIONS: Seed weight was effectively incorporated into association studies for biomass traits in maize seedlings, whereas the incorporation into genomic predictions, particularly in water-stressed plants, was not worthwhile.

Distribution of ß-Glucan, Phenolic Acids, and Proteins as Functional Phytonutrients of Hull-Less Barley Grain.

Two hull-less barley varieties were roller-milled, and breaks (B) and reduction flours (C), shorts, and bran were collected. Shorts, which mainly originate from endosperm cells with a smaller amount of the outer layers, had the largest yield (48.87-51.54%). Ash (0.82-3.10%) and protein (9.95-14.8%) increased from flours toward shorts and bran, while starch decreased (82.31-48.69%). In contrast to clear distribution differences in protein content (bran > shorts > C > B), albumins/globulins content was lowest in bran (0.78-0.90 g/100 gdw), and their distribution between fractions was uneven and genotype dependent. Distribution of hordeins (6.69-10.49 g/100 gdw) was more distinct and generally decreased in order from bran > B > shorts > C. The proportion of nutritionally poor C-hordeins in total hordeins varied from 28.33% to 30.24%, without significant differences between fractions. The ß-glucan content varied from 0.80% to 7.49% with decreasing content in the order bran, shorts > C > B. Shorts and bran could be classified as moderate and high ß-glucan flour (5.70-7.22%). The total phenolic and antioxidant activities ranged from 0.91 to 2.21 mg GAE/gdw and 28.81-72.06%, respectively. Ferulic and sinapic acids determined by high-performance liquid chromatography (HPLC) were major contributors to the antioxidant activity (45.16-1026.91 ug/gdw and 18.93-206.52 ug/gdw), respectively. The yield and high content of phytonutrients make hull-less barley shorts suitable for the production of health-promoting food and food supplements.

Changes in Antioxidant System during Grain Development of Wheat (Triticum aestivum L.) and Relationship with Protein Composition under FHB Stress.

Fusarium head blight (FHB) is found in both temperate and semi-tropical regions and causes losses in wheat (Triticum aestivum L.) resulting in reduced yield, deteriorated grain quality, and contamination of the grains with mycotoxins, primarily deoxynivalenol (DON). In this study, we focused on the identification of protein components in Fusarium-inoculated and non-inoculated wheat samples along with the major antioxidant enzymes that Fusarium can encounter during FHB infection process in six winter wheat varieties when FHB symptoms started to occur. Our hypothesis was that wheat antioxidants and H2O2 may play a role against Fusarium infections, consequently preserving protein grain components. Results showed that in more resistant varieties (Olimpija and Vulkan) DON content of inoculated flour was less accumulated and thus the major gluten network was not significantly attacked by Fusarium spp. The maximum increase in guaiacol peroxidase (POD) activity in response to FHB treatment was detected in the FHB-resistant varieties Olimpija and Vulkan, while the lowest increase in H2O2 content was detected in the FHB-susceptible variety Golubica. A particular reduction in the content of both total glutenin and high-molecular-weight glutenin subunits was detected in susceptible wheat varieties after serious artificial infection, along with increased DON accumulation. The defense mechanism in wheat varieties differed, where resistant varieties' protein composition depended on POD activity as a detoxification agent.

Changes in Protein Composition in the Grain and Malt after Fusarium Infection Dependently of Wheat Resistance.

The grain yield, as well as the quality and safety of the wheat grains and corresponding malt can be compromised by Fusarium spp. infection. The protein content of the grain affects the chemical composition and enzyme levels of the finished malt. The malting industry demands varieties with good malting and brewing performance, as well as good agronomic performance and disease resistance. The best method of disease control is breeding and selection for resistant varieties. Due to higher requirements for malting wheat worldwide, the goal of this investigation was to explore changes in protein distribution in wheat grains and corresponding malt, which are under higher pressure of Fusarium head blight (FHB) infestation in field conditions. The present study provides new knowledge on the impact of the FHB on the distribution of protein components of naturally Fusarium-infected (control) and Fusarium-inoculated wheat varieties in the grain and the corresponding malt in two consecutive years (2015/2016 and 2016/2017). The results showed that Fusarium infection of the susceptible variety Golubica, decreased total glutenins (5.9%), and both high and low molecular weight glutenin subunits (2.5% and 3.5%, respectively) in wheat grains, compared to control, in 2016. In contrast, gliadins and α-gliadins increased significantly (+7.6% and +5.1%, respectively) in the same variety. Wheat grains of the more resistant variety Vulkan showed an increase of the total glutenins content (+4.3%), and of high and of low molecular weight glutenin subunits (+1.2% and +3.2%, respectively) after Fusarium-inoculation, compared to naturally infected grains in 2016. Susceptible variety Golubica increased total glutenins (+9.1%), and both high and low molecular weight glutenin subunits (+3.5% and +5.6%, respectively) after Fusarium-inoculation in wheat malt, compared to naturally infected malt in 2016. In 2017, when disease pressure was higher than in 2016, there was a tendency in all varieties to increase gliadins and its sub fractions after malting, and to decrease glutenins and its sub fractions in Fusarium-inoculated treatment. In conclusion, FHB dramatically depressed grain yield (up to 37%) and quality (glutenins and high molecular weight subunits) in the susceptible Fusarium variety, which makes it inconvenient for malting.

Genetic Correlations Between Photosynthetic and Yield Performance in Maize Are Different Under Two Heat Scenarios During Flowering.

Chlorophyll fluorescence (ChlF) parameters are reliable early stress indicators in crops, but their relations with yield are still not clear. The aims of this study are to examine genetic correlations between photosynthetic performance of JIP-test during flowering and grain yield (GY) in maize grown under two heat scenarios in the field environments applying quantitative genetic analysis, and to compare efficiencies of indirect selection for GY through ChlF parameters and genomic selection for GY. The testcrosses of 221 intermated recombinant inbred lines (IRILs) of the IBM Syn4 population were evaluated in six environments at two geographically distinctive locations in 3 years. According to day/night temperatures and vapor pressure deficit (VPD), the two locations in Croatia and Turkey may be categorized to the mild heat and moderate heat scenarios, respectively. Mild heat scenario is characterized by daytime temperatures often exceeding 33°C and night temperatures lower than 20°C while in moderate heat scenario the daytime temperatures often exceeded 33°C and night temperatures were above 20°C. The most discernible differences among the scenarios were obtained for efficiency of electron transport beyond quinone A (QA) [ET/(TR-ET)], performance index on absorption basis (PIABS) and GY. Under the moderate heat scenario, there were tight positive genetic correlations between ET/(TR-ET) and GY (0.73), as well as between PIABS and GY (0.59). Associations between the traits were noticeably weaker under the mild heat scenario. Analysis of quantitative trait loci (QTL) revealed several common QTLs for photosynthetic and yield performance under the moderate heat scenario corroborating pleiotropy. Although the indirect selection with ChlF parameters is less efficient than direct selection, ET/(TR-ET) and PIABS could be efficient secondary breeding traits for selection under moderate heat stress since they seem to be genetically correlated with GY in the stressed environments and not associated with yield performance under non-stressed conditions predicting GY during flowering. Indirect selection through PIABS was also shown to be more efficient than genomic selection in moderate heat scenario.

The Pressure of Fusarium Disease and Its Relation with Mycotoxins in The Wheat Grain and Malt.

Fusarium head blight (FHB) is one of the most destructive wheat fungal diseases, causing yield loss, quality reduction, and accumulation of mycotoxins. The aim of this research was to summarize the occurrence of major Fusarium mycotoxins: deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-AcDON), nivalenol (NIV), and zearalenone (ZEN) in two consecutive years to search the relationship between disease incidence and severity with mycotoxins found in control and inoculated grains and corresponding malt. In addition, deoxynivalenol-3-glucoside (D3G) in one-year research was measured. Tested wheat varieties showed infection scores of 3% ('U1' and 'Sirban Prolifik') to 79% ('Golubica') for Type I resistance evaluation. There were few moderately resistant varieties in view of their areas under the disease progress curve, which can be considered Type III resistance ('Sirban Prolifik' and 'U1'). According to the data quantified by LC-MS/MS, DON decreased in infected malt in comparison to corresponding grain, while ZEN occurred only in infected malt samples. Both 3-AcDON and NIV increased in inoculated malt in comparison to corresponding grain, due to a combination of plant metabolism and de novo synthesis by molds during malting. Based on the results, we can draw a few conclusions: the resistance to Fusarium decreased quantified concentrations of DON; ZEN gets synthetized during malting; unregulated 3-AcDON and NIV increase during malting; more resistant varieties have converted DON to D3G more successfully. Modified mycotoxins should be also included to legislation, since they could be transformed back to the corresponding mycotoxins under food processing conditions or during digestion.

Performance Indices in Wheat Chlorophyll a Fluorescence and Protein Quality Influenced by FHB.

Very little is known about the physiological interactions between wheat quality and Fusarium head blight (FHB), which substantially reduces wheat grain yield and quality worldwide. In order to investigate stress-induced changes in flag leaves from plants artificially inoculated with Fusarium, we screened for chlorophyll a fluorescence transient at 1, 2, 4, 7 and 14 days after Fusarium inoculation. Our results indicate that the maximum quantum yield of photochemistry (Fv/Fm) and the performance index (PI) were not affected by FHB, but there were significant differences in those two traits between different varieties and measurement times. FHB caused a significant reduction in the percentage of glutenins (GLU), high-molecular-weight (HMW), and low-molecular-weight (LMW) subunits in 'Kraljica' and 'Golubica', unlike 'Vulkan', where the percentage of GLU increased.

Quantitative trait loci mapping of metal concentrations in leaves of the maize IBM population.

Characterizing concentrations of several beneficiary and toxic metals in maize leaves is of importance for ionomic studies and for silage production. The intermated B73 × Mo17 maize population (IBM) was evaluated for concentrations of eight metals (cadmium - Cd, copper - Cu, iron - Fe, potassium - K, magnesium - Mg, manganese - Mn, strontium - Sr and zinc - Zn) in ear-leaf to map quantitative trait loci (QTL) with 2161 molecular markers across the genome. QTL analysis revealed nine significant QTLs for concentrations of Cd, Cu, Fe, K, Mg and Sr combined over two environments. Median resolution for the QTL interval was less than 1 cM on a regular F2 map, which is a big improvement compared with the prior mapping (8 cM). The highest LOD scores of 15.52 and 15.31 were detected for K and Cd concentrations, respectively, explaining more than 20 percent of the phenotypic variance. No QTLs were found to be colocalized. QTL mapping in the IBM population did not confirm our earlier QTL results demonstrating considerable QTL ×genetic background interaction. The only exception is confirmation of the major QTL for Cd accumulation on chromosome 2. Our results could facilitate further genetic and physical mapping of genes for metal accumulation in maize.

Quantitative trait loci for biofortification traits in maize grain.

Detecting genes that influence biofortification traits in cereal grain could help increase the concentrations of bioavailable mineral elements in crops to solve the global mineral malnutrition problem. The aims of this study were to detect the quantitative trait loci (QTLs) for phosphorus (P), iron (Fe), zinc (Zn), and magnesium (Mg) concentrations in maize grain in a mapping population, as well as QTLs for bioavailable Fe, Zn, and Mg, by precalculating their respective ratios with P. Elemental analysis of grain samples was done by coupled plasma-optical emission spectrometry in 294 F(4) lines of a biparental population taken from field trials of over 3 years. The population was mapped using sets of 121 polymorphic markers. QTL analysis revealed 32 significant QTLs detected for 7 traits, of which some were colocalized. The Additive-dominant model revealed highly significant additive effects, suggesting that biofortification traits in maize are generally controlled by numerous small-effect QTLs. Three QTLs for Fe/P, Zn/P, and Mg/P were colocalized on chromosome 3, coinciding with simple sequence repeats marker bnlg1456, which resides in close proximity to previously identified phytase genes (ZM phys1 and phys2). Thus, we recommend the ratios as bioavailability traits in biofortification research.