Early Antioxidative Response to Desiccant-Stimulated Drought Stress in Field-Grown Traditional Wheat Varieties.

Extended drought affects the production and quality of wheat (Triticum aestivum L.), one of the world's most important food crops. Breeding for increased drought resistance is becoming increasingly important due to the rising demand for food production. Four old traditional Croatian wheat cultivars were used in the present study to examine the early antioxidant response of flag leaves to desiccant-stimulated drought stress and to identify drought-tolerant cultivars accordingly. The results indicate that the enzymatic antioxidant system plays the most significant role in the early response of adult wheat plants to drought stress and the removal of excessive H2O2, particularly GPOD and APX. Nada and Dubrava cultivars revealed the strongest activation of the enzymatic defense mechanism, which prevented H2O2 accumulation and lipid peroxidation. Additionally, the Nada cultivar also showed increased synthesis of proline and specific phenolic compounds, which both contribute to the increased stress tolerance. Among the cultivars investigated, cultivar Nada has the broadest genetic base, which may explain why it possesses the ability to activate both enzymatic and non-enzymatic defense mechanisms in an early response to drought stress. This suggests that old traditional wheat cultivars with broad genetic bases can be a valuable source of drought tolerance, which is especially important given the current climate change.

Characterization of Forage Quality, Phenolic Profiles, and Antioxidant Activity in Alfalfa (Medicago sativa L.).

Alfalfa (Medicago sativa L.) is one of the most important forage species and is recently more in focus for human consumption mainly due to its content of bioactive phenolics. Samples of the seventeen alfalfa cultivars/populations were collected at the Agricultural Institute Osijek with the aim to evaluate their forage quality, phenolic profiles, and antioxidant potential. Significant differences (p < 0.05) existed among studied alfalfa in all analyzed traits. The cultivar OS 99 and populations L7 and L20 were characterized by high crude protein content (22.5−24.7%) and the lowest neutral (40.2−42.9%) and acid detergent fibres (33−35.5%). The soluble-free phenolics from alfalfa leaves were extracted by methanol while insoluble cell-wall bound phenolics were released by alkaline hydrolysis. The bound phenolic extract showed a stronger DPPH scavenging capacity (20.8 mg TE/g dm) than the soluble (11.4 mg TE/g dm). The HPLC data revealed that more phenolics were found in the bound (3638.0 µg/g dm) than in the soluble form (912.3 µg/g dm). In the soluble extract of the alfalfa leaves, the major compound was catechin (338.3 µg/g dm), while rutin, epicatechin, and ferulic acid were minor ones. In the bound phenolic extract, the most abundant was ferulic (2198.2 µg/g dm) and p-coumaric acid (983.7 µg/g dm), followed by myricetin, apigenin, and quercetin. The principal component analysis revealed that alfalfa cultivars/populations were better discriminated based on the data on phenolics, rather than on forage quality. The cultivars/populations Florida 66, OS 66, L 40, L 42, Seed Force 4, and Torlesse were the most interesting in terms of phenolic health-promoting characteristics.

Photosynthetic Variability of Oblacinska Sour Cherry Ecotypes under Drought.

The selection of drought-tolerant sour cherry genotypes is essential for developing sustainable fruit production in today's climate-change conditions. The phenotypic heterogenic population of sour cherry Oblacinska, with high and regular yield suitable for mechanical harvesting and industrial processing, is a traditional and predominant cultivar in northern Croatia (Pannonian region) and Serbia commercial orchards. In this context, 2-year old virus-free sour cherry plants of 4 isolated Oblacinska sour cherry ecotypes (OS, 18, D6, and BOR) produced by micropropagation were exposed to severe drought in a greenhouse under semi-controlled conditions to evaluate its photosynthetic intra-varietal variability. Relative water content (RWC), chlorophyll fluorescence (ChlF), and photosynthetic pigments were evaluated during the ten days of the experiment. As a visible symptom of stress, the withering of plants was followed by a diminution of RWC and photosynthetic pigments in the drought exposed leaves of sour cherry ecotypes compared to the control treatment. ChlF elucidated variability in the photosynthetic efficiency within studied sour cherry ecotypes, highlighting PIABS, PItotal, and ψE0 as the most sensitive and thus the most informative JIP parameters for drought screening. Among the investigated ecotypes, BOR proved to be the most sensitive. The Oblacinska sour cherry ecotype OS showed the highest tolerance to drought conditions and, therefore, can be used as a source of tolerance in sour cherry breeding programs.

Correlation of elongation factor 1A accumulation with photosynthetic pigment content and yield in winter wheat varieties under heat stress conditions.

Heat stress is one of the most important environmental factors that influences wheat growth and development, leading to significant losses in grain yield and has become a significant detrimental factor for worldwide wheat production. In recent years, several studies suggested that eukaryotic elongation factor 1A (eEF1A), may contribute to heat tolerance in plants, therefore the aim of this study was: to investigate the accumulation of eEF1A in wheat under conditions of moderate and high air temperatures; to determine the amount of photosynthetic pigments and to determine the yield traits; and to examine whether there is a correlation between eEF1A accumulation, photosynthetic pigments, and yield in different wheat varieties. The results showed that heat stress induced accumulation of eEF1A significantly different among wheat varieties and showed that varieties with a higher accumulation of eEF1A under heat stress are characterized by a smaller decrease in the photosynthetic pigments. A correlation between higher accumulation of eEF1A under heat stress and yield traits was found. Analyzed parameters from two growing seasons, indicated that the higher accumulation of eEF1A and a smaller decrease in photosynthetic pigments distinguishes the varieties more resistant to heat stress. The analysis of the molecular mechanisms by immunoblot, under conditions of high and moderate air temperatures in two growing seasons, aims to develop agricultural strategy and develop wheat varieties tolerant to heat stress.

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.

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.

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 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.

Forage Legumes as Sources of Bioactive Phytoestrogens for Use in Pharmaceutics: A Review.

BACKGROUND: Alfalfa and red clover are the most widespread and most important perennial legumes, primarily used as a high-quality feed for livestock. Both alfalfa and red clover, as well as some other plant species from Fabaceae family, are a rich natural source of phytoestrogens, nonsteroidal compounds with an estrogenic activity whose beneficial effects in the prevention and treatment of many diseases are demonstrated in numerous scientific studies. OBJECTIVES: Nowadays, various herbal preparations are present on the world market and their use is constantly increasing, as well as the growing demands of consumers for environmentally sound and health-safe production of plant species used as sources of active substances. Because of their widespread distribution, the possibility of organic breeding, and also significant increases in surface area under genetically modified crops in most EU countries, alfalfa and red clover have become more interesting alternative sources of phytoestrogens. The most common phytoestrogens in these forage legumes are genistein, daidzein, glycitein, formononetin, biochanin, coumestrol, kaempferol and apigenin. The content of these substances is significantly influenced by a number of factors including genotype, environment, production technology, plant maturity stage, and individual plant parts. CONCLUSION: Available evidence suggests that forage legumes represent high promising sources of health-promoting phytoestrogens. Due to numerous favorable features, they can find commercial application in different industries such as pharmaceutical, nutraceutical, cosmetic, and agriculture.

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.