Due to climate change in recent years, there has been an increasing water deficit during the winter wheat sowing period. This study evaluated six Croatian winter wheat varieties’ physiological, biochemical, and molecular responses under two drought stress levels at the germination/seedling growth stage. Lipid peroxidation was mainly induced under both drought stress treatments, while the antioxidative response was variety-specific. The most significant role in the antioxidative response had glutathione along with the ascorbate-glutathione pathway. Under drought stress, wheat seedlings responded in proline accumulation that was correlated with the P5CS gene expression. Expression of genes encoding dehydrins (DHN5, WZY2) was highly induced under the drought stress in all varieties, while genes encoding transcription factors were differentially regulated. Expression of DREB1 was upregulated under severe drought stress in most varieties, while the expression of WRKY2 was downregulated or revealed control levels. Different mechanisms were shown to contribute to the drought tolerance in different varieties, which was mainly associated with osmotic adjustment and dehydrins expression. Identifying different mechanisms in drought stress response would advance our understanding of the complex strategies contributing to wheat tolerance to drought in the early growth stage and could contribute to variety selection useful for developing new drought-tolerant varieties.

Fusarium head blight (FHB) is one of the most studied fungal diseases of wheat, causing massive grain yield and quality losses. This study aimed to extend previous studies on the physiological and biochemical responses of winter wheat to FHB stress in a controlled environment by focusing on the ascorbate-glutathione pathway (AsA-GSH), photosynthetic efficiency, and stress hormone levels, thus providing insight into the possible interactions of different defense mechanisms during infection. The activity of AsA-GSH metabolism was increased in FHB resistant varieties, maintaining the redox state of spikes, and consequently preserving functional photosystem II. Furthermore, carotenoids (Car) were shown to be the major pigments in the photosystem assembly, as they decreased in FHB-stressed spikes of resistant and moderately resistant varieties, compared to controls. Car are also the substrate for the synthesis of abscisic acid (ABA), which acts as a fungal effector and its elevated content leads to increased FHB susceptibility in inoculated spikes. The results of this study contributed to the knowledge of FHB resistance mechanisms and can be used to improve the breeding of FHB resistant varieties, which is considered to be the most effective control measure.

Fusarium head blight (FHB) is an important disease of wheat and production of mycotoxins makes it a major threat in most wheat-producing areas worldwide. This study aimed to identify the impact of epidemic FHB conditions (usage of artificial Fusarium inoculation) on mycotoxin levels in unprocessed wheat. Fusarium levels were monitored at two locations in two treatments (natural infection and inoculation with Fusarium graminearum and F. culmorum) where 13 mycotoxins were evaluated by LC/MS-MS in six winter wheat varieties. Due to favorable conditions for infection with Fusarium fungi during the flowering period at location Tovarnik, wheat varieties had higher disease severity and increased mycotoxin accumulation, compared to Osijek. The most abundant mycotoxins in treatment with inoculation with Fusarium fungi were deoxynivalenol (DON), culmorin (CUL) and hydroxyculmorins. In treatment with natural infection, DON did not exceed maximum limits set by EU. Varieties with lower initial resistance accumulated DON even in naturally infected samples at Tovarnik. These results highlighted the impact of environment variation in the production of Fusarium mycotoxins where FHB initial resistance had a higher impact on the accumulation of mycotoxins than general resistance. Furthermore, wheat samples with higher DON concentration also contained elevated levels of CUL and hydroxyculmorins, showing that CUL can have a possible role in Fusarium virulence. The FHB evaluations provide important information about the genetic resistance of wheat varieties, as well as risk assessment considering mycotoxin accumulation in epidemic conditions.

Wheat is one of the three most important cereals in the world, along with rice and maize. It serves as the primary food and source of energy for about 30–40% of the world’s population. However, the low levels of micronutrients in wheat grains can lead to deficiencies of those micronutrients in people whose dietary habits are mostly based on cereals such as wheat. Apart from iron (Fe) and zinc (Zn), a lack of selenium (Se) is also one of the biggest problems in the world. The essentiality of Se has been confirmed for all animals and humans, and the lack of this micronutrient can cause serious health issues. Wheat dominates the world’s cereal production, so it is one of the best plants for biofortification. Due to the fact that agronomic biofortification is not an economical or environmentally acceptable approach, genetic improvement of cereals such as wheat for the enhanced content of micronutrients in the grain represents the most efficient biofortification approach.

Fusarium head blight (FHB) is one of the most dangerous diseases of winter wheat, resulting in reduced grain yield and quality, and production of mycotoxins by the Fusarium fungi. In the present study, changes in the grain metabolomics of winter wheat samples infected with Fusarium spp. and corresponding non-infected samples from two locations in Croatia were investigated by GC-MS. A Mann–Whitney test revealed that 24 metabolites detected were significantly separated between Fusarium-inoculated and non-infected samples during the variety by treatment interactions. The results confirmed that in grains of six FHB-resistant varieties, ten metabolites were identified as possible resistance-related metabolites. These metabolites included heptadecanoic acid, 9-(Z)-hexadecenoic acid, sophorose, and secolaganin in grains of FHB-resistant varieties at the Osijek location, as well as 2-methylaminomethyltartronic acid, maleamic acid, 4-hydroxyphenylacetonitrile, 1,4-lactonearabinonic acid, secolaganin, and alanine in grains of FHB-resistant varieties at the Tovarnik location. Moreover, on the PCA bi-plot, FHB-susceptible wheat varieties were closer to glycyl proline, decanoic acid, and lactic acid dimer that could have affected other metabolites, and thus, suppressed resistance to FHB. Although defense reactions were genetically conditioned and variety specific, resulting metabolomics changes may give insight into defense-related pathways that could be manipulated to engineer plants with improved resistance to the pathogen.