Agronomic strategies to enhance the early vigor and yield of maize part II: the role of seed applied biostimulant, hybrid, and starter fertilization on crop performance.

Maize cropping systems need to be re-designed, within a sustainable intensification context, by focusing on the application of high-use efficiency crop practices, such as those that are able to enhance an early plant vigor in the first critical growth stages; such practices could lead to significant agronomic and yield benefits. The aim of this study has been to evaluate the effects of the cultivation of hybrids with superior early vigor, of the distribution of starter fertilizers at sowing, and of the seed application of biostimulants on promoting plant growth and grain yield in full factorial experiments carried out in both a growth chamber and in open fields. The greatest benefits, in terms of plant growth enhancement (plant height, biomass, leaf area) and cold stress mitigation, were detected for the starter fertilization, followed by the use of an early vigor hybrid and a biostimulant seed treatment. The starter fertilization and the early vigor hybrid led to earlier flowering dates, that is, of 2.1 and 2.8 days, respectively, and significantly reduced grain moisture at harvest. Moreover, the early vigor hybrid, the starter NP fertilization, and the biostimulant treatment increased grain yield by 8.5%, 6.0%, and 5.1%, respectively, compared to the standard hybrid and the untreated controls. The combination of all the considered factors resulted in the maximum benefits, compared to the control cropping system, with an increase in the plant growth of 124%, a reduction of the sowing-flowering period of 5 days, and a gain in grain yield of 14%. When choosing the most suitable crop practice, the diversity of each cropping system should be considered, according to the pedo-climatic conditions, the agronomic background, the yield potential, and the supply chain requirements.

Multi-Mycotoxin Long-Term Monitoring Survey on North-Italian Maize over an 11-Year Period (2011-2021): The Co-Occurrence of Regulated, Masked and Emerging Mycotoxins and Fungal Metabolites.

Maize is considered one of the most susceptible crops to mycotoxin-producing fungi throughout the world, mainly belonging to the Fusarium spp. and Aspergillus spp. Maize is mainly used as animal feeds in Italy, as well as for human consumption, being essential for all the protected designation of origin (DOP) products. Our study investigated the occurrence of regulated mycotoxins in 3769 maize grain samples collected from 88 storage centers by the National Monitoring Network over an 11-year period (2011-2021). Moreover, an in-depth survey over a 4-year period, characterized by extremely different meteorological conditions, was conducted to investigate the co-occurrence of regulated, masked, and emerging mycotoxins. The survey confirmed that Fusarium spp. was the most frequent fungi and fumonisins were the main mycotoxins that were constantly detected in the different years and areas. Moreover, the areas characterized by high fumonisin levels were also the most prone to contamination by emerging mycotoxins produced by the same Fusarium species of the Liseola section. On the other hand, as a result of climatic changes, maize grains have also been affected by the increased frequency of aflatoxin accumulation. Deoxynivalenol, zearalenone, and other emerging mycotoxins produced by the same Fusarium species as the Discolor section occurred more abundantly in some areas in Northern Italy and in years characterized by predisposing meteorological conditions.

The Effect of Foliar Fungicide and Insecticide Application on the Contamination of Fumonisins, Moniliformin and Deoxynivalenol in Maize Used for Food Purposes.

The fungal ear rot of maize cultivated in temperate areas is mainly due to the Fusarium species. The use of insecticides against European Corn Borer (ECB) reduces the severity of fungal ear rot as well as the fumonisin (FB) and moniliformin (MON) levels in maize kernels at harvest, which in turn results in a lowering of their effect on deoxynivalenol (DON) control. However, the direct fungicidal control of ear rot has rarely been implemented for maize, and the first studies reported conflicting results on the reduction of mycotoxins. In the present experiment, field trials were carried out in North Italy over three growing seasons to study the effect of fungicide application timings on maize to control mycotoxins, considering the interaction of the application with the insecticide treatment, according to a full factorial split plot design. The mycotoxin content was determined through LC-MS/MS analysis. The field trials showed a significant reduction in ECB severity (75%), fungal ear rot severity (68%), Fusarium Liseola section infection (46%), FBs (75%) and MON (79%) as a result of the insecticide application for all the years, while the DON content increased by 60%. On the other hand, a fungicide application alone or applied in plots protected by an insecticide was never effective for the fungal symptoms, infection or mycotoxin content. The results confirm that a correct insecticide application to control ECB damage is the most effective agrochemical solution for the control of fungal ear rot, FBs and MON.

The Role of Nitrogen Fertilization on the Occurrence of Regulated, Modified and Emerging Mycotoxins and Fungal Metabolites in Maize Kernels.

The European Food Safety Authority is currently evaluating the risks related to the presence of emerging mycotoxins in food and feeds. The aim of this study was to investigate the role of soil fertility, resulting from different nitrogen fertilization rates, on the contamination of regulated mycotoxins and emerging fungal metabolites in maize grains. The trial was carried out in the 2012-2013 growing seasons as part of a long-term (20-year) experimental platform area in North-West Italy, where five different N rates, ranging from 0 to 400 kg N ha-1, were applied to maize each year. Maize samples were analyzed by means of a dilute-and-shoot multi-mycotoxin LC-MS/MS method, and more than 25 of the most abundant mycotoxins and fungal metabolites were detected. Contamination by fumonisins and other fungal metabolites produced by Fusarium spp. of the section Liseola was observed to have increased in soils that showed a poor fertility status. On the other hand, an overload of nitrogen fertilization was generally associated with higher deoxynivalenol and zearalenone contamination in maize kernels, as well as a higher risk of other fungal metabolites produced by Fusarium spp. sections Discolor and Roseum. A balanced application of N fertilizer, in accordance with maize uptake, generally appears to be the best solution to guarantee an overall lower contamination by regulated mycotoxins and emerging fungal metabolites.

Characterization and selection of functional yeast strains during sourdough fermentation of different cereal wholegrain flours.

The increasing demand for healthy baked goods boosted studies on sourdough microbiota with beneficial metabolic traits, to be used as potential functional starters. Here, 139 yeasts isolated from cereal-based fermented foods were in vitro characterized for their phytase and antioxidant activities. The molecular characterization at strain level of the best 39 performing isolates showed that they did not derive from cross contamination by baker's yeast. Afterwards, the 39 isolates were in vivo analyzed for their leavening ability, phytase activity and polyphenols content using five different wholegrain flours, obtained from conventional and pigmented common wheat, emmer and hull-less barley. Combining these findings, through multivariate permutation analysis, we identified the 2 best performing strains, which resulted diverse for each flour. Doughs singly inoculated with the selected strains were further analyzed for their antioxidant capacity, phenolic acids, xanthophylls and anthocyanins content. All the selected yeasts significantly increased the total antioxidant activity, the soluble, free and conjugated, forms of phenolic acids and anthocyanins of fermented doughs. This study revealed the importance of a specific selection of yeast strains for wholegrain flours obtained from different cereals or cultivars, in order to enhance the pro-technological, nutritional and nutraceutical traits of fermented doughs.

Distribution of bioactive compounds in pearled fractions of tritordeum.

Hexaploid tritordeum is the amphidiploid cereal derived from the cross between wild barley and durum wheat. The present study compares two cultivars of tritordeum with other cereals grown in the same experimental area to weigh up its potential use as ingredient for health-valued foods. Tritordeum shows 2.5-fold higher concentration of lutein than common wheat and barley, and 1.2-fold higher than durum wheat, while the concentration of ß-glucans is 5 folds lower than the one observed for barley. Based on the distribution of bioactive compounds in pearled fractions, the use of whole-grain flours seems the best way to exploit the antioxidant potential of tritordeum. Nevertheless, the internal layers of the kernel of this cereal are characterized on average by high concentrations of antioxidants (32.0 mg/kg and 518 mg/kg soluble and cell wall-bound phenolic acids, respectively), making tritordeum interesting also for the production of refined flours rich in bioactive compounds.

Development and Comparison of Two Multiresidue Methods for the Determination of 17 Aspergillus and Fusarium Mycotoxins in Cereals Using HPLC-ESI-TQ-MS/MS.

Cereals can be contaminated by several mycotoxins, whose co-presence may represent an undervalued risk for humans and animals. Maize and wheat are the most contaminated cereals and in temperate areas could be affected in field conditions by several Fusarium and Aspergillus infections. To date, only B-fumonisins (FBs), aflatoxins (AFs), zearalenone (ZEA), deoxynivalenol (DON) and T-2 and HT-2 toxins have been regulated in cereals in European Union. The other fungal metabolites, are commonly referred to as "emerging" and "masked" mycotoxins, and more information on their occurrence in combination with the regulated mycotoxins, are needed to design combined toxicological and exposure assessments. This research intends to develop and compare two multiresidue HPLC-ESI-TQ-MS/MS methods for the simultaneous determination of the main regulated, emerging and masked mycotoxins in maize and wheat, among which: FB1, FB2, DON, ZEA, AFB1, AFB2, AFG1, AFG2, moniliformin (MON), deoxynivalenol-3-glucoside (DON-3-G), 3-acetyldeoxynivalenol (3-ADON), 15-acetyldeoxynivalenol (15-ADON), nivalenol (NIV), enniatins A, A1, B, B1 (ENNA, ENNA1, ENNB, ENNB1). The extraction was performed for both methods using a mixture of CH3CN/H2O/CH3COOH (79/20/1, v/v/v), while the dilution/purification was carried out through two different procedures: (1) by the "dilute-and-shoot" technique diluting 1:2 the filtered extract with CH3CN/H2O/CH3COOH (20/79/1, v/v/v) to reduce the matrix effect; (2) using the Oasis® PRiME HLB clean-up columns. The analysis was carried out using CH3OH and H2O both acidified with 0.1% of CH3COOH as eluents. The injection volume was 20 µL and the flow rate 200 µL min-1. The analysis of two reference material (maize and wheat), was performed to evaluate the trueness and precision of the two methods by matrix-matched calibration curves. For all the regulated mycotoxins analyzed by both methods, the range of recovery percentage established by the Regulation (EC) No. 401/2006 was respected, except for ZEA by using the Oasis® PRiME HLB clean-up columns. Nevertheless, the results suggest that the Oasis® PRiME HLB clean-up columns, could be a valid alternative to the dilute-and-shoot method, although an additional cost for the clean-up has to be considered. In conclusion, both two analytical methods considerably reduce the analytical time and costs and therefore result to be promising and applicable for high-throughput routine multi-mycotoxins analysis by the use of a TQ.

Bioactive compound and antioxidant activity distribution in roller-milled and pearled fractions of conventional and pigmented wheat varieties.

In this study, the chemical composition of pigmented wheats (yellow, purple and blue types), and the distribution of the bioactive compounds in their roller-milled and pearled fractions, were compared with conventional wheats (red and white types). Roller-milling promoted the recovery of total dietary fiber, ß-glucans, phenolic acids and anthocyanins in the bran fraction, which resulted also in a higher total antioxidant activity than the refined flour. Conversely, lutein resulted mainly concentrated in the refined flour. In the same way, the distribution pattern in the pearled fractions differ depending on the bioactive considered. The study highlights that a careful selection of the most appropriate fractionation process should be performed to produce flours rich in bioactive compounds. Roller-milling resulted useful for the production of refined flours rich in xanthophylls, with particular emphasis to the yellow-grained wheats. Contrarily, pearling could be more useful in the valorization of the health potential of anthocyanin-pigmented varieties.

Folate distribution in barley (Hordeum vulgare L.), common wheat (Triticum aestivum L.) and durum wheat (Triticum turgidum durum Desf.) pearled fractions.

BACKGROUND: Wholegrain cereals are an important source of folates. In this study, total folate was analysed in pearled fractions of barley and wheat cultivars employing AOAC Official Method 2004.05. In particular, the distribution of folate in the kernels was evaluated in three barley cultivars (two hulled types and a hulless one as well as two- and six-row types) and in a common and a durum wheat cultivar. RESULTS: A noticeable variation in the folate content was observed between the barley [653-1033 ng g(-1) dry matter (DM)] and wheat cultivars (1024-1119 ng g(-1) DM). The highest folate content was detected in the hulless barley cultivar (1033 ng g(-1) DM). A significant reduction in total folate, from 63% to 86%, was observed in all cultivars from the outermost to the innermost pearled fractions. CONCLUSION: Results proved that folates are mainly present in the germ and in the outer layers of the kernel. This is the first study reporting the folate distribution in kernels of both common and durum wheat and in a hulless barley cultivar. Results suggest that the pearling process could be useful for the selection of intermediate fractions that could be used in order to develop folate-enhanced ingredients and products.

Progressive Pearling of Barley Kernel: Chemical Characterization of Pearling Fractions and Effect of Their Inclusion on the Nutritional and Technological Properties of Wheat Bread.

Two hulled barley varieties have been sequentially pearled for one to eight cycles, each with 5% removal. The derived fractions were analyzed for their bioactive compound content. The dietary fiber (DF) decreased from the external to the internal layers, whereas ß-glucans showed an inverse trend. Deoxynivalenol contamination was concentrated in the outer layers. The total antioxidant activity (TAA) was higher in the 15-25% fractions, which were used to prepare bread. Five mixtures of refined wheat flour, with an increasing replacement of this pearled barley fraction, were compared with a control for the bioactive compound content, as well as for the rheological and physical bread properties. The inclusion of pearled fractions with up to a 10% substitution leads to a clear enhancement of the DF and TAA, with only minor detrimental effects on the physical parameters. Selected byproducts of barley pearling could be proposed as functional ingredients for bakery products rich in DF and TAA.