Lignocellulolytic enzymes play a crucial role in efficiently converting lignocellulose into valuable platform molecules in various industries. However, they are limited by their production yields, costs, and stability. Consequently, their production by producers adapted to local environments and the choice of low-cost raw materials can address these limitations. Due to the large amounts of olive stones (OS) generated in Morocco which are still undervalued, Penicillium crustosum , Fusarium nygamai , Trichoderma capillare , and Aspergillus calidoustus , are cultivated under different fermentation techniques using this by-product as a local lignocellulosic substrate. Based on a multilevel factorial design, their potential to produce lignocellulolytic enzymes during 15 days of dark incubation was evaluated. The results revealed that P. crustosum expressed a maximum total cellulase activity of 10.9 IU/ml under sequential fermentation (SF) and 3.6 IU/ml of beta-glucosidase activity under submerged fermentation (SmF). F. nygamai recorded the best laccase activity of 9 IU/ml under solid-state fermentation (SSF). Unlike T. capillare, SF was the inducive culture for the former activity with 7.6 IU/ml. A. calidoustus produced, respectively, 1,009 mu g/ml of proteins and 11.5 IU/ml of endoglucanase activity as the best results achieved. Optimum cellulase production took place after the 5th day under SF, while ligninases occurred between the 9th and the 11th days under SSF. This study reports for the first time the lignocellulolytic activities of F. nygamai and A. calidoustus . Furthermore, it underlines the potential of the four fungi as biomass decomposers for environmentally -friendly applications, emphasizing the efficiency of OS as an inducing substrate for enzyme production.

BACKGROUND:Gaining insight into crop diversity, both at the genetic and phenotypic levels, is of prime importance for onion breeding with an enhanced yield and quality in combination with improved resistance to biotic and abiotic stresses. In the current study, 192 different onion plants, representing 16 ecotypes, were characterized using ISSR markers.RESULTS:Based on the ISSR marker profile, there was a clear grouping of the plants into 16 different ecotypes. Though the 16 populations originated from the same geographic region in Morocco, a significant genetic diversity was detected. After a genomic characterization, field trials in three different environments in Morocco were laid out. The phenotypic characterization showed that there were always significant differences between ecotypes, and for most traits, there was also a significant environmental effect and a significant interaction between environment and ecotype. The broad-sense heritability (H2) for the phenotypic traits associated with color (L*, a*, and b*) was the largest (84.2%, 80.6%, 79.2%), demonstrating that color is conditioned primarily by genetic factors. In contrast, the H2 for yield was the lowest (41.8%), indicating that the environment has a substantial effect on yield. In addition, there was a significant association between the presence/absence of certain bands and various phenotypic traits.CONCLUSION:ISSR markers are a powerful tool in distinguishing onion ecotypes. In addition, significant associations between marker scores and phenotypic traits could be detected, representing particular importance for future breeding programs.

As the awareness on the ecological impact of chemical phosphate fertilizers grows, research turns to sustainable alternatives such as the implementation of phosphate solubilizing bacteria (PSB), which make largely immobile phosphorous reserves in soils available for uptake by plants. In this review, we introduce the mechanisms by which plants facilitate P-uptake and illustrate how PSB improve the bioavailability of this nutrient. Next, the effectiveness of PSB on increasing plant biomass and P-uptake is assessed using a meta-analysis approach. Our review demonstrates that improved P-uptake does not always translate in improved plant height and biomass. We show that the effect of PSB on plants does not provide an added benefit when using bacterial consortia compared to single strains. Moreover, the commonly reported species for P-solubilization, Bacillus spp. and Pseudomonas spp., are outperformed by the scarcely implemented Burkholderia spp. Despite the similar responses to PSB in monocots and eudicots, species responsiveness to PSB varies within both clades. Remarkably, the meta-analysis challenges the common belief that PSB are less effective under field conditions compared to greenhouse conditions. This review provides innovative insights and identifies key questions for future research on PSB to promote their implementation in agriculture.

The unique ecosystem of the Congolese rainforest has only scarcely been explored for its plant-fungal interactions. Here, we characterized the root fungal communities of field-grown maize and of Panicum from adjacent borders in the Congo Basin and assessed parameters that could shape them. The soil properties indicated that comparable poor soil conditions prevailed in fields and borders, illustrating the low input character of local subsistence farming. The rhizosphere fungal communities, dominated by ascomycetous members, were structured by plant species, slash-and-burn practices and soil P, pH and C/N ratio. Examining fungi with potential plant growth-promoting abilities, the glomeromycotan communities appeared to be affected by the same parameters, whereas the inconspicuous symbionts of the order Sebacinales seemed less susceptible to environmental and anthropogenic factors. Notwithstanding the low abundances at which they were detected, sebacinoids occurred in 87% of the field samples, implying that they represent a consistent taxon within indigenous fungal populations across smallholder farm sites. Pending further insight into their ecosystem functionality, these data suggest that Sebacinales are robust root inhabitants that might be relevant for on-farm inoculum development within sustainable soil fertility management in the Sub-Saharan region.

Sugarcane juice is a traditional drink consumed on a daily basis in Egypt. The current work was conducted to (1) evaluate the multi-mycotoxin contamination in sugarcane juice, (2) identify the profiles of the mycotoxigenic fungi, and (3) assess the risk of mycotoxin exposure among the adult population. Sugarcane juice samples (n = 89) were randomly collected in two seasons from Egypt. Quantification of 13 mycotoxins was done through a validated LC-MS/MS method. The isolated fungal species were identified using the morphological and molecular approaches and a risk assessment was performed using @Risk software. In general, 63% (n = 56) of the samples were contaminated with aflatoxin B-1 (AFB(1)) and fumonisin B-1 (FB1). Juice collected during winter had a contamination range of 0.1-3 mu g L-1 for AFBI and 2.8-57.9 mu g L-1 for FB1, while samples from the summer season had a contamination range of 0.3-1.3 mu g L-1 for AFB(1) only. A multiple locus characterization showed the presence of Aspergillus, especially A. novoparasiticus, and Penicillium species. A remarkable difference in the levels of juice consumption was observed between the two seasons and between male and female consumer groups. However, the exposure levels using the probabilistic approach were not alarming.

Sugarcane juice is a traditional beverage consumed on a daily basis, by millions of people, in different countries such as Brazil, Egypt, India and Pakistan. Detection of mycotoxigenic fungi in sugarcane fields has been documented, however, very little is known about mycotoxin contamination in the other sugarcane-based products, especially sugarcane juice (1). The current study investigated the co-contamination of several mycotoxins (AFB1, AFB2, AFG1, AFG2, DAS, DON, FB1, FB2, FB3, OTA, STERIG, T-2, and ZEN). Further, a polyphasic approach was applied in order to identify and classify the isolated fungal species (2). Mycotoxin contamination results have been used to estimate the dietary exposure of the Egyptian population to mycotoxins through juice consumption. Samples (n=89) were randomly collected from local vendors and juice shops during two seasons (summer 2016 and winter 2017) from Assiut City, Upper Egypt. Extraction and quantification of mycotoxins were performed by liquid-liquid extraction followed by a validated UPLC-MS/MS analytical method. Morphological and molecular identification of mycotoxigenic fungi and other microorganisms was performed according to Samson et al., 2010 (2). Dereplication strategy was also applied to unravel the diversity of the produced secondary metabolites using an in-house screening library with a fast DDA UHPLC–TOF-MS profiling method that can screen for hundreds of mycotoxins and other metabolites. Risk assessment of mycotoxins using probabilistic and deterministic approaches at various scenarios for adult male and female Egyptian juice consumers was performed. So far the obtained results for the targeted analysis, exhibit the contamination of sugarcane juice samples with AFB1 and FB1 mycotoxins in 63% (n=56) of the samples. Juice collected during winter had a contamination range of 0.1-3 µg.L−1 for AFB1 and 4-58 µg.L−1 for FB1, while samples from the summer season had a contamination range of 0.3-1.3 µg.L−1 for AFB1 only. Furthermore, the risk assessment using probabilistic and deterministic approaches for adult male and female Egyptian juice consumers pinpointed a remarkable difference in levels of exposure to mycotoxins between the two seasons and between males & females. Other results will be presented during the workshop. References: 1. Abdallah MF, Krska R, Sulyok M (2016). Mycotoxin Contamination in Sugarcane Grass and Juice: First Report on Detection of Multiple Mycotoxins and Exposure Assessment for Aflatoxins B₁ and G₁ in Humans. 2. Samson RA, Houbraken J, Thrane U, et al. (2010). Food and indoor fungi. CBS KNAW Biodiversity Center, Utrecht.

Fusarium verticillioides is the most common fungal pathogen associated with maize ear rot in Tanzania. In a two-year trial, we investigated the efficacy of crop protection (insecticide and/or fungicide) and fertilizer (nitrogen and/or phosphorus) treatments in reducing the occurrence of F. verticillioides and its mycotoxins in maize grown in Tanzania. Seasonal differences were seen to have a substantial influence on the incidence and severity of insect infestation, Fusarium ear and kernel rot, biomass of F. verticillioides and contamination with fumonisins. With regard to the application of fertilizers, it was concluded that the impact on maize stalk borer injury, Fusarium symptoms and fumonisin levels was not significant, whereas crop protection significantly reduced maize damage. The application of an insecticide was most effective in reducing insect injury and as a result of the reduced insect injury the insecticide treatment also resulted in a significant decrease in Fusarium symptoms. In 2014, fumonisin levels were also significantly lower in maize treated with an insecticide. Additionally, significant positive correlations between insect damage and Fusarium symptoms were observed. In conclusion, this study clearly shows that application of an insecticide alone or in combination with a fungicide at anthesis significantly reduces insect damage and consequently reduces F. verticillioides infection and associated fumonisin contamination.

Maize cultivation in Tanzania is hampered by the presence of maize stalk borer (Busseola fusca) and Fusarium verticillioides, causing ear, kernel and stalk damage. Furthermore, accumulation of fumonisin mycotoxins deteriorates grain quality. Optimized agronomic practices remain the primary line of defense against pests and diseases. Therefore, the aim of this study was to gain insight into the effects of hybrid, sowing and harvest date on injuries due to maize stalk borer, Fusarium verticillioides symptoms and fumonisin contamination in maize grown in Tanzania. The analyses of the data collected during two growing seasons (2013 and 2014) showed early sowing and early harvesting as important measures for minimizing these problems. Although the impact of hybrid was not always significant, the early maturing hybrid STUKA M1 was consistently less susceptible compared to the hybrid STAHA. Correlation analysis revealed that stalk borer injuries, Fusarium symptoms and fumonisin levels were positively associated. The associations between yield and stalk borer injuries, Fusarium symptoms and fumonisin levels were negative, indicating that besides the mycotoxin issues, the insect and fungal pathogen also contribute to yield losses.

Fusarium head blight is a disease caused by a complex of Fusarium species. F. poae is omnipresent throughout Europe in spite of its low virulence. In this study, we assessed a geographically diverse collection of F. poae isolates for its genetic diversity using AFLP (Amplified Fragment Length Polymorphism). Furthermore, studying the mating type locus and chromosomal insertions, we identified hallmarks of both sexual recombination and clonal spread of successful genotypes in the population. Despite the large genetic variation found, all F. poae isolates possess the nivalenol chemotype based on Tri7 sequence analysis. Nevertheless, Tri gene clusters showed two layers of genetic variability. Firstly, the Tri1 locus was highly variable with mostly synonymous mutations and mutations in introns pointing to a strong purifying selection pressure. Secondly, in a subset of isolates, the main trichothecene gene cluster was invaded by a transposable element between Tri5 and Tri6. To investigate the impact of these variations on the phenotypic chemotype, mycotoxin production was assessed on artificial medium. Complex blends of type A and type B trichothecenes were produced but neither genetic variability in the Tri genes nor variability in the genome or geography accounted for the divergence in trichothecene production. In view of its complex chemotype, it will be of utmost interest to uncover the role of trichothecenes in virulence, spread and survival of F. poae.

Global food safety depends on continuous monitoring of food contaminants such as mycotoxins in cereals and cereal-derived products. Here, we combine this type of investigation with quantitative occurrence data on Fusarium infestation of these products in extensive correlation studies. Finally, this contributes to a thorough understanding of the presence, origin and physiology of Fusarium Head Blight (FHB) related mycotoxins and the correlations within their ranks.Two hundred and thirty-seven samples were analyzed from diverse cereal matrices, representing the most important stages of the cereal food and feed chain in Belgium. Food, feed and non-processed field samples were investigated, with a strong emphasis on whole-grain food products. Two approaches were pursued to estimate the full scope of FHB and its repercussions: UPLC–MS/MS was applied to detect twelve different mycotoxins, and Q-PCR was used to measure the presence of ten Fusarium species. We found that different matrices have different characteristic contamination profiles, and extensive correlation studies identified certain mycotoxins for future assessment (e.g. moniliformin produced by the Fusarium avenaceum/Fusarium tricinctum species group). The investigated harvest year of 2012 yielded many non-processed field materials containing elevated levels of deoxynivalenol (DON), while even in a so-called DON-year less prevalent toxins such as T-2 and HT-2 might be considered problematic due to their consistent co-occurrence with related mycotoxins.Our data illustrate complex interactions between the many Fusarium species that are responsible for FHB and their mycotoxins. Correlation studies demonstrate that consistent co-occurrence of mycotoxins is not to be neglected, and pinpoint issues for future surveillance and legislation.

Triticale is the intergeneric hybrid between wheat and rye. With the expansion of the triticale growing area, powdery mildew has emerged and become a significant disease on this new host. Recent research demonstrated that this new' powdery mildew on triticale has emerged through a host range expansion of powdery mildew of wheat. Moreover, isolates sampled from triticale still infect their previous host, wheat, but isolates sampled from wheat hardly infect triticale. Race-specific and adult-plant resistance have been identified in triticale cultivars. The main objective of this study was to characterize the cellular basis of powdery mildew resistance in triticale. Commonalities with resistance responses in other cereals such as wheat, barley and oat are discussed. A detailed comparative histological study of various resistance responses during cross-inoculation of either virulent or avirulent wheat and triticale isolates on both hosts was carried out. The present data provide evidence that for incompatible interactions, the formation of non-penetrated papillae is the predominant resistance response, while the hypersensitive response (HR) acts as a second line of defence, to cut the fungus off from nutrients, if penetration resistance fails. It is not clear yet what causes the slower growth and reduced colony size of triticale isolates when inoculated on wheat. Possibly, post-penetration resistance mechanisms, other than HR, are switched on during these (semi-) compatible interactions. Molecular studies on gene expression and gene function of defence-related genes might reveal further insights into the genetic basis of these resistance responses.

The development of new crop species and their associated agro-ecosystems led simultaneously to the emergence of new pathogens (Stukenbrock and McDonald, 2008). This research focused on the recent emergence of powdery mildew (Blumeria graminis) on triticale (x Triticosecale Wittmack). In a first part, we aimed to gain insights into the evolutionary origin of this pathogen on its new host. A secondary aim was to investigate the presence of powdery mildew resistance in current commercial triticale cultivars, including its cellular basis of resistance. To address these research goals, we have pursued a molecular, pathological and cytological approach. This discussion will reflect on the experimental findings described in this research and their impact for future management of powdery mildew on triticale and other cereals.

Fusarium head blight (FHB) is a devastating disease on small cereal crops and is primarily caused by Fusarium graminearum. FHB typically is associated with a contamination of the infected grains with mycotoxins, which can act as virulence factors for the advancing fungus. Some wheat plants have evolved a typical resistance designated Fhb1 resistance, which is associated with a glucosyltransferase function, capable of detoxifying the deposited deoxynivalenol (DON), a mycotoxin abundantly produced by F. graminearum. However, the resulting 'masked mycotoxin', deoxynivalenol-3-glucoside (DON-3G), may be metabolized in the digestive tract of animals, releasing the native toxin anew. The present study aims to assess the glucosylation capacity in the Belgian commercial wheat pool and to determine the importance of DON glucosylation in the complex background of natural field circumstances. Clear indications were found that several Belgian commercial wheat cultivars, to some extent, do possess a glucosyltransferase function capable of detoxifying significant amounts of DON. However, the level of glucosylation in the field did not correlate well with disease severity. The present research provides the first large-scale assessment of this glucosylation capacity in Belgian commercial wheat cultivars. (C) 2013 Elsevier Ltd. All rights reserved.

Triticale (×Triticosecale) is the intergeneric hybrid between the female parent wheat and the male parent rye. With the expansion of the triticale growing area, powdery mildew emerged on this new host and has become a significant disease on triticale. Recent research demonstrated that this "new" powdery mildew on triticale has emerged through a host range expansion of powdery mildew of wheat. Moreover, this expansion occurred recently and multiple times at different locations in Europe. An effective and environmentally sensitive approach to controlling powdery mildew involves breeding crop plants for resistance. The main goal of this study was to identify the presence of powdery mildew resistance in commercial triticale cultivars. First, the avirulence (AVR) genes and gene complexity carried by this new powdery mildew population on triticale were characterized. Virulence was identified for all the resistance genes evaluated in the present study, and virulence frequencies higher than 50% were recorded on the genes Pm3f, Pm5b, Pm6, Pm7, Pm8, and Pm17. Using molecular markers, the presence of resistance genes Pm3f and Pm17 was identified in certain triticale cultivars. The triticale cultivars were also evaluated for the presence of quantitative resistance at adult plant growth stages in a 2-year field experiment. Despite the high disease pressure, cultivars highly resistant at the adult-plant growth stages were identified. Because 'Grenado' also showed effective race-specific resistance, this cultivar could be of high value for breeding for durable resistance to powdery mildew. Altogether, this study reveals valuable information on the presence of powdery mildew resistance in commercial triticale cultivars, which can be used in breeding programs in triticale. Additionally, this study underscores the need to broaden the base of powdery mildew resistance in triticale through introgression and deployment of new sources of mildew resistance, including quantitative resistance.

BACKGROUND:Powdery mildew, caused by the obligate biotrophic fungus Blumeria graminis, is a major problem in cereal production as it can reduce quality and yield. B. graminis has evolved eight distinct formae speciales (f.sp.) which display strict host specialization. In the last decade, powdery mildew has emerged on triticale, the artificial intergeneric hybrid between wheat and rye. This emergence is probably triggered by a host range expansion of the wheat powdery mildew B. graminis f.sp. tritici. To gain more precise information about the evolutionary processes that led to this host range expansion, we pursued a combined pathological and genetic approach.RESULTS:B. graminis isolates were sampled from triticale, wheat and rye from different breeding regions in Europe. Pathogenicity tests showed that isolates collected from triticale are highly pathogenic on most of the tested triticale cultivars. Moreover, these isolates were also able to infect several wheat cultivars (their previous hosts), although a lower aggressiveness was observed compared to isolates collected from wheat. Phylogenetic analysis of nuclear gene regions identified two statistically significant clades, which to a certain extent correlated with pathogenicity. No differences in virulence profiles were found among the sampled regions, but the distribution of genetic variation demonstrated to be geography dependent. A multilocus haplotype network showed that haplotypes pathogenic on triticale are distributed at different sites in the network, but always clustered at or near the tips of the network.CONCLUSIONS:This study reveals a genetic structure in B. graminis with population differentiation according to geography and host specificity. In addition, evidence is brought forward demonstrating that the host range expansion of wheat isolates to the new host triticale occurred recently and multiple times at different locations in Europe.