Lipid Signaling Modulates the Response to Fumonisin Contamination and Its Source, Fusarium verticillioides, in Maize
ABSTRACT: Fumonisin-contaminated maize (Zea mays L.) products are a major health concern because of their toxic effects in humans and animals. Breeding maize for increased mycotoxin resistance is one of the key sustainable strategies for mitigating the effects of fumonisin contamination. Recent studies suggest a link between fumonisin accumulation and plant lipid and oxylipin profiles. However, the data collected so far do not reveal a cause-and-effect relationship. In this study, to decipher the multifactorial nature of mycotoxin resistance and plant–pathogen interaction mechanisms, we examined the oxylipin and complex lipid profiles of two maize hybrids (H21 and H22, the latter showing significantly lower FBs content) grown in the open field in two locations over 3years. Untargeted ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight (UHPLC-Q-TOF), together with chemometrics analysis, successfully distinguished between the two hybrids as having low- and high-level fumonisin contamination. Considering that H21 and H22 were exposed to the same environmental factors, the higher activation of lipid signaling systems in H22 suggests that other routes are enabled in the less susceptible hybrids to limit fumonisin B (FB) accumulation. Our results highlighted the crucial role played by oxylipin and sphingolipid signaling in modulating the complex maize response to F. verticillioides infection. Overall, our results returned a global view on the changes in lipid metabolites related to fumonisin accumulation under open field conditions, and revealed a strong activation of the lipid signaling cascade in maize in the presence of FB1.
Project description:Fumonisins are mycotoxins (MTs) produced mainly by the fungus Fusarium verticillioides, the main pathogens of maize which cause ear rot. The aim of this work was to evaluate some factors that may lead to high fumonisin production by F. verticillioides in maize grains, correlating the pathogen inoculation method with different genotypes grown in four Brazilian states. Experiments were conducted in 2015-2016 in maize crops from experimental maize fields located in four distinct states of Brazil. Results showed that contamination by fumonisin mycotoxins occurred even on symptomatic or asymptomatic grains. In all municipalities, the samples showed levels of fumonisin B1 that were higher than would be tolerable for the human consumption of corn products (the current tolerance limit for fumonisin is 1.5 ?g g-1). High severity of grains infected with F. verticillioides does not always show high concentrations of fumonisins. Environments with higher temperatures may influence the production of high concentrations of fumonisin in maize hybrids. Spray inoculation methods and inoculation at the center of spikes did not influence fumonisin concentrations. Results showed that the hybrids P3630H, P32R48 and P3250 presented higher disease severity, as well as higher mycotoxin levels in the studied locations with higher temperatures.
Project description:Maize is by quantity the most important C4 cereal crop; however, future climate changes are expected to increase maize susceptibility to mycotoxigenic fungal pathogens and reduce productivity. While rising atmospheric [CO2 ] is a driving force behind the warmer temperatures and drought, which aggravate fungal disease and mycotoxin accumulation, our understanding of how elevated [CO2 ] will effect maize defences against such pathogens is limited. Here we report that elevated [CO2 ] increases maize susceptibility to Fusarium verticillioides proliferation, while mycotoxin levels are unaltered. Fumonisin production is not proportional to the increase in F.?verticillioides biomass, and the amount of fumonisin produced per unit pathogen is reduced at elevated [CO2 ]. Following F.?verticillioides stalk inoculation, the accumulation of sugars, free fatty acids, lipoxygenase (LOX) transcripts, phytohormones and downstream phytoalexins is dampened in maize grown at elevated [CO2 ]. The attenuation of maize 13-LOXs and jasmonic acid production correlates with reduced terpenoid phytoalexins and increased susceptibility. Furthermore, the attenuated induction of 9-LOXs, which have been suggested to stimulate mycotoxin biosynthesis, is consistent with reduced fumonisin per unit fungal biomass at elevated [CO2 ]. Our findings suggest that elevated [CO2 ] will compromise maize LOX-dependent signalling, which will influence the interactions between maize and mycotoxigenic fungi.
Project description:Mycotoxin contamination is one of the main problems affecting corn production, due to its significant risk to human and animal health. The Fusarium and Aspergillus species are the main producers of mycotoxins in maize, infecting both pre-harvest and during storage. In this work, we evaluated the presence of mycotoxins and their producing species along maize production cycles in three different stages (anthesis, harvest, and storage) during three consecutive seasons (2016-2018). Fungal occurrences were studied using species-specific PCR protocols, whereas mycotoxin levels were determined by LC-MS/MS. Fumonisin-producing Fusarium species (F. verticillioides and F. proliferatum), as well as the aflatoxin producer Aspergillus flavus, were the most predominant species at all stages; although, during some seasons, the presence of F. graminearum and A. niger aggregate species were also identified. Contrastingly, fumonisins were the only mycotoxins detected and levels were always under legal regulations. The results presented here demonstrate that even when fungal contamination occurs at the early stages of the maize production cycle, the application of good agricultural and storage practices might be crucial to ensure mycotoxin-free grains.
Project description:The mycotoxin fumonisin (FB) has become a major problem in maize products in southeastern Asia. Fumonisin can affect the health of humans and many animals. Fumonisin contamination can be reduced by detoxifying microbial enzyme. Screening of 95 potent natural sources resulted in 5.3% of samples yielding a total of five bacterial isolates that were a promising solution, reducing approximately 10.0-30.0% of fumonisin B1 (FB1). <i>Serratia marcescens</i>, one of the dominant degrading bacteria, was identified with Gram staining, 16S rRNA gene, and MALDI-TOF/TOF MS. Cell-free extract showed the highest fumonisin reduction rates, 30.3% in solution and 37.0% in maize. Crude proteins from bacterial cells were analyzed with a label-free quantification technique. The results showed that hydrolase enzymes and transferase enzymes that can cooperate in the fumonisin degradation process were highly expressed in comparison to their levels in a control. These studies have shown that <i>S. marcescens</i> 329-2 is a new potential bacterium for FB1 reduction, and the production of FB1-reducing enzymes should be further explored.
Project description:Infection by the fumonisin-producing Fusarium spp. and subsequent fumonisin contamination of maize adversely affect international trade and economy with deleterious effects on human and animal health. In developed countries high standards of the major food suppliers and retailers are upheld and regulatory controls deter the importation and local marketing of fumonisin-contaminated food products. In developing countries regulatory measures are either lacking or poorly enforced, due to food insecurity, resulting in an increased mycotoxin exposure. The lack and poor accessibility of effective and environmentally safe control methods have led to an increased interest in practical and biological alternatives to reduce fumonisin intake. These include the application of natural resources, including plants, microbial cultures, genetic material thereof, or clay minerals pre- and post-harvest. Pre-harvest approaches include breeding for resistant maize cultivars, introduction of biocontrol microorganisms, application of phenolic plant extracts, and expression of antifungal proteins and fumonisin degrading enzymes in transgenic maize cultivars. Post-harvest approaches include the removal of fumonisins by natural clay adsorbents and enzymatic degradation of fumonisins through decarboxylation and deamination by recombinant carboxylesterase and aminotransferase enzymes. Although, the knowledge base on biological control methods has expanded, only a limited number of authorized decontamination products and methods are commercially available. As many studies detailed the use of natural compounds in vitro, concepts in reducing fumonisin contamination should be developed further for application in planta and in the field pre-harvest, post-harvest, and during storage and food-processing. In developed countries an integrated approach, involving good agricultural management practices, hazard analysis and critical control point (HACCP) production, and storage management, together with selected biologically based treatments, mild chemical and physical treatments could reduce fumonisin contamination effectively. In rural subsistence farming communities, simple, practical, and culturally acceptable hand-sorting, maize kernel washing, and dehulling intervention methods proved to be effective as a last line of defense for reducing fumonisin exposure. Biologically based methods for control of fumonisin-producing Fusarium spp. and decontamination of the fumonisins could have potential commercial application, while simple and practical intervention strategies could also impact positively on food safety and security, especially in rural populations reliant on maize as a dietary staple.
Project description:Mycotoxins are toxic secondary fungal metabolites that can negatively affect animal productivity when ingested through feed. In order to assess mycotoxin contamination of poultry feed and feed ingredients vis-a-vis source tracking of feed contamination in Nigeria, 102 samples of feed (n?=?30) and feed ingredients (n?=?72) were collected from in-house mills of poultry farms across 12 states of Nigeria and analyzed for multiple mycotoxins using LC/MS-MS. One hundred and forty microbial metabolites were detected in the feed and feed ingredients. The most frequent mycotoxin in the feed was fumonisin B<sub>1</sub>, occurring in 97% of the samples at mean concentration of 1014 ?g kg<sup>-1</sup>. AFB<sub>1</sub> occurred in 83% of the feed samples at mean concentration of 74 ?g kg<sup>-1</sup> and in all feed ingredients except fish meal and other cereals (millet and rice). Feed samples analyzed in this study were contaminated with at least four mycotoxins: aflatoxins and fumonisin co-occurring in 80% of the samples. Peanut cake and maize contributed the most to the levels of aflatoxin and fumonisin, respectively, in the feed. Consequently, there is a need to explore other cereal- and protein-based ingredients for compounding feeds in order to reduce the risk associated with high mycotoxin (e.g. aflatoxin) intake in poultry.
Project description:Fusarium verticillioides is a common maize pathogen causing ear rot (FER) and contamination of the grains with the fumonisin B1 (FB1) mycotoxin. Resistance to FER and FB1 contamination are quantitative traits, affected by environmental conditions, and completely resistant maize genotypes to the pathogen are so far unknown. In order to uncover genomic regions associated to reduced FER and FB1 contamination and identify molecular markers for assisted selection, an F2:3 population of 188 progenies was developed crossing CO441 (resistant) and CO354 (susceptible) genotypes. FER severity and FB1 contamination content were evaluated over 2 years and sowing dates (early and late) in ears artificially inoculated with F. verticillioides by the use of either side-needle or toothpick inoculation techniques.Weather conditions significantly changed in the two phenotyping seasons and FER and FB1 content distribution significantly differed in the F3 progenies according to the year and the sowing time. Significant positive correlations (P?<?0.01) were detected between FER and FB1 contamination, ranging from 0.72 to 0.81. A low positive correlation was determined between FB1 contamination and silking time (DTS). A genetic map was generated for the cross, based on 41 microsatellite markers and 342 single nucleotide polymorphisms (SNPs) derived from Genotyping-by-Sequencing (GBS). QTL analyses revealed 15 QTLs for FER, 17 QTLs for FB1 contamination and nine QTLs for DTS. Eight QTLs located on linkage group (LG) 1, 2, 3, 6, 7 and 9 were in common between FER and FB1, making possible the selection of genotypes with both low disease severity and low fumonisin contamination. Moreover, five QTLs on LGs 1, 2, 4, 5 and 9 located close to previously reported QTLs for resistance to other mycotoxigenic fungi. Finally, 24 candidate genes for resistance to F. verticillioides are proposed combining previous transcriptomic data with QTL mapping.This study identified a set of QTLs and candidate genes that could accelerate breeding for resistance of maize lines showing reduced disease severity and low mycotoxin contamination determined by F. verticillioides.
Project description:Mycotoxins occur worldwide in the major grains, and producers, traders and processors are all challenged to prevent serious health problems for consumers. The challenges originate with pre-harvest fungi infections in the grain fields, increased contamination during improper storage and, finally, the mycotoxin accumulation in commercial food and feed products. Little is known about the multi-mycotoxin occurrence in maize and wheat commercially produced in South Africa. This is the first comprehensive study that reports on the multi-mycotoxin occurrence in South African produced maize and wheat crops after harvest, over four production seasons, in all the production regions of the country. The study was made possible with the development of a fit-for-purpose, cost-effective LC-MS/MS multi-mycotoxin method, validated for 13 "regulated" mycotoxins. A low mycotoxin risk was found in South African produced wheat, with only deoxynivalenol (DON) in 12.5% of the 160 samples at levels well below the 2000 µg/kg South African (SA) regulatory level. It was concluded that aflatoxin B1 (AFB1) is seldom present in South African produced commercial maize. The concentrations, regional variation and seasonal trends of deoxynivalenol and fumonisins, the two most prevalent mycotoxins, and of zearalenone (ZON), are reported for white and yellow maize in all the production provinces, based on the analytical results of 1400 maize samples. A threefold to eightfold increase in deoxynivalenol mean concentrations in white maize was observed in the main production regions in the fourth season, with 8.9% samples above 2000 µg/kg. A strong correlation was found between higher deoxynivalenol concentrations and the presence of 15-acetyl-deoxynivalenol (15-ADON). The mean fumonisin concentrations were well below the 4000 µg/kg South African regulatory value. A possible shift in the incidence and severity of mycotoxigenic Fusarium spp. in the provinces must be investigated. The variations and trends highlight the importance of a continuous monitoring of multi-mycotoxins in South Africa along the grain value chain.
Project description:The spatiotemporal trends in aflatoxin B1 (AFB1), fumonisin B1 (FB1), and deoxynivalenol (DON) accumulation were analyzed in a range of food commodities (maize, groundnut, pearl millet, rice, and wheat) in village settings in Unnao, Uttar Pradesh, India. Samples (n = 1549) were collected across six communities and six time points spanning a calendar year and were analyzed for mycotoxins using enzyme-linked immunosorbent assays. AFB1 and FB1 were common across surveyed villages, with moderate to high detection rates (45-75%) observed across commodities. AFB1 levels in maize and groundnuts and FB1 levels in maize and pearl millet frequently exceeded regulatory threshold levels of 15 μg/kg (AFB1) and 2 μg/g (FB1). DON was analyzed in wheat, with 3% of samples yielding detectable levels and none exceeding 1 μg/g. In rice, AFB1 levels were highest in the bran and husk and lower in the kernel. Commodity type significantly influenced AFB1 detection status, while commodity type, season, and visual quality influenced samples' legal status. Storage characteristics and household socioeconomic status indicators did not have significant effects on contamination. No significant effects of any variables on FB1 detection or legal status were observed. Data on mycotoxin contamination, combined with data on local dietary intake, were used to estimate spatiotemporal mycotoxin exposure profiles. Estimated seasonal <i>per capita</i> exposure levels for AFB1 (5.4-39.3 ng/kg body weight/day) and FB1 (~0-2.4 μg/kg body weight/day) exceeded provisional maximum tolerable daily intake levels (1 ng/kg body weight/day for AFB1 and 2 μg/kg body weight/day for FB1) in some seasons and locations. This study demonstrates substantial dietary mycotoxin exposure risk in Unnao food systems and serves as an evidentiary foundation for participatory food safety intervention in the region.
Project description:The presence of mycotoxins in cereal grain is a very important food safety issue with the occurrence of masked mycotoxins extensively investigated in recent years. This study investigated the variation of different Fusarium metabolites (including the related regulated, masked, and emerging mycotoxin) in maize from various agriculture regions of South Africa. The relationship between the maize producing regions, the maize type, as well as the mycotoxins was established. A total of 123 maize samples was analyzed by a LC-MS/MS multi-mycotoxin method. The results revealed that all maize types exhibited a mixture of free, masked, and emerging mycotoxins contamination across the regions with an average of 5 and up to 24 out of 42 investigated Fusarium mycotoxins, including 1 to 3 masked forms at the same time. Data obtained show that fumonisin B1, B2, B3, B4, and A1 were the most prevalent mycotoxins and had maximum contamination levels of 8908, 3383, 990, 1014, and 51.5 µg/kg, respectively. Deoxynivalenol occurred in 50% of the samples with a mean concentration of 152 µg/kg (max 1380 µg/kg). Thirty-three percent of the samples were contaminated with zearalenone at a mean concentration of 13.6 µg/kg (max 146 µg/kg). Of the masked mycotoxins, DON-3-glucoside occurred at a high incidence level of 53%. Among emerging toxins, moniliformin, fusarinolic acid, and beauvericin showed high occurrences at 98%, 98%, and 83%, and had maximum contamination levels of 1130, 3422, and 142 µg/kg, respectively. Significant differences in the contamination pattern were observed between the agricultural regions and maize types.