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1.
Curr Opin Lipidol ; 33(4): 257-263, 2022 08 01.
Article in English | MEDLINE | ID: mdl-35942820

ABSTRACT

PURPOSE OF REVIEW: The role of lipoprotein (a) in atherogenesis has been the subject of argument for many years. Evidence that it is raised in familial hypercholesterolaemia has been disputed not least because a mechanism related to low density lipoprotein (LDL) receptor mediated catabolism has been lacking. Whether lipoprotein (a) increases the already raised atherosclerotic cardiovascular disease (ASCVD) risk in familial hypercholesterolaemia is also more dubious than is often stated. We review the evidence in an attempt to provide greater clarity. RECENT FINDINGS: Lipoprotein (a) levels are raised as a consequence of inheriting familial hypercholesterolaemia. The mechanism for this is likely to involve increased hepatic production, probably mediated by PCSK9 augmented by apolipoprotein E. The extent to which raised lipoprotein (a) contributes to the increased ASCVD risk in familial hypercholesterolaemia remains controversial.Unlike, for example, statins which are effective across the whole spectrum of LDL concentrations, drugs in development to specifically lower lipoprotein (a) are likely to be most effective in people with the highest levels of lipoprotein (a). People with familial hypercholesterolaemia may therefore be in the vanguard of those in whom theses agents should be exhibited. SUMMARY: Inheritance of familial hypercholesterolaemia undoubtedly increases the likelihood that lipoprotein (a) will be raised. However, in familial hypercholesterolaemia when ASCVD incidence is already greatly increased due to high LDL cholesterol, whether lipoprotein (a) contributes further to this risk cogently needs to be tested with drugs designed to specifically lower lipoprotein (a).


Subject(s)
Atherosclerosis , Hydroxymethylglutaryl-CoA Reductase Inhibitors , Hypercholesterolemia , Hyperlipoproteinemia Type II , Atherosclerosis/complications , Atherosclerosis/epidemiology , Atherosclerosis/genetics , Humans , Hydroxymethylglutaryl-CoA Reductase Inhibitors/therapeutic use , Hypercholesterolemia/complications , Hyperlipoproteinemia Type II/complications , Hyperlipoproteinemia Type II/epidemiology , Hyperlipoproteinemia Type II/genetics , Lipoprotein(a) , Proprotein Convertase 9
3.
Data Brief ; 42: 108033, 2022 Jun.
Article in English | MEDLINE | ID: mdl-35330736

ABSTRACT

Information on the transcriptomic changes that occur within sclerotia of Aspergillus flavus during its sexual cycle is very limited and warrants further research. The findings will broaden our knowledge of the biology of A. flavus and can provide valuable insights in the development or deployment of non-toxigenic strains as biocontrol agents against aflatoxigenic strains. This article presents transcriptomic datasets included in our research article entitled, "Development of sexual structures influences metabolomic and transcriptomic profiles in Aspergillus flavus" [1], which utilized transcriptomics to identify possible genes and gene clusters associated with sexual reproduction and fertilization in A. flavus. RNA was extracted from sclerotia of a high fertility cross (Hi-Fert-Mated), a low fertility cross (Lo-Fert-Mated), and unmated strains (Hi-Fert-Unmated and Lo-Fert-Unmated) of A. flavus collected immediately after crossing and at every two weeks until eight weeks of incubation on mixed cereal agar at 30 °C in continuous darkness (n = 4 replicates from each treatment for each time point; 80 total). Raw sequencing reads obtained on an Illumina NovaSeq 6000 were deposited in NCBI's Sequence Read Archive (SRA) repository under BioProject accession number PRJNA789260. Reads were mapped to the A. flavus NRRL 3357 genome (assembly JCVI-afl1-v2.0; GCA_000006275.2) using STAR software. Differential gene expression analyses, functional analyses, and weighted gene co-expression network analysis were performed using DESeq2 R packages. The raw and analyzed data presented in this article could be reused for comparisons with other datasets to obtain transcriptional differences among strains of A. flavus or closely related species. The data can also be used for further investigation of the molecular basis of different processes involved in sexual reproduction and sclerotia fertility in A. flavus.

4.
Fungal Biol ; 126(3): 187-200, 2022 03.
Article in English | MEDLINE | ID: mdl-35183336

ABSTRACT

Sclerotium (female) fertility, the ability of a strain to produce ascocarps, influences internal morphological changes during sexual reproduction in Aspergillus flavus. Although sclerotial morphogenesis has been linked to secondary metabolite (SM) biosynthesis, metabolic and transcriptomic changes within A. flavus sclerotia during sexual development are not known. Successful mating between compatible strains may result in relatively high or low numbers of ascocarps being produced. Sclerotia from a high fertility cross (Hi-Fert-Mated), a low fertility cross (Lo-Fert-Mated), unmated strains (Hi-Fert-Unmated and Lo-Fert-Unmated) were harvested immediately after crosses were made and every two weeks until 8 weeks of incubation, then subjected to targeted metabolomics (n = 106) and transcriptomics analyses (n = 80). Aflatoxin B1 production varied between Hi-Fert-Mated and Hi-Fert-Unmated sclerotia, while it remained low or was undetected in Lo-Fert-Mated and Lo-Fert-Unmated sclerotia. Profiling of 14 SMs showed elevated production of an aflavazole analog, an aflavinine isomer, and hydroxyaflavinine in Hi-Fert-Mated sclerotia at 4 to 8 weeks. Similarly, genes ayg1, hxtA, MAT1, asd-3, preA and preB, and genes in uncharacterized SM gene clusters 30 and 44 showed increased expression in Hi-Fert-Mated sclerotia at these time points. These results broaden our knowledge of the biochemical and transcriptional processes during sexual development in A. flavus.


Subject(s)
Aflatoxins , Aspergillus flavus , Aflatoxins/metabolism , Gene Expression Profiling , Metabolomics , Reproduction/genetics , Transcriptome
6.
Mycologia ; 112(5): 908-920, 2020.
Article in English | MEDLINE | ID: mdl-32821029

ABSTRACT

Aspergillus flavus contaminates agricultural products worldwide with carcinogenic aflatoxins that pose a serious health risk to humans and animals. The fungus survives adverse environmental conditions through production of sclerotia. When fertilized by a compatible conidium of an opposite mating type, a sclerotium transforms into a stroma within which ascocarps, asci, and ascospores are formed. However, the transition from a sclerotium to a stroma during sexual reproduction in A. flavus is not well understood. Early events during the interaction between sexually compatible strains of A. flavus were visualized using conidia of a green fluorescent protein (GFP)-labeled MAT1-1 strain and sclerotia of an mCherry-labeled MAT1-2 strain. Both conidia and sclerotia of transformed strains germinated to produce hyphae within 24 h of incubation. Hyphal growth of these two strains produced what appeared to be a network of interlocking hyphal strands that were observed at the base of the mCherry-labeled sclerotia (i.e., region in contact with agar surface) after 72 h of incubation. At 5 wk following incubation, intracellular green-fluorescent hyphal strands were observed within the stromatal matrix of the mCherry-labeled strain. Scanning electron microscopy of stromata from a high- and low-fertility cross and unmated sclerotia was used to visualize the formation and development of sexual structures within the stromatal and sclerotial matrices, starting at the time of crossing and thereafter every 2 wk until 8 wk of incubation. Morphological differences between sclerotia and stromata became apparent at 4 wk of incubation. Internal hyphae and croziers were detected inside multiple ascocarps that developed within the stromatal matrix of the high-fertility cross but were not detected in the matrix of the low-fertility cross or the unmated sclerotia. At 6 to 8 wk of incubation, hyphal tips produced numerous asci, each containing one to eight ascospores that emerged out of an ascus following the breakdown of the ascus wall. These observations broaden our knowledge of early events during sexual reproduction and suggest that hyphae from the conidium-producing strain may be involved in the early stages of sexual reproduction in A. flavus. When combined with omics data, these findings could be useful in further exploration of the molecular and biochemical mechanisms underlying sexual reproduction in A. flavus.


Subject(s)
Aspergillus flavus/cytology , Aspergillus flavus/growth & development , Fruiting Bodies, Fungal/cytology , Fruiting Bodies, Fungal/growth & development , Reproduction/physiology , Spores, Fungal/cytology , Spores, Fungal/growth & development , Aspergillus flavus/genetics , Fertility , Food Contamination , Fruiting Bodies, Fungal/genetics , Genetic Variation , Genotype , Humans , Mycotoxins , Plant Development/genetics , Plant Development/physiology , Reproduction/genetics , Spores, Fungal/genetics
7.
G3 (Bethesda) ; 10(10): 3515-3531, 2020 10 05.
Article in English | MEDLINE | ID: mdl-32817124

ABSTRACT

Efforts in genome sequencing in the Aspergillus genus have led to the development of quality reference genomes for several important species including A. nidulans, A. fumigatus, and A. oryzae However, less progress has been made for A. flavus As part of the effort of the USDA-ARS Annual Aflatoxin Workshop Fungal Genome Project, the isolate NRRL3357 was sequenced and resulted in a scaffold-level genome released in 2005. Our goal has been biologically driven, focusing on two areas: isolate variation in aflatoxin production and drought stress exacerbating aflatoxin production by A. flavus Therefore, we developed two reference pseudomolecule genome assemblies derived from chromosome arms for two isolates: AF13, a MAT1-2, highly stress tolerant, and highly aflatoxigenic isolate; and NRRL3357, a MAT1-1, less stress tolerant, and moderate aflatoxin producer in comparison to AF13. Here, we report these two reference-grade assemblies for these isolates through a combination of PacBio long-read sequencing and optical mapping, and coupled them with comparative, functional, and phylogenetic analyses. This analysis resulted in the identification of 153 and 45 unique genes in AF13 and NRRL3357, respectively. We also confirmed the presence of a unique 310 Kb insertion in AF13 containing 60 genes. Analysis of this insertion revealed the presence of a bZIP transcription factor, named atfC, which may contribute to isolate pathogenicity and stress tolerance. Phylogenomic analyses comparing these and other available assemblies also suggest that the species complex of A. flavus is polyphyletic.


Subject(s)
Aflatoxins , Aspergillus flavus , Aspergillus flavus/genetics , Base Sequence , Genome, Fungal , Phylogeny
8.
Front Microbiol ; 11: 853, 2020.
Article in English | MEDLINE | ID: mdl-32582038

ABSTRACT

The interaction between Aspergillus flavus and Zea mays is complex, and the identification of plant genes and pathways conferring resistance to the fungus has been challenging. Therefore, the authors undertook a systems biology approach involving dual RNA-seq to determine the simultaneous response from the host and the pathogen. What was dramatically highlighted in the analysis is the uniformity in the development patterns of gene expression of the host and the pathogen during infection. This led to the development of a "stage of infection index" that was subsequently used to categorize the samples before down-stream system biology analysis. Additionally, we were able to ascertain that key maize genes in pathways such as the jasmonate, ethylene and ROS pathways, were up-regulated in the study. The stage of infection index used for the transcriptomic analysis revealed that A. flavus produces a relatively limited number of transcripts during the early stages (0 to 12 h) of infection. At later stages, in A. flavus, transcripts and pathways involved in endosomal transport, aflatoxin production, and carbohydrate metabolism were up-regulated. Multiple WRKY genes targeting the activation of the resistance pathways (i.e., jasmonate, phenylpropanoid, and ethylene) were detected using causal inference analysis. This analysis also revealed, for the first time, the activation of Z. mays resistance genes influencing the expression of specific A. flavus genes. Our results show that A. flavus seems to be reacting to a hostile environment resulting from the activation of resistance pathways in Z. mays. This study revealed the dynamic nature of the interaction between the two organisms.

9.
Rev. argent. microbiol ; 51(4): 292-301, dic. 2019. graf
Article in English | LILACS | ID: biblio-1057392

ABSTRACT

Abstract Aflatoxin is a carcinogenic secondary metabolite produced mainly by Aspergillus flavus and Aspergillus parasiticus, which can seriously endanger the health of humans and animals. Oxidative stress is a common defense response, and it is known that reactive oxygen species (ROS) can induce the synthesis of a series of secondary metabolites, including aflatoxin. By using mutants lacking the afap 1 gene, the role of afap 1 gene in oxidative stress and aflatoxin synthesis was assessed. The growth of the mutant strains was significantly inhibited by the increase in the concentration of H2O2, inhibition was complete at 40mmol/l. However, in the quantitative analysis by HPLC, the concentration of AFB1 increased with the increased H 2O 2 until 10mmol/l. Following an analysis based on the information provided by the NCBI BLAST analysis, it was assumed that Afap1, a basic leucine zipper (bZIP) transcription factor, was associated with the oxidative stress in this fungus. Treatment with 5mmol/l H 2O 2 completely inhibited the growth of the mutant strains in afap 1 but did not affect the growth of the CA14PTs strain (non-mutant strain). In addition, the concentration of AFB 1 in the mutant strains was approximately V of that observed in the CA14PTs strain. These results suggested that Afap1 plays a key role in the regulation of oxidative stress and aflatoxin production in A. flavus. ©2018 Published by Elsevier España, S.L.U. on behalf of Asociación Argentina de Microbiología. This is an open access article under the CC BY-NC-ND license (https://creativecommons.org/ licenses/by-nc-nd/4.0/).


Resumen La aflatoxina es un metabolito secundario cancerígeno producido principalmente por Aspergillus flavus y Aspergillus parasiticus, que pone en riesgo grave a la salud de los humanos y los animales. El estrés oxidativo es una respuesta de defensa común, y es sabido que las especies reactivas de oxígeno (ROS) pueden inducir la síntesis de una serie de metabolitos secundarios, incluida la aflatoxina. Empleando mutantes carentes del gen afap1 se evaluó el papel de Afap1 en el estrés oxidativo y la síntesis de aflatoxinas. El crecimiento de las cepas mutadas se vio significativamente inhibido con el aumento de la concentración de H 2O 2, la inhibición fue completa a 40mmol/l. Sin embargo, en el análisis cuantitativo por HPLC, la concentración de la aflatoxina AFBi aumentó con el aumento de la concentración de H 2O 2 hasta 10mmol/l. Tras un análisis apoyado en la información provista por la herramienta NCBI BLAST, se supuso que Afap1, un factor de transcripción de la cremallera de leucina básica (bZIP), estaba asociado con el estrés oxidativo en este hongo. El tratamiento con 5mmol/l de H 2O 2 inhibió completamente el crecimiento de las cepas mutantes en afap1, pero no afectó el crecimiento de la cepa CA14PTs (cepa no mutada). Además, la concentración de AFB 1 en las cepas mutadas fue de aproximadamente 1/4 de la observada en CA14PTs. Estos resultados sugieren que Afap1 juega un papel clave en la regulación del estrés oxidativo y la producción de aflatoxinas en A. flavus.


Subject(s)
Aspergillus flavus/pathogenicity , Aflatoxins/biosynthesis , Transcription Factors/analysis , Oxidative Stress/physiology
10.
J Agric Food Chem ; 67(18): 5230-5239, 2019 May 08.
Article in English | MEDLINE | ID: mdl-30986348

ABSTRACT

Conventional methods for detecting aflatoxigenic fungus and aflatoxin contamination are generally time-consuming, sample-destructive, and require skilled personnel to perform, making them impossible for large-scale nondestructive screening detection, real-time, and on-site analysis. Therefore, the potential of visible-near-infrared (Vis-NIR) spectroscopy over the 400-2500 nm spectral range was examined for determination of aflatoxigenic fungus infection and the corresponding aflatoxin contamination on corn kernels in a rapid and nondestructive manner. The two A. flavus strains, AF13 and AF38, were used to represent the aflatoxigenic fungus and nonaflatoxigenic fungus, respectively, for artificial inoculation on corn kernels. The partial least-squares discriminant analysis (PLS-DA) models based on different combinations of spectral range (I: 410-1070 nm; II: 1120-2470 nm), corn side (endosperm or germ side), spectral variable number (full spectra or selected variables), modeling approach (two-step or one-step), and classification threshold (20 or 100 ppb) were developed and their performances were compared. The first study focusing on detection of aflatoxigenic fungus-infected corn kernels showed that, in classifying the "control+AF38-inoculated" and AF13-inoculated corn kernels, the full spectral PLS-DA models using the preprocessed spectra over range II and one-step approach yielded more accurate prediction results than using the spectra over range I and the two-step approach. The advantage of the full spectral PLS-DA models established using one corn side than the other side were not consistent in the explored combination cases. The best full spectral PLS-DA model obtained was obtained using the germ-side spectra over range II with the one-step approach, which achieved an overall accuracy of 91.11%. The established CARS-PLSDA models performed better than the corresponding full-spectral PLS-DA models, with the better model achieved an overall accuracy of 97.78% in separating the AF13-inoculated corn kernels and the uninfected control and AF38-inoculated corn kernels. The second study focusing on the detection of aflatoxin-contaminated corn kernels showed that, based on the aflatoxin threshold of 20 and 100 ppb, the best overall accuracy in classifying the aflatoxin-contaminated and healthy corn kernels attained 86.67% and 84.44%, respectively, using the CARS-PLSDA models. The quantitative modeling results using partial least-squares regression (PLSR) obtained the correlation coefficient of prediction set ( RP) of 0.91, which indicated the possibility of using Vis-NIR spectroscopy to quantify aflatoxin concentration in aflatoxigenic fungus-infected corn kernels.


Subject(s)
Aflatoxins/chemistry , Aspergillus/isolation & purification , Food Contamination/analysis , Spectroscopy, Near-Infrared/methods , Zea mays/microbiology , Aflatoxins/metabolism , Aspergillus/chemistry , Aspergillus/classification , Aspergillus/metabolism , Seeds/chemistry , Seeds/microbiology , Zea mays/chemistry
11.
Rev Argent Microbiol ; 51(4): 292-301, 2019.
Article in English | MEDLINE | ID: mdl-30905507

ABSTRACT

Aflatoxin is a carcinogenic secondary metabolite produced mainly by Aspergillus flavus and Aspergillus parasiticus, which can seriously endanger the health of humans and animals. Oxidative stress is a common defense response, and it is known that reactive oxygen species (ROS) can induce the synthesis of a series of secondary metabolites, including aflatoxin. By using mutants lacking the afap 1 gene, the role of afap1 gene in oxidative stress and aflatoxin synthesis was assessed. The growth of the mutant strains was significantly inhibited by the increase in the concentration of H2O2, inhibition was complete at 40mmol/l. However, in the quantitative analysis by HPLC, the concentration of AFB1 increased with the increased H2O2 until 10mmol/l. Following an analysis based on the information provided by the NCBI BLAST analysis, it was assumed that Afap1, a basic leucine zipper (bZIP) transcription factor, was associated with the oxidative stress in this fungus. Treatment with 5mmol/l H2O2 completely inhibited the growth of the mutant strains in afap 1 but did not affect the growth of the CA14PTs strain (non-mutant strain). In addition, the concentration of AFB1 in the mutant strains was approximately » of that observed in the CA14PTs strain. These results suggested that Afap1 plays a key role in the regulation of oxidative stress and aflatoxin production in A. flavus.


Subject(s)
Aflatoxins/biosynthesis , Aspergillus flavus/physiology , Basic-Leucine Zipper Transcription Factors/physiology , Oxidative Stress/physiology , Aspergillus flavus/metabolism
12.
Appl Spectrosc ; 73(4): 415-423, 2019 Apr.
Article in English | MEDLINE | ID: mdl-30700102

ABSTRACT

Current methods for detecting aflatoxin contamination of agricultural and food commodities are generally based on wet chemical analyses, which are time-consuming, destructive to test samples, and require skilled personnel to perform, making them impossible for large-scale nondestructive screening and on-site detection. In this study, we utilized visible-near-infrared (Vis-NIR) spectroscopy over the spectral range of 400-2500 nm to detect contamination of commercial, shelled peanut kernels (runner type) with the predominant aflatoxin B1 (AFB1). The artificially contaminated samples were prepared by dropping known amounts of aflatoxin standard dissolved in 50:50 (v/v) methanol/water onto peanut kernel surface to achieve different contamination levels. The partial least squares discriminant analysis (PLS-DA) models established using the full spectra over different ranges achieved good prediction results. The best overall accuracy of 88.57% and 92.86% were obtained using the full spectra when taking 20 and 100 parts per billion (ppb), respectively, as the classification threshold. The random frog (RF) algorithm was used to find the optimal characteristic wavelengths for identifying the surface AFB1-contamination of peanut kernels. Using the optimal spectral variables determined by the RF algorithm, the simplified RF-PLS-DA classification models were established. The better RF-PLS-DA models attained the overall accuracies of 90.00% and 94.29% with the 20 ppb and 100 ppb thresholds, respectively, which were improved compared to using the full spectral variables. Compared to using the full spectral variables, the employed spectral variables of the simplified RF-PLS-DA models were decreased by at least 94.82%. The present study demonstrated that the Vis-NIR spectroscopic technique combined with appropriate chemometric methods could be useful in identifying AFB1 contamination of peanut kernels.


Subject(s)
Aflatoxin B1/analysis , Arachis/chemistry , Aspergillus flavus/metabolism , Food Contamination , Arachis/microbiology , Spectroscopy, Near-Infrared/methods
13.
Crit Rev Food Sci Nutr ; 59(1): 173-180, 2019.
Article in English | MEDLINE | ID: mdl-28846441

ABSTRACT

Mycotoxins are the foremost naturally occurring contaminants of food products such as corn, peanuts, tree nuts, and wheat. As the secondary metabolites, mycotoxins are mainly synthesized by many species of the genera Aspergillus, Fusarium and Penicillium, and are considered highly toxic and carcinogenic to humans and animals. Most mycotoxins are detected and quantified by analytical chemistry-based methods. While mycotoxigenic fungi are usually identified and quantified by biological methods. However, these methods are time-consuming, laborious, costly, and inconsistent because of the variability of the grain-sampling process. It is desirable to develop rapid, non-destructive and efficient methods that objectively measure and evaluate mycotoxins and mycotoxigenic fungi in food. In recent years, some spectroscopy-based technologies such as hyperspectral imaging (HSI), Raman spectroscopy, and Fourier transform infrared spectroscopy have been extensively investigated for their potential use as tools for the detection, classification, and sorting of mycotoxins and toxigenic fungal contaminants in food. HSI integrates both spatial and spectral information for every pixel in an image, making it suitable for rapid detection of large quantities of samples and more heterogeneous samples and for in-line sorting in the food industry. In order to track the latest research developments in HSI, this paper gives a brief overview of the theories and fundamentals behind the technology and discusses its applications in the field of rapid detection and sorting of mycotoxins and toxigenic fungi in food products. Additionally, advantages and disadvantages of HSI are compared, and its potential use in commercial applications is reported.


Subject(s)
Food Contamination/analysis , Mycotoxins/chemistry , Spectrum Analysis/methods , Animals , Fungi/chemistry , Fungi/metabolism , Humans
14.
Front Microbiol ; 10: 3152, 2019.
Article in English | MEDLINE | ID: mdl-32038584

ABSTRACT

In an effort to control aflatoxin contamination in food and/or feed grains, a segment of research has focused on host resistance to eliminate aflatoxin from susceptible crops, including maize. To this end, screening tools are key to identifying resistant maize genotypes. The traditional field screening techniques, the kernel screening laboratory assay (KSA), and analytical methods (e.g., ELISA) used for evaluating corn lines for resistance to fungal invasion, all ultimately require sample destruction. A technological advancement on the basic BGYF presumptive screening test, fluorescence hyperspectral imaging offers an option for non-destructive and rapid image-based screening. The present study aimed to differentiate fluorescence spectral signatures of representative resistant and susceptible corn hybrids infected by a toxigenic (SRRC-AF13) and an atoxigenic (SRRC-AF36) strain of Aspergillus flavus, at several time points (5, 7, 10, and 14 days), in order to evaluate fluorescence hyperspectral imaging as a viable technique for early, non-invasive aflatoxin screening in resistant and susceptible corn lines. The study utilized the KSA to promote fungal growth and aflatoxin production in corn kernels inoculated under laboratory conditions and to provide actual aflatoxin values to relate with the imaging data. Each time point consisted of 78 kernels divided into four groups (30-susceptible, 30-resistant, 9-susceptible control, and 9-resistant control), per inoculum. On specified days, kernels were removed from the incubator and dried at 60°C to terminate fungal growth. Dry kernels were imaged with a VNIR hyperspectral sensor (image spectral range of 400-1000 nm), under UV excitation centered at 365 nm. Following imaging, kernels were submitted for the chemical AflaTest assay (VICAM). Fluorescence emissions were compared for all samples over 14 days. Analysis of strain differences separating the fluorescence emission peaks of resistant from the susceptible strain indicated that the emission peaks of the resistant strain and the susceptible strains differed significantly (p < 0.01) from each other, and there was a significant difference in fluorescence intensity between the treated and control kernels of both strains. These results indicate a viable role of fluorescence hyperspectral imaging for non-invasive screening of maize lines with divergent resistance to invasion by aflatoxigenic fungi.

16.
Anal Chem ; 90(24): 14331-14338, 2018 12 18.
Article in English | MEDLINE | ID: mdl-30444348

ABSTRACT

Aflatoxins, highly toxic and carcinogenic to humans, are synthesized via multiple intermediates by a complex pathway in several Aspergilli, including Aspergillus flavus. Few analytical methods are available for monitoring the changes in metabolite profiles of the aflatoxin biosynthesis pathway under different growth and environmental conditions. In the present study, we developed by a D-optimal mixture design a solvent system, methanol/dichloromethane/ethyl acetate/formic acid (0.36/0.31/0.32/0.01), that was suitable for extracting the pathway metabolites. The matrix effect from dilution of cell extracts was negligible. To facilitate the identification of these metabolites, we constructed a fragmentation ion library. We further employed liquid chromatography coupled with high-resolution mass spectroscopy (UHPLC-HRMS) for simultaneous quantification of the metabolites. The limit of detection (LOD) and limit of quantitation (LOQ) were 0.002-0.016 and 0.008-0.05 µg/kg, respectively. The spiked recovery rates ranged from 81.3 to 100.3% with intraday and interday precision less than 7.6%. Using the method developed to investigate the time-course aflatoxin biosynthesis, we found that precursors, including several possible toxins (with a carcinogenic group similar to aflatoxin B1), occurred together with aflatoxin, and that production increased rapidly at the early growth stage, peaked on day four, and then decreased substantially. The maximum production of aflatoxin B1 and aflatoxin B2 occurred 1 day later. Moreover, the dominant branch pathway was the one for aflatoxin B1 formation. We revealed that the antiaflatoxigenicity mechanism of Leclercia adecarboxylata WT16 was associated with a factor upstream of the aflatoxin biosynthesis pathway. The design strategies can be applied to characterize or detect other secondary metabolites to provide a snapshot of the dynamic changes during their biosynthesis.


Subject(s)
Aflatoxins/biosynthesis , Aspergillus flavus/metabolism , Mass Spectrometry , Aflatoxins/chemistry , Aflatoxins/isolation & purification , Chromatography, High Pressure Liquid , Food Contamination , Solvents/chemistry
17.
PLoS One ; 13(7): e0199169, 2018.
Article in English | MEDLINE | ID: mdl-29966003

ABSTRACT

Aspergillus flavus is a saprophytic fungus that infects corn, peanuts, tree nuts and other agriculturally important crops. Once the crop is infected the fungus has the potential to secrete one or more mycotoxins, the most carcinogenic of which is aflatoxin. Aflatoxin contaminated crops are deemed unfit for human or animal consumption, which results in both food and economic losses. Within A. flavus, two morphotypes exist: the S strains (small sclerotia) and L strains (large sclerotia). Significant morphological and physiological differences exist between the two morphotypes. For example, the S-morphotypes produces sclerotia that are smaller (< 400 µm), greater in quantity, and contain higher concentrations of aflatoxin than the L-morphotypes (>400 µm). The morphotypes also differ in pigmentation, pH homeostasis in culture and the number of spores produced. Here we report the first full genome sequence of an A. flavus S morphotype, strain AF70. We provide a comprehensive comparison of the A. flavus S-morphotype genome sequence with a previously sequenced genome of an L-morphotype strain (NRRL 3357), including an in-depth analysis of secondary metabolic clusters and the identification SNPs within their aflatoxin gene clusters.


Subject(s)
Aspergillus flavus/genetics , Genome, Fungal/genetics , Plant Diseases/genetics , Spores, Fungal/genetics , Aflatoxins/genetics , Aflatoxins/toxicity , Arachis/microbiology , Aspergillus flavus/classification , Aspergillus flavus/pathogenicity , Crops, Agricultural/genetics , Crops, Agricultural/microbiology , Nuts/microbiology , Plant Diseases/microbiology , Spores, Fungal/pathogenicity , Zea mays/microbiology
19.
Toxins (Basel) ; 10(4)2018 04 04.
Article in English | MEDLINE | ID: mdl-29617309

ABSTRACT

Mycotoxins are major food contaminants affecting global food security, especially in low and middle-income countries. The European Union (EU) funded project, MycoKey, focuses on “Integrated and innovative key actions for mycotoxin management in the food and feed chains” and the right to safe food through mycotoxin management strategies and regulation, which are fundamental to minimizing the unequal access to safe and sufficient food worldwide. As part of the MycoKey project, a Mycotoxin Charter (charter.mycokey.eu) was launched to share the need for global harmonization of mycotoxin legislation and policies and to minimize human and animal exposure worldwide, with particular attention to less developed countries that lack effective legislation. This document is in response to a demand that has built through previous European Framework Projects—MycoGlobe and MycoRed—in the previous decade to control and reduce mycotoxin contamination worldwide. All suppliers, participants and beneficiaries of the food supply chain, for example, farmers, consumers, stakeholders, researchers, members of civil society and government and so forth, are invited to sign this charter and to support this initiative.


Subject(s)
Environmental Exposure/prevention & control , Food Contamination/prevention & control , International Cooperation , Mycotoxins , Global Health , Humans
20.
Physiol Mol Biol Plants ; 24(3): 513-519, 2018 May.
Article in English | MEDLINE | ID: mdl-29692558

ABSTRACT

A comparative transcriptome analysis was performed using the genes significantly differentially expressed in cotton, corn and peanut in response to aflatoxin producing fungus Aspergillus flavus with an objective of identifying candidate resistance genes in cotton. Two-way analyses identified 732 unique genes to be differentially regulated by the fungus with only 26 genes common across all three crops that were considered candidate A. flavus resistance genes with an assumption that these genes have specific roles in conferring the resistance trait. Genes of membrane cellular component involved in DNA binding with involvement in defense responses were highly represented among the differentially expressed unique genes. Most (six) of these genes coded for 2-oxoglutarate (2OG) and Fe(II)-dependent oxygenase superfamily proteins. Genes encoding helix loop helix protein, alcohol dehydrogenase and UDP glycosylation transferase which were upregulated in response to both atoxigenic and toxigenic strains of A. flavus, could be potential resistance candidate genes for downstream functional manipulation to confer resistance.

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