Resistance to Aflatoxin Contamination in Corn as Influenced by Relative Humidity and Kernel Germination

1996 ◽  
Vol 59 (3) ◽  
pp. 276-281 ◽  
Author(s):  
B. Z. GUO ◽  
J. S. RUSSIN ◽  
R. L. BROWN ◽  
T. E. CLEVELAND ◽  
N. W. WIDSTROM

Kernels of corn population GT-MAS:gk, resistant to aflatoxin B1 production by Aspergillus flavus, and susceptible Pioneer hybrid 3154 were tested for aflatoxin when incubated under different relative humidities (RH). High aflatoxin levels were not detected in either genotype at RH < 91%. Resistance in GT-MAS:gk was consistent across all RH levels (91 to 100%) at which significant aflatoxin accumulation was detected. Aflatoxin levels in GT-MAS:gk averaged about 98% less than those in susceptible Pioneer 3154, which suggests that storage of this or other genotypes with similar resistance mechanisms may be possible under moisture conditions less exacting than are required with susceptible hybrids. Results for fungus growth and sporulation ratings on kernel surfaces were similar to those for aflatoxin levels. When kernels of both genotypes were preincubated 3 days at 100% RH prior to inoculation with A. flavus, germination percentages increased to very high levels compared to those of kernels that were not preincubated. In preincubated kernels aflatoxin levels remained consistently low in GT-MAS:gk but decreased markedly (61%) in Pioneer 3154. When eight susceptible hybrids were evaluated for aflatoxin accumulation in preincubated kernels, seven of these supported significantly lower toxin levels than kernels not subjected to preincubation. Average reduction across hybrids was 83%, and reductions within hybrids ranged from 68 to 96%. Preincubated kernels of one susceptible hybrid (Deltapine G-4666) supported aflatoxin levels comparable to those in resistant GT-MAS:gk. Data suggest that an inhibitor of aflatoxin biosynthesis may be induced during kernel germination. Possible mechanisms for embryo effects on resistance to aflatoxin accumulation are discussed.

Toxins ◽  
2019 ◽  
Vol 11 (2) ◽  
pp. 70 ◽  
Author(s):  
Mingguan Yang ◽  
Laifeng Lu ◽  
Shuhua Li ◽  
Jing Zhang ◽  
Zhenjing Li ◽  
...  

Aspergillus flavus is a soilborne pathogenic fungus that poses a serious public health threat due to it contamination of food with carcinogenic aflatoxins. Our previous studies have demonstrated that benzenamine displayed strong inhibitory effects on the mycelial growth of A. flavus. In this study, we systematically investigated the inhibitory effects of benzenamine on the development, aflatoxin biosynthesis, and virulence in A. flavus, as well as the underlying mechanism. The results indicated that benzenamine exhibited great capacity to combat A. flavus at a concentration of 100 µL/L, leading to significantly decreased aflatoxin accumulation and colonization capacity in maize. The transcriptional profile revealed that 3589 genes show altered mRNA levels in the A. flavus after treatment with benzenamine, including 1890 down-regulated and 1699 up-regulated genes. Most of the differentially expressed genes participated in the biosynthesis and metabolism of amino acid, purine metabolism, and protein processing in endoplasmic reticulum. Additionally, the results brought us to a suggestion that benzenamine affects the development, aflatoxin biosynthesis, and pathogenicity of A. flavus via down-regulating related genes by depressing the expression of the global regulatory factor leaA. Overall, this study indicates that benzenamine have tremendous potential to act as a fumigant against pathogenic A. flavus. Furthermore, this work offers valuable information regarding the underlying antifungal mechanism of benzenamine against A. flavus at the level of transcription, and these potential targets may be conducive in developing new strategies for preventing aflatoxin contamination.


2008 ◽  
Vol 1 (3) ◽  
pp. 333-340 ◽  
Author(s):  
H. Abbas ◽  
R. Zablotowicz ◽  
H. Bruns

To successfully exploit biological control it is desirable to understand how the introduced agent colonises the host and interferes with establishment of the pest. This study assessed field colonisation of maize by Aspergillus flavus strains as biological control agents to reduce aflatoxin contamination. Maize (corn, Zea mays L.) ears were inoculated with A. flavus using a pin-bar technique in 2004 and 2005. Non-aflatoxigenic strains K49 (NRRL 30797) & CT3 (NRRL 30798) and toxigenic F3W4 (NRRL 30798) were compared against a carrier control (0.2% aqueous Tween 20). Ten ears were sampled over 12 to 20 days, visually assessed, and curves fit to a three compartment Gompertz equation or other best appropriate regressions. Aflatoxin was determined by HPLC and cyclopiazonic acid (CPA) by LC/MS. The Gompertz model describes growth parameters, e.g. growth constant, lag phase and maximum colonisation characterised patterns of maize colonisation for most inoculated treatments. Aflatoxin accumulation in maize inoculated with F3W4 was about 35,000 ng/g in 2004 and 2005, with kinetics of aflatoxin accumulation in 2005 well described by the Gompertz equation. Less than 200 ng/g was observed in maize inoculated with strains CT3 & K49 and accumulation was described by a linear or logistic model. Maize inoculated with strains CT3 and F3W4 accumulated a maximum of 220 and 169 µg/kg CPA, respectively, compared to 22 and 0.2 µg/kg in the control and K49 inoculated, respectively. This technique can be used to elucidate colonisation potential of non-toxigenic A. flavus in maize in relation to biological control of aflatoxin. The greatest reduction of aflatoxin and CPA in maize inoculated with strain K49 and Gompertz parameters on colonisation indicates its superiority to CT3 as a biological control agent. The dynamics of maize colonisation by A. flavus strains and subsequent mycotoxin accumulation generated by using the pin-bar technique has implications for characterising the competence of biocontrol strains for reducing aflatoxin contamination.


Author(s):  
Shyam L. Kandel ◽  
Rubaiya Jesmin ◽  
Brian M. Mack ◽  
Rajtilak Majumdar ◽  
Matthew K. Gilbert ◽  
...  

Aspergillus flavus is an opportunistic pathogen of oilseed crops such as maize, peanut, cottonseed, and tree nuts and produces carcinogenic secondary metabolites known as aflatoxins during seed colonization. Aflatoxin contamination not only reduces the value of the produce but also is a health hazard to humans and animals. Previously, we observed inhibition of A. flavus aflatoxin biosynthesis upon exposure to the marine bacterium, Vibrio gazogenes (Vg). In this study, we used RNA sequencing to examine the transcriptional profiles of A. flavus treated with both live and heat-inactivated dead Vg and control samples. Fungal biomass, total accumulated aflatoxins, and expression profiles of genes constituting secondary metabolite biosynthetic gene clusters were determined at 24, 30, and 40 h after treatment. Statistically significant reductions in total aflatoxins were detected in Vg-treated samples as compared to control samples at 40 h. But no statistical difference in fungal biomass was observed upon these treatments. The Vg treatments were most effective on aflatoxin biosynthesis as was reflected in significant downregulation of majority of the genes in the aflatoxin gene cluster including the aflatoxin pathway regulator gene, aflR. Along with aflatoxin genes, we also observed significant downregulation in some other secondary metabolite gene clusters including cyclopiazonic acid and aflavarin, suggesting that the treatment may inhibit other secondary metabolites as well. Finally, a weighted gene correlation network analysis identified an upregulation of ten genes that were most strongly associated with Vg-dependent aflatoxin inhibition and provide a novel start-point in understanding the mechanisms that result in this phenomenon.


1999 ◽  
Vol 65 (6) ◽  
pp. 2738-2740 ◽  
Author(s):  
Sui-Sheng T. Hua ◽  
James L. Baker ◽  
Melanie Flores-Espiritu

ABSTRACT The nor mutant of Aspergillus flavus has a defective norsolorinic acid reductase, and thus the aflatoxin biosynthetic pathway is blocked, resulting in the accumulation of norsolorinic acid, a bright red-orange pigment. We developed a visual agar plate assay to monitor yeast strains for their ability to inhibit aflatoxin production by visually scoring the accumulation of this pigment of the nor mutant. We identified yeast strains that reduced the red-orange pigment accumulation in the normutant. These yeasts also reduced aflatoxin accumulation by a toxigenic strain of A. flavus. These yeasts may be useful for reducing aflatoxin contamination of food commodities.


2007 ◽  
Vol 73 (22) ◽  
pp. 7268-7276 ◽  
Author(s):  
Ludmila V. Roze ◽  
Randolph M. Beaudry ◽  
Anna E. Arthur ◽  
Ana M. Calvo ◽  
John E. Linz

ABSTRACT Aspergillus parasiticus is one primary source of aflatoxin contamination in economically important crops. To prevent the potential health and economic impacts of aflatoxin contamination, our goal is to develop practical strategies to reduce aflatoxin synthesis on susceptible crops. One focus is to identify biological and environmental factors that regulate aflatoxin synthesis and to manipulate these factors to control aflatoxin biosynthesis in the field or during crop storage. In the current study, we analyzed the effects of aspergillus volatiles on growth, development, aflatoxin biosynthesis, and promoter activity in the filamentous fungus A. parasiticus. When colonies of Aspergillus nidulans and A. parasiticus were incubated in the same growth chamber, we observed a significant reduction in aflatoxin synthesis and asexual sporulation by A. parasiticus. Analysis of the headspace gases demonstrated that A. nidulans produced much larger quantities of 2-buten-1-ol (CA) and 2-ethyl-1-hexanol (EH) than A. parasiticus. In its pure form, EH inhibited growth and increased aflatoxin accumulation in A. parasiticus at all doses tested; EH also stimulated aflatoxin transcript accumulation. In contrast, CA exerted dose-dependent up-regulatory or down-regulatory effects on aflatoxin accumulation, conidiation, and aflatoxin transcript accumulation. Experiments with reporter strains carrying nor-1 promoter deletions and mutations suggested that the differential effects of CA were mediated through separate regulatory regions in the nor-1 promoter. The potential efficacy of CA as a tool for analysis of transcriptional regulation of aflatoxin biosynthesis is discussed. We also identify a novel, rapid, and reliable method to assess norsolorinic acid accumulation in solid culture using a Chroma Meter CR-300 apparatus.


2004 ◽  
Vol 31 (2) ◽  
pp. 134-141 ◽  
Author(s):  
H. Q. Xue ◽  
T. G. Isleib ◽  
H. T. Stalker ◽  
G. A. Payne ◽  
G. OBrian

Abstract Anatoxins are carcinogenic and extremely toxic secondary metabolites produced primarily by two fungi, Aspergillus flavus Link ex Fries and A. parasiticus Speare. Elimination of aflatoxin contamination in peanut (Arachis hypogaea L.) is a high priority of the peanut industry. Resistant cultivars should be an effective and low-cost part of an integrated aflatoxin management program. To date, no cultivated peanut has been reported with stable high levels of resistance to aflatoxin production. Arachis species and interspecific tetraploid lines have been evaluated for resistance to several peanut diseases and insect pests, and highly resistant accessions have been reported. Seven accessions of A. cardenasii Krapov. and W.C. Gregory, 29 of A. duranensis Krapov. and W.C. Gregory, and 17 interspecific tetraploid lines derived from A. hypogaea × A. cardenasii were inoculated with A. flavus strain NRRL 3357 and analyzed for aflatoxin content after incubation. On average, A. duranensis and A. cardenasii accumulated significantly less aflatoxin than A. hypogaea checks. The mean difference between the two wild species was not significant. Arachis duranensis accessions PI 468319 (GKBSPSc 30073), PI 468200 (GKBSPSc 30064), and PI 262133 (GKP 10038 sl.); and A. cardenasii accessions PI 262141 (GKP 10017) and PI 475997 (KSSc 36018) had reduced levels of aflatoxin accumulation and should be valuable sources of resistance to aflatoxin contamination. Of the interspecific tetraploid lines, only GP-NC WS 2 supported aflatoxin production not significantly different from resistant parent A. cardenasii GKP 10017, and it appears to be a line with reduced capacity for aflatoxin accumulation.


2013 ◽  
Vol 1 (04) ◽  
pp. 148-151 ◽  
Author(s):  
P. N. Rajarajan ◽  
K. M. Rajasekaran ◽  
N. K. Asha Devi

Aflatoxin is a naturally occurring Mycotoxin produced by Aspergillus flavus and Aspergillus parasiticus. Aspergillus flavus is common and widespread in nature and is most often found when certain grains are grown under stressful conditions such as draught. The mold occurs in soil, decaying vegetation, hay and grains undergoing microbiological deterioration and invades all types of organic substrates whenever and wherever the conditions are favourable for its growth. Favourable conditions include high moisture content and high temperature.The aflatoxin group is comprised of aflatoxin B1,B2,G1 and G2. In addition , aflatoxin M1 (AFM1), a hydroxylated metabolite of AFB1, is excreted in the milk of dairy cows consuming an AFB1-contaminated ration. Aflatoxin B1 a prototype of the aflatoxins, is widely recognized as the most potent hepato carcinogenic compound and along with other certain members of the group, possess additional toxic properties including mutagenicity, tetrogenicity, acute cellular toxicity and it suppresses the immune system. Aflatoxin contamination of food and feed has gained global significance as a result of its deleterious effects on human as well as animal health. The marketability of food products is adversely affected by aflatoxin contamination.


2018 ◽  
Vol 6 (4) ◽  
pp. 600-605 ◽  
Author(s):  
Mohamed Mahmoud Deabes ◽  
Wagdy Khalil Bassaly Khalil ◽  
Ashraf Gamil Attallah ◽  
Tarek Ahmed El-Desouky ◽  
Khayria Mahmoud Naguib

AIM: In this study, we evaluated the effect of silver nanoparticles (AgNPs) on the production of aflatoxin B1 (AFB1) through assessment the transcription activity of aflatoxin biosynthesis pathway genes in Aspergillus flavus ATCC28542.MATERIAL AND METHODS: The mRNAs were quantitative by Real Time-polymerase chain reaction (qRT-PCR) of A. flavus grown in yeast extract sucrose (YES) medium containing AgNPs. Specific primers that are involved in the AFB1 biosynthesis which highly specific to A. flavus, O-methyltransferase gene (omt-A), were designed and used to detect the fungus activity by quantitative PCR assay. The AFB1 production (from A. flavus growth) which effected by AgNPs were measured in YES medium by high-pressure liquid chromatography (HPLC).RESULTS: The AFB1 produced by A. flavus have the highest reduction with 1.5 mg -100 ml of AgNPs were added in media those records 88.2%, 67.7% and 83.5% reduction by using AgNP HA1N, AgNP HA2N and AgNP EH, respectively. While on mycelial growth give significantly inhibitory effect. These results have been confirmed by qRT-PCR which showed that culture of A. flavus with the presence of AgNPs reduced the expression levels of omt-A gene.CONCLUSION: Based on the results of the present study, AgNPs inhibit growth and AFB1 produced by Aspergillus flavus ATCC28542. This was confirmed through RT-PCR approach showing the effect of AgNPs on omt-A gene involved in aflatoxin biosynthesis.


Toxins ◽  
2019 ◽  
Vol 11 (1) ◽  
pp. 41 ◽  
Author(s):  
Elisabeth Tumukunde ◽  
Ding Li ◽  
Ling Qin ◽  
Yu Li ◽  
Jiaojiao Shen ◽  
...  

Aspergillus flavus is one of the fungi from the big family of Aspergillus genus and it is capable of colonizing a large number of seed/crops and living organisms such as animals and human beings. SakA (also called hogA/hog1) is an integral part of the mitogen activated protein kinase signal of the high osmolarity glycerol pathway. In this study, the AfsakA gene was deleted (∆AfsakA) then complemented (∆AfsakA::AfsakA) using homologous recombination and the osmotic stress was induced by 1.2 mol/L D-sorbital and 1.2 mol/L sodium chloride. The result showed that ∆AfsakA mutant caused a significant influence on conidial formation compared to wild-type and ∆AfsakA::AfsakA strains. It was also found that AfsakA responds to both the osmotic stress and the cell wall stress. In the absence of osmotic stress, ∆AfsakA mutant produced more sclerotia in contrast to other strains, whereas all strains failed to generate sclerotia under osmotic stress. Furthermore, the deletion of AfsakA resulted in the increase of Aflatoxin B1 production compared to other strains. The virulence assay on both maize kernel and peanut seeds showed that ∆AfsakA strain drastically produced more conidia and Aflatoxin B1 than wild-type and complementary strains. AfSakA-mCherry was located to the cytoplasm in the absence of osmotic stress, while it translocated to the nucleus upon exposure to the osmotic stimuli. This study provides new insights on the development and evaluation of aflatoxin biosynthesis and also provides better understanding on how to prevent Aspergillus infections which would be considered the first step towards the prevention of the seeds damages caused by A. flavus.


2011 ◽  
Vol 343-344 ◽  
pp. 1168-1171 ◽  
Author(s):  
Liang Bin Hu ◽  
Hong Bo Li ◽  
Jun Liang Sun ◽  
Jie Zeng

Control of aflatoxin contamination has been a worldwide problem. Laminarin from Laminaria digitata is one kind of polysaccharides with multiple biological activities. In this paper, the inhibitory effects of Laminarin on the growth and toxin production of A. flavus was studied. The results indicated that 150 and 200 µg/mL of Laminarin ccould significantly inhibit the aflatoxin production in Sabouraud liquid medium (Sab), without affecting mycelium growth. In addition, the results also showed that certain concentration Laminaria could decrease the infection of peanut seeds by A. flavus as well as the contamination by aflatoxin B1.


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