Modelling the colonisation of maize by toxigenic and non-toxigenic Aspergillus flavus strains: implications for biological control

2008 ◽  
Vol 1 (3) ◽  
pp. 333-340 ◽  
Author(s):  
H. Abbas ◽  
R. Zablotowicz ◽  
H. Bruns
Keyword(s):  
Biological Control ◽  
Aspergillus Flavus ◽  
Growth Parameters ◽  
Biological Control Agent ◽  
Cyclopiazonic Acid ◽  
Aflatoxin Contamination ◽  
Tween 20 ◽  
Lag Phase ◽  
Gompertz Equation ◽  
Aflatoxin Accumulation

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.

scholarly journals Interactions of Saprophytic Yeasts with anor Mutant of Aspergillus flavus

1999 ◽  
Vol 65 (6) ◽  
pp. 2738-2740 ◽  
Author(s):  
Sui-Sheng T. Hua ◽  
James L. Baker ◽  
Melanie Flores-Espiritu
Keyword(s):  
Aspergillus Flavus ◽  
Agar Plate ◽  
Aflatoxin Production ◽  
Aflatoxin Contamination ◽  
Orange Pigment ◽  
Yeast Strains ◽  
Norsolorinic Acid ◽  
Aflatoxin Accumulation ◽  
Food Commodities ◽  
Pigment Accumulation

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.

Identification of Aspergillus flavus Isolates as Potential Biocontrol Agents of Aflatoxin Contamination in Crops

2013 ◽  
Vol 76 (6) ◽  
pp. 1051-1055 ◽  
Author(s):  
L. J. ROSADA ◽  
J. R. SANT'ANNA ◽  
C. C. S. FRANCO ◽  
G. N. M. ESQUISSATO ◽  
P. A. S. R. SANTOS ◽  
...  
Keyword(s):  
Biological Control ◽  
Aspergillus Flavus ◽  
Filamentous Fungus ◽  
Microscopic Examination ◽  
Genetic Recombination ◽  
Aflatoxin Contamination ◽  
Biocontrol Agents ◽  
Biological Control Agents ◽  
Nit Mutants ◽  
The Stability

Aspergillus flavus, a haploid organism found worldwide in a variety of crops, including maize, cottonseed, almond, pistachio, and peanut, causes substantial and recurrent worldwide economic liabilities. This filamentous fungus produces aflatoxins (AFLs) B1 and B2, which are among the most carcinogenic compounds from nature, acutely hepatotoxic and immunosuppressive. Recent efforts to reduce AFL contamination in crops have focused on the use of nonaflatoxigenic A. flavus strains as biological control agents. Such agents are applied to soil to competitively exclude native AFL strains from crops and thereby reduce AFL contamination. Because the possibility of genetic recombination in A. flavus could influence the stability of biocontrol strains with the production of novel AFL phenotypes, this article assesses the diversity of vegetative compatibility reactions in isolates of A. flavus to identify heterokaryon self-incompatible (HSI) strains among nonaflatoxigenic isolates, which would be used as biological controls of AFL contamination in crops. Nitrate nonutilizing (nit) mutants were recovered from 25 A. flavus isolates, and based on vegetative complementation between nit mutants and on the microscopic examination of the number of hyphal fusions, five nonaflatoxigenic (6, 7, 9 to 11) and two nontoxigenic (8 and 12) isolates of A. flavus were phenotypically characterized as HSI. Because the number of hyphal fusions is reduced in HSI strains, impairing both heterokaryon formation and the genetic exchanges with aflatoxigenic strains, the HSI isolates characterized here, especially isolates 8 and 12, are potential agents for reducing AFL contamination in crops.

scholarly journals Variation in Occurrence and Aflatoxigenicity of Aspergillus flavus from Two Climatically Varied Regions in Kenya

Toxins ◽  
2020 ◽  
Vol 12 (1) ◽  
pp. 34 ◽  
Author(s):  
Ethel Monda ◽  
Joel Masanga ◽  
Amos Alakonya
Keyword(s):  
Biological Control ◽  
Aspergillus Flavus ◽  
Plant Pathogens ◽  
Cropping Systems ◽  
Bacterial Species ◽  
Climatic Conditions ◽  
Aflatoxin Contamination ◽  
Maize Cultivation ◽  
Bacillus Spp ◽  
Study Sites

Aflatoxins are carcinogenic chemical metabolites produced by Aspergillus spp. of the section Flavi. In Kenya, Aspergillus flavus is the most prevalent and has been associated with several acute and chronic aflatoxin outbreaks in the past. In this study, we evaluated the occurrence of A. flavus in soils from two agro-ecological regions with contrasting climatic conditions, aflatoxin contamination histories and cropping systems. Aspergillus spp. were first isolated from soils before the identification and determination of their aflatoxigenicity. Further, we determined the occurrence of Pseudomonas and Bacillus spp. in soils from the two regions. These bacterial species have long been associated with biological control of several plant pathogens including Aspergillus spp. Our results show that A. flavus occurred widely and produced comparatively higher total aflatoxin levels in all (100%) study sites from the eastern to the western regions of Kenya. For the western region, A. flavus was detected in 4 locations (66.7%) that were previously under maize cultivation with the isolates showing low aflatoxigenicity. A. flavus was not isolated from soils under sugarcane cultivation. Distribution of the two bacterial species varied across the regions but we detected a weak relationship between occurrence of bacterial species and A. flavus. We discuss these findings in the context of the influence of climate, microbial profiles, cropping systems and applicability in the deployment of biological control remedies against aflatoxin contamination.

Atoxigenic Aspergillus flavus biological control of aflatoxin contamination: what is the mechanism?

2015 ◽  
Vol 8 (2) ◽  
pp. 235-244 ◽  
Author(s):  
K.E. Damann Jr.
Keyword(s):  
Biological Control ◽  
Aspergillus Flavus ◽  
Population Biology ◽  
Aflatoxin Production ◽  
Infection Process ◽  
Aflatoxin Contamination ◽  
Signal Molecules ◽  
Nutrient Competition ◽  
Toxigenic Strain ◽  
Aflatoxin Inhibition

The term ‘competitive exclusion’ involving physical blockage of growth or access of the toxigenic strain to the seed target has been used to describe the mechanism of biological control of aflatoxin contamination. However, recent evidence suggests that a form of intraspecific aflatoxin inhibition requiring growth of the competing strains together during the infection process in such a way that hyphae physically interact or touch is the trigger for preventing induction of aflatoxin synthesis. This direct touch-based inhibition of aflatoxin synthesis is posited to be the mechanistic basis of biological control in this system. Evidence for this idea comes from the published observations that co-culture of toxigenic and atoxigenic strains in a suspended disc system, in which the hyphae physically interact, prevents aflatoxin production. However, growth of the same strains in the same medium in the two compartments of a filter insert plate well system, separating the atoxigenic and toxigenic strains with a 0.4 μm or 3.0 μm filter, allows aflatoxin production approaching that of the toxigenic strain alone. When the strains are mixed and placed in both the insert and the well compartments, the intraspecific aflatoxin inhibition occurs as it did in the suspended disc culture system. This further suggests that neither nutrient competition nor soluble signal molecules, which should pass through the filter, are involved in intraspecific aflatoxin inhibition. When the two strains are separated by a 12 μm filter that would allow some passage of conidia or hyphae between the compartments the aflatoxin synthesis is approximately half that of the toxigenic strain alone. This phenomenon could be termed ‘competitive inclusion’ or ‘competitive phenotype conversion’. Work of others that relates to understanding the phenomenon is discussed, as well as an Aspergillus flavus population biology study from the Louisiana maize agro-ecosystem which has biological control implications.

scholarly journals Evaluation of Arachis Species and Interspecific Tetraploid Lines for Resistance to Aflatoxin Production by Aspergillus flavus

2004 ◽  
Vol 31 (2) ◽  
pp. 134-141 ◽  
Author(s):  
H. Q. Xue ◽  
T. G. Isleib ◽  
H. T. Stalker ◽  
G. A. Payne ◽  
G. OBrian
Keyword(s):  
Aspergillus Flavus ◽  
Insect Pests ◽  
Low Cost ◽  
Aflatoxin Production ◽  
Management Program ◽  
Aflatoxin Contamination ◽  
Sources Of Resistance ◽  
Resistant Parent ◽  
Arachis Species ◽  
Aflatoxin Accumulation

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.

Aflatoxin Reduction in Corn Through Field Application of Competitive Fungi†

1999 ◽  
Vol 62 (6) ◽  
pp. 650-656 ◽  
Author(s):  
JOE W. DORNER ◽  
RICHARD J. COLE ◽  
DONALD T. WICKLOW
Keyword(s):  
Biological Control ◽  
Aspergillus Flavus ◽  
Weather Conditions ◽  
Type A ◽  
Field Application ◽  
Aflatoxin Contamination ◽  
Applied Strain ◽  
Wild Type ◽  
Separate Part ◽  
Aflatoxin Concentration

Soil in corn plots was inoculated with nonaflatoxigenic strains of Aspergillus flavus and A. parasiticus during crop years 1994 to 1997 to determine the effect of application of the nontoxigenic strains on preharvest aflatoxin contamination of corn. Corn plots in a separate part of the field were not inoculated and served as controls. Inoculation resulted in significant increases in the total A. flavus/parasiticus soil population in treated plots, and that population was dominated by the applied strain of A. parasiticus (NRRL 21369). In the years when weather conditions favored aflatoxin contamination (1996 and 1997), corn was predominately colonized by A. flavus as opposed to A. parasiticus. In 1996, colonization by wild-type A. flavus was significantly reduced in treated plots compared with control plots, but total A. flavus/parasiticus colonization was not different between the two groups. A change to a more aggressive strain of A. flavus (NRRL 21882) as part of the biocontrol inoculum in 1997 resulted in a significantly (P < 0.001) higher colonization of corn by the applied strain. Weather conditions did not favor aflatoxin contamination in 1994 and 1995. In 1996, the aflatoxin concentration in corn from treated plots averaged 24.0 ppb, a reduction of 87% compared with the aflatoxin in control plots that averaged 188.4 ppb. In 1997, aflatoxin was reduced by 66% in treated corn (29.8 ppb) compared with control corn (87.5 ppb). Together, the data indicated that although the applied strain of A. parasiticus dominated in the soil, the nonaflatoxigenic strains of A. flavus were more responsible for the observed reductions in aflatoxin contamination. Inclusion of a nonaflatoxigenic strain of A. parasiticus in a biological control formulation for aflatoxin contamination may not be as important for airborne crops, such as corn, as for soilborne crops, such as peanuts.

Reduction in Aflatoxin Content of Maize by Atoxigenic Strains of Aspergillus flavus

1991 ◽  
Vol 54 (8) ◽  
pp. 623-626 ◽  
Author(s):  
ROBERT L. BROWN ◽  
PETER J. COTTY ◽  
THOMAS E. CLEVELAND
Keyword(s):  
Biological Control ◽  
Aspergillus Flavus ◽  
Aflatoxin Contamination ◽  
Biological Control Agents ◽  
Field Plot ◽  
Toxigenic Strain ◽  
Aflatoxin Content ◽  
Potential Use

In field plot experiments, an atoxigenic strain of Aspergillus flavus interfered with preharvest aflatoxin contamination of corn when applied either simultaneously with or one day prior to a toxigenic strain. The atoxigenic strain reduced preharvest aflatoxin contamination 80 to 95%. The atoxigenic strain was also effective in reducing postharvest aflatoxin contamination caused by both an introduced toxigenic strain and by strains resident on the kernels. The results suggest that atoxigenic strains of A. flavus may have potential use as biological control agents directed at reducing both preharvest and postharvest aflatoxin contamination of corn.

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
Keyword(s):  
Relative Humidity ◽  
Aspergillus Flavus ◽  
Aflatoxin B1 ◽  
Resistance Mechanisms ◽  
Aflatoxin Contamination ◽  
Aflatoxin Biosynthesis ◽  
Aflatoxin Accumulation ◽  
Moisture Conditions ◽  
Very High ◽  
Fungus Growth

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.

Production of the 14 kDa trypsin inhibitor protein is important for maize resistance against Aspergillus flavus infection/aflatoxin accumulation

2016 ◽  
Vol 9 (2) ◽  
pp. 215-228 ◽  
Author(s):  
Z.-Y. Chen ◽  
M.L. Warburton ◽  
L. Hawkins ◽  
Q. Wei ◽  
Y. Raruang ◽  
...  
Keyword(s):  
Aspergillus Flavus ◽  
Trypsin Inhibitor ◽  
Aflatoxin Production ◽  
Field Resistance ◽  
Aflatoxin Contamination ◽  
Direct Role ◽  
Independent Events ◽  
Sequence Polymorphisms ◽  
Transgenic Lines ◽  
Aflatoxin Accumulation

Maize (Zea mays L.) is one of the major crops susceptible to Aspergillus flavus Link ex. Fries infection and subsequent aflatoxin contamination. Previous studies found the production of an antifungal 14 kDa trypsin inhibitor (TI) was associated with maize aflatoxin resistance. To further investigate whether the TI plays any direct role in resistance, a TI gene silencing vector was constructed and transformed into maize. Mature kernels were produced from 66 transgenic lines representing 18 independent events. A final total of twelve lines representing four independent events were confirmed positive for transformation, five of which showed significant reduction (63 to 88%) in TI transcript abundance in seedling leaf tissue and seven of which showed significant TI protein reduction (39-85%) in mature kernels. Six of the seven silenced transgenic lines supported higher levels of aflatoxin production compared to negative controls. To further confirm the role of TI in field resistance to aflatoxin accumulation, DNA sequence polymorphisms from within the gene or linked simple sequence repeats were tested in four quantitative trait loci (QTL) mapping populations for QTL effect, and three QTL with log of the odds scores of 11, 4.5, and 3.0 and possibly caused by the TI protein encoding gene were found. Sequence polymorphisms were also tested for association to aflatoxin levels in an association mapping panel, and three single nucleotide polymorphisms were found associated with aflatoxin accumulation (P<0.01). The data from both RNAi and genetic mapping studies demonstrated that production of the TI in maize is important for its resistance to A. flavus infection and/or aflatoxin production.

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