scholarly journals Identification of a Maize Kernel Stress-Related Protein and Its Effect on Aflatoxin Accumulation

2004 ◽  
Vol 94 (9) ◽  
pp. 938-945 ◽  
Author(s):  
Z.-Y. Chen ◽  
R. L. Brown ◽  
K. E. Damann ◽  
T. E. Cleveland

Aflatoxins are carcinogens produced mainly by Aspergillus flavus during infection of susceptible crops such as maize. Through proteomic comparisons of maize kernel embryo proteins of resistant and susceptible genotypes, several protein spots previously were found to be unique or upregulated in resistant embryos. In the present study, one of these protein spots was sequenced and identified as glyoxalase I (GLX-I; EC 4.4.1.5). The full-length cDNA of the glyoxalase I gene (glx-I) was cloned. GLX-I constitutive activity was found to be significantly higher in the resistant maize lines compared with susceptible ones. After kernel infection by A. flavus, GLX-I activity remained lower in susceptible genotypes than in resistant genotypes. However, fungal infection significantly increased methylglyoxal (MG) levels in two of three susceptible genotypes. Further, MG was found to induce aflatoxin production in A. flavus culture at a concentration as low as 5.0 μM. The mode of action of MG may be to stimulate the expression of aflR, an aflatoxin biosynthesis regulatory gene, which was found to be significantly upregulated in the presence of 5 to 20 μM MG. These data suggest that GLX-I may play an important role in controlling MG levels inside kernels, thereby contributing to the lower levels of aflatoxins found in resistant maize genotypes.

2001 ◽  
Vol 64 (11) ◽  
pp. 1785-1792 ◽  
Author(s):  
ZHI-YUAN CHEN ◽  
ROBERT L. BROWN ◽  
THOMAS E. CLEVELAND ◽  
KENNETH E. DAMANN ◽  
JOHN S. RUSSIN

Maize genotypes resistant or susceptible to aflatoxin production or contamination were compared for differences in both constitutive and inducible proteins. Five additional constitutive proteins were found to be associated with resistance in over 8 of the 10 genotypes examined. Among these, the 58- and 46-kDa proteins were identified as globulin-1 and globulin-2, respectively. Differences in the ability to induce specific antifungal proteins, such as the higher synthesis of the 22-kDa zeamatin in resistant genotypes, were also observed between resistant and susceptible kernels incubated under germinating conditions (31°C, 100% humidity). Both constitutive and inducible proteins appear to be necessary for kernel resistance. Embryo-killed kernels (unable to synthesize new proteins) supported the highest level of aflatoxins, whereas imbibed kernels (to hasten protein induction) supported the lowest among all treatments. This suggests that the synthesis of new proteins by the embryo plays an important role in conferring resistance. However, significantly lower levels of aflatoxin production in embryo-killed resistant kernels than in susceptible ones suggest that, in reality, high levels of constitutive antifungal proteins are indispensable to kernel resistance.


1995 ◽  
Vol 58 (3) ◽  
pp. 296-300 ◽  
Author(s):  
BAO Z. GUO ◽  
JOHN S. RUSSIN ◽  
THOMAS E. CLEVELAND ◽  
ROBERT L. BROWN ◽  
NEIL W. WIDSTROM

Thirteen maize hybrids and one maize population, MAS:gk, were screened for susceptibility to aflatoxin production by Aspergillus flavus. Marked differences in aflatoxin B1 production were detected among the maize genotypes tested. Most commercial hybrids consistently supported high levels of aflatoxin accumulation. Aflatoxin levels did not differ between intact and wounded kernels of these genotypes. However, different results were obtained from 4 of the 13 hybrids and the maize population MAS:gk. Levels of aflatoxin accumulation in intact kernels of these genotypes were lower than in the previous susceptible group of genotypes. In addition, aflatoxin levels were higher in wounded than in intact kernels. MAS:gk not only supported the lowest levels of aflatoxin production in intact kernels, but aflatoxin levels in endosperm-wounded kernels also were significantly lower in MAS:gk than in wounded kernels of all tested hybrids. Treatment with KOH to remove cutin from intact kernels prior to inoculation with A. flavus effected substantial increases in aflatoxin accumulation in MAS:gk, but only marginal increases in the susceptible hybrid Pioneer 3154. Removing wax from the surface of MAS:gk kernels greatly increased the susceptibility of this genotype to aflatoxin accumulation. When wax removal was combined with treatment with potassium hydroxide (KOH) or purified cutinase, aflatoxin levels in kernels were equal to those in wounded control kernels in both genotypes. These results indicated that wax and cutin layers of maize kernel pericarps may play a role in resistance to aflatoxin accumulation in MAS:gk and some other genotypes. However, results suggest further that resistance in MAS:gk also may be due to other preformed compounds as well.


Plant Disease ◽  
1998 ◽  
Vol 82 (3) ◽  
pp. 281-284 ◽  
Author(s):  
G. L. Windham ◽  
W. P. Williams

Field studies were conducted for 2 years in Mississippi to monitor maize kernel infection and aflatoxin accumulation caused by Aspergillus flavus at various times during the growing season. Hybrids resistant and susceptible to A. flavus were compared to determine temporal differences in infection and aflatoxin levels. The resistant hybrids tested were Mo18W × Mp313E, Mp420 × Tx601, and SC54 × SC76; and the susceptible hybrids tested were GA209 × Mp339, Mp307 × Mp428, and Mp68:616 × SC212M. The top ear of each plant was inoculated with a suspension containing A. flavus conidia at 7 days after midsilk (50% of the plants in a plot had silks emerged) using the side needle technique. Inoculated ears were harvested 35, 42, 49, 56, and 63 days after midsilk to determine kernel infection by A. flavus and aflatoxin contamination. Differences in aflatoxin levels between resistant and susceptible hybrids occurred in all harvest dates. However, significant differences between resistant and susceptible hybrids for kernel infection were not observed until 42 days after midsilk. Differences between resistant and susceptible hybrids occurred for kernel infection and aflatoxin concentrations 49, 56, and 63 days after midsilk. Incidence of kernel infection (8.1% for GA209 × Mp339) was the highest 49 days after midsilk, and aflatoxin concentrations (510 ng/g for Mp307 × Mp428) were the highest 63 days after midsilk. Maximum differences between resistant and susceptible hybrids for aflatoxin levels were observed 63 days after midsilk. Two of the resistant hybrids, Mo18W × Mp313E and Mp420 × Tx601, had significantly less aflatoxin than the three susceptible hybrids 63 days after midsilk.


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.


1980 ◽  
Vol 43 (5) ◽  
pp. 381-384 ◽  
Author(s):  
M. F. DUTTON ◽  
M. S. ANDERSON

The effect of a range of organophosphorus and various other compounds on production of aflatoxin by Aspergillus flavus was investigated. Five organophosphorus compounds - Chlormephos, Ciodrin, Naled, Phosdrin and Trichlorphon- at concentrations of 20 and 100 μg/ml of culture fluid were found to have activity similar to Dichlorvos, in that they lowered the level of aflatoxin produced and caused formation of several anthraquinone pigments. Two of these pigments have not previously been described, one was named Versicol and a suggested structure is presented, whilst the other compound was shown to be its acetate derivative. A rationale is suggested for the required elements of structure, which are necessary for an organophosphorus compound to have Dichlorvos-type activity. Two unrelated compounds, ammonium nitrate and Tridecanone were also found to elicit Dichlorvos-type activity. It is likely that tridecanone or its breakdown products competitively inhibit enzymes involved in aflatoxin biosynthesis. It is possible that this inhibition effect explains the lowering of aflatoxin production in lipid-rich commodities infected by A. flavus.


2020 ◽  
Vol 18 (1) ◽  
pp. 297-302 ◽  
Author(s):  
Hamideh Mahmoodzadeh Hosseini ◽  
Siavash Hamzeh Pour ◽  
Jafar Amani ◽  
Sima Jabbarzadeh ◽  
Mostafa Hosseinabadi ◽  
...  

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.


2012 ◽  
Vol 11 (9) ◽  
pp. 1104-1111 ◽  
Author(s):  
Jeffrey W. Cary ◽  
Pamela Y. Harris-Coward ◽  
Kenneth C. Ehrlich ◽  
Brian M. Mack ◽  
Shubha P. Kale ◽  
...  

ABSTRACT The transcription factors NsdC and NsdD are required for sexual development in Aspergillus nidulans . We now show these proteins also play a role in asexual development in the agriculturally important aflatoxin (AF)-producing fungus Aspergillus flavus . We found that both NsdC and NsdD are required for production of asexual sclerotia, normal aflatoxin biosynthesis, and conidiophore development. Conidiophores in nsdC and nsdD deletion mutants had shortened stipes and altered conidial heads compared to those of wild-type A. flavus . Our results suggest that NsdC and NsdD regulate transcription of genes required for early processes in conidiophore development preceding conidium formation. As the cultures aged, the Δ nsdC and Δ nsdD mutants produced a dark pigment that was not observed in the wild type. Gene expression data showed that although AflR is expressed at normal levels, a number of aflatoxin biosynthesis genes are expressed at reduced levels in both nsd mutants. Expression of aflD , aflM , and aflP was greatly reduced in nsdC mutants, and neither aflatoxin nor the proteins for these genes could be detected. Our results support previous studies showing that there is a strong association between conidiophore and sclerotium development and aflatoxin production in A. flavus.


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.


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