scholarly journals Peanut Seed Cultivars with Contrasting Resistance to Aspergillus parasiticus Colonization Display Differential Temporal Response of Protease Inhibitors

2017 ◽  
Vol 107 (4) ◽  
pp. 474-482 ◽  
Author(s):  
Virginia Müller ◽  
Gustavo Bonacci ◽  
Carlos Batthyany ◽  
María V. Amé ◽  
Fernando Carrari ◽  
...  

Significant efforts are being made to minimize aflatoxin contamination in peanut seeds and one possible strategy is to understand and exploit the mechanisms of plant defense against fungal infection. In this study we have identified and characterized, at biochemical and molecular levels, plant protease inhibitors (PPIs) produced in peanut seeds of the resistant PI 337394 and the susceptible Forman cultivar during Aspergillus parasiticus colonization. With chromatographic methods and 2D-electrophoresis-mass spectrometry we have isolated and identified four variants of Bowman-Birk trypsin inhibitor (BBTI) and a novel Kunitz-type protease inhibitor (KPI) produced in response to A. parasiticus colonization. KPI was detected only in the resistant cultivar, while BBTI was produced in the resistant cultivar in a higher concentration than susceptible cultivar and with different isoforms. The kinetic expression of KPI and BBTI genes along with trypsin inhibitory activity was analyzed in both cultivars during infection. In the susceptible cultivar an early PPI activity response was associated with BBTI occurrence. Meanwhile, in the resistant cultivar a later response with a larger increase in PPI activity was associated with BBTI and KPI occurrence. The biological significance of PPI in seed defense against fungal infection was analyzed and linked to inhibitory properties on enzymes released by the fungus during infection, and to the antifungal effect of KPI.

2015 ◽  
Vol 22 (2) ◽  
pp. 149-163 ◽  
Author(s):  
Maria Macedo ◽  
Caio de Oliveira ◽  
Poliene Costa ◽  
Elaine Castelhano ◽  
Marcio Silva-Filho

1979 ◽  
Vol 62 (5) ◽  
pp. 1076-1079 ◽  
Author(s):  
Lawrence M Lenovich ◽  
W Jeffrey Hurst

Abstract Aflatoxin was produced in both non-autoclaved and autoclaved Ivory Coast cocoa beans inoculated with Aspergillus parasiticus NRRL 2999 under optimum laboratory growth conditions. Total aflatoxin levels ranged from 213 to 5597 ng/g substrate. Aflatoxin was quantitated by using high pressure liquid chromatography (HPLC). Raw, non-autoclaved cocoa beans, also inoculated with aspergilli, produced 6359 ng aflatoxin/g substrate. Variation in aflatoxin production between bean varieties was observed. Total aflatoxin levels of 10,446 and 23,076 ng/g substrate were obtained on Ivory Coast beans inoculated with A. parasiticus NRRL 2999 and NRRL 3240, respectively. Aflatoxin production on Trinidad and Malaysian beans was 28 and 65 ng aflatoxin/g substrate. These data support previously reported low level natural aflatoxin contamination in cocoa.


2021 ◽  
Vol 3 (1) ◽  
Author(s):  
Abhishek Kumar ◽  
Hardik Pathak ◽  
Seema Bhadauria ◽  
Jebi Sudan

AbstractMycotoxins are secondary metabolites produced by several fungal species and molds. Under favorable conditions like high temperature and moisture, they contaminate a large number of food commodities and regional crops during pre and post-harvesting. Aflatoxin is the main mycotoxin that harm animal and human health due to its carcinogenic nature. Aflatoxins are mainly released by Aspergillus flavus and Aspergillus parasiticus. AFB1 constitutes the most harmful type of aflatoxins and is a potent hepato-carcinogenic, mutagenic, teratogenic and it suppresses the immune system. To maintain food safety and to prevent aflatoxin contamination in food crops, combined approaches of using resistant varieties along with recommended farming practices should be followed. This review concentrates on various aspects of mycotoxin contamination in crops and recent methods to prevent or minimize the contamination.


2011 ◽  
Vol 4 (1) ◽  
pp. 37-42 ◽  
Author(s):  
A. Rosas-Taraco ◽  
E. Sanchez ◽  
S. García ◽  
N. Heredia ◽  
D. Bhatnagar

Toxigenic fungi invade crops prior to harvest as well as during storage and produce harmful, even carcinogenic toxins such as aflatoxins. Since consumers demand safe commodities, and due to enhanced public awareness of the dangers of many synthetic fungicides, the importance of investigating alternative, natural products to control these toxigenic fungi is clear. This study investigated the effect of aqueous extracts of Agave americana on growth, conidia and aflatoxin production. Aspergillus parasiticus strains SRRC 148, SRRC 143 (Su-1), and A. parasiticus SRRC 162, a mutant (nor-) that accumulates norsolorinic acid (NOR, an orange-coloured intermediate of the aflatoxin pathway), were first inoculated into Adye and Mateles liquid medium, then plant extracts were added, and incubated at 28 °C for 7 days. Aflatoxin and norsolorinic acid were assayed by HPLC and spectrophotometry, respectively. While the extract of A. americana stimulated growth of the studied fungi, conidiogenesis, norsolorinic acid accumulation (in the nor- mutant), and aflatoxin production were significantly affected. The reduction was produced by the extracts at concentrations higher than 5-10 mg/ml, where all types of total aflatoxin analysed (aflatoxins B1, B2, G1 and G2) were reduced from 64% to >99% in the whole culture, and a reduction of 75% of norsolorinic acid. The results of the present work indicate that extracts of A. americana may be promising safe alternatives to harmful fungicides for controlling aflatoxin contamination.


2013 ◽  
Vol 6 (2) ◽  
pp. 151-158 ◽  
Author(s):  
K. Rajasekaran ◽  
C.M. Sickler ◽  
R.L. Brown ◽  
J.W. Cary ◽  
D. Bhatnagar

Resistance or susceptibility of maize inbreds to infection by Aspergillus flavus was evaluated by the kernel screening assay. A green fluorescent protein-expressing strain of A. flavus was used to measure fungal spread and aflatoxin levels in real-time following fungal infection of kernels. Among the four inbreds tested, MI82 showed the most resistance and Ga209 the least. TZAR101 was also resistant to fungal infection, whereas Va35 was susceptible to fungal infection. However, Va35 produced lower aflatoxin levels compared to the susceptible line Ga209. Fluorescence microscopy indicated that the site of entry of the fungus into the kernel was consistently through the pedicel. Entry through the pericarp was never observed in undamaged kernels. In view of these results, incorporation or overexpression of antifungal proteins should be targeted to the pedicel and basal endosperm region in developing kernels. Once the fungus has entered through the pedicel, it spreads quickly through the open spaces between the pericarp and the aleurone layer, ultimately colonising the endosperm and scutellum and, finally, the embryo. A clear correlation was established between fungal fluorescence and aflatoxin levels. This method provides a quick, reliable means of evaluating resistance to A. flavus in undamaged kernels and provides breeders with a rapid method to evaluate maize germplasm.


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.


2017 ◽  
Vol 11 (1) ◽  
pp. e1368599 ◽  
Author(s):  
Sachin Rustgi ◽  
Edouard Boex-Fontvieille ◽  
Christiane Reinbothe ◽  
Diter von Wettstein ◽  
Steffen Reinbothe

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