Distribution, Genetic Diversity and Biocontrol of Aflatoxigenic Aspergillus flavus in Serbian Maize Fields

Maize is one of the leading export products in the Republic of Serbia. As a country where economic development depends on agriculture, maize production plays a critical role as a crop of strategic importance. Potential aflatoxin contamination of maize poses a risk to food and feed safety and tremendous economic losses. No aflatoxin contamination of maize samples harvested in 2019 and 2020 in different localities in the Republic of Serbia was detected by the Enzyme-Linked Immunosorbent Assay (ELISA) test and High-Performance Liquid Chromatography (HPLC) method. On the other hand, the Cluster Amplification Patterns (CAP) analyses of the isolated Aspergillus flavus strains from 2019 maize samples confirmed the presence of key biosynthesis genes responsible for aflatoxin production. Artificial inoculation and subsequent HPLC analysis of the inoculated maize samples confirmed the high capacity of the A. flavus strains for aflatoxin production, pointing to a high risk of contamination under favorable conditions. Prevention of aflatoxin contamination is primarily based on A. flavus control, where biocontrol agents play a significant role as sustainable disease management tools. In this study, antagonistic activity screening of the novel strains belonging to the Bacillus genus indicated superior suppression of A. flavus strains by two Bacillus strains isolated from the rhizosphere of Phaseolus vulgaris.

[1]  F. M. Ghazali,et al.  Biocontrol of Aflatoxins Using Non-Aflatoxigenic Aspergillus flavus: A Literature Review , 2021, Journal of fungi.

[2]  Marta Wyzińska,et al.  The Most Important Fungal Diseases of Cereals—Problems and Possible Solutions , 2021, Agronomy.

[3]  S. Yusoff,et al.  Revamping Ecosystem Services through Agroecology—The Case of Cereals , 2021 .

[4]  R. Krska,et al.  The impact of management practices to prevent and control mycotoxins in the European food supply chain: MyToolBox project results , 2021, World Mycotoxin Journal.

[5]  A. Shade,et al.  Cross-continental biogeography of the common bean rhizosphere microbiome reveals a persistent core membership , 2019, bioRxiv.

[6]  J. Grahovac,et al.  Xanthomonas campestris biocontrol agent: Selection, medium formulation and bioprocess kinetic analysis , 2021, Chemical Industry and Chemical Engineering Quarterly.

[7]  Fatang Jiang,et al.  Bacillus species as potential biocontrol agents against citrus diseases , 2020 .

[8]  Said M S Massomo,et al.  Aspergillus flavus and aflatoxin contamination in the maize value chain and what needs to be done in Tanzania , 2020 .

[9]  Rahim Khan,et al.  Morphological Characterization and Determination of Aflatoxigenic and Non-Aflatoxigenic Aspergillus flavus Isolated from Sweet Corn Kernels and Soil in Malaysia , 2020, Agriculture.

[10]  J. Grahovac,et al.  Pepper Bacterial Spot Control by Bacillus velezensis: Bioprocess Solution , 2020, Microorganisms.

[11]  B. Gichimu,et al.  Morphological and Molecular Characterization of Toxigenic Aspergillus flavus from Groundnut Kernels in Kenya , 2020, International journal of microbiology.

[12]  R. Arias,et al.  Genetic diversity of aflatoxin-producing Aspergillus flavus isolated from selected groundnut growing agro-ecological zones of Uganda , 2020, BMC Microbiology.

[13]  Moncef Mrabet,et al.  Potential of common bean (Phaseolus vulgaris L.) root microbiome in the biocontrol of root rot disease and traits of performance , 2020, Journal of Plant Diseases and Protection.

[14]  R. Krska,et al.  Biological Control of Aflatoxin in Maize Grown in Serbia , 2020, Toxins.

[15]  I. Oswald,et al.  Aflatoxin Biosynthesis and Genetic Regulation: A Review , 2020, Toxins.

[16]  Lihua Li,et al.  Antifungal activity of endophytic Bacillus safensis B21 and its potential application as a biopesticide to control rice blast. , 2020, Pesticide biochemistry and physiology.

[17]  D. Tsitsigiannis,et al.  Effective Biopesticides and Biostimulants to Reduce Aflatoxins in Maize Fields , 2019, Front. Microbiol..

[18]  Pradeep Kumar,et al.  Aflatoxins in Food and Feed: An Overview on Prevalence, Detection and Control Strategies , 2019, Front. Microbiol..

[19]  S. Mokrani,et al.  Growth Stimulation of Phaseolus Vulgaris L Plantules by Strain Bacillus Amyloliquefaciens Hla Producer of Beneficial Agricultural Enzymes , 2019 .

[20]  Bojana Šarić,et al.  Aflatoxins in maize harvested in the Republic of Serbia over the period 2012–2016 , 2018, Food additives & contaminants. Part B, Surveillance.

[21]  E. Nabti.  Growth Stimulation of Phaseolus vulgaris L Plantules by Strain Bacillus amyloliquefaciens Hla Producer of Beneficial Agricultural Enzymes , 2018, JOJ Horticulture & Arboriculture.

[22]  I. Jajić,et al.  Aflatoxin in Maize Silage Collected from AP Vojvodina, Serbia , 2018 .

[23]  Y. Grosse,et al.  Cyclopiazonic acid: 50th anniversary of its discovery , 2018 .

[24]  S. Koutroubas,et al.  Current Status and Recent Developments in Biopesticide Use , 2018 .

[25]  D. Spadaro,et al.  Unraveling the mode of antifungal action of Bacillus subtilis and Bacillus amyloliquefaciens as potential biocontrol agents against aflatoxigenic Aspergillus parasiticus , 2017, Food Control.

[26]  Mahadevaswamy,et al.  Bioefficacy of Bacillus subtilis against Aspergillus flavus, the cause of aflatoxin contamination in chilli , 2018 .

[27]  A. Bharose,et al.  Antifungal Activity and Metabolites Study of Bacillus Strain Against Aflatoxin Producing Aspergillus , 2018 .

[28]  E. F. Abd_Allah,et al.  Bacillus: A Biological Tool for Crop Improvement through Bio-Molecular Changes in Adverse Environments , 2017, Front. Physiol..

[29]  J. Eloff,et al.  THE USE OF PLANTS TO PROTECT PLANTS AND FOOD AGAINST FUNGAL PATHOGENS: A REVIEW , 2017, African journal of traditional, complementary, and alternative medicines : AJTCAM.

[30]  Joachim Müller,et al.  Innovative technologies to manage aflatoxins in foods and feeds and the profitability of application – A review , 2017, Food control.

[31]  Jamil Shafi,et al.  Bacillus species as versatile weapons for plant pathogens: a review , 2017 .

[32]  A. B. Moura,et al.  Bacterial selection for biological control of plant disease: criterion determination and validation , 2016, Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology].

[33]  Bishwo N. Adhikari,et al.  Degeneration of aflatoxin gene clusters in Aspergillus flavus from Africa and North America , 2016, AMB Express.

[34]  H. J. van der Fels-Klerx,et al.  Aflatoxin B1 contamination in maize in Europe increases due to climate change , 2016, Scientific Reports.

[35]  C. Hurburgh,et al.  Potential economic losses to the US corn industry from aflatoxin contamination , 2016, Food additives & contaminants. Part A, Chemistry, analysis, control, exposure & risk assessment.

[36]  P. Cotty,et al.  Genetic Analysis of the Aspergillus flavus Vegetative Compatibility Group to Which a Biological Control Agent That Limits Aflatoxin Contamination in U.S. Crops Belongs , 2015, Applied and Environmental Microbiology.

[37]  P. Cotty,et al.  Method for monitoring deletions in the aflatoxin biosynthesis gene cluster of Aspergillus flavus with multiplex PCR , 2015, Letters in applied microbiology.

[38]  Naresh Magan,et al.  Effect of climate change on Aspergillus flavus and aflatoxin B1 production , 2014, Front. Microbiol..

[39]  M. Mangal,et al.  Macro and micromorphological characterization of different Aspergillus isolates. , 2014 .

[40]  I. Mandic,et al.  Presence of aflatoxins in cereals from Serbia , 2014 .

[41]  Bojana Šarić,et al.  Natural occurrence of aflatoxins in maize harvested in Serbia during 2009–2012 , 2013 .

[42]  B. Grujić,et al.  AFLATOXIN STANDARDS AND MAIZE TRADE , 2013 .

[43]  Jiujiang Yu,et al.  Current Understanding on Aflatoxin Biosynthesis and Future Perspective in Reducing Aflatoxin Contamination , 2012, Toxins.

[44]  Pankaj Kumar,et al.  Bacillus strains isolated from rhizosphere showed plant growth promoting and antagonistic activity against phytopathogens. , 2012, Microbiological research.

[45]  M. C. Orozco-Mosqueda,et al.  Mechanisms of biocontrol and plant growth-promoting activity in soil bacterial species of Bacillus and Pseudomonas: a review , 2012 .

[46]  C. Probst,et al.  Relationships between in vivo and in vitro aflatoxin production: reliable prediction of fungal ability to contaminate maize with aflatoxins. , 2012, Fungal biology.

[47]  W. Wade,et al.  Bergey’s Manual of Systematic Bacteriology , 2012 .

[48]  P. Fickers,et al.  Bacillus-based biological control of plant diseases , 2011 .

[49]  J. Hellin,et al.  Crops that feed the world 6. Past successes and future challenges to the role played by maize in global food security , 2011, Food Security.

[50]  A. de Vicente,et al.  Plant protection and growth stimulation by microorganisms: biotechnological applications of Bacilli in agriculture. , 2011, Current opinion in biotechnology.

[51]  J. Dorner.  Efficacy of a biopesticide for control of aflatoxins in corn. , 2010, Journal of food protection.

[52]  C. Balachandran,et al.  Histopathological changes in broiler chickens fed aflatoxin and cyclopiazonic acid. , 2009 .

[53]  N. Lima,et al.  A polyphasic approach to the identification of aflatoxigenic and non-aflatoxigenic strains of Aspergillus Section Flavi isolated from Portuguese almonds. , 2009, International journal of food microbiology.

[54]  A. M. Fernandes,et al.  Determination of Aflatoxins in Peanut Products in the Northeast Region of São Paulo, Brazil , 2009, International journal of molecular sciences.

[55]  C. Fininsa,et al.  Evaluation of rhizosphere bacterial antagonists for their potential to bioprotect potato (Solanum tuberosum) against bacterial wilt (Ralstonia solanacearum) , 2008 .

[56]  M. Klich.  Aspergillus flavus: the major producer of aflatoxin. , 2007, Molecular plant pathology.

[57]  K. Padmalatha,et al.  Optimization of DNA isolation and PCR protocol for RAPD analysis of selected medicinal and aromatic plants of conservation concern from Peninsular India , 2006 .

[58]  Deepak Bhatnagar,et al.  The aflatoxin pathway regulator AflR induces gene transcription inside and outside of the aflatoxin biosynthetic cluster. , 2006, FEMS microbiology letters.

[59]  A. Allameh,et al.  A Survey on Distribution of Aspergillus Section Flavi in Corn Field Soils in Iran: Population Patterns Based on Aflatoxins, Cyclopiazonic Acid and Sclerotia Production , 2006, Mycopathologia.

[60]  P. Cotty.  Comparison of four media for the isolation ofAspergillus flavus group fungi , 1994, Mycopathologia.

[61]  S. Kern,et al.  Sodium boric acid: a Tris-free, cooler conductive medium for DNA electrophoresis. , 2004, BioTechniques.

[62]  E. Montesinos.  Development, registration and commercialization of microbial pesticides for plant protection , 2003, International microbiology : the official journal of the Spanish Society for Microbiology.

[63]  P. Cotty.  Influence of field application of an atoxigenic strain of Aspergillus flavus on the populations of A. flavus infectiing cotton bolls and on the aflatoxin content of cottonseed , 1994 .

[64]  R. Harvey,et al.  Toxicological evaluation of aflatoxin and cyclopiazonic acid in broiler chickens. , 1992, Poultry science.

[65]  P. Cotty,et al.  Virulence and cultural characteristics of two Aspergillus flavus strains pathogenic on cotton. , 1989 .