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Toxins (Basel). 2019 Mar 25;11(3). pii: E179. doi: 10.3390/toxins11030179.

The Development of a qPCR Assay to Measure Aspergillus flavus Biomass in Maize and the Use of a Biocontrol Strategy to Limit Aflatoxin Production.

Author information

1
Plant Stress Laboratory 204/207, Department of Molecular and Cell Biology, MCB Building, Upper Campus, University of Cape Town, Private bag X3, Rondebosch, Cape Town 7701, South Africa. alfmite@yahoo.com.
2
Department of Botany, School of Biological Sciences, University of Nairobi, P.O. Box 30197, Nairobi 00100, Kenya. alfmite@yahoo.com.
3
Department of Botany, School of Biological Sciences, University of Nairobi, P.O. Box 30197, Nairobi 00100, Kenya. dorisokoth@yahoo.com.
4
Plant Stress Laboratory 204/207, Department of Molecular and Cell Biology, MCB Building, Upper Campus, University of Cape Town, Private bag X3, Rondebosch, Cape Town 7701, South Africa. Suhail.Rafudeen@uct.ac.za.

Abstract

Aspergillus flavus colonisation of maize can produce mycotoxins that are detrimental to both human and animal health. Screening of maize lines, resistant to A. flavus infection, together with a biocontrol strategy, could help minimize subsequent aflatoxin contamination. We developed a qPCR assay to measure A. flavus biomass and showed that two African maize lines, GAF4 and KDV1, had different fungal loads for the aflatoxigenic isolate (KSM014), fourteen days after infection. The qPCR assay revealed no significant variation in A. flavus biomass between diseased and non-diseased maize tissues for GAF4, while KDV1 had a significantly higher A. flavus biomass (p < 0.05) in infected shoots and roots compared to the control. The biocontrol strategy using an atoxigenic isolate (KSM012) against the toxigenic isolate (KSM014), showed aflatoxin production inhibition at the co-infection ratio, 50:50 for both maize lines (KDV1 > 99.7% and GAF ≥ 69.4%), as confirmed by bioanalytical techniques. As far as we are aware, this is the first report in Kenya where the biomass of A. flavus from maize tissue was detected and quantified using a qPCR assay. Our results suggest that maize lines, which have adequate resistance to A. flavus, together with the appropriate biocontrol strategy, could limit outbreaks of aflatoxicoses.

KEYWORDS:

Aspergillus flavus; Pathogenesis; aflatoxins; fungal biomass; maize lines; qPCR assay

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