Molecular profile of aflatoxigenic and non-aflatoxigenic isolates of Aspergillus flavus isolated from stored maize

Abstract

Maize is a significant staple crop and utilized in Saudi Arabia as food and feed, but maize is often infected with Aspergillus flavus in tropical and subtropical climates, especially during storage. This study intended at a polyphasic approach, consisting of microscopic morphological, biochemical, and molecular characterizations that were applied to 29 of A. flavus isolates of stored maize, with the goal of characterization and identification of aflatoxigenic and non-aflatoxigenic A. flavus isolates. The technique of real-time PCR (RTi-PCR) was used to detection of A. flavus in stored maize samples, the findings have been very accurate. Centered on macroscopic morphological (primarily colony color and morphology of conidia) and microscopic (morphology of conidia and size) characteristics. Results have shown 23 A. flavus isolates (80%) were categorized as the dark green of colonies also all isolates were rough conidia. The isolates have been two different groups, 16 isolates (62%) had sclerotium-forming and the remaining 13 isolates (38%) had no sclerotium-forming isolates. To the identification of aflatoxigenic isolates of A. flavus in stored maize, we utilized the qualitative methods (easy and inexpensive) like UV test, yellow pigmentation, and ammonia vapor and quantitative method as HPLC (accurate and expensive). the accuracy methods to the identification aflatoxigenicity isolates, vary, and classified in the following descending order: HPLC (100%) > UV method (81%) > yellow pigmentation (YP) and ammonia vapor (AV) (63%). The profile of Aflatoxigenicity of A. flavus isolates by HPLC has been involved in two types first of 11 isolates (38%) have been aflatoxigenic isolates while 18 isolates (62%) were non-aflatoxigenic isolates. The expression of six aflatoxins (AFs) genes (aflD, aflM, aflO, aflP, aflR, and aflQ) was estimated using PCR and RT-PCR. PCR of all genes did not correspond to the aflatoxigenic isolates. The transcriptional analysis of aflO and aflQ was a beneficial marker for discriminating aflatoxigenic from non-aflatoxigenic A. flavus isolates. Also, qRT-PCR indicated that non-aflatoxigenic isolates had a high incidence of defect or downregulation in late AF-genes contrast with early AF-genes. therefore, these non-aflatoxigenic isolates could be critical factors for an efficient and competent strategy for the control of aflatoxin contamination pre-harvest can be considered.

Keywords

• Maize
• Aspergillus flavus
• Aflatoxins
• HPLC
• Aflatoxin genes