ABSTRACT
Aspergillus flavus colonization in developing kernels of maize single-cross hybrids resistant (Mp313E × Mp717) and susceptible (GA209 × T173) to aflatoxin accumulation was determined in the field over three growing seasons (2012-2014). Plants were hand pollinated, and individual kernels were inoculated with a needle dipped in a suspension of A. flavus conidia 21 days after pollination. Kernels were harvested at 1- to 2-day intervals from 1 to 21 days after inoculation (DAI). Kernels were placed in FAA fixative, dehydrated, embedded in paraffin, sectioned, and stained with toluidine blue. Kernels were also collected additional kernels for aflatoxin analyses in 2013 and 2014. At 2 DAI, A. flavus hyphae were observed among endosperm cells in the susceptible hybrid, but colonization of the endosperm in the resistant hybrid was limited to the wound site of the resistant hybrid. Sections of the scutellum of the susceptible hybrid were colonized by A. flavus by 5 DAI. Fungal growth was slower in the resistant hybrid compared to the susceptible hybrid. By 10 DAI, A. flavus had colonized a large section of the embryo in the susceptible hybrid; whereas in the resistant hybrid, approximately half of the endosperm had been colonized and very few cells in the embryo were colonized. Fungal colonization in some of the kernels of the resistant hybrid was slowed in the aleurone layer or at the endosperm-scutellum interface. In wounded kernels with intact aleurone layers, the fungus spread around the kernel between the pericarp and aleurone layer with minimal colonization of the endosperm. Aflatoxin B1 was first detected in susceptible kernel tissues 8 DAI in 2013 (14 µg/kg) and 2014 (18 µg/kg). The resistant hybrid had significantly lower levels of aflatoxin accumulation compared to the susceptible hybrid at harvests 10, 21, and 28 DAI in 2013, and 20 and 24 DAI in 2014. Our study found differential A. flavus colonization of susceptible and resistant kernel tissues, and that the aleurone and the outer layer of the scutellum slowed the rate of colonization by A. flavus.
ABSTRACT
BACKGROUND: Aspergillus flavus Link:Fr, an opportunistic fungus that produces aflatoxin, is pathogenic to maize and other oilseed crops. Aflatoxin is a potent carcinogen, and its presence markedly reduces the value of grain. Understanding and enhancing host resistance to A. flavus infection and/or subsequent aflatoxin accumulation is generally considered an efficient means of reducing grain losses to aflatoxin. Different proteomic, genomic and genetic studies of maize (Zea mays L.) have generated large data sets with the goal of identifying genes responsible for conferring resistance to A. flavus, or aflatoxin. RESULTS: In order to maximize the usage of different data sets in new studies, including association mapping, we have constructed a relational database with web interface integrating the results of gene expression, proteomic (both gel-based and shotgun), Quantitative Trait Loci (QTL) genetic mapping studies, and sequence data from the literature to facilitate selection of candidate genes for continued investigation. The Corn Fungal Resistance Associated Sequences Database (CFRAS-DB) (http://agbase.msstate.edu/) was created with the main goal of identifying genes important to aflatoxin resistance. CFRAS-DB is implemented using MySQL as the relational database management system running on a Linux server, using an Apache web server, and Perl CGI scripts as the web interface. The database and the associated web-based interface allow researchers to examine many lines of evidence (e.g. microarray, proteomics, QTL studies, SNP data) to assess the potential role of a gene or group of genes in the response of different maize lines to A. flavus infection and subsequent production of aflatoxin by the fungus. CONCLUSIONS: CFRAS-DB provides the first opportunity to integrate data pertaining to the problem of A. flavus and aflatoxin resistance in maize in one resource and to support queries across different datasets. The web-based interface gives researchers different query options for mining the database across different types of experiments. The database is publically available at http://agbase.msstate.edu.
