Reference Gene Selection for RT-qPCR Analysis in Maize Kernels Inoculated with Aspergillus flavus

Dafne Alves Oliveira; Juliet D. Tang; Marilyn L. Warburton

doi:10.3390/toxins13060386

Treesearch

Reference Gene Selection for RT-qPCR Analysis in Maize Kernels Inoculated with Aspergillus flavus

Formally Refereed

Authors:

Dafne Alves Oliveira, Juliet D. Tang, Marilyn L. Warburton

Year:

2021

Type:

Scientific Journal

Station:

Forest Products Laboratory

DOI:

https://doi.org/10.3390/toxins13060386

Source:

Toxins

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Abstract

Resistance against infection by the fungus Aspergillus flavus Link in commercial maize (Zea mays L.) is the topic of many studies, but few studies have investigated the effects of A. flavus infection on gene expression levels in ear kernels. A crucial component of gene expression profiling by RT-qPCR is having a reliable set of reference genes that show relatively constant expression across the treatments and phenotypes under study. Currently, however, there is no published information on reference genes suitable for measuring changes in kernel gene expression levels after infection with A. flavus. Thus, in this study, six candidate reference genes (ACT1, β-Tub2, eIF4A2, TATA, EFIα, and GAPDH) were evaluated and ranked according to their expression stability. The genes were amplifed from first-strand cDNA samples synthesized from kernels of two susceptible and two resistant maize lines that were either inoculated with A. flavus or water or not inoculated. Three software packages were used to calculate and rank the stability of expression for these genesgeNorm, NormFinder, and BestKeeper. The analysis revealed that the most stable genes to normalize expression levels from maize kernels responding to A. flavus inoculation and wounding were ACT1, EFIα, and eIF4A2.

Keywords

candidate gene, gene expression, afatoxin

Citation

Alves Oliveira, Dafne; Tang, Juliet D.; Warburton, Marilyn L. 2021. Reference Gene Selection for RT-qPCR Analysis in Maize Kernels Inoculated with Aspergillus flavus. Toxins. 13(6): 386. https://doi.org/10.3390/toxins13060386.