Jong-Hwa Kim

In eukaryotes, the MAP kinase signaling pathway plays pivotal roles in regulating the expression of genes required for growth, development, and stress response. Here, we deleted the mpkB gene (AFLA_034170), an ortholog of the Saccharomyces cerevisiae FUS3 gene, to characterize its function in Aspergillus flavus, a cosmopolitan, pathogenic, and aflatoxin-producing fungus. Previous studies revealed that MpkB positively regulates sexual and asexual differentiation in Aspergillus nidulans. In A. flavus, mpkB deletion resulted in an approximately 60% reduction in conidia production compared to the wild type without mycelial growth defects. Moreover, the mutant produced immature and abnormal conidiophores exhibiting vesicular dome-immaturity in the conidiophore head, decreased phialide numbers, and very short stalks. Interestingly, the ΔmpkB mutant could not produce sclerotia but produced aflatoxin B1 normally. Taken together, these results suggest that the A. flavus MpkB MAP kinase positively regulates conidiation and sclerotia formation but is not involved in the production of secondary metabolites such as aflatoxin B1.

Afla-Guard^® is a commercial non-toxigenic Aspergillus flavus strain used to decrease aflatoxin contamination level in field. Its mitochondrial genome was sequenced, showing that its length is 29,208 bp with typical configuration of Aspergillus mitochondrial genome. 17 SNPs and 27 INDELs were identified by comparing with previous A. flavus mitochondrial genome. Phylogenetic trees present that A. flavus of Afla-Guard^® was clustered with the previous A. flavus mitochondrial genome.

The mitogenome of Aspergillus flavus SRRC1009 was sequenced to investigate intraspecific variations on mitochondrial genomes of A. flavus. It shows 29,202 bp with a typical configuration of Aspergillus mitogenome. Sixteen SNPs and 22 INDELs and 17 SNPs and 27 INDELs were identified against AflaGuard® and JQ355000, respectively. Phylogenetic trees present in the three A. flavus mitochondrial genomes were clustered with A. oryzae mitochondrial genome in one clade.

The glycogen-associated protein phosphatase (PP1G/ R-GL) may play a central role in the hormonal control of glycogen metabolism in the skeletal muscle. Here, we investigated the in vivo epinephrine effect of glycogen metabolism in the skeletal muscle of the wild-type and R-GL knockout mice. The administration of epinephrine increased blood glucose levels from 200 +/- 20 to 325 +/- 20 mg/dl in both wild-type and knockout mice. Epinephrine decreased the glycogen synthase -/+ G6P ratio from 0.24 +/- 0.04 to 0.10 +/- 0.02 in the wild-type, and from 0.17 +/- 0.02 to 0.06 +/- 0.01 in the knockout mice. Conversely, the glycogen phosphorylase activity ratio increased from 0.21 +/- 0.04 to 0.65 +/- 0.07 and from 0.30 +/- 0.04 to 0.81 +/- 0.06 in the epinephrine treated wild-type and knockout mice respectively. The glycogen content of the knockout mice was substantially lower (27%) than that of both wild-type mice; and epinephrine decreased glycogen content in the wild-type and knockout mice. Also, in Western blot analysis there was no compensation of the other glycogen targeting components PTG/R5 and R6 in the knockout mice compared with the wild-type. Therefore, R-GL is not required for the epinephrine stimulation of glycogen metabolism, and rather another phosphatase and/or regulatory subunit appears to be involved.