Laccases, as a kind of multicopper oxidase, play an important role in pigment synthesis and growth in fungi and are involved in their interactions with host plants.  In Setosphaeria turcica, 9 laccase-like multicopper oxidases have been identified, and StLAC2 is involved in the synthesis of the melanin that accumulates in the cell wall.  The function of another major laccase gene, StLAC6, was studied here.  The knockout of StLAC6 had no effect on the growth, morphology or invasion ability of S. turcica, but the morphology and function of peroxisomes of knockout mutants were abnormal.  The knockout of the StLAC6 gene resulted in increased contents of phenolic compounds and melanin and the sensitivity to fungicides increased compared with wild type strains.  In the mutants of StLAC6, there is a significant change of the expression levels of other laccase genes.  This study provides a new insight into laccase functions and the relationship of the laccase gene family in plant pathogenic fungi.   

Setosphaeria turcica (syn. Exserohilum turcicum) is the pathogenic fungus of maize (Zea mays) that causes northern leaf blight, which is a major maize disease worldwide.  Melanized appressoria are highly specialized infection structures formed by germinated conidia of S. turcica that infect maize leaves.  The appressorium penetrates the plant cuticle by generating turgor, and glycerol is known to be the main source of the turgor.  Here, the infection position penetrated by the appressorium on maize leaves was investigated, most of the germinated conidia entered the leaf interior by directly penetrating the epidermal cells, and the appressorium structure was necessary for the infection, whether it occurred through epidermal cells or stomata.  Then, to investigate the effects of key factors in the development of the appressorium, we studied the effects of three inhibitors, including a melanin inhibitor (tricyclazole, TCZ), a DNA replication inhibitor (hydroxyurea, HU), and an autophagy inhibitor (3-methyladenine, 3-MA), on appressorium turgor and glycerol content.  As results, appressorium turgor pressure and glycerol concentration in the appressorium reached their highest levels at the mature stage of the appressorium under the control and inhibitor treatments.  The three inhibitors had the greatest effects on appressorium turgor pressure at this stage.  Glycogen and liposomes are the main substances producing glycerol.  It was also found inhibitors affected the distribution of glycogen and liposomes, which were detected in the conidia, the germ tube, and the appressorium during appressorium development.  This study provides profound insight into the relationship between appressorium turgor pressure and glycerol content, which was affected by the synthesis of melanin, DNA replication, and autophagy in the developing appressorium during a S. turcica infection.