MoLEU1 , MoLEU2 , and MoLEU4 regulated by MoLEU3 are involved in leucine biosynthesis, fungal development, and pathogenicity in Magnaporthe oryzae

Amino acids are vital components in cell metabolism. Leucine is a regulatory factor that generates significant impact on protein synthesis/turnover, modulates diverse cellular signalling pathways and participates in oxidative processes and immune responses. Here, we identified and characterized the functions of a leucine-associated Zn2 Cys6 -type transcription factor, MoLeu3. Disruption of MoLEU3 resulted in significantly reduced pathogenicity in barley and rice. Quantitative RT-PCR showed that the expression levels of the putative leucine biosynthesis-related genes, MoLEU1, MoLEU2 and MoLEU4 were downregulated in the DeltaMoleu3 mutant. We used high-throughput gene knockout method to generate the null mutants of MoLEU1, MoLEU2 and MoLEU4 respectively. The DeltaMoleu1, DeltaMoleu2 and DeltaMoleu4 mutants are leucine auxotroph and showed similar phenotypic characterizations, including reduced conidiation, delayed mobilization and degradation of glycogen and lipid droplets, limited appressorium-mediated penetration, and restricted invasive hyphae growth within host cells. Collectively, MoLEU1, MoLEU2, and MoLEU4 regulated by MoLEU3 play crucial roles in fungal development and infectious processes through modulation of leucine biosynthesis in Magnaporthe oryzae.