The C3HC type zinc-finger protein (ZFC3) interacting with Lon/MAP1 is important for mitochondrial gene regulation, infection hypha development and longevity of Magnaporthe oryzae

Abstract

Background: The rice blast is a typical fungal disease caused by Magnaporthe oryzae, and the mitochondrial ATPdependent Lon protease (MAP1) has been proven to be involved in blast development. We previously screened a C3HC type Zinc-finger domain protein (ZFC3), which is interacted with MAP1. The purpose of this research was to study the biological function of ZFC3 protein in M. oryzae. Results: We first confirmed that the ZFC3-RFP fusion protein is localized within the mitochondria. The deleted mutant strains of ZFC3 (ΔZFC3) showed the enhanced expression level of mtATP6, particularly mtATP8, and almost unchanged nATP9. ΔZFC3 produces more conidia and more tolerance to multiple stressors. The knockout strain shows more melanin accumulation suggests the susceptibility to aging. ΔZFC3 displays faster early-stage hypha infiltration involved in MAP1-mediated pathogenicity in host rice. Conclusion: These results support the view that ZFC3 is a key regulator involved in gene regulation, stress response, cell wall integrity, longevity, conidiation, infection hypha development and MAP1-mediated pathogenicity in M. oryzae.

Contribution of the Mitochondrial Carbonic Anhydrase (MoCA1) to Conidiogenesis and Pathogenesis in Magnaporthe oryzae

“…The result showed that the content of ATP in mycelia and conidia of Δ MoCA1 strain was lower than wild-type and Δ MoCA1/MoCA1 strains ( Figure 5E ), indicating that the loss of MoCA1 affects the energy metabolism in the mutant strain, which, in turn, affects the synthesis of intracellular ATP. Furthermore, when ATP synthesis genes expression level of mtATP6 (GenBank: MGG_21007), mtATP8 (GenBank: MGG_21008), and nATP9 (GenBank: MGG_00892) was checked using qRT-PCR ( Liu et al, 2020 ), the results showed that ATP synthesis genes were significantly downregulated in Δ MoCA1 mutant ( Figure 5F ). From these results we confer, that MoCA1 is involved as a positive regulator in ATP synthesis and energy metabolism in mitochondria of M. oryzae .…”

Cleavage of PrePL by Lon promotes growth and pathogenesis in Magnaporthe oryzae

“…High levels of reactive oxygen species (ROS) generally damage cells causing premature ageing (Harman, 1972; Tsilibaris et al ., 2006; Cui et al ., 2017). Lon proteases and/or their substrates are also associated with senescence or ageing in fungi (Cui et al ., 2015; Liu et al ., 2020). By measuring the continuous growth of hyphae on solid media plates, the lifespan and senescence of the four transgenic strains (Δ PrePL/PrePL , Δ PrePL/Pc , Δ PrePL and Δ PrePL/PrePL ΔTL/NQ ) were compared with the wild‐type strain using 20‐day culture cycles.…”

“…Lon proteases are highly conserved ATP‐dependent proteases that are important for various cellular processes, including growth, reproduction and pathogenicity in prokaryotes and eukaryotes (Tsilibaris et al ., 2006; Coleman et al ., 2009; Li et al ., 2014; Cui et al ., 2015; Liu et al ., 2020). In bacteria, the type III secretion system translocates effector proteins into the cytoplasm of host cells.…”

“…We previously showed that Lon/MAP1 (the M. oryzae homologue of Lon) plays important roles in fungal stress resistance and pathogenic development (Li et al ., 2014). Additionally, Lon/MAP1 physically interacts with a series of proteins, most of which are involved in M. oryzae growth, development and pathogenicity (Cui et al ., 2015; Liu et al ., 2020). Among them, PrePL (a predicted protein with low similarity to the leucine‐rich repeat motif) is the smallest Lon/MAP1‐binding protein.…”

Contribution of the Tyrosinase (MoTyr) to Melanin Synthesis, Conidiogenesis, Appressorium Development, and Pathogenicity in Magnaporthe oryzae

“…oryzae , MoCA1 participates in appressorium maturation and morphogenesis; it was found that the loss of the gene reduced melanin synthesis and infection ability [ 61 ]. C3HC type Zinc-finger protein (ZFC3) and deletion of this gene led to increased melanin synthesis and its ability to infect rice cells [ 71 ]. These results suggest that M .…”