Abstract
Magnaporthe oryzae is the causal agent of rice blast outbreaks. L-ascorbic acid (ASC) is a famous antioxidant found in nature. However, while ASC is rare or absent in fungi, a five-carbon analog, D-erythroascorbic acid (EASC), seems to appear to be a substitute for ASC. Although the antioxidant function of ASC has been widely described, the specific properties and physiological functions of EASC remain poorly understood. In this study, we identified a D-arabinono-1,4-lactone oxidase (ALO) domain-containing protein, MoAlo1, and found that MoAlo1 was localized to mitochondria. Disruption of MoALO1 (ΔMoalo1) exhibited defects in vegetative growth as well as conidiogenesis. The ΔMoalo1 mutant was found to be more sensitive to exogenous H2O2. Additionally, the pathogenicity of conidia in the ΔMoalo1 null mutant was reduced deeply in rice, and defective penetration of appressorium-like structures (ALS) formed by the hyphal tips was also observed in the ΔMoalo1 null mutant. When exogenous EASC was added to the conidial suspension, the defective pathogenicity of the ΔMoalo1 mutant was restored. Collectively, MoAlo1 is essential for growth, conidiogenesis, and pathogenicity in M. oryzae.
Highlights
Introduction published maps and institutional affilMagnaporthe oryzae is a pathogen that threatens all crops around the world
All strains used in this study were cultured in complete medium (CM), Yeast glucose (YG) medium, V8 medium, and oatmeal agar (OMA) medium at 25 ◦ C under a 16 h light/8 h dark photoperiod for 8 days according to a previous study [20,21]
There are many resuggesting that ASC is rare or even absent in microorganisms, but a five-carbon analog, ports suggesting that ASC is rare or even absent in microorganisms, but a five-carbon
Summary
All strains used in this study (including the wild-type Guy and ∆Moalo mutant, the complementary strain MoAlo1C, ∆Moalo1::ALO, and ∆Moalo1::FAD) were cultured in complete medium (CM), Yeast glucose (YG) medium, V8 medium, and oatmeal agar (OMA) medium at 25 ◦ C under a 16 h light/8 h dark photoperiod for 8 days according to a previous study [20,21]. All PCR products and linearized vectors were verified by agarose gel electrophoresis and purified using a DNA gel extraction kit (Axygen, Hangzhou, China) Both the upstream and downstream fragments of. Since the housekeeping genes are stably expressed in cells, in order to verify whether the null mutant has only one copy of the gene-deletion cassette, we used the β-tubulin gene (only one copy in the M. oryzae genome) as a control, and the number of copies of BAR was determined by Real-Time Quantitative. In order to measure the mycelial growth and conidiation, a 5-mm agar block of 8-dayold wild-type ∆Moalo mutant and the complementary strain MoAlo1C were inoculated in the center of CM agar plates at 25 ◦ C with a 16 h light and 8 h dark phase, respectively.
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