Image_2_Glycerol-3-Phosphate Shuttle Is Involved in Development and Virulence in the Rice Blast Fungus Pyricularia oryzae.PDF

<p>The glycerol-3-phosphate (G-3-P) shuttle is an important pathway for delivery of cytosolic reducing equivalents into mitochondrial oxidative phosphorylation, and plays essential physiological roles in yeast, plants, and animals. However, its role has been unclear in filamentous and pathogenic fungi. Here, we characterize the function of the G-3-P shuttle in Pyricularia oryzae by genetic and molecular analyses. In P. oryzae, a glycerol-3-phosphate dehydrogenase 1 (PoGpd1) is involved in NO production, conidiation, and utilization of several carbon sources (pyruvate, sodium acetate, glutamate, and glutamine). A glycerol-3-phosphate dehydrogenase 2 (PoGpd2) is essential for glycerol utilization and fungal development. Deletion of PoGPD2 led to delayed aerial hyphal formation, accelerated aerial hyphal collapse, and reduced conidiation on complete medium (CM) under a light–dark cycle. Aerial mycelial surface hydrophobicity to water and Tween 20 was decreased in ΔPogpd2. Melanin synthesis genes required for cell wall construction and two transcription factor genes (COS1 and CONx2) required for conidiation and/or aerial hyphal differentiation were down-regulated in the aerial mycelia of ΔPogpd2 and ΔPogpd1. Culturing under continuous dark could complement the defects of aerial hyphal differentiation of ΔPogpd2 observed in a light–dark cycle. Two light-sensitive protein genes (PoSIR2 encoding an NAD⁺-dependent deacetylase and TRX2 encoding a thioredoxin 2) were up-regulated in ΔPogpd2 cultured on CM medium in a light–dark cycle. ΔPogpd2 showed an increased intracellular NAD⁺/NADH ratio and total NAD content, and alteration of intracellular ATP production. Culturing on minimal medium also could restore aerial hyphal differentiation of ΔPogpd2, which is deficient on CM medium in a light–dark cycle. Two glutamate synthesis genes, GDH1 and PoGLT1, which synthesize glutamate coupled with oxidation of NADH to NAD⁺, were significantly up-regulated in ΔPogpd2 in a light–dark cycle. Moreover, deletion of PoGpd1 or PoGpd2 led to reduced virulence of conidia or hyphae on rice. The glycerol-3-phosphate shuttle is involved in cellular redox, fungal development, and virulence in P. oryzae.</p

Table_1_Glycerol-3-Phosphate Shuttle Is Involved in Development and Virulence in the Rice Blast Fungus Pyricularia oryzae.XLSX

<p>The glycerol-3-phosphate (G-3-P) shuttle is an important pathway for delivery of cytosolic reducing equivalents into mitochondrial oxidative phosphorylation, and plays essential physiological roles in yeast, plants, and animals. However, its role has been unclear in filamentous and pathogenic fungi. Here, we characterize the function of the G-3-P shuttle in Pyricularia oryzae by genetic and molecular analyses. In P. oryzae, a glycerol-3-phosphate dehydrogenase 1 (PoGpd1) is involved in NO production, conidiation, and utilization of several carbon sources (pyruvate, sodium acetate, glutamate, and glutamine). A glycerol-3-phosphate dehydrogenase 2 (PoGpd2) is essential for glycerol utilization and fungal development. Deletion of PoGPD2 led to delayed aerial hyphal formation, accelerated aerial hyphal collapse, and reduced conidiation on complete medium (CM) under a light–dark cycle. Aerial mycelial surface hydrophobicity to water and Tween 20 was decreased in ΔPogpd2. Melanin synthesis genes required for cell wall construction and two transcription factor genes (COS1 and CONx2) required for conidiation and/or aerial hyphal differentiation were down-regulated in the aerial mycelia of ΔPogpd2 and ΔPogpd1. Culturing under continuous dark could complement the defects of aerial hyphal differentiation of ΔPogpd2 observed in a light–dark cycle. Two light-sensitive protein genes (PoSIR2 encoding an NAD⁺-dependent deacetylase and TRX2 encoding a thioredoxin 2) were up-regulated in ΔPogpd2 cultured on CM medium in a light–dark cycle. ΔPogpd2 showed an increased intracellular NAD⁺/NADH ratio and total NAD content, and alteration of intracellular ATP production. Culturing on minimal medium also could restore aerial hyphal differentiation of ΔPogpd2, which is deficient on CM medium in a light–dark cycle. Two glutamate synthesis genes, GDH1 and PoGLT1, which synthesize glutamate coupled with oxidation of NADH to NAD⁺, were significantly up-regulated in ΔPogpd2 in a light–dark cycle. Moreover, deletion of PoGpd1 or PoGpd2 led to reduced virulence of conidia or hyphae on rice. The glycerol-3-phosphate shuttle is involved in cellular redox, fungal development, and virulence in P. oryzae.</p

Image_3_Glycerol-3-Phosphate Shuttle Is Involved in Development and Virulence in the Rice Blast Fungus Pyricularia oryzae.pdf

<p>The glycerol-3-phosphate (G-3-P) shuttle is an important pathway for delivery of cytosolic reducing equivalents into mitochondrial oxidative phosphorylation, and plays essential physiological roles in yeast, plants, and animals. However, its role has been unclear in filamentous and pathogenic fungi. Here, we characterize the function of the G-3-P shuttle in Pyricularia oryzae by genetic and molecular analyses. In P. oryzae, a glycerol-3-phosphate dehydrogenase 1 (PoGpd1) is involved in NO production, conidiation, and utilization of several carbon sources (pyruvate, sodium acetate, glutamate, and glutamine). A glycerol-3-phosphate dehydrogenase 2 (PoGpd2) is essential for glycerol utilization and fungal development. Deletion of PoGPD2 led to delayed aerial hyphal formation, accelerated aerial hyphal collapse, and reduced conidiation on complete medium (CM) under a light–dark cycle. Aerial mycelial surface hydrophobicity to water and Tween 20 was decreased in ΔPogpd2. Melanin synthesis genes required for cell wall construction and two transcription factor genes (COS1 and CONx2) required for conidiation and/or aerial hyphal differentiation were down-regulated in the aerial mycelia of ΔPogpd2 and ΔPogpd1. Culturing under continuous dark could complement the defects of aerial hyphal differentiation of ΔPogpd2 observed in a light–dark cycle. Two light-sensitive protein genes (PoSIR2 encoding an NAD⁺-dependent deacetylase and TRX2 encoding a thioredoxin 2) were up-regulated in ΔPogpd2 cultured on CM medium in a light–dark cycle. ΔPogpd2 showed an increased intracellular NAD⁺/NADH ratio and total NAD content, and alteration of intracellular ATP production. Culturing on minimal medium also could restore aerial hyphal differentiation of ΔPogpd2, which is deficient on CM medium in a light–dark cycle. Two glutamate synthesis genes, GDH1 and PoGLT1, which synthesize glutamate coupled with oxidation of NADH to NAD⁺, were significantly up-regulated in ΔPogpd2 in a light–dark cycle. Moreover, deletion of PoGpd1 or PoGpd2 led to reduced virulence of conidia or hyphae on rice. The glycerol-3-phosphate shuttle is involved in cellular redox, fungal development, and virulence in P. oryzae.</p

Characterization of Coke Formed during Thermal Reaction of Tar

Coking of volatiles generated from coal in pyrolysis has been a focal issue in coal pyrolysis and upgrading of coal tar, but limited work can be found in the literature on evolution of coke in composition and structure under the pyrolysis conditions. This work characterizes the coke formed in reaction of a subbituminous coal tar at 300, 400, and 500 °C in 40 min in a semibatch system which allows natural evaporation of light fractions. The coke is categorized into two types, the one suspended in tetrahydrofuran (THF), coke-S, and the one deposited on the wall of tube reactor, coke-D. It is found that coke-D accounts for 70–85% of total coke. With increasing tar reaction temperature and time the quantity of coke increases from 1.0% to 16.3 wt % and the particle size of coke-S increases from a most probable size of approximately 0.1 to 700–800 μm. This change is accompanied by reduction in alkyl side chains and heteroatoms (O, N, and S), as well as the enrichment in the aromatic C_ar–C_ar bond, which lead to a decrease in H/C ratio from 0.9 to 0.6 and increase in aromaticity <i>f</i>_a from 0.70 to 0.86. The carbon distribution in coke-S is similar to that in bituminous coals and is composed of 3–7 fused aromatic rings. The changes in coke-S also include increase in radical concentration and decreases in the radicals’ <i>g</i> value and line width, indicating continued pyrolysis and condensation of the coke due to the removal of oxygen atoms and side chains on the aromatic structure. When compared with coke-S, coke-D formed under the same conditions is more condensed as indicated by higher radical concentration and lower <i>g</i> value and line width. The morphological change in coke-D includes transformation of small irregular particles to spherical-like particles and to coke film that crack in 30 min at 300 °C or 10 min at 500 °C