Corrigendum: Dicer-Like Genes Are Required for H2O2 and KCl Stress Responses, Pathogenicity and Small RNA Generation in Valsa mali.
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ABSTRACT:
SUBMITTER: Feng H
PROVIDER: S-EPMC5532425 | biostudies-other | 2017
REPOSITORIES: biostudies-other
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Project description:Valsa mali (V. mali) is the causative agent of apple tree Valsa canker, which heavily damages the production of apples in China. However, the biological roles of the RNA interfering (RNAi) pathway in the pathogenicity of V. mali remain unknown. Dicer-like proteins (DCLs) are important components that control the initiation of the RNAi pathway. In this study, VmDCL1 and VmDCL2 were isolated and functionally characterized in V. mali. VmDCL1 and VmDCL2 are orthologous in evolution to the DCLs in Cryphonectria parasitica. The deletion of VmDCL1 and VmDCL2 did not affect vegetative growth when the mutants (ΔVmDCL1, ΔVmDCL2 and ΔVmDCL1DCL2) and wild type strain 03-8 were grown on a PDA medium at 25°C in the dark. However, the colony of ΔVmDCL1 increased by 37.1% compared to the 03-8 colony in a medium containing 0.05% H2O2 3 days after inoculation, and the growth of ΔVmDCL1 was significantly inhibited in a medium containing 0.5 M KCl at a ratio of 25.7%. Meanwhile, in the presence of 0.05% H2O2, the growth of ΔVmDCL2 decreased by 34.5% compared with the growth of 03-8, but ΔVmDCL2 grew normally in the presence of 0.5 M KCl. More importantly, the expression of VmDCL2 was up-regulated 125-fold during the pathogen infection. In the infection assays using apple twigs, the pathogenicity of ΔVmDCL2 and ΔVmDCL1DCL2 was significantly reduced compared with that of 03-8 at a ratio of 24.7 and 41.3%, respectively. All defective phenotypes could be nearly rescued by re-introducing the wild type VmDCL1 and VmDCL2 alleles. Furthermore, the number and length distribution of unique small RNAs (unisRNAs) in the mutants and 03-8 were analyzed using deep sequencing. The number of unisRNAs was obviously lower in ΔVmDCL1, ΔVmDCL2 and ΔVmDCL1DCL2 than that in 03-8, and the length distribution of the sRNAs also markedly changed after the VmDCLs were deleted. These results indicated that VmDCLs function in the H2O2 and KCl stress response, pathogenicity and generation of sRNAs.
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Project description:Apple Valsa canker, caused by the fungus Valsa mali, is one of the most destructive diseases of apple trees in East Asia. Feruloyl esterases (ferulic acid esterases, FAEs), which belong to a subclass of carboxylic esterases, can cleave ester bonds that crosslink hydroxycinnamic acids and arabinoxylans or certain pectins in plant cell walls. However, a pathogenic role of FAE has not been demonstrated in plant-pathogenic fungi. In this study, the FAE gene family, including one type A, one type B, three type C and two type D FAE genes, was identified in V. mali. Five of the seven FAE genes had highly elevated transcript levels in V. mali-apple tree bark interactions compared with mycelia grown in axenic culture. Signal peptides of the VmFAEs were confirmed using yeast signal sequence trap assays. To examine whether FAEs are required for the pathogenicity of V. mali, seven single- and six double-gene deletion mutants were generated. Compared with the wild-type, three of the seven FAE single-deletion mutants showed significantly reduced pathogenicity and three of the six FAE double-deletion mutants exhibited greater reductions in pathogenicity, suggesting the joint action of FAEs in the V. mali-apple tree interaction. Most of the FAE mutants that exhibited a significant reduction in pathogenicity had significantly lower FAE activity than the wild-type fungus. These results indicate that secreted FAEs are required for the full pathogenicity of the phytopathogenic fungus V. mali.
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Project description:Valsa mali is the causative agent of apple tree valsa canker, which causes significant losses in apple production. It produces various toxic compounds that kill plant cells, facilitating infection. Among these, protocatechuic acid exhibits the highest phytotoxic activity. However, those genes involved in toxin production have not been studied. In this study we identified four hydroxybenzoate hydroxylase genes (VmHbh1, VmHbh2, VmHbh3, and VmHbh4) from the transcriptome of V. mali. The VmHbh protein had high enzymatic activities of hydroxybenzoate hydroxylase, which could convert 4-hydroxybenzoate to protocatechuic acid. These four VmHbh genes all had highly elevated transcript levels during the V. mali infection process, especially VmHbh1 and VmHbh4, with 26.0- and 53.4-fold increases, respectively. Mutants of the four genes were generated to study whether VmHbhs are required for V. mali pathogenicity. Of the four genes, the VmHbh1 and VmHbh4 deletion mutants considerably attenuated V. mali virulence in apple leaves and in twigs, coupled with much reduced toxin levels. The VmHbh2 and VmHbh3 deletion mutants promoted the transcript levels of the other VmHbhs, suggesting functional redundancies of VmHbhs in V. mali virulence. The results provide insights into the functions of VmHbhs in the production of protocatechuic acid by V. mali during its infection of apple trees.
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Project description:In order to find some basis of salinity resistance in the chloroplastic metabolism, a halophytic Thellungiella salsuginea was compared with glycophytic Arabidopsis thaliana. In control T.s. plants the increased ratios of chlorophyll a/b and of fluorescence emission at 77 K (F730 /F685 ) were documented, in comparison to A.t.. This was accompanied by a higher YII and lower NPQ (non-photochemical quenching) values, and by a more active PSI (photosystem I). Another prominent feature of the photosynthetic electron transport (PET) in T.s. was the intensive production of H2 O2 from PQ (plastoquinone) pool. Salinity treatment (0.15 and 0.30 M NaCl for A.t. and T.s., respectively) led to a decrease in ratios of chl a/b and F730 /F685 . In A.t., a salinity-driven enhancement of YII and NPQ was found, in association with the stimulation of H2 O2 production from PQ pool. In contrast, in salinity-treated T.s., these variables were similar as in controls. The intensive H2 O2 generation was accompanied by a high activity of PTOX (plastid terminal oxidase), whilst inhibition of this enzyme led to an increased H2 O2 formation. It is hypothesized, that the intensive H2 O2 generation from PQ pool might be an important element of stress preparedness in Thellungiella plants. In control T.s. plants, a higher activation state of carboxylase ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco, EC 4.1.1.39) was also documented in concert with the attachment of Rubisco activase (RCA) to the thylakoid membranes. It is supposed, that a closer contact of RCA with PSI in T.s. enables a more efficient Rubisco activation than in A.t.
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Project description:Unbalanced tumor necrosis factor (TNF)-α production is associated with pathogenesis of a variety of human diseases. However, the molecular pathways maintaining TNF-α homeostasis remain elusive. Here, we report that NF-κB/p65-DICER-miRs axis negatively regulates TNF-α production. We demonstrated that NF-κB bound to DICER promoter and transcriptionally regulated DICER expression. In addition, the NF-κB/DICER signaling suppresses TNF-α expression by generating mature forms of miR-125b and miR-130a which negatively regulate TNF-α mRNA. Furthermore, we showed that the hepatocyte-specific depletion of Dicer in mice resulted in TNF-α overproduction and sensitized the mice to endotoxin, which could be corrected by administration of miR-125b mimics. These data suggest that NF-κB/p65-DICER-miRs axis involved in maintaining of TNF-α homeostasis, and injection of miR-125b as a potential therapeutic method for septic shock.
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Project description:PURPOSE:Oxidative stress (OS) is believed to be a major contributor to age-related cataract and other age-related diseases. METHODS:cDNA microarrays were used to identify the spectrum and range of genes with transcript levels that are altered in response to acute H(2)O(2)-induced OS in human lens epithelial (HLE) cells. HLE cells were treated with 50 microM H(2)O(2) for 1 hour in the absence of serum, followed by a return to complete medium. RNAs were prepared from treated and untreated cells at 0, 1, 2, and 8 hours after H(2)O(2) treatment. RESULTS:The data showed 1171 genes that were significantly up- and downregulated in response to H(2)O(2) treatment. Several functional subcategories of genes were identified, including those encoding DNA repair proteins, antioxidant defense enzymes, molecular chaperones, protein biosynthesis enzymes, and trafficking and degradation proteins. Differential expression of selected genes was confirmed at the level of RNA and/or protein. Many of the identified genes (e.g., glutathione S-transferase [MGST2], thioredoxin reductase beta, and peroxiredoxin 2) have been identified as participants in OS responses in the lens and other systems. Some genes induced by OS in the current study (e.g., oxygen regulated protein [ORP150] and heat shock protein [HSP40]) are better known to respond to other forms of stress. Two genes (receptor tyrosine kinase [AXL/ARK] and protein phosphatase 2A) are known to be differentially expressed in cataract. Most of the genes point to a novel pathways associated with OS. CONCLUSIONS:The present data provide a global perspective on those genes that respond to acute OS, point to novel genes and pathways associated with OS, and set the groundwork for understanding the functions of OS-related genes in lens protection and disease.
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