RESUMO
Allitol is a hexitol produced by reducing the rare sugar D-allulose with a metal catalyst under hydrogen gas. To confirm the safe level of allitol, we conducted a series of safety assessments. From the results of Ames mutagenicity assay using Salmonella typhimurium strains TA98, TA100, TA1535, and TA1537, Escherichia coli strain WP2uvrA, and an in vitro chromosomal aberration test on cultured Chinese hamster cells, allitol did not show any significant genotoxic effect. No significant effects on general condition, urinalysis, hematology, physiology, histopathology, or at necropsy were observed at a dose of 1500 mg/kg body weight of allitol in the acute and 90-day subchronic oral-toxicity assessments for rats. A further study performed on healthy adult humans showed that the acute use level of allitol for diarrhea was 0.2 g/kg body weight for both men and women. The results of current safety assessment studies suggest that allitol is safe for human consumption.
Assuntos
Aberrações Cromossômicas , Escherichia coli , Masculino , Cricetinae , Ratos , Humanos , Feminino , Animais , Ratos Sprague-Dawley , Testes de Mutagenicidade/métodos , Cricetulus , Escherichia coli/genética , Peso Corporal , Ingestão de AlimentosRESUMO
In Thailand, four systemic fungicides-carbendazim (Car), azoxystrobin (Azo), difenoconazole (Dif), and penthiopyrad (Pen)-are commonly used to control soybean anthracnose caused by Colletotrichum truncatum; however, the pathogen has developed resistance. From 2019 to 2020, fungicide resistance in C. truncatum from fields in Chiang Rai and Chiang Mai was monitored. In tests of 85 C. truncatum isolates for resistance to multiple fungicides, 15.3% were CarRAzoR, 34.1% were triple resistant (CarRAzoRDifR or CarRAzoRPenR), and 50.6% were CarRAzoRDifRPenR. Surprisingly, all isolates tested had lost their sensitivity to one or more of the fungicides tested. The carbendazim-resistant isolates carried a point mutation in the ß-tubulin gene at codon 198 (E198A) or 200 (F200Y), and all azoxystrobin-resistant isolates had a mutation in the cytochrome b gene at codon 143 (G143A) or 129 (F129L). Moreover, a novel mutation at codon 208 (S208Y) in the gene encoding succinate dehydrogenase subunit B was detected in all of the isolates highly resistant to penthiopyrad. No mutation linked with difenoconazole resistance was detected in the genes encoding cytochrome P450 sterol 14α-demethylase. To the best of our knowledge, this is the first report of C. truncatum isolates resistant to multiple fungicides and serves as a warning to take measures to prevent the occurrence and distribution of these multiple-fungicide-resistant populations in soybean fields.
Assuntos
Fungicidas Industriais , Fungicidas Industriais/farmacologia , Glycine max , Tailândia , CódonRESUMO
D-Allose is classified as a 'rare sugar,' i.e., part of the group of monosaccharides that are present in low quantities in the natural world. D-Allose has been demonstrated to exert many physiological functions. The effects of the rare sugars on immune responses are largely unexplored. Here, we investigated the physiological effects of D-allose on murine dendritic cells' cytokine production. When plasmacytoid dendritic cells (pDCs) were stimulated with a Toll-like receptor 7 (TLR7) ligand, a single-stranded RNA (ssRNA), or a TLR9 ligand, CpG DNA, in the medium containing D-allose, the productions of both interferon-alpha (IFN-α) and interleukin (IL)-12p40 were severely decreased. In contrast, a normal production of these cytokines was observed when pDCs were stimulated with other TLR7 ligands, an imidazoquinoline, or a guanosine analog. In contrast to the pDCs, conventional dendritic cells (cDCs) produced IL-12p40 and tumor necrosis factor-alpha (TNF-α) in response to an imidazoquinoline or CpG DNA even though D-allose was present in the medium. D-Allose did not induce pDC death, and not inhibit the endocytic uptake of fluorophore-labeled CpG DNA into pDCs. These results suggested that D-allose exerts its inhibitory effects after CpG DNA is internalized. We analyzed the TLR7/9 signal-induced activation of downstream signaling molecules in pDCs and observed that when pDCs were stimulated with a ssRNA or CpG DNA, the phosphorylation status of the MAPK family, which includes Erk1/2, JNK/SAPK, and p38 MAPK, was attenuated in the presence of D-allose compared to D-glucose controls. The stimulation of pDCs with an imidazoquinoline induced a strong phosphorylation of these MAPK family members even in the presence of D-allose. These findings reveal that D-allose can inhibit the cytokine production by pDCs stimulated with ssRNA or CpG DNA via an attenuation of the phosphorylation of MAPK family members.
Assuntos
Receptor 7 Toll-Like , Receptor Toll-Like 9 , Animais , Citocinas , DNA , Células Dendríticas , Glucose/farmacologia , Imunidade , Ligantes , CamundongosRESUMO
Previously, we succeeded to produce the core structure of the host-selective ACR toxin (1) on brown leaf spot on rough lemon when the polyketide synthase ACRTS2 gene was heterologously expressed in Aspergillus oryzae (AO). To confirm the production of 1 in AO, the detection limit and suppressing decarboxylation were improved, and these efforts led us to conclude the direct production of 1 instead of its decarboxylation product. During this examination, minor ACR-toxin-related metabolites were found. Their structure determination enabled us to propose a decarboxylation mechanism and a novel branching route forming byproducts from the coupling of the dihydropyrone moiety of 1 with the acetaldehyde and kojic acid abundant in AO. The involvement of putative cyclase ACRTS3 in the chain release of linear polyketide was excluded by the coexpression analysis of ACRTS2 and ACRTS3. Taken together, we concluded that the production of 1 in AO is solely responsible for ACRTS2.
Assuntos
Aspergillus oryzaeRESUMO
D-allose is a rare sugar that has been reported to up-regulate thioredoxin-interacting protein (TXNIP) expression and affect the production of intracellular reactive oxygen species (ROS). However, the antitumor effect of D-allose is unknown. This study aimed to determine whether orally administered D-allose could be a candidate drug against bladder cancer (BC). To this end, BC cell lines were treated with varying concentrations of D-allose (10, 25, and 50 mM). Cell viability and intracellular ROS levels were assessed using cell viability assay and flow cytometry. TXNIP expression was evaluated using Western blotting. The antitumor effect of orally administered D-allose was assessed using a xenograft mouse model. D-allose reduced cell viability and induced intracellular ROS production in BC cells. Moreover, D-allose stimulated TXNIP expression in a dose-dependent manner. Co-treatment of D-allose and the antioxidant L-glutathione canceled the D-allose-induced reduction in cell viability and intracellular ROS elevation. Furthermore, oral administration of D-allose inhibited tumor growth without adverse effects (p < 0.05). Histopathological findings in tumor tissues showed that D-allose decreased the nuclear fission rate from 4.1 to 1.1% (p = 0.004). Oral administration of D-allose suppressed BC growth in a preclinical mouse model, possibly through up-regulation of TXNIP expression followed by an increase in intracellular ROS. Therefore, D-allose is a potential therapeutic compound for the treatment of BC.
Assuntos
Açúcares , Neoplasias da Bexiga Urinária , Animais , Linhagem Celular Tumoral , Glucose/metabolismo , Humanos , Camundongos , Espécies Reativas de Oxigênio , Neoplasias da Bexiga Urinária/tratamento farmacológicoRESUMO
Highly reducing polyketide synthases (HR-PKSs) produce structurally diverse polyketides (PKs). The PK diversity is constructed by a variety of factors, including the ß-keto processing, chain length, methylation pattern, and relative and absolute configurations of the substituents. We examined the stereochemical course of the PK processing for the synthesis of polyhydroxy PKs such as phialotides, phomenoic acid, and ACR-toxin. Heterologous expression of a HR-PKS gene, a trans-acting enoylreductase gene, and a truncated non-ribosomal peptide synthetase gene resulted in the formation of a linear PK with multiple stereogenic centers. The absolute configurations of the stereogenic centers were determined by chemical degradation followed by comparison of the degradation products with synthetic standards. A stereochemical rule was proposed to explain the absolute configurations of other reduced PKs and highlights an error in the absolute configurations of a reported structure. The present work demonstrates that focused functional analysis of functionally related HR-PKSs leads to a better understanding of the stereochemical course.
Assuntos
Proteínas Fúngicas/química , Policetídeo Sintases/química , Policetídeos/síntese química , Ascomicetos/enzimologia , Proteínas Fúngicas/genética , Mutação , Oxirredução , Policetídeo Sintases/genética , EstereoisomerismoRESUMO
In Arabidopsis thaliana, a mitogen-activated protein kinase pathway, MEKK1-MKK1/MKK2-MPK4, is important for basal resistance and disruption of this pathway results in dwarf, autoimmune phenotypes. To elucidate the complex mechanisms activated by the disruption of this pathway, we have previously developed a mutant screening system based on a dwarf autoimmune line that overexpressed the N-terminal regulatory domain of MEKK1. Here, we report that the second group of mutants, smn2, had defects in the SMN2 gene, encoding a DEAD-box RNA helicase. SMN2 is identical to HEN2, whose function is vital for the nuclear RNA exosome because it provides non-ribosomal RNA specificity for RNA turnover, RNA quality control and RNA processing. Aberrant SMN1/RPS6 transcripts were detected in smn2 and hen2 mutants. Disease resistance against Pseudomonas syringae pv. tomato DC3000 (hopA1), which is conferred by SMN1/RPS6, was decreased in smn2 mutants, suggesting a functional connection between SMN1/RPS6 and SMN2/HEN2. We produced double mutants mekk1smn2 and mpk4smn2 to determine whether the smn2 mutations suppress the dwarf, autoimmune phenotypes of the mekk1 and mpk4 mutants, as the smn1 mutations do. As expected, the mekk1 and mpk4 phenotypes were suppressed by the smn2 mutations. These results suggested that SMN2 is involved in the proper function of SMN1/RPS6. The Gene Ontology enrichment analysis using RNA-seq data showed that defense genes were downregulated in smn2, suggesting a positive contribution of SMN2 to the genome-wide expression of defense genes. In conclusion, this study provides novel insight into plant immunity via SMN2/HEN2, an essential component of the nuclear RNA exosome.
Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/genética , RNA Helicases DEAD-box/genética , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas/genética , Genes de Plantas/genética , Arabidopsis/imunologia , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/fisiologia , RNA Helicases DEAD-box/metabolismo , RNA Helicases DEAD-box/fisiologia , Estudo de Associação Genômica AmplaRESUMO
KEY MESSAGE: OsNINJA1-interacting protein, OsSRO1a, acts as a mediator that suppresses OsMYC2 activity in response to JA. Jasmonic acid (JA) is an important plant hormone for the stable growth and development of higher plants. The rice gene NOVEL INTERACTOR OF JAZ1 (OsNINJA1) interacts with Jasmonate ZIM-domain (JAZ) proteins and is a repressor of JA signaling. In this study, we identified several OsNINJA1-interacting proteins in rice from a yeast two-hybrid screen. Among the newly identified genes, we focused on SIMILAR TO RCD ONE1a (OsSRO1a) and investigated its role in JA signaling. Full-length OsSRO1a interacted with OsNINJA1 in plant cells but not in yeast cells. OsSRO1a also interacted with OsMYC2, a positive transcription factor in JA signaling, in both plant and yeast cells. The expression of OsSRO1a was upregulated at a late phase after JA treatment. Transgenic rice plants overexpressing OsSRO1a exhibited JA-insensitive phenotypes. In wild-type plants, JA induces resistance against rice bacterial blight, but this phenotype was suppressed in the OsSRO1a-overexpressing plants. Furthermore, the degradation of chlorophyll under dark-induced senescence conditions and the JA-induced upregulation of OsMYC2-responsive genes were suppressed in the OsSRO1a-overexpressing plants. These results suggest that OsSRO1a is a negative regulator of the OsMYC2-mediated JA signaling pathway in rice.
Assuntos
Ciclopentanos/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Oryza/metabolismo , Oxilipinas/metabolismo , Proteínas de Plantas/metabolismo , Proteínas Proto-Oncogênicas c-myc/metabolismo , Transdução de Sinais/genética , Senescência Celular/genética , Senescência Celular/efeitos da radiação , Regulação da Expressão Gênica de Plantas/efeitos da radiação , Oryza/genética , Fenótipo , Folhas de Planta/genética , Folhas de Planta/metabolismo , Folhas de Planta/efeitos da radiação , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Domínios Proteicos , Proteínas Proto-Oncogênicas c-myc/genética , Transdução de Sinais/efeitos da radiação , Técnicas do Sistema de Duplo-Híbrido , Regulação para CimaRESUMO
Mitogen-activated protein kinase (MAPK) pathways have a pivotal role in innate immunity signaling in plants. In Arabidopsis, the MAPK pathway that consists of MEKK1, MKK1/MKK2 and MPK4 is involved in pattern-triggered immunity signaling upstream of defense gene expression. This pathway is partly guarded by SUMM2, a nucleotide-binding domain leucine-rich repeat (NLR) protein, which is activated by disruption of the MAPK pathway. To identify other components required for the guard mechanism, here we developed a new mutant screening system utilizing a dwarf autoimmune line that overexpressed the N-terminal regulatory domain of MEKK1. Mutants with suppression of the dwarf, autoimmune phenotypes were identified, and one locus responsible for the phenotype was designated as suppressor of MEKK1N overexpression-induced dwarf 1 (SMN1). MutMap analysis revealed that SMN1 encodes the Toll/Interleukin-1 receptor (TIR)-class NLR protein RPS6, a previously identified resistant protein against bacterial pathogen Pseudomonas syringae pv. tomato expressing the HopA1 effector. Importantly, mutations in SMN1/RPS6 also partially suppressed the dwarf, autoimmune phenotypes of mekk1 and mpk4 plants. Our results suggest that the two structurally distinct NLR proteins, SMN1/RPS6 and SUMM2, monitor integrity of the MEKK1-MKK1/MKK2-MPK4 pathway.
Assuntos
Autoimunidade/fisiologia , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Proteínas Serina-Treonina Quinases/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Arabidopsis/microbiologia , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Autoimunidade/genética , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas NLR/genética , Proteínas NLR/metabolismo , Imunidade Vegetal/genética , Imunidade Vegetal/fisiologia , Proteínas Serina-Treonina Quinases/genética , Pseudomonas syringae/patogenicidade , Transdução de SinaisRESUMO
Jasmonic acid (JA) is a plant hormone that plays an important role in the defense response and stable growth of rice. In this study, we investigated the role of the JA-responsive valine-glutamine (VQ)-motif-containing protein OsVQ13 in JA signaling in rice. OsVQ13 was primarily located in the nucleus and cytoplasm. The transgenic rice plants overexpressing OsVQ13 exhibited a JA-hypersensitive phenotype and increased JA-induced resistance to Xanthomonas oryzae pv. oryzae (Xoo), which is the bacteria that causes rice bacterial blight, one of the most serious diseases in rice. Furthermore, we identified a mitogen-activated protein kinase, OsMPK6, as an OsVQ13-associating protein. The expression of genes regulated by OsWRKY45, an important WRKY-type transcription factor for Xoo resistance that is known to be regulated by OsMPK6, was upregulated in OsVQ13-overexpressing rice plants. The grain size of OsVQ13-overexpressing rice plants was also larger than that of the wild type. These results indicated that OsVQ13 positively regulated JA signaling by activating the OsMPK6-OsWRKY45 signaling pathway in rice.
Assuntos
Ciclopentanos/metabolismo , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Oryza/genética , Oxilipinas/metabolismo , Proteínas de Plantas/metabolismo , Transdução de Sinais , Fatores de Transcrição/metabolismo , Resistência à Doença/genética , Grão Comestível/genética , Grão Comestível/crescimento & desenvolvimento , Grão Comestível/metabolismo , Oryza/crescimento & desenvolvimento , Oryza/metabolismo , Oryza/microbiologia , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Xanthomonas/patogenicidadeRESUMO
MAIN CONCLUSION: The jasmonic acid (JA)-responsive transcription factor OsMYC2 acts as a positive regulator of leaf senescence by direct regulation of some senescence-associated genes in rice. OsMYC2, a transcription factor (TF), acts as a positive regulator of jasmonic acid (JA) signaling involved in development and defense in rice. Here, we report that OsMYC2 plays an important role in leaf senescence under dark-induced senescence (DIS) conditions. Overexpression of OsMYC2 significantly promoted leaf senescence, indicated by reduction of chlorophyll content under DIS conditions in rice. Leaf senescence under the DIS conditions was negatively regulated by OsJAZ8, a rice jasmonate ZIM-domain protein involved in the JA signaling pathway. OsMYC2 upregulated the expression of some senescence-associated genes (SAGs) and selectively bound to the G-box/G-box-like motifs in the promoters of some SAGs in vivo. These results suggest that OsMYC2 acts as a positive regulator of leaf senescence by direct- or indirect-regulation of SAGs in rice.
Assuntos
Oryza/metabolismo , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Ciclopentanos/metabolismo , Regulação da Expressão Gênica de Plantas , Oryza/genética , Oxilipinas/metabolismo , Folhas de Planta/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Fatores de Transcrição/genéticaRESUMO
JASMONATE ZIM-domain (JAZ) proteins act as transcriptional repressors of jasmonic acid (JA) responses and play a crucial role in the regulation of host immunity in plants. Here, we report that OsMYC2, a JAZ-interacting transcription factor in rice (Oryza sativa L.), plays an important role in the resistance response against rice bacterial blight, which is one of the most serious diseases in rice, caused by Xanthomonas oryzae pv. oryzae (Xoo). The results showed that OsMYC2 interacted with some OsJAZ proteins in a JAZ-interacting domain (JID)-dependent manner. The up-regulation of OsMYC2 in response to JA was regulated by OsJAZ8. Transgenic rice plants overexpressing OsMYC2 exhibited a JA-hypersensitive phenotype and were more resistant to Xoo. A large-scale microarray analysis revealed that OsMYC2 up-regulated OsJAZ10 as well as many other defense-related genes. OsMYC2 selectively bound to the G-box-like motif of the OsJAZ10 promoter in vivo and regulated the expression of early JA-responsive genes, but not of late JA-responsive genes. The nuclear localization of OsMYC2 depended on a nuclear localization signal within JID. Overall, we conclude that OsMYC2 acts as a positive regulator of early JA signals in the JA-induced resistance against Xoo in rice.
Assuntos
Oryza/genética , Oryza/microbiologia , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Xanthomonas/patogenicidade , Núcleo Celular/metabolismo , Ciclopentanos/metabolismo , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas , Oxilipinas/metabolismo , Doenças das Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Regulação para CimaRESUMO
The filamentous fungus Alternaria alternata includes seven pathogenic variants (pathotypes), which produce different host-selective toxins and cause disease on different plants. The Japanese pear, strawberry and tangerine pathotypes produce AK-toxin, AF-toxin and ACT-toxin, respectively, which have a common structural moiety, 9,10-epoxy-8-hydroxy-9-methyl-decatrienoic acid (EDA). Here, we identified a new gene, AKT7 (AK-toxin biosynthetic gene 7), from the Japanese pear pathotype, which encodes a cytochrome P450 monooxygenase and functions to limit AK-toxin production. AKT7 homologs were found in the strawberry pathotype, but not the tangerine pathotype. However, the strawberry pathotype homolog appeared to include a premature stop codon. Although the Japanese pear pathotype strain has multiple copies of AKT7, a single-copy disruption resulted in mutants with increased production of AK-toxin and EDA. AKT7 overexpression in the three pathotypes caused marked reductions of toxin and EDA production, suggesting that Akt7 catalyzes a side reaction of EDA or its precursor. AKT7 overexpression caused reduced virulence in these pathotypes. We also found that AKT7 transcripts predominantly include misspliced mRNAs, which have premature stop codons. Our observations suggest that the AK-toxin production required for full virulence is regulated in a complex way by the copy number and intron information content of AKT7.
Assuntos
Alternaria/metabolismo , Proteínas Fúngicas/genética , Micotoxinas/biossíntese , Doenças das Plantas/microbiologia , Alternaria/crescimento & desenvolvimento , Alternaria/patogenicidade , Sequência de Bases , Proteínas Fúngicas/metabolismo , Dosagem de Genes , Expressão Gênica , Íntrons/genética , Dados de Sequência Molecular , Micotoxinas/química , Micotoxinas/genética , Folhas de Planta/microbiologia , Pyrus/microbiologia , Splicing de RNA , Metabolismo Secundário , Análise de Sequência de DNA , VirulênciaRESUMO
Jasmonic acid (JA) is involved in the regulation of host immunity in plants. Recently, we demonstrated that JA signalling has an important role in resistance to rice bacterial blight caused by Xanthomonas oryzae pv. oryzae (Xoo) in rice. Here, we report that many volatile compounds accumulate in response to exogenous application of JA, including the monoterpene linalool. Expression of linalool synthase was up-regulated by JA. Vapour treatment with linalool induced resistance to Xoo, and transgenic rice plants overexpressing linalool synthase were more resistance to Xoo, presumably due to the up-regulation of defence-related genes in the absence of any treatment. JA-induced accumulation of linalool was regulated by OsJAZ8, a rice jasmonate ZIM-domain protein involving the JA signalling pathway at the transcriptional level, suggesting that linalool plays an important role in JA-induced resistance to Xoo in rice.
Assuntos
Ciclopentanos/farmacologia , Resistência à Doença , Monoterpenos/metabolismo , Oryza/metabolismo , Oxilipinas/farmacologia , Doenças das Plantas/microbiologia , Monoterpenos Acíclicos , Ciclopentanos/metabolismo , Redes e Vias Metabólicas , Dados de Sequência Molecular , Oryza/microbiologia , Oxilipinas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Transdução de Sinais , Transcriptoma , XanthomonasRESUMO
BACKGROUND: Peritoneal dialysis (PD) is a crucial dialysis method for treating end-stage kidney disease. However, its use is restricted due to high glucose-induced peritoneal injury and hyperglycaemia, particularly in patients with diabetes mellitus. In this study, we investigated whether partially replacing d-glucose with the rare sugar d-allose could ameliorate peritoneal injury and hyperglycaemia induced by peritoneal dialysis fluid (PDF). METHODS: Rat peritoneal mesothelial cells (RPMCs) were exposed to a medium containing d-glucose or d-glucose partially replaced with different concentrations of d-allose. Cell viability, oxidative stress and cytokine production were evaluated. Sprague-Dawley (SD) rats were administrated saline, a PDF containing 4% d-glucose (PDF-G4.0%) or a PDF containing 3.6% d-glucose and 0.4% d-allose (PDF-G3.6%/A0.4%) once a day for 4 weeks. Peritoneal injury and PD efficiency were assessed using immuno-histological staining and peritoneal equilibration test, respectively. Blood glucose levels were measured over 120 min following a single injection of saline or PDFs to 24-h fasted SD rats. RESULTS: In RPMCs, the partial replacement of d-glucose with d-allose increased cell viability and decreased oxidative stress and cytokine production compared to d-glucose alone. Despite the PDF-G3.6%/A0.4% having a lower d-glucose concentration compared to PDF-G4.0%, there were no significant changes in osmolality. When administered to SD rats, the PDF-G3.6%/A0.4% suppressed the elevation of peritoneal thickness and blood d-glucose levels induced by PDF-G4.0%, without impacting PD efficiency. CONCLUSIONS: Partial replacement of d-glucose with d-allose ameliorated peritoneal injury and hyperglycaemia induced by high concentration of d-glucose in PDF, indicating that d-allose could be a potential treatment option in PD.
Assuntos
Hiperglicemia , Diálise Peritoneal , Humanos , Ratos , Animais , Diálise Peritoneal/efeitos adversos , Diálise Peritoneal/métodos , Hiperglicemia/patologia , Ratos Sprague-Dawley , Soluções para Diálise/efeitos adversos , Peritônio/patologia , Glucose , CitocinasRESUMO
Metallothionein is a small cysteine-rich protein known to have a metal-binding function. We isolated three different lengths of rough lemon cDNAs encoding a metallothionein (RlemMT1, RlemMT2 and RlemMT3), and only RlemMT1-recombinant protein had zinc-binding activity. Appropriate concentration of zinc is an essential micronutrient for living organisms, while excess zinc is toxic. Zinc also stimulates the production of host-selective ACR-toxin for citrus leaf spot pathogen of Alternaria alternata rough lemon pathotype. Trapping of zinc by RlemMT1-recombinant protein or by a zinc-scavenging agent in the culture medium caused suppression of ACR-toxin production by the fungus. Since ACR-toxin is the disease determinant for A. alternata rough lemon pathotype, addition of RlemMT1 to the inoculum suspension led to a significant decrease in symptoms on rough lemon leaves as a result of reduced ACR-toxin production from the zinc trap around infection sites. RlemMT1-overexpression mutant of A. alternata rough lemon pathotype also produced less ACR-toxin and reduced virulence on rough lemon. This suppression was caused by an interruption of zinc absorption by cells from the trapping of the mineral by RlemMT1 and an excess supplement of ZnSO(4) restored toxin production and pathogenicity. Based on these results, we propose that zinc adsorbents including metallothionein likely can act as a plant defense factor by controlling toxin biosynthesis via inhibition of zinc absorption by the pathogen.
Assuntos
Alternaria/patogenicidade , Proteínas de Transporte/metabolismo , Citrus/metabolismo , Citrus/microbiologia , Metalotioneína/metabolismo , Micotoxinas/biossíntese , Proteínas de Plantas/metabolismo , Alternaria/genética , Alternaria/metabolismo , Sequência de Aminoácidos , Sequência de Bases , Proteínas de Transporte/genética , Citrus/genética , Clonagem Molecular , DNA de Plantas/genética , Genes Fúngicos , Genes de Plantas , Interações Hospedeiro-Patógeno/genética , Interações Hospedeiro-Patógeno/fisiologia , Metalotioneína/genética , Dados de Sequência Molecular , Mutação , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Virulência , Zinco/metabolismoRESUMO
We previously reported that a rare sugar D-allose, which is the D-glucose epimer at C3, inhibits the gibberellin-dependent responses such as elongation of the second leaf sheath and induction of α-amylase in embryo-less half seeds in rice (Fukumoto et al. 2011). D-Allose suppresses expressions of gibberellin-responsive genes downstream of SLR1 protein in the gibberellin-signaling through hexokinase (HXK)-dependent pathway. In this study, we discovered that D-allose induced expression of ABA-related genes including OsNCED1-3 and OsABA8ox1-3 in rice. Interestingly, D-allose also up-regulated expression of OsABF1, encoding a conserved bZIP transcription factor in ABA signaling, in rice. The D-allose-induced expression of OsABF1 was diminished by a hexokinase inhibitor, D-mannoheptulose (MNH). Consistently, D-allose also inhibited Arabidopsis growth, but failed to trigger growth retardation in the glucose-insensitive2 (gin2) mutant, which is a loss-of-function mutant of the glucose sensor AtHXK1. D-Allose activated AtABI5 expression in transgenic gin2 over-expressing wild-type AtHXK1 but not in gin2 over-expressing the catalytic mutant AtHXK1(S177A), indicating that the D-allose phosphorylation by HXK to D-allose 6-phosphate (A6P) is the first step for the up-regulation of AtABI5 gene expression as well as D-allose-induced growth inhibition. Moreover, overexpression of OsABF1 showed increased sensitivity to D-allose in rice. These findings indicated that the phosphorylation of D-allose at C6 by hexokinase is essential and OsABF1 is involved in the signal transduction for D-allose-induced growth inhibition.
Assuntos
Glucose/metabolismo , Glucose/farmacologia , Hexoquinase/metabolismo , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Hexoquinase/genética , Oryza/efeitos dos fármacos , Oryza/genética , Fosforilação , Proteínas de Plantas/genéticaRESUMO
Only D-allose, among various rare monosaccharides tested, induced resistance to Xanthomonas oryzae pv. oryzae in susceptible rice leaves with defence responses: reactive oxygen species, lesion mimic formation, and PR-protein gene expression. These responses were suppressed by ascorbic acid or diphenylene iodonium. Transgenic rice plants overexpressing OsrbohC, encoding NADPH oxidase, were enhanced in sensitivity to D-allose. D-Allose-mediated defence responses were suppressed by the presence of a hexokinase inhibitor. 6-Deoxy-D-allose, a structural derivative of D-allose unable to be phosphorylated, did not confer resistance. Transgenic rice plants expressing Escherichia coli AlsK encoding D-allose kinase to increase D-allose 6-phosphate synthesis were more sensitive to D-allose, but E. coli AlsI encoding D-allose 6-phosphate isomerase expression to decrease D-allose 6-phosphate reduced sensitivity. A D-glucose 6-phosphate dehydrogenase-defective mutant was also less sensitive, and OsG6PDH1 complementation restored full sensitivity. These results reveal that a monosaccharide, D-allose, induces rice resistance to X. oryzae pv. oryzae by activating NADPH oxidase through the activity of D-glucose 6-phosphate dehydrogenase, initiated by hexokinase-mediated conversion of D-allose to D-allose 6-phosphate, and treatment with D-allose might prove to be useful for reducing disease development in rice.
Assuntos
Glucose/imunologia , Oryza/genética , Espécies Reativas de Oxigênio/imunologia , Regulação da Expressão Gênica de Plantas , NADPH Oxidases/genética , NADPH Oxidases/imunologia , Oryza/metabolismo , Oryza/microbiologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Proteínas de Plantas/imunologia , Xanthomonas/fisiologiaRESUMO
Crystal structures were obtained for the two C2 epimeric azido-γ-lactones 2-azido-2-deoxy-3,5:6,7-di-O-isopropylidene-d-glycero-d-ido-heptono-1,4-lactone and 2-azido-2-deoxy-3,5:6,7-di-O-isopropylidene-d-glycero-d-gulo-heptono-1,4-lactone prepared from kinetic and thermodynamic azide displacements of a triflate derived from d-glucoheptonolactone. Azido-γ-lactones are very useful intermediates in the synthesis of iminosugars and polyhydroxylated amino acids. In this study two epimeric azido-heptitols allow biotechnological transformations via Izumoring techniques to 8 of the 16 possible homonojirimycin analogues, 5 of which were isolated pure because of the lack of stereoselectivity of the final reductive amination. A side-by-side glycosidase inhibition profile of 11 of the possible 16 HNJ stereoisomers derived from d-glucose and d-mannose is presented.
Assuntos
1-Desoxinojirimicina/análogos & derivados , Azidas/química , Glucose/química , Lactonas/química , Termodinâmica , 1-Desoxinojirimicina/química , Cinética , Modelos Moleculares , Conformação Molecular , EstereoisomerismoRESUMO
Recent studies have shown that D-allose, a rare sugar, elicits antitumor effects on different types of solid cancers, such as hepatocellular carcinoma, non-small-cell lung cancer, and squamous cell carcinoma of the head and neck. In this study, we examined the effects of D-allose on the proliferation of human glioblastoma (GBM) cell lines (i.e., U251MG and U87MG) in vitro and in vivo and the underlying mechanisms. D-allose treatment inhibited the proliferation of U251MG and U87MG cells in a dose-dependent manner (3-50 mM). However, D-allose treatment did not affect cell cycles or apoptosis in these cells but significantly decreased the cell division frequency in both GBM cell lines. In a subcutaneous U87MG cell xenograft model, intraperitoneal injection of D-allose (100 mg/kg/day) significantly reduced the tumor volume in 28 days. These data indicate that D-allose-induced reduction in cell proliferation is associated with a subsequent decrease in the number of cell divisions, independent of cell-cycle arrest and apoptosis. Thus, D-allose could be an attractive additive to therapeutic strategies for GBM.