RESUMO
Receptor-like cytoplasmic kinases (RLCKs) mediate the intracellular signaling downstream of pattern-recognition receptors (PRRs). Several RLCKs from subfamily VII of rice (Oryza sativa) have important roles in plant immunity, but the role of RLCK VII-4 in pattern-triggered immune (PTI) signaling and resistance to pathogens has not yet been investigated. Here, we generated by multiplex clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9-mediated genome editing rice sextuple mutant lines where the entire RLCK VII-4 subfamily is inactivated and then analyzed the resulting lines for their response to chitin and flg22 and for their immunity to Xanthomonas oryzae pv. oryzae (Xoo) and Magnaporthe oryzae. Analysis of the rlckvii-4 mutants revealed that they have an impaired reactive oxygen system burst and reduced defense gene expression in response to flg22 and chitin. This indicates that members of the rice RLCK VII-4 subfamily are required for immune signaling downstream of multiple PRRs. Furthermore, we found that the rice RLCK VII-4 subfamily is important for chitin-induced callose deposition and mitogen-activated protein kinase activation and that it is crucial for basal resistance against Xoo and M. oryzae pathogens. This establishes that the RLCK VII-4 subfamily has critical functions in the regulation of multiple PTI pathways in rice and opens the way for deciphering the precise role of its members in the control of rice PTI.
Assuntos
Oryza , Xanthomonas , Oryza/metabolismo , Reconhecimento da Imunidade Inata , Imunidade Vegetal/genética , Transdução de Sinais , Xanthomonas/metabolismo , Receptores de Reconhecimento de Padrão/genética , Receptores de Reconhecimento de Padrão/metabolismo , Quitina/metabolismo , Doenças das Plantas/microbiologia , Regulação da Expressão Gênica de PlantasRESUMO
KEY MESSAGE: OsICS1 but not OsICS1-L mediates the rice response to Xoo inoculation, with its overexpression increasing resistance against this pathogen. OsICS1 but not OsICS-L is directly upregulated by OsWRKY6. Rice (Oryza sativa) is a staple crop for about half of the global population and is particularly important in the diets of people living in Asia, Latin America, and Africa. This crop is continually threatened by bacterial leaf blight disease caused by Xanthomonas oryzae pv. oryzae (Xoo), which drastically reduces yields; therefore, it is needed to elucidate the plant's resistance mechanisms against Xoo. Isochorismate synthase (ICS1) generates salicylic acid (SA) and increases resistance against bacterial disease. The OsICS1 is differently annotated in rice genome databases and has not yet been functionally characterized in the context of Xoo infection. Here, we report that the expression of the OsICS1 is directly regulated by OsWRKY6 and increases plant resistance against Xoo. Inoculation with Xoo increased the expression of OsICS1 but not that of the long variant of OsICS1 (OsICS1-L). OsWRKY6 directly activated the OsICS1 promoter but not the OsICS1-L promoter. OsICS1 overexpression in rice increased resistance against Xoo through the induction of SA-dependent bacterial defense genes. These data show that OsICS1 promotes resistance against Xoo infection.
Assuntos
Oryza , Xanthomonas , Humanos , Ásia , Oryza/genética , Regiões Promotoras Genéticas/genética , Ácido SalicílicoRESUMO
Plants have intricate mechanisms that tailor their defence responses to pathogens. WRKY transcription factors play a pivotal role in plant immunity by regulating various defence signalling pathways. Many WRKY genes are transcriptionally activated upon pathogen attack, but how their functions are regulated after transcription remains elusive. Here, we show that OsWRKY7 functions as a crucial positive regulator of rice basal immunity against Xanthomonas oryzae pv. oryzae (Xoo). The activity of OsWRKY7 was regulated at both translational and post-translational levels. Two translational products of OsWRKY7 were generated by alternative initiation. The full-length OsWRKY7 protein is normally degraded by the ubiquitin-proteasome system but was accumulated following elicitor or pathogen treatment, whereas the alternate product initiated from the downstream in-frame start codon was stable. Both the full and alternate OsWRKY7 proteins have transcriptional activities in yeast and rice cells, and overexpression of each form enhanced resistance to Xoo infection. Furthermore, disruption of the main AUG in rice increased the endogenous translation of the alternate stabilized form of OsWRKY7 and enhanced bacterial blight resistance. This study provides insights into the coordination of alternative translation and protein stability in the regulation of plant growth and basal defence mediated by the OsWRKY7 transcription factor, and also suggests a promising strategy to breed disease-resistant rice by translation initiation control.
Assuntos
Oryza , Xanthomonas , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo , Oryza/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Melhoramento Vegetal , Resistência à Doença/genética , Imunidade Vegetal/genética , Doenças das Plantas/microbiologia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
Enhancing carbohydrate export from source to sink tissues is considered to be a realistic approach for improving photosynthetic efficiency and crop yield. The rice sucrose transporters OsSUT1, OsSWEET11a and OsSWEET14 contribute to sucrose phloem loading and seed filling. Crucially, Xanthomonas oryzae pv. oryzae (Xoo) infection in rice enhances the expression of OsSWEET11a and OsSWEET14 genes, and causes leaf blight. Here we show that co-overexpression of OsSUT1, OsSWEET11a and OsSWEET14 in rice reduced sucrose synthesis and transport leading to lower growth and yield but reduced susceptibility to Xoo relative to controls. The immunity-related hypersensitive response (HR) was enhanced in the transformed lines as indicated by the increased expression of defence genes, higher salicylic acid content and presence of HR lesions on the leaves. The results suggest that the increased expression of OsSWEET11a and OsSWEET14 in rice is perceived as a pathogen (Xoo) attack that triggers HR and results in constitutive activation of plant defences that are related to the signalling pathways of pathogen starvation. These findings provide a mechanistic basis for the trade-off between plant growth and immunity because decreased susceptibility against Xoo compromised plant growth and yield.
Assuntos
Regulação da Expressão Gênica de Plantas , Proteínas de Membrana Transportadoras , Oryza , Doenças das Plantas , Imunidade Vegetal , Proteínas de Plantas , Plantas Geneticamente Modificadas , Ácido Salicílico , Sacarose , Xanthomonas , Oryza/microbiologia , Oryza/genética , Oryza/imunologia , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Xanthomonas/fisiologia , Doenças das Plantas/microbiologia , Doenças das Plantas/imunologia , Sacarose/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Proteínas de Membrana Transportadoras/genética , Ácido Salicílico/metabolismo , Folhas de Planta/metabolismo , Folhas de Planta/imunologiaRESUMO
Many ubiquitin E3 ligases function in plant immunity. Here, we show that Oryza sativa (rice) DDB1 binding WD (OsDWD1) suppresses immune responses by targeting O. sativa non-expresser of pathogenesis-related gene 1 (OsNPR1) for degradation. Knock-down and overexpression experiments in rice plants showed that OsDWD1 is a negative regulator of the immune response and that OsNPR1 is a substrate of OsDWD1 and a substrate receptor of OsCRL4. After constructing the loss-of-function mutant OsDWD1R239A , we showed that the downregulation of OsNPR1 seen in rice lines overexpressing wild-type (WT) OsDWD1 (OsDWD1WT -ox) was compromised in OsDWD1R239A -ox lines, and that OsNPR1 upregulation enhanced resistance to pathogen infection, confirming that OsCRL4OsDWD1 regulates OsNPR1 protein levels. The enhanced disease resistance seen in OsDWD1 knock-down (OsDWD1-kd) lines contrasted with the reduced disease resistance in double knock-down (OsDWD1/OsNPR1-kd) lines, indicating that the enhanced disease resistance of OsDWD1-kd resulted from the accumulation of OsNPR1. Moreover, an in vivo heterologous protein degradation assay in Arabidopsis thaliana ddb1 mutants confirmed that the CUL4-based E3 ligase system can also influence OsNPR1 protein levels in Arabidopsis. Although OsNPR1 was degraded by the OsCRL4OsDWD1 -mediated ubiquitination system, the phosphodegron-motif-mutated NPR1 was partially degraded in the DWD1-ox protoplasts. This suggests that there might be another degradation process for OsNPR1. Taken together, these results indicate that OsDWD1 regulates OsNPR1 protein levels in rice to suppress the untimely activation of immune responses.
Assuntos
Arabidopsis , Oryza , Oryza/metabolismo , Ubiquitina-Proteína Ligases/genética , Ubiquitina-Proteína Ligases/metabolismo , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/metabolismo , Resistência à Doença , Arabidopsis/genéticaRESUMO
Bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae (Xoo) has a significant impact on rice yield and quality worldwide. Traditionally, bactericide application has been commonly used to control this devastating disease. However, the overuse of fungicides has led to a number of problems such as the development of resistance and environmental pollution. Therefore, the development of new methods and approaches for disease control are still urgent. In this paper, a series of cinnamic acid derivatives were designed and synthesized, and three novel T3SS inhibitors A10, A12 and A20 were discovered. Novel T3SS inhibitors A10, A12 and A20 significantly inhibited the hpa1 promoter activity without affecting Xoo growth. Further studies revealed that the title compounds A10, A12 and A20 significantly impaired hypersensitivity in non-host plant tobacco leaves, while applications on rice significantly reduced symptoms of bacterial leaf blight. RT-PCR showed that compound A20 inhibited the expression of T3SS-related genes. In summary, this work exemplifies the potential of the title compound as an inhibitor of T3SS and its efficacy in the control of bacterial leaf blight.
Assuntos
Oryza , Xanthomonas , Sistemas de Secreção Tipo III/genética , Sistemas de Secreção Tipo III/metabolismo , Cinamatos/farmacologia , Cinamatos/metabolismo , Xanthomonas/metabolismo , Oryza/metabolismoRESUMO
MAIN CONCLUSION: The rice protein OsWRKY6 directly activates OsWRKY45 and OsWRKY47 expression, and also activates OsPR1a and OsPR1b through the two OsWRKYs, and this transcriptional module participates in Xa1-mediated defense against the pathogen Xanthomonas oryzae pv. oryzae. Biotic stress, the pathogen Xanthomonas oryzae pv. oryzae (Xoo) in particular, negatively impacts worldwide productivity and yield in the staple crop rice (Oryza sativa). OsWRKY transcription factors are involved in various biotic stress responses in rice, and OsWRKY6 specifically acts as an important defense regulator against Xoo. However, the relationship between OsWRKY6 and other OsWRKYs, as well as its role in resistance (R) gene-mediated defense, have yet to be studied in depth. Here, we characterized a transcriptional cascade triggered by OsWRKY6 that regulated defense against Xoo infection mediated by the NBS-LRR protein Xa1. OsWRKY45 and OsWRKY47 were identified as direct transcriptional targets of OsWRKY6, and their two gene products reciprocally activated their two genes. Furthermore, OsWRKY6 activated OsPR1a and OsPR1b via the OsWRKY45 and OsWRKY47. Two OsWRKY6 RNAi knockdown lines showed significantly reduced defense even against an incompatible Xoo infection, and the expression of OsWRKY6 was not regulated by OsWRKY51 and OsWRKY88. This study reveals that a novel downstream transcriptional pathway activated by OsWRKY6 is involved in Xa1-mediated defense against Xoo.
Assuntos
Oryza , Xanthomonas , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas , Oryza/genética , Oryza/metabolismo , Doenças das Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Xanthomonas/metabolismoRESUMO
Many TGA transcription factors participate in immune responses in the SA-mediated signaling pathway in Arabidopsis. This study identified a transcription factor OsTGAL1, which is induced upon infection by Xoo. Overexpression of OsTGAL1 increased the susceptibility of rice to Xoo. Plants overexpressing OsTGAL1 could affect the expression of many SA signaling-related genes. OsTGAL1 was able to interact with the promoter of OsSGT1, which encodes a key enzyme for SA metabolism. The transcript of OsSGT1 was induced by Xoo and this responsive expression was further increased in plants overexpressing OsTGAL1. OsSGT1 knockout lines had enhanced resistance to Xoo, and knocking out OsSGT1 in plants overexpressing OsTGAL1 blocked the susceptibility caused by OsTGAL1. Altered expression levels of several OsPRs in all the transgenic plants demonstrated that SA-mediated signaling had been affected. Furthermore, we identified an oxidoreductase of CC-type glutaredoxin, OsGRX17, which interacted with OsTGAL1. OsGRX17 reduced the regulation of OsTGAL1 on OsSGT1, and this may be due to its redox modulation. Thus, our results demonstrate that OsTGAL1 negatively regulates resistance to Xoo by its effects on SA metabolism via the activation of OsSGT1, which provides valuable targets for plant breeders in developing new cultivars that are resistant to Xoo.
Assuntos
Arabidopsis , Oryza , Xanthomonas , Arabidopsis/genética , Arabidopsis/metabolismo , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas , Glucosiltransferases/metabolismo , Oryza/metabolismo , Doenças das Plantas/microbiologia , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Ácido Salicílico/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismoRESUMO
The emergence of phytopathogenic bacteria resistant to antibacterial agents has rendered previously manageable plant diseases intractable, highlighting the need for safe and environmentally responsible agrochemicals. Inhibition of bacterial cell division by targeting bacterial cell division protein FtsZ has been proposed as a promising strategy for developing novel antibacterial agents. We previously identified 4'-demethylepipodophyllotoxin (DMEP), a naturally occurring substance isolated from the barberry species Dysosma versipellis, as a novel chemical scaffold for the development of inhibitors of FtsZ from the rice blight pathogen Xanthomonas oryzae pv. oryzae (Xoo). Therefore, constructing structure-activity relationship (SAR) studies of DMEP is indispensable for new agrochemical discovery. In this study, we performed a structure-activity relationship (SAR) study of DMEP derivatives as potential XooFtsZ inhibitors through introducing the structure-based virtual screening (SBVS) approach and various biochemical methods. Notably, prepared compound B2, a 4'-acyloxy DMEP analog, had a 50% inhibitory concentration of 159.4 µM for inhibition of recombinant XooFtsZ GTPase, which was lower than that of the parent DMEP (278.0 µM). Compound B2 potently inhibited Xoo growth in vitro (minimum inhibitory concentration 153 mg L-1) and had 54.9% and 48.4% curative and protective control efficiencies against rice blight in vivo. Moreover, compound B2 also showed low toxicity for non-target organisms, including rice plant and mammalian cell. Given these interesting results, we provide a novel strategy to discover and optimize promising bactericidal compounds for the management of plant bacterial diseases.
Assuntos
Oryza , Xanthomonas , Antibacterianos/química , Proteínas de Bactérias/metabolismo , Divisão Celular , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Podofilotoxina/metabolismo , Podofilotoxina/farmacologia , Relação Estrutura-AtividadeRESUMO
Over half of the earth's population consumes rice as the primary food crop for dietary calories. However, severe loss of rice yield occurs due to bacterial blight caused by Xanthomonas oryzae pv. oryzae (Xoo) and bakanae disease caused by Fusarium fujikuroi (F. fujikuroi). Therefore, broad-spectrum resistance (BSR) to these pathogens is essential for rice cultivation. The Nonexpressor of Pathogenesis-Related Genes1 (NPR1), which is related to the signal molecule salicylic acid (SA) and the expression of pathogenesis-related (PR) genes, is a key regulator of systemic acquired resistance (SAR). Although five NPR1 homologs (NHs) have been identified in rice thus far, their cellular and biological functions remain largely unexplored. In this study, we identified a novel rice NH gene from Oryza sativa L. cv. Dongjin. The genetic variation of single nucleotide polymorphisms in OsNH5 caused a single amino acid substitution of asparagine for serine at residue 16. OsNH5N16 was mainly located in the nucleus, and its transcription was induced by Xoo. We generated transgenic rice lines constitutively expressing OsNH5N16 to investigate its function. Plants that overexpressed OsNH5N16 displayed enhanced BSR to Xoo and F. fujikuroi compared with wild varieties, and the transcription of PR genes such as OsPR1, GLUC, and CHIT2 was considerably upregulated. Moreover, we revealed that SA increases the transcription of OsNH5N16 and the promoter activity of OsPR1 regulated by OsNH5N16. These results showed that OsNH5N16 enhances BSR by regulating the expression of PR genes related to SAR and it is controlled by SA at the transcriptional and post-translational levels. This is the first report on the innate immune response conferring BSR associated with NH5.
Assuntos
Resistência à Doença , Genes de Plantas , Oryza/genética , Oryza/microbiologia , Doenças das Plantas/microbiologia , Proteínas de Plantas/metabolismo , Homologia de Sequência do Ácido Nucleico , Sequência de Aminoácidos , Fusarium/fisiologia , Regulação da Expressão Gênica de Plantas , Proteínas de Plantas/química , Proteínas de Plantas/genética , Regiões Promotoras Genéticas/genética , Ácido Salicílico/farmacologia , Transcrição Gênica , Regulação para Cima/genética , Xanthomonas/fisiologiaRESUMO
A few reports have indicated that a single gene confers resistance to bacterial blight, sheath blight and rice blast. In this study, we identified a novel disease resistance mutant gene, methyl esterase-like (osmesl) in rice. Mutant rice with T-DNA insertion displayed significant resistance to bacterial blight caused by Xanthomonas oryzae pv. oryzae (Xoo), sheath blight caused by Rhizoctonia solani and rice blast caused by Magnaporthe oryzae. Additionally, CRISPR-Cas9 knockout mutants and RNAi lines displayed resistance to these pathogens. Complementary T-DNA mutants demonstrated a phenotype similar to the wild type (WT), thereby indicating that osmesl confers resistance to pathogens. Protein interaction experiments revealed that OsMESL affects reactive oxygen species (ROS) accumulation by interacting with thioredoxin OsTrxm in rice. Moreover, qRT-PCR results showed significantly reduced mRNA levels of multiple ROS scavenging-related genes in osmesl mutants. Nitroblue tetrazolium staining showed that the pathogens cause ROS accumulation, and quantitative detection revealed significantly increased levels of H2 O2 in the leaves of osmesl mutants and RNAi lines after infection. The abundance of JA, a hormone associated with disease resistance, was significantly more in osmesl mutants than in WT plants. Overall, these results suggested that osmesl enhances disease resistance to Xoo, R. solani and M. oryzae by modulating the ROS balance.
Assuntos
Oryza , Xanthomonas , Ascomicetos , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas , Oryza/genética , Doenças das Plantas/genética , Espécies Reativas de Oxigênio , RhizoctoniaRESUMO
Nucleotide-binding site-leucine-rich repeat (NB-LRR) resistance proteins are critical for plant resistance to pathogens; however, their mechanism of activation and signal transduction is still not well understood. We identified a mutation in an as yet uncharacterized rice coiled-coil (CC)-NB-LRR, Oryza sativa RPM1-like resistance gene 1 (OsRLR1), which leads to hypersensitive response (HR)-like lesions on the leaf blade and broad-range resistance to the fungal pathogen Pyricularia oryzae (syn. Magnaporthe oryzae) and the bacterial pathogen Xanthomonas oryzae pv. oryzae, together with strong growth reduction. Consistently, OsRLR1-overexpression lines showed enhanced resistance to both pathogens. Moreover, we found that OsRLR1 mediates the defence response through direct interaction in the nucleus with the transcription factor OsWRKY19. Down-regulation of OsWRKY19 in the rlr1 mutant compromised the HR-like phenotype and resistance response, and largely restored plant growth. OsWRKY19 binds to the promoter of OsPR10 to activate the defence response. Taken together, our data highlight the role of a new residue involved in the NB-LRR activation mechanism, allowing identification of a new NB-LRR downstream signalling pathway.
Assuntos
Oryza , Xanthomonas , Ascomicetos , Sítios de Ligação , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas , Nucleotídeos , Oryza/genética , Oryza/metabolismo , Doenças das Plantas , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismoRESUMO
The bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae (Xoo) is one of the most serious rice diseases, causing huge yield losses worldwide. Several technologies and approaches have been opted to reduce the damage; however, these have had limited success. Recently, scientists have been focusing their efforts on developing efficient and environmentally friendly nanobactericides for controlling bacterial diseases in rice fields. In the present study, a scanning electron microscope (SEM), transmission electron microscope (TEM), and a confocal laser scanning microscope (CLSM) were utilized to investigate the mode of actions of ginger EOs on the cell structure of Xoo. The ginger EOs caused the cells to grow abnormally, resulting in an irregular form with hollow layers, whereas the dimethylsulfoxide (DMSO) treatment showed a typical rod shape for the Xoo cell. Ginger EOs restricted the growth and production of biofilms by reducing the number of biofilms generated as indicated by CLSM. Due to the instability, poor solubility, and durability of ginger EOs, a nanoemulsions approach was used, and a glasshouse trial was performed to assess their efficacy on BLB disease control. The in vitro antibacterial activity of the developed nanobactericides was promising at different concentration (50-125 µL/mL) tested. The efficacy was concentration-dependent. There was significant antibacterial activity recorded at higher concentrations. A glasshouse trial revealed that developed nanobactericides managed to suppress BLB disease severity effectively. Treatment at a concentration of 125 µL/mL was the best based on the suppression of disease severity index, AUDPC value, disease reduction (DR), and protection index (PI). Furthermore, findings on plant growth, physiological features, and yield parameters were significantly enhanced compared to the positive control treatment. In conclusion, the results indicated that ginger essential oils loaded-nanoemulsions are a promising alternative to synthetic antibiotics in suppressing Xoo growth, regulating the BLB disease, and enhancing rice yield under a glasshouse trial.
Assuntos
Óleos Voláteis , Oryza , Doenças das Plantas/microbiologia , Xanthomonas/crescimento & desenvolvimento , Zingiber officinale/química , Óleos Voláteis/química , Óleos Voláteis/farmacologia , Oryza/química , Oryza/crescimento & desenvolvimento , Oryza/microbiologia , Oryza/ultraestrutura , Xanthomonas/ultraestruturaRESUMO
Rice is a particularly widely consumed food crop globally, but its yield is seriously damaged by bacterial blight due to Xanthomonas oryzae pv. oryzae (Xoo) and bakanae disease due to Fusarium fujikuroi (F. fujikuroi). However, broad-spectrum resistance (BSR) to both Xoo and F. fujikuroi remains largely elusive. In this study, we showed that rice monothiol glutaredoxin GRXS15 localizes in mitochondria and the nucleus, and its transcription is induced by Xoo. Transgenic rice lines constitutively expressing OsGRXS15 showed enhanced disease resistance to Xoo and F. fujikuroi, while CRISPR/Cas9-based knockout mutants showed reduced resistance compared with the wild-type plants. The transcription of pathogenesis-related (PR) genes was significantly induced in OsGRXS15-expressing plants. The rice transcription factor OsWRKY65 was identified as a binding partner, and it directly interacted with OsGRXS15 in the nucleus. Moreover, we revealed that the interaction of OsGRXS15 and OsWRKY65 results in the upregulation of OsPR1. These results suggested that OsGRXS15 interacts with transcription factors, and it confers BSR through regulating the expression of genes related to pathogen response. This is the first report on the nuclear function associated with the monothiol glutaredoxin GRXS15.
Assuntos
Fusarium/patogenicidade , Glutarredoxinas/metabolismo , Oryza/genética , Oryza/microbiologia , Xanthomonas/patogenicidade , Núcleo Celular/metabolismo , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas , Glutarredoxinas/genética , Interações Hospedeiro-Patógeno/genética , Imunidade Inata , Mitocôndrias/metabolismo , Oryza/citologia , Oryza/imunologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Imunidade Vegetal/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente ModificadasRESUMO
Rice (Oryza sativa L.) is a global staple crop, but its yield is severely threatened by bacterial leaf blight (BLB) caused by Xanthomonas oryzae pv. oryzae (Xoo). The OsWRKY transcription factor family play a central role in innate plant immunity against Xoo, but the various biological functions of a large number of OsWRKYs remain to be understood. We characterized the role of OsWRKY114 against Xoo. OsWRKY114 has transcriptional activity in yeast and localizes in the nucleus. When OsWRKY114 is overexpressed in transgenic plants they show enhanced disease resistance against Xoo compared with wild types. By using genetic- and cell-based functional analyses, we showed OsWRKY114 directly associates with the promoters of OsPR1a and Chitinase and increases the promoter activities. These results suggest that OsWRKY114 enhances the innate immunity of Asian rice against Xoo through direct activation of defense genes that include OsPR1a and chtinase. This is the first report to functionally characterize OsWRKY114 in Xoo infection.
Assuntos
Regulação da Expressão Gênica de Plantas , Oryza/genética , Doenças das Plantas/microbiologia , Proteínas de Plantas/metabolismo , Transativadores/metabolismo , Xanthomonas , Quitinases/genética , Resistência à Doença/genética , Oryza/microbiologia , Proteínas de Plantas/química , Regiões Promotoras Genéticas , Alinhamento de Sequência , Análise de Sequência de Proteína , Transativadores/química , Ativação TranscricionalRESUMO
The Gram-negative bacterium Pseudomonas taiwanensis is a novel bacterium that uses shrimp shell waste as its sole sources of carbon and nitrogen. It is a versatile bacterium with potential for use in biological control, with activities including toxicity toward insects, fungi, and the rice pathogen Xanthomonas oryzae pv.oryzae (Xoo). In this study, the complete 5.08-Mb genome sequence of P. taiwanensis CMS was determined by a combination of NGS/Sanger sequencing and optical mapping. Comparison of optical maps of seven Pseudomonas species showed that P. taiwanensis is most closely related to P. putida KT 2400. We screened a total of 11,646 individual Tn5-transponson tagged strains to identify genes that are involved in the production and regulation of the iron-chelator pyoverdine in P. taiwanensis, which is a key anti-Xoo factor. Our results indicated that the two-component system (TCS) EnvZ/OmpR plays a positive regulatory role in the production of pyoverdine, whereas the sigma factor RpoS functions as a repressor. The knowledge of the molecular basis of the regulation of pyoverdine by P. taiwanensis provided herein will be useful for its development for use in biological control, including as an anti-Xoo agent.
Assuntos
Elementos de DNA Transponíveis , Mutagênese Insercional , Oryza/microbiologia , Doenças das Plantas/microbiologia , Pseudomonas , Controle Biológico de Vetores , Pseudomonas/genética , Pseudomonas/metabolismo , Sequenciamento Completo do Genoma , Xanthomonas/crescimento & desenvolvimentoRESUMO
BACKGROUND: Cell wall degrading enzymes (CWDEs) induce plant immune responses and E3 ubiquitin ligases are known to play important roles in regulating plant defenses. Expression of the rice E3 ubiquitin ligase, OsPUB41, is enhanced upon treatment of leaves with Xanthomonas oryzae pv. oryzae (Xoo) secreted CWDEs such as Cellulase and Lipase/Esterase. However, it is not reported to have a role in elicitation of immune responses. RESULTS: Expression of the rice E3 ubiquitin ligase, OsPUB41, is induced when rice leaves are treated with either CWDEs, pathogen associated molecular patterns (PAMPs), damage associated molecular patterns (DAMPs) or pathogens. Overexpression of OsPUB41 leads to induction of callose deposition, enhanced tolerance to Xoo and Rhizoctonia solani infection in rice and Arabidopsis respectively. In rice, transient overexpression of OsPUB41 leads to enhanced expression of PR genes and SA as well as JA biosynthetic and response genes. However, in Arabidopsis, ectopic expression of OsPUB41 results in upregulation of only JA biosynthetic and response genes. Transient overexpression of either of the two biochemically inactive mutants (OsPUB41C40A and OsPUB41V51R) of OsPUB41 in rice and stable transgenics in Arabidopsis ectopically expressing OsPUB41C40A failed to elicit immune responses. This indicates that the E3 ligase activity of OsPUB41 protein is essential for induction of plant defense responses. CONCLUSION: The results presented here suggest that OsPUB41 is possibly involved in elicitation of CWDE triggered immune responses in rice.
Assuntos
Arabidopsis/genética , Regulação da Expressão Gênica de Plantas/imunologia , Oryza/genética , Imunidade Vegetal/genética , Proteínas de Plantas/genética , Ubiquitina-Proteína Ligases/genética , Xanthomonas/fisiologia , Arabidopsis/imunologia , Parede Celular/imunologia , Oryza/imunologia , Folhas de Planta/enzimologia , Folhas de Planta/microbiologia , Proteínas de Plantas/imunologia , Ubiquitina-Proteína Ligases/imunologia , Xanthomonas/enzimologiaRESUMO
Targeting virulence factors of bacterial without affecting their growth and survival, has been an initiative strategy for the development of novel anti-microbial agents. The type III secretion system (T3SS), one of essential and highly conserved virulence factors in most Gram-negative pathogenic bacteria, has been regarded as an effective target that developed new anti-microbial drugs. Xanthomonas oryzae pv. oryzae (Xoo) is one of the most important bacterial pathogens on rice, which causes leaf blight disease. To discover potential anti-virulence agents against the pathogens, a new series of 1,3-thiazolidine-2-thione derivatives containing 5-phenyl-2-furan were designed and synthesized. Their structures were characterized by 1H NMR, 13C NMR, MS, and elemental analysis. All the title compounds inhibited the promoter activity of a harpin gene hpa1, significantly, that were further checked for the impact on bacterial growth. The results indicated that treatment of Xoo with the title compound III-7 did not affect bacterial growth or survival. Moreover, quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis showed that the expression of the Xoo T3SS was suppressed by treatment with the inhibitor. The mRNA levels of representative genes in the hrp (hypersensitive response and pathogenicity) cluster, as well as the regulatory genes hrpG and hrpX, were reduced. Finally, the in vivo test demonstrated that the compounds could reduce the disease symptoms of Xoo on the rice cultivar (Oryza sativa) IR24.
Assuntos
Antibacterianos/farmacologia , Tiazolidinas/farmacologia , Tionas/farmacologia , Sistemas de Secreção Tipo III/antagonistas & inibidores , Xanthomonas/efeitos dos fármacos , Antibacterianos/síntese química , Antibacterianos/química , Relação Dose-Resposta a Droga , Testes de Sensibilidade Microbiana , Estrutura Molecular , Relação Estrutura-Atividade , Tiazolidinas/síntese química , Tiazolidinas/química , Tionas/síntese química , Tionas/química , Sistemas de Secreção Tipo III/genética , Sistemas de Secreção Tipo III/metabolismo , Xanthomonas/crescimento & desenvolvimento , Xanthomonas/metabolismoRESUMO
Xanthomonas oryzae pv. oryzae (Xoo) infection directly leads to a severe disease known as leaf blight, which is a major cause of yield loss of rice. Use of traditional bactericides has resulted in severe resistance in pathogenic bacteria. A new approach screening compounds that target the virulence factors rather than killing bacterial pathogens is imperative. In gram-negative bacteria, the type III secretion system (T3SS) is a conserved and significant virulence factor considered as a target for drug development. Therefore, we designed and synthesized a new series of 5-phenyl-2-furan carboxylic acid derivatives stitched with 2-mercapto-1,3,4-thiadiazole. Bioassays revealed that the title candidates attenuated the hypersensitive response through suppressing the promoter activity of a harpin gene hpa1 without affecting bacterial growth. Quantitative real time polymerase chain reaction (qRT-PCR) analysis demonstrated reduced the expression of several genes associated with T3SS, when title compounds were applied. Additionally, hrp gene cluster members, including hrpG and hrpX, had reduced mRNA levels. In vivo greenhouse tests showed that candidate compounds could alleviate the effects of Xoo infection in rice (Oryza sativa) and possess better protective activity against rice bacterial leaf blight than bismerthiazol and thiodiazole copper. All tested compounds were safe to rice. This work suggests there are new safe options for Xoo control in rice from these 1,3,4-thiadiazole derivatives.
Assuntos
Tiadiazóis/síntese química , Tiadiazóis/farmacologia , Sistemas de Secreção Tipo III/efeitos dos fármacos , Xanthomonas/efeitos dos fármacos , Antibacterianos/farmacologia , Oryza/microbiologiaRESUMO
The initiative strategy for the development of novel anti-microbial agents usually uses the virulence factors of bacteria as a target, without affecting their growth and survival. The type III secretion system (T3SS), one of the essential virulence factors in most Gram-negative pathogenic bacteria because of its highly conserved construct, has been regarded as an effective target that developed new anti-microbial drugs. Xanthomonas oryzae pv. oryzae (Xoo) causes leaf blight diseases and is one of the most important pathogens on rice. To find potential anti-virulence agents against this pathogen, a number of natural compounds were screened for their effects on the T3SS of Xoo. Three of 34 compounds significantly inhibited the promoter activity of the harpin gene, hpa1, and were further checked for their impact on bacterial growth and on the hypersensitive response (HR) caused by Xoo on non-host tobacco plants. The results indicated that treatment of Xoo with CZ-1, CZ-4 and CZ-9 resulted in an obviously attenuated HR without affecting bacterial growth and survival. Moreover, quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis showed that the expression of the Xoo T3SS was suppressed by treatment with the three inhibitors. The mRNA levels of representative genes in the hypersensitive response and pathogenicity (hrp) cluster, as well as the regulatory genes hrpG and hrpX, were reduced. Finally, the in vivo test demonstrated that the compounds could reduce the disease symptoms of Xoo on the rice cultivar (Oryza sativa) IR24.