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1.
Cell ; 186(15): 3196-3207.e17, 2023 07 20.
Artigo em Inglês | MEDLINE | ID: mdl-37369204

RESUMO

Pathogens produce diverse effector proteins to manipulate host cellular processes. However, how functional diversity is generated in an effector repertoire is poorly understood. Many effectors in the devastating plant pathogen Phytophthora contain tandem repeats of the "(L)WY" motif, which are structurally conserved but variable in sequences. Here, we discovered a functional module formed by a specific (L)WY-LWY combination in multiple Phytophthora effectors, which efficiently recruits the serine/threonine protein phosphatase 2A (PP2A) core enzyme in plant hosts. Crystal structure of an effector-PP2A complex shows that the (L)WY-LWY module enables hijacking of the host PP2A core enzyme to form functional holoenzymes. While sharing the PP2A-interacting module at the amino terminus, these effectors possess divergent C-terminal LWY units and regulate distinct sets of phosphoproteins in the host. Our results highlight the appropriation of an essential host phosphatase through molecular mimicry by pathogens and diversification promoted by protein modularity in an effector repertoire.


Assuntos
Monoéster Fosfórico Hidrolases , Phytophthora , Monoéster Fosfórico Hidrolases/metabolismo , Proteínas/metabolismo , Phytophthora/química , Phytophthora/metabolismo , Plantas/metabolismo , Processamento de Proteína Pós-Traducional , Proteína Fosfatase 2/metabolismo , Doenças das Plantas
2.
PLoS Pathog ; 20(4): e1012138, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38640110

RESUMO

Proper transcription orchestrated by RNA polymerase II (RNPII) is crucial for cellular development, which is rely on the phosphorylation state of RNPII's carboxyl-terminal domain (CTD). Sporangia, developed from mycelia, are essential for the destructive oomycetes Phytophthora, remarkable transcriptional changes are observed during the morphological transition. However, how these changes are rapidly triggered and their relationship with the versatile RNPII-CTD phosphorylation remain enigmatic. Herein, we found that Phytophthora capsici undergone an elevation of Ser5-phosphorylation in its uncanonical heptapeptide repeats of RNPII-CTD during sporangia development, which subsequently changed the chromosomal occupation of RNPII and primarily activated transcription of certain genes. A cyclin-dependent kinase, PcCDK7, was highly induced and phosphorylated RNPII-CTD during this morphological transition. Mechanistically, a novel DCL1-dependent microRNA, pcamiR1, was found to be a feedback modulator for the precise phosphorylation of RNPII-CTD by complexing with PcAGO1 and regulating the accumulation of PcCDK7. Moreover, this study revealed that the pcamiR1-CDK7-RNPII regulatory module is evolutionarily conserved and the impairment of the balance between pcamiR1 and PcCDK7 could efficiently reduce growth and virulence of P. capsici. Collectively, this study uncovers a novel and evolutionary conserved mechanism of transcription regulation which could facilitate correct development and identifies pcamiR1 as a promising target for disease control.


Assuntos
MicroRNAs , Phytophthora , RNA Polimerase II , Transcrição Gênica , RNA Polimerase II/metabolismo , RNA Polimerase II/genética , Fosforilação , MicroRNAs/metabolismo , MicroRNAs/genética , Phytophthora/patogenicidade , Phytophthora/genética , Phytophthora/metabolismo , Quinases Ciclina-Dependentes/metabolismo , Quinases Ciclina-Dependentes/genética
3.
Plant Cell ; 35(4): 1186-1201, 2023 03 29.
Artigo em Inglês | MEDLINE | ID: mdl-36625683

RESUMO

Elicitins are a large family of secreted proteins in Phytophthora. Clade 1 elicitins were identified decades ago as potent elicitors of immune responses in Nicotiana species, but the mechanisms underlying elicitin recognition are largely unknown. Here we identified an elicitin receptor in Nicotiana benthamiana that we named REL for Responsive to ELicitins. REL is a receptor-like protein (RLP) with an extracellular leucine-rich repeat (LRR) domain that mediates Phytophthora resistance by binding elicitins. Silencing or knocking out REL in N. benthamiana abolished elicitin-triggered cell death and immune responses. Domain deletion and site-directed mutagenesis revealed that the island domain (ID) located within the LRR domain of REL is crucial for elicitin recognition. In addition, sequence polymorphism in the ID underpins the genetic diversity of REL homologs in various Nicotiana species in elicitin recognition and binding. Remarkably, REL is phylogenetically distant from the elicitin response (ELR) protein, an LRR-RLP that was previously identified in the wild potato species Solanum microdontum and REL and ELR differ in the way they bind and recognize elicitins. Our findings provide insights into the molecular basis of plant innate immunity and highlight a convergent evolution of immune receptors towards perceiving the same elicitor.


Assuntos
Phytophthora , Solanum , Proteínas/metabolismo , Plantas/metabolismo , Phytophthora/genética , Phytophthora/metabolismo , Nicotiana/metabolismo , Solanum/metabolismo , Doenças das Plantas
4.
Proc Natl Acad Sci U S A ; 118(10)2021 03 09.
Artigo em Inglês | MEDLINE | ID: mdl-33658365

RESUMO

Oomycete pathogens such as Phytophthora secrete a repertoire of effectors into host cells to manipulate host immunity and benefit infection. In this study, we found that an RxLR effector, Avr1d, promoted Phytophthora sojae infection in soybean hairy roots. Using a yeast two-hybrid screen, we identified the soybean E3 ubiquitin ligase GmPUB13 as a host target for Avr1d. By coimmunoprecipitation (Co-IP), gel infiltration, and isothermal titration calorimetry (ITC) assays, we confirmed that Avr1d interacts with GmPUB13 both in vivo and in vitro. Furthermore, we found that Avr1d inhibits the E3 ligase activity of GmPUB13. The crystal structure Avr1d in complex with GmPUB13 was solved and revealed that Avr1d occupies the binding site for E2 ubiquitin conjugating enzyme on GmPUB13. In line with this, Avr1d competed with E2 ubiquitin conjugating enzymes for GmPUB13 binding in vitro, thereby decreasing the E3 ligase activity of GmPUB13. Meanwhile, we found that inactivation of the ubiquitin ligase activity of GmPUB13 stabilized GmPUB13 by blocking GmPUB13 degradation. Silencing of GmPUB13 in soybean hairy roots decreased P. sojae infection, suggesting that GmPUB13 acts as a susceptibility factor. Altogether, this study highlights a virulence mechanism of Phytophthora effectors, by which Avr1d competes with E2 for GmPUB13 binding to repress the GmPUB13 E3 ligase activity and thereby stabilizing the susceptibility factor GmPUB13 to facilitate Phytophthora infection. This study unravels the structural basis for modulation of host targets by Phytophthora effectors and will be instrumental for boosting plant resistance breeding.


Assuntos
Complexos Multiproteicos/química , Phytophthora/química , Ubiquitina-Proteína Ligases/química , Complexos Multiproteicos/metabolismo , Phytophthora/metabolismo , Doenças das Plantas/microbiologia , Ligação Proteica , Ubiquitina-Proteína Ligases/metabolismo
5.
Mol Plant Microbe Interact ; 36(5): 283-293, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-37022145

RESUMO

The oomycete Pythium myriotylum is a necrotrophic pathogen that infects many crop species worldwide, including ginger, soybean, tomato, and tobacco. Here, we identified a P. myriotylum small cysteine-rich protein, PmSCR1, that induces cell death in Nicotiana benthamiana by screening small, secreted proteins that were induced during infection of ginger and did not have a predicted function at the time of selection. Orthologs of PmSCR1 were found in other Pythium species, but these did not have cell death-inducing activity in N. benthamiana. PmSCR1 encodes a protein containing an auxiliary activity 17 family domain and triggers multiple immune responses in host plants. The elicitor function of PmSCR1 appears to be independent of enzymatic activity, because the heat inactivation of PmSCR1 protein did not affect PmSCR1-induced cell death or other defense responses. The elicitor function of PmSCR1 was also independent of BAK1 and SOBIR1. Furthermore, a small region of the protein, PmSCR186-211, is sufficient for inducing cell death. A pretreatment using the full-length PmSCR1 protein promoted the resistance of soybean and N. benthamiana to Phytophthora sojae and Phytophthora capsici infection, respectively. These results reveal that PmSCR1 is a novel elicitor from P. myriotylum, which exhibits plant immunity-inducing activity in multiple host plants. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.


Assuntos
Phytophthora , Pythium , Cisteína , Proteínas/metabolismo , Phytophthora/metabolismo , Imunidade Vegetal , Nicotiana , Doenças das Plantas
6.
Mol Biol Rep ; 50(8): 6493-6504, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37326749

RESUMO

BACKGROUND: Phytophthora cinnamomi is an Oomycetes associated with soil, this Oomycete is one of the most destructive species of Phytophthora, being responsible for the decline of more than 5000 ornamental, forest, or fruit plants. It can secrete a class of protein NPP1 (Phytophthora necrosis inducing protein 1), responsible for inducing necrosis in leaves and roots of plants, leading to their death. OBJECTIVE: This work will report the characterization of the Phytophthora cinnamomi NPP1 gene responsible for the infection of Castanea sativa roots and will characterize the mechanisms of interaction between Phytophthora cinnamomi and Castanea sativa, by gene silencing NPP1 from Phytophthora cinnamomi mediated by RNAi. METHODS AND RESULTS: For silencing a part of the coding region of the NPP1 gene, was placed in the sense and antisense directions between an intron and ligated to the integrative vector pTH210. Cassette integration was confirmed by PCR and sequencing on the hygromycin-resistant Phytophthora cinnamomi transformants. Transformants obtained with the silenced gene was used to infect Castanea sativa. CONCLUSIONS: Plants infected with these transformants showed a great reduction in disease symptoms, confirming iRNA as a potential alternative biological tool in the study of molecular factors, and in the control and management of Phytophthora cinnamomi.


Assuntos
Phytophthora , Phytophthora/genética , Phytophthora/metabolismo , Interferência de RNA , Técnicas de Amplificação de Ácido Nucleico , Plantas/genética , Necrose/genética , Doenças das Plantas/genética
7.
Biotechnol Lett ; 45(1): 115-124, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36450976

RESUMO

OBJECTIVE: To examine the influence of widely used protein affinity tags and the tobacco PR1a signal peptide (SP) on detection, purification and bioactivity analyses of the small oomycete apoplastic effector SCR96 in planta. RESULTS: Through agroinfiltration, the phytotoxic effector SCR96 of Phytophthora cactorum was expressed in Nicotiana benthamiana leaf apoplast as a fusion protein carrying single affinity tag (His, HA or FLAG) at either C- or N-terminus. Leaf necrosis caused by different affinity-tagged SCR96 varied among tags and replicates. All of tagged proteins can be detected by antibodies against SCR96. All of SCR96 fusions except N-terminally fused 6His-tagged protein were detected using tag antibodies, indicating that 6His tag may be degraded when fused at N-terminus. Interestingly, C-terminal His- and FLAG-tagged SCR96 maintained the biological activity after purification. In the substitution assay of SCR96 SP, we observed that PR1a SP can lead chimeric SCR96 expression in N. benthamiana, but the replacement totally disrupted its bioactivity. CONCLUSION: C-terminal His or FLAG tag, along with its original SP, is efficient enough to enable detection and purification of functional SCR96 from N. benthamiana leaf apoplast, which would facilitate plant-pathogen interaction studies.


Assuntos
Nicotiana , Phytophthora , Nicotiana/genética , Nicotiana/metabolismo , Sinais Direcionadores de Proteínas/genética , Proteínas/metabolismo , Phytophthora/genética , Phytophthora/metabolismo , Anticorpos/metabolismo , Cromatografia de Afinidade
8.
PLoS Genet ; 16(3): e1008646, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-32150559

RESUMO

Centromeres are chromosomal regions that serve as platforms for kinetochore assembly and spindle attachments, ensuring accurate chromosome segregation during cell division. Despite functional conservation, centromere DNA sequences are diverse and often repetitive, making them challenging to assemble and identify. Here, we describe centromeres in an oomycete Phytophthora sojae by combining long-read sequencing-based genome assembly and chromatin immunoprecipitation for the centromeric histone CENP-A followed by high-throughput sequencing (ChIP-seq). P. sojae centromeres cluster at a single focus at different life stages and during nuclear division. We report an improved genome assembly of the P. sojae reference strain, which enabled identification of 15 enriched CENP-A binding regions as putative centromeres. By focusing on a subset of these regions, we demonstrate that centromeres in P. sojae are regional, spanning 211 to 356 kb. Most of these regions are transposon-rich, poorly transcribed, and lack the histone modification H3K4me2 but are embedded within regions with the heterochromatin marks H3K9me3 and H3K27me3. Strikingly, we discovered a Copia-like transposon (CoLT) that is highly enriched in the CENP-A chromatin. Similar clustered elements are also found in oomycete relatives of P. sojae, and may be applied as a criterion for prediction of oomycete centromeres. This work reveals a divergence of centromere features in oomycetes as compared to other organisms in the Stramenopila-Alveolata-Rhizaria (SAR) supergroup including diatoms and Plasmodium falciparum that have relatively short and simple regional centromeres. Identification of P. sojae centromeres in turn also advances the genome assembly.


Assuntos
Centrômero/genética , Oomicetos/genética , Phytophthora/genética , Alveolados/genética , Centrômero/metabolismo , Proteína Centromérica A/genética , Cromatina/genética , Imunoprecipitação da Cromatina/métodos , Proteínas Cromossômicas não Histona/genética , Segregação de Cromossomos/genética , Heterocromatina/genética , Histonas/genética , Cinetocoros/metabolismo , Cinetocoros/fisiologia , Phytophthora/metabolismo , Rhizaria/genética , Estramenópilas/genética
9.
Proc Natl Acad Sci U S A ; 117(44): 27685-27693, 2020 11 03.
Artigo em Inglês | MEDLINE | ID: mdl-33082226

RESUMO

Hosts and pathogens are engaged in a continuous evolutionary struggle for physiological dominance. A major site of this struggle is the apoplast. In Phytophthora sojae-soybean interactions, PsXEG1, a pathogen-secreted apoplastic endoglucanase, is a key focal point of this struggle, and the subject of two layers of host defense and pathogen counterdefense. Here, we show that N-glycosylation of PsXEG1 represents an additional layer of this coevolutionary struggle, protecting PsXEG1 against a host apoplastic aspartic protease, GmAP5, that specifically targets PsXEG1. This posttranslational modification also attenuated binding by the previously described host inhibitor, GmGIP1. N-glycosylation of PsXEG1 at N174 and N190 inhibited binding and degradation by GmAP5 and was essential for PsXEG1's full virulence contribution, except in GmAP5-silenced soybeans. Silencing of GmAP5 reduced soybean resistance against WT P. sojae but not against PsXEG1 deletion strains of P. sojae. The crucial role of N-glycosylation within the three layers of defense and counterdefense centered on PsXEG1 highlight the critical importance of this conserved apoplastic effector and its posttranslational modification in Phytophthora-host coevolutionary conflict.


Assuntos
Ácido Aspártico Endopeptidases/metabolismo , Celulase/metabolismo , Glycine max/microbiologia , Phytophthora/patogenicidade , Doenças das Plantas/microbiologia , Proteínas de Plantas/metabolismo , Ácido Aspártico Endopeptidases/genética , Celulase/genética , Resistência à Doença/genética , Técnicas de Silenciamento de Genes , Glicosilação , Interações Hospedeiro-Patógeno/genética , Phytophthora/metabolismo , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Ligação Proteica , Processamento de Proteína Pós-Traducional , Proteólise , Glycine max/enzimologia , Glycine max/genética , Virulência
10.
Chem Biodivers ; 20(3): e202201103, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36683342

RESUMO

The plant diseases caused by a variety of pathogens such as viruses, bacteria and fungi pose a great threat to global food production and food safety. Therefore, the search for green, efficient and pollution-free pesticides has become an important task. In this article, 23 myricetin derivatives containing thiazolebisamides active groups have been designed and synthesized. Their activities were evaluated by performing in vitro antibacterial and in vivo antiviral assays, microscale thermophoresis (MST) and molecular docking assays. The results of in vivo antiviral assays showed that compounds A4 and A23 exhibited good antiviral activity with EC50 values of 79.0 and 54.1 µg/mL for therapeutic activity and 103.3 and 91.2 µg/mL for protective activity, respectively. The dissociation constants (Kd) values of compounds A4 and A23 against TMV-CP were 0.021 and 0.018 µM, respectively, determined by microscale thermophoresis (MST), which were much smaller than those of the commercial drug ningnanmycin (NNM), which were 2.84 µM. The interaction of compounds A4, A23 with TMV-CP was further verified at the molecular level. In addition, in vitro antifungal assays of this series of compounds showed that they exhibited some inhibitory activity against a variety of fungi, especially against the phytophthora capsici. Among them, A13 and A20 showed similar inhibitory activity to the control drug azoxystrobin at 100 µg/mL against the phytophthora capsici.


Assuntos
Antifúngicos , Antivirais , Flavonoides , Antifúngicos/química , Antifúngicos/farmacologia , Antivirais/química , Antivirais/farmacologia , Desenho de Fármacos , Flavonoides/química , Flavonoides/farmacologia , Fungos/efeitos dos fármacos , Fungos/metabolismo , Testes de Sensibilidade Microbiana , Simulação de Acoplamento Molecular , Phytophthora/efeitos dos fármacos , Phytophthora/metabolismo , Relação Estrutura-Atividade , Tiazóis/química , Tiazóis/farmacologia , Vírus do Mosaico do Tabaco/química , Vírus do Mosaico do Tabaco/metabolismo
11.
Int J Mol Sci ; 24(1)2023 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-36614226

RESUMO

Major latex-like proteins (MLPs) play crucial roles in abiotic and biotic stresses. However, little was known about this gene family in cucumbers. In this study, a total of 37 putative cucumber MLP genes were identified on a genome-wide level and classified into three groups by sequence homologous comparison with Arabidopsis thaliana. Chromosome mapping suggested that only tandem duplication occurred in evolution. The multiple regulatory cis-elements related to stress, hormone, light and growth response were found in the promoter region of these CsMLP genes, indicating that CsMLPs might be widely involved in the process of plant growth, development and various stress conditions. Transcriptome analysis indicated a strong reprogramming of MLPs expression in response to Phytophthora melonis infection in cucumber. Knockdown of CsMLP1 reduced the P. melonis tolerance, while transient overexpression of CsMLP1 improved disease tolerance in cucumber. Conversely, the silence of CsMLP5 decreased the lesion area caused by P. melonis in the cotyledons, and overexpression of CsMLP5 promoted lesion expansion. Taken together, our results provide a comprehensive basis for further mining the function of CsMLP members and will also be significant for elucidating the evolutionary relationship in cucumber.


Assuntos
Arabidopsis , Cucumis sativus , Phytophthora , Cucumis sativus/genética , Cucumis sativus/metabolismo , Látex/metabolismo , Genoma de Planta , Phytophthora/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Arabidopsis/genética , Filogenia , Regulação da Expressão Gênica de Plantas
12.
Int J Mol Sci ; 24(6)2023 Mar 07.
Artigo em Inglês | MEDLINE | ID: mdl-36982179

RESUMO

Downy mildew caused by oomycete pathogen Plasmopara viticola is a devastating disease of grapevine. P. viticola secretes an array of RXLR effectors to enhance virulence. One of these effectors, PvRXLR131, has been reported to interact with grape (Vitis vinifera) BRI1 kinase inhibitor (VvBKI1). BKI1 is conserved in Nicotiana benthamiana and Arabidopsis thaliana. However, the role of VvBKI1 in plant immunity is unknown. Here, we found transient expression of VvBKI1 in grapevine and N. benthamiana increased its resistance to P. viticola and Phytophthora capsici, respectively. Furthermore, ectopic expression of VvBKI1 in Arabidopsis can increase its resistance to downy mildew caused by Hyaloperonospora arabidopsidis. Further experiments revealed that VvBKI1 interacts with a cytoplasmic ascorbate peroxidase, VvAPX1, an ROS-scavenging protein. Transient expression of VvAPX1 in grape and N. benthamiana promoted its resistance against P. viticola, and P. capsici. Moreover, VvAPX1 transgenic Arabidopsis is more resistant to H. arabidopsidis. Furthermore, both VvBKI1 and VvAPX1 transgenic Arabidopsis showed an elevated ascorbate peroxidase activity and enhanced disease resistance. In summary, our findings suggest a positive correlation between APX activity and resistance to oomycetes and that this regulatory network is conserved in V. vinifera, N. benthamiana, and A. thaliana.


Assuntos
Arabidopsis , Oomicetos , Phytophthora , Vitis , Ascorbato Peroxidases/metabolismo , Arabidopsis/genética , Arabidopsis/metabolismo , Phytophthora/metabolismo , Proteínas/metabolismo , Resistência à Doença/genética , Vitis/genética , Vitis/metabolismo , Doenças das Plantas/genética , Regulação da Expressão Gênica de Plantas
13.
New Phytol ; 235(4): 1599-1614, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35491752

RESUMO

Pathogens and pests secrete proteins (effectors) to interfere with plant immunity through modification of host target functions and disruption of immune signalling networks. The extent of convergence between pathogen and herbivorous insect virulence strategies is largely unexplored. We found that effectors from the oomycete pathogen, Phytophthora capsici, and the major aphid pest, Myzus persicae target the host immune regulator SIZ1, an E3 SUMO ligase. We used transient expression assays in Nicotiana benthamiana as well as Arabidopsis mutants to further characterize biological role of effector-SIZ1 interactions in planta. We show that the oomycete and aphid effector, which both contribute to virulence, feature different activities towards SIZ1. While M. persicae effector Mp64 increases SIZ1 protein levels in transient assays, P. capsici effector CRN83_152 enhances SIZ1-E3 SUMO ligase activity in vivo. SIZ1 contributes to host susceptibility to aphids and an oomycete pathogen. Knockout of SIZ1 in Arabidopsis decreased susceptibility to aphids, independent of SNC1, PAD4 and EDS1. Similarly SIZ1 knockdown in N. benthamiana led to reduced P. capsici infection. Our results suggest convergence of distinct pathogen and pest virulence strategies on an E3 SUMO ligase to enhance host susceptibility.


Assuntos
Afídeos , Proteínas de Arabidopsis , Arabidopsis , Phytophthora , Animais , Afídeos/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Regulação da Expressão Gênica de Plantas , Herbivoria , Ligases/metabolismo , Phytophthora/metabolismo , Ubiquitina-Proteína Ligases/metabolismo , Virulência
14.
Mol Biol Rep ; 49(2): 1463-1473, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-34751913

RESUMO

Phytophthora cinnamomi is classified as one of the most devastating plant pathogens in the world. It has a destructive effect on more than 5000 horticultural and forestry species in the world, and especially on Castanea sativa. The genus Phytophthora belongs to the Class Oomycetes, a group of fungus like organisms which provoke plant diseases via motile zoospores. Control of this organism is considered very challenging because of the limited range of effective chemical inhibitors. The development of sustainable control measures for the future management of P. cinnamomi requires in-depth knowledge of the cellular and molecular bases of development and metabolism. The aim of this review was to identify molecular factors associated with the metabolism of P. cinnamomi by studying the genes implicated in fundamental metabolism using tools of bioinformatics. Also, some genes involved in pathogenicity will be cited and characterized, such as genes coding for transglycosylases. Genomic sequences of P. cinnamomi were analyzed using an open reading frame (ORF) finder. The identified ORFs products (proteins) were compared to sequences already described and with known functions present in databases such as NCBI and fungi database. In this way, homologous proteins were found, with the respective specific domains, to proteins involved in the metabolism and pathogenicity of Phytophthora ssp.


Assuntos
Phytophthora/genética , Phytophthora/metabolismo , Phytophthora/patogenicidade , Biologia Computacional/métodos , Simulação por Computador , Genômica/métodos , Doenças das Plantas/microbiologia , Raízes de Plantas/microbiologia , Virulência/genética
15.
Int J Mol Sci ; 23(14)2022 Jul 21.
Artigo em Inglês | MEDLINE | ID: mdl-35887401

RESUMO

Vitis vinifera plants are disease-susceptible while Vitis pseudoreticulata plants are disease-resistant; however, the molecular mechanism remains unclear. In this study, the single-stranded DNA- and RNA-binding protein gene Whirly (VvWhy1 and VpWhy1) were cloned from V. vinifera "Cabernet Sauvignon" and V. pseudoreticulata "HD1". VvWhy1 and VpWhy1 promoter sequences (pVv and pVp) were also isolated; however, the identity of the promoter sequences was far lower than that between the Why1 coding sequences (CDSs). Both Why1 gene sequences had seven exons and six introns, and they had a C-terminal Whirly conserved domain and N-terminal chloroplast transit peptide, which was then verified to be chloroplast localization. Transcriptional expression showed that VpWhy1 was strongly induced by Plasmopara viticola, while VvWhy1 showed a low expression level. Further, the GUS activity indicated pVp had high activity involved in response to Phytophthora capsici infection. In addition, Nicotiana benthamiana transiently expressing pVp::VvWhy1 and pVp::VpWhy1 enhanced the P. capsici resistance. Moreover, Why1, PR1 and PR10 were upregulated in pVp transgenic N. benthamiana leaves. This research presented a novel insight into disease resistance mechanism that pVp promoted the transcription of Why1, which subsequently regulated the expression of PR1 and PR10, further enhancing the resistance to P. capsici.


Assuntos
Phytophthora , Vitis , DNA de Cadeia Simples/metabolismo , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas , Phytophthora/metabolismo , Doenças das Plantas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Vitis/genética , Vitis/metabolismo
16.
Biosci Biotechnol Biochem ; 85(8): 1802-1808, 2021 Jul 23.
Artigo em Inglês | MEDLINE | ID: mdl-34057177

RESUMO

The heterothallic group of the plant pathogen Phytophthora can sexually reproduce between the cross-compatible mating types A1 and A2. The mating hormone α2, produced by A2 mating type and utilized to promote the sexual reproduction of the partner A1 type, is known to be biosynthesized from phytol. In this study, we identified 2 biosynthetic intermediates, 11- and 16-hydroxyphytols (1 and 2), for α2 by administering the synthetic intermediates to an A2-type strain to produce α2 and by administering phytol to A2 strains to detect the intermediates in the mycelia. The results suggest that α2 is biosynthesized by possibly 2 cytochrome P450 oxygenases via 2 hydroxyphytol intermediates (1 and 2) in A2 hyphae and secreted outside.


Assuntos
Fator de Acasalamento/biossíntese , Phytophthora/metabolismo , Fator de Acasalamento/química , Análise Espectral/métodos , Estereoisomerismo
17.
Pestic Biochem Physiol ; 172: 104757, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33518050

RESUMO

Although phosphite (Phi)-based fertilizers are used in large quantities in agriculture, the use of Phi-based fungicides against soybean root rot caused by Phytophthora sojae are limited. While, their low toxicity are of high ecological and economic focus. Limited attention has been paid to Phi translocation efficiency in soybeans and the efficacy of Phi as a fungicide against P. sojae. In this study, we evaluated the efficiency of Phi translocation in the Williams soybean cultivar by determining the Phi concentrations in roots, stems, and leaves using high-performance ion chromatography after the application of Phi to the roots. Phi was translocated from roots to leaves within 1 h and its concentration increased significantly in leaves within 36 h after Phi application. Results of an in vitro growth inhibition assay and an in vivo infection assay showed that Phi inhibited P. sojae. Additionally, we examined the activation of the salicylic acid (SA) and ethylene (ET) defense pathways by Phi. The expression of SA and ET pathway-related genes was upregulated in most soybean tissues after Phi application. Our results provide evidence that Phi translocation suppresses root rot caused by P. sojae in soybean.


Assuntos
Fosfitos , Phytophthora , Regulação da Expressão Gênica de Plantas , Fosfitos/farmacologia , Phytophthora/metabolismo , Doenças das Plantas , Proteínas de Plantas/genética , Glycine max/metabolismo
18.
Genomics ; 112(3): 2309-2317, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-31923618

RESUMO

Root and collar rot disease caused by Phytophthora capsici (Leonian) is one of the most serious diseases in pepper, Capsicum annuum L. Knowledge about resistant genes is limited in pepper accessions to P. capsici. In this study, a diverse collection of 37 commercial edible and ornamental genotypes, and implication of seven novel candidate DEGs genes (XLOC_ 021757, XLOC_021821, XLOC_012788, XLOC_011295, XLOC_021928, XLOC_015473 and XLOC_000341) were up-regulated on resistant and susceptible pepper cultivars, through real-time polymerase chain reaction (qPCR) at transplanting and maturing stages. All seven related defense-gene candidates were up-regulated in all inoculated accessions to P. capsici, but these genes were highly expressed in resistant ones, 19OrnP-PBI, 37ChillP-Paleo, and 23CherryP-Orsh. The transcriptional levels of the seven related candidate DEGs were 5.90, 5.64, 5.62, 5.18, 3.94, 3.69, 3.16 folds higher in the resistant pepper genotypes, than the control ones, non-inoculated genotypes respectively. The candidate genes expressed herein, will provide a basis for further gene cloning and functional verification studies, and also will aid in an understanding of the regulatory mechanism of pepper resistance to P. capsici.


Assuntos
Capsicum , Resistência à Doença/genética , Phytophthora/genética , Doenças das Plantas/genética , Capsicum/anatomia & histologia , Capsicum/genética , Capsicum/crescimento & desenvolvimento , Fenótipo , Filogenia , Phytophthora/classificação , Phytophthora/isolamento & purificação , Phytophthora/metabolismo , Doenças das Plantas/etiologia , Reação em Cadeia da Polimerase em Tempo Real
19.
Molecules ; 26(11)2021 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-34071493

RESUMO

In this study, the antifungal potential of chemical constituents from Piper pesaresanum and some synthesized derivatives was determined against three phytopathogenic fungi associated with the cocoa crop. The methodology included the phytochemical study on the aerial part of P. pesaresanum, the synthesis of some derivatives and the evaluation of the antifungal activity against the fungi Moniliophthora roreri, Fusarium solani and Phytophthora sp. The chemical study allowed the isolation of three benzoic acid derivatives (1-3), one dihydrochalcone (4) and a mixture of sterols (5-7). Seven derivatives (8-14) were synthesized from the main constituents, of which compounds 9, 10, 12 and 14 are reported for the first time. Benzoic acid derivatives showed strong antifungal activity against M. roreri, of which 11 (3.0 ± 0.8 µM) was the most active compound with an IC50 lower compared with positive control Mancozeb® (4.9 ± 0.4 µM). Dihydrochalcones and acid derivatives were active against F. solani and Phytophthora sp., of which 3 (32.5 ± 3.3 µM) and 4 (26.7 ± 5.3 µM) were the most active compounds, respectively. The preliminary structure-activity relationship allowed us to establish that prenylated chains and the carboxyl group are important in the antifungal activity of benzoic acid derivatives. Likewise, a positive influence of the carbonyl group on the antifungal activity for dihydrochalcones was deduced.


Assuntos
Antifúngicos/farmacologia , Cacau/metabolismo , Piper/metabolismo , Agaricales/metabolismo , Ácido Benzoico/química , Chalconas/química , Cromatografia , Fusarium/metabolismo , Concentração Inibidora 50 , Íons , Espectroscopia de Ressonância Magnética , Estrutura Molecular , Compostos Fitoquímicos , Phytophthora/metabolismo , Relação Estrutura-Atividade
20.
J Integr Plant Biol ; 63(7): 1382-1396, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-33586843

RESUMO

Plant pathogens rely on effector proteins to suppress host innate immune responses and facilitate colonization. Although the Phytophthora sojae RxLR effector Avh241 promotes Phytophthora infection, the molecular basis of Avh241 virulence remains poorly understood. Here we identified non-race specific disease resistance 1 (NDR1)-like proteins, the critical components in plant effector-triggered immunity (ETI) responses, as host targets of Avh241. Avh241 interacts with NDR1 in the plasma membrane and suppresses NDR1-participated ETI responses. Silencing of GmNDR1s increases the susceptibility of soybean to P. sojae infection, and overexpression of GmNDR1s reduces infection, which supports its positive role in plant immunity against P. sojae. Furthermore, we demonstrate that GmNDR1 interacts with itself, and Avh241 probably disrupts the self-association of GmNDR1. These data highlight an effective counter-defense mechanism by which a Phytophthora effector suppresses plant immune responses, likely by disturbing the function of NDR1 during infection.


Assuntos
Proteínas de Arabidopsis/metabolismo , Phytophthora/metabolismo , Fatores de Transcrição/metabolismo , Proteínas de Arabidopsis/genética , Membrana Celular/metabolismo , Doenças das Plantas/parasitologia , Imunidade Vegetal/genética , Imunidade Vegetal/fisiologia , Glycine max/parasitologia , Fatores de Transcrição/genética , Virulência/fisiologia
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