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1.
Front Plant Sci ; 14: 1096831, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37342142

RESUMO

Sclerotinia sclerotiorum, the causal agent of Sclerotinia stem rot (SSR) on more than 450 plant species, is a notorious fungal pathogen. Nitrate reductase (NR) is required for nitrate assimilation that mediates the reduction of nitrate to nitrite and is the major enzymatic source for NO production in fungi. To explore the possible effects of nitrate reductase SsNR on the development, stress response, and virulence of S. sclerotiorum, RNA interference (RNAi) of SsNR was performed. The results showed that SsNR-silenced mutants showed abnormity in mycelia growth, sclerotia formation, infection cushion formation, reduced virulence on rapeseed and soybean with decreased oxalic acid production. Furthermore SsNR-silenced mutants are more sensitive to abiotic stresses such as Congo Red, SDS, H2O2, and NaCl. Importantly, the expression levels of pathogenicity-related genes SsGgt1, SsSac1, and SsSmk3 are down-regulated in SsNR-silenced mutants, while SsCyp is up-regulated. In summary, phenotypic changes in the gene silenced mutants indicate that SsNR plays important roles in the mycelia growth, sclerotia development, stress response and fungal virulence of S. sclerotiorum.

2.
Front Microbiol ; 12: 632575, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34122358

RESUMO

The northern corn leaf blight (NCLB) pathogen Setosphaeria turcica (Luttrell) Leonard and Suggs is one of the main biotic constraints on sweet corn (Zea mays L.) yield and quality in Fujian Province, China. Currently, however, there is comparatively little information available regarding the distribution of mating types, population genetics, and reproductive strategies of this pathogen in Fujian. In this study, we investigated the distribution of mating types and population genetics of 117 isolates of S. turcica collected from seven of the main sweet corn-growing regions in Fujian Province, based on multiple polymerase chain reaction analyses using two mating type-specific primer pairs and 11 inter-simple sequence repeat markers. Furthermore, we examined the mode of reproduction of Fujian S. turcica populations. Both MAT1-1 and MAT1-2 mating types were detected throughout all seven sampling locations. The majority of MAT1-2 isolates were detected from Dongyou, Jian'ou, Pingnan, Songxi, and Longyan, whereas a large proportion of the detected MAT1-1 isolates were among those collected from Dongfeng and Nanjing. Furthermore, we detected five shared multi-locus haplotypes among S. turcica isolates from Dongyou, Jian'ou, Pingnan, Nanjing, and Songxi, whereas no shared haplotypes were observed between the Dongfeng (or Longyan) population and these five populations. Pairwise comparisons of the indices ΦPT and Nm, and population structure and principal coordinate analyses indicated genetic differentiation between both the regional and the mating type populations of S. turcica in Fujian. The skewed mating type ratio associated with low a haplotypic diversity and evident linkage disequilibrium reveals a mixed reproductive strategy for S. turcica populations in Fujian Province. The findings of this study advance our current understanding of the genetic diversity, population structure, and reproductive strategies of S. turcica populations infecting sweet corn in Fujian Province, and will potentially contribute to further resistance breeding efforts.

3.
Gene ; 764: 145081, 2021 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-32860897

RESUMO

Metalaxyl is one of the main fungicides used to control pepper blight caused by Phytophthora capsici. Metalaxyl resistance of P. capsici, caused by the long-term intense use of this fungicide, has become one of the most serious challenges facing pest management. In this study, a conserved domain RPOLA-N of the RPA190 gene of P. capsici (RPA190-pc) was identified from the P. capsici SD1-9 strain. The role of the RPA190-pc underlying the metalaxyl resistance of P. capsici was investigated. Three P. capsici mutants, two with downregulated RPA190-pc (SD1-9C-3 and C-4) expression and one showing upregulation (OESD1-9-1), were obtained by Polyethylene Glycol (PEG) mediated protoplast transformations of P. capsici SD1-9. Quantitative real-time reverse transcription PCR results showed that RPA190-pc was downregulated by more than 60% in SD1-9C-3/C-4 and upregulated 3-fold in OESD1-9-1 compared with that of the control strain SD1-9. Evaluation of the metalaxyl resistance of these three transformants showed that the EC50 values of metalaxyl against SD1-9C-3, SD1-9C-4, and OESD1-9-1 were 120.0 µg·mL-1, 24.4 µg·mL-1, and 15573.0 µg·mL-1, respectively, corresponding to 63.3% decrease, 92.5% decrease, and 47.7-fold increase relative to the EC50 value in SD1-9. Compared with SD1-9, the mycelia of transformants SD1-9C-3, SD1-9C-4, and OESD1-9-1 showed more branches and shorter branches; and the transformants had different pathogenicity to different hosts plants. The expression of the candidate gene RPA190-pc during 10 life-history stages was further studied, the results showed that expression level reached a maximum at the zoospores stage, and it gradually increased with the increase of SD1 and SD1-9 infection time of pepper leaves, indicated that RPA190-pc may be related to the growth and pathogenicity of P. capsici. These results indicate that the expression of RPA190-pc is involved in the regulation of P. capsici resistance to metalaxyl.


Assuntos
Farmacorresistência Fúngica/genética , Proteínas Fúngicas/genética , Fungicidas Industriais/farmacologia , Phytophthora/genética , RNA Polimerase I/genética , Alanina/análogos & derivados , Alanina/farmacologia , Capsicum/microbiologia , Regulação da Expressão Gênica no Desenvolvimento , Regulação Fúngica da Expressão Gênica , Genes Fúngicos , Estágios do Ciclo de Vida/genética , Mutação , Micélio/genética , Micélio/crescimento & desenvolvimento , Phytophthora/efeitos dos fármacos , Phytophthora/crescimento & desenvolvimento , Phytophthora/patogenicidade , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Folhas de Planta/microbiologia , Domínios Proteicos/genética , Esporângios/genética , Esporângios/crescimento & desenvolvimento , Virulência/genética
4.
Mol Plant Pathol ; 22(1): 92-107, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33191557

RESUMO

Chitin, a fungal microbial-associated molecular pattern, triggers various defence responses in several plant systems. Although it induces stomatal closure, the molecular mechanisms of its interactions with guard cell signalling pathways are unclear. Based on screening of public microarray data obtained from the ATH1 Affymetrix and Arabidopsis eFP browser, we isolated a cDNA encoding a Ras-related nuclear protein 1 AtRAN1. AtRAN1 expression was enriched in guard cells in a manner consistent with involvement in the control of the stomatal movement. AtRAN1 mutation impaired chitin-induced stomatal closure and accumulation of reactive oxygen species and nitric oxide in guard cells. In addition, Atran1 mutant plants exhibited compromised chitin-enhanced plant resistance to both bacterial and fungal pathogens due to changes in defence-related genes. Furthermore, Atran1 mutant plants were hypersensitive to drought stress compared to Col-0 plants, and had lower levels of stress-responsive genes. These data demonstrate a previously uncharacterized signalling role for AtRAN1, mediating chitin-induced signalling.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/genética , Quitina/metabolismo , Resistência à Doença/genética , Proteínas Monoméricas de Ligação ao GTP/metabolismo , Doenças das Plantas/imunologia , Proteínas de Ligação a RNA/metabolismo , Transdução de Sinais , Proteína ran de Ligação ao GTP/metabolismo , Arabidopsis/imunologia , Arabidopsis/microbiologia , Arabidopsis/fisiologia , Proteínas de Arabidopsis/genética , Secas , Proteínas Monoméricas de Ligação ao GTP/genética , Óxido Nítrico/metabolismo , Doenças das Plantas/microbiologia , Estômatos de Plantas/genética , Estômatos de Plantas/imunologia , Estômatos de Plantas/microbiologia , Estômatos de Plantas/fisiologia , Proteínas de Ligação a RNA/genética , Espécies Reativas de Oxigênio/metabolismo , Proteína ran de Ligação ao GTP/genética
5.
J Agric Food Chem ; 68(31): 8151-8162, 2020 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-32633954

RESUMO

Phytophthora sojae is a serious soil-borne pathogen, and the major control measures undertaken include the induction of soybean-resistance genes, fungicides, and scientific and reasonable planting management. Owing to the safety and resistance of fungicides, it is of great importance to screen new control alternatives. In a preliminary study, we observed that propyl gallate (PG) exerts a considerable inhibitory effect on P. sojae and can effectively prevent and cure soybean diseases, although the underlying mechanism remains unclear. To explore the inhibitory mechanism of PG on P. sojae, we analyzed the differences in the protein profile of P. sojae before and after treatment with PG using tandem mass tag (TMT) proteomics. Proteomic analysis revealed that the number of differentially expressed proteins (DEPs) was 285, of which 75 were upregulated and 210 were downregulated, and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways primarily comprised glycolysis, tricarboxylic acid cycle, fatty acid metabolism, secondary metabolite generation, and other pathways. Among the DEPs involved in PG inhibition of P. sojae are two closely related uncharacterized proteins encoded by PHYSODRAFT_522340 and PHYSODRAFT_344464, denoted PsFACL and PsCPT herein. The CRISPR/Cas9 knockout technique revealed that PsFACL and PsCPT were involved in the growth rate and pathogenicity. In addition, the results of gas chromatography-mass spectrometry (GC-MS) showed that there were differences in fatty acid levels between wild-type (WT) and CRISPR/Cas9 knockout transformants. Knocking out PsFACL and PsCPT resulted in the restriction of the synthesis and ß-oxidation of long-chain fatty acids, respectively. These suggest that PsFACL and PsCPT were also involved in the regulation of the fatty acid metabolism. Our results aid in understanding the mechanism underlying the inhibition of P. sojae growth by PG.


Assuntos
Fungicidas Industriais/farmacologia , Phytophthora/efeitos dos fármacos , Phytophthora/genética , Galato de Propila/farmacologia , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Phytophthora/química , Phytophthora/metabolismo , Doenças das Plantas/microbiologia , Proteômica , Glycine max/microbiologia
6.
Microorganisms ; 8(2)2020 Feb 18.
Artigo em Inglês | MEDLINE | ID: mdl-32085491

RESUMO

Metalaxyl is one of the main fungicides used to control pepper blight caused by Phytophthora capsici. Metalaxyl resistance of P. capsici, caused by the long-term intense use of this fungicide, has become one of the most serious challenges facing pest management. To reveal the potential resistance mechanism of P. capsici to fungicide metalaxyl, a metalaxyl-resistant mutant strain SD1-9 was obtained under laboratory conditions. The pathogenicity test showed that mutant strain SD1-9 had different pathogenicity to different host plants with or without the treatment of metalaxyl compared with that of the wild type SD1. Comparative transcriptome sequencing of mutant strain SD1-9 and wild type SD1 led to the identification of 3845 differentially expressed genes, among them, 517 genes were upregulated, while 3328 genes were down-regulated in SD1-9 compared to that in the SD1. The expression levels of 10 genes were further verified by real-time RT-PCR. KEGG analysis showed that the differentially expressed genes were enriched in the peroxisome, endocytosis, alanine and tyrosine metabolism. The expression of the candidate gene XLOC_020226 during 10 life history stages was further studied, the results showed that expression level reached a maximum at the zoospores stage and basically showed a gradually increasing trend with increasing infection time in pepper leaves in SD1-9 strain, while its expression gradually increased in the SD1 strain throughout the 10 stages, indicated that XLOC_020226 may be related to the growth and pathogenicity of P. capsici. In summary, transcriptome analysis of plant pathogen P. capsici strains with different metalaxyl resistance not only provided database of the genes involved in the metalaxyl resistance of P. capsici, but also allowed us to gain novel insights into the potential resistance mechanism of P. capsici to metalaxyl in peppers.

7.
Int J Mol Sci ; 20(12)2019 Jun 14.
Artigo em Inglês | MEDLINE | ID: mdl-31207889

RESUMO

With the improper application of fungicides, Phytophthora sojae begins to develop resistance to fungicides, and biological control is one of the potential ways to control it. We screened two strains of Bacillus; Bacillus amyloliquefaciens JDF3 and Bacillus subtilis RSS-1, which had an efficient inhibitory effect on P. sojae. They could inhibit mycelial growth, the germination of the cysts, and the swimming of the motile zoospores. To elucidate the response of P. sojae under the stress of B. amyloliquefaciens and B. subtilis, and the molecular mechanism of biological control, comparative transcriptome analysis was applied. Transcriptome analysis revealed that the expression gene of P. sojae showed significant changes, and a total of 1616 differentially expressed genes (DEGs) were detected. They participated in two major types of regulation, namely "specificity" regulation and "common" regulation. They might inhibit the growth of P. sojae mainly by inhibiting the activity of ribosome. A pot experiment indicated that B. amyloliquefaciens and B. subtilis enhanced the resistance of soybean to P. sojae, and their control effects of them were 70.7% and 65.5%, respectively. In addition, B. amyloliquefaciens fermentation broth could induce an active oxygen burst, NO production, callose deposition, and lignification. B. subtilis could also stimulate the systemic to develop the resistance of soybean by lignification, and phytoalexin.


Assuntos
Antibiose , Bacillus amyloliquefaciens/fisiologia , Bacillus subtilis/fisiologia , Phytophthora/fisiologia , Imunidade Vegetal , Transcriptoma , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Glucanos/metabolismo , Lignina/metabolismo , Óxido Nítrico/metabolismo , Phytophthora/metabolismo , Phytophthora/patogenicidade , Espécies Reativas de Oxigênio/metabolismo , Glycine max/imunologia , Glycine max/microbiologia
8.
Appl Microbiol Biotechnol ; 102(22): 9867-9869, 2018 11.
Artigo em Inglês | MEDLINE | ID: mdl-30328492

RESUMO

There is an error in the Original Publication. Two images were mistakenly edited in Fig.6 (panel (a)) and Fig.7 (panel (a). Please find below the corrected figures.

9.
Plant Sci ; 270: 37-46, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29576085

RESUMO

The filamentous fungus Sclerotinia sclerotiorum is an important plant pathogen with a worldwide distribution. It can infect a wide variety of plants, causing serious disease in many types of crops, such as rapeseed, sunflower and soybean. Sclerotinia stem rot caused by this fungus affects main crops and has led to great economic loss. Elicitors are a group of compounds that inspire the host plant to produce an immune response against invading pathogens. This study describes a protein that has high homology with the Trichoderma elicitor Sm1 and was found in the genome of S. sclerotiorum. We named this protein SsSm1. To determine whether this protein has an elicitor function like its homology protein, we constructed a heterologous expression vector for SsSm1 and expressed it in Escherichia coli. The protein of heterologous expression led to the formation of lesions in tobacco that closely resemble hypersensitive response lesions. Transient expression of the encoding gene of SsSm1 in tobacco leaves also caused hypersensitive response. Then, RNA silencing was used to identify the function of SsSm1. The hyphal growth and pathogenicity of silenced transformants were shown to be obviously lagging and branched abnormally. Transformants produced less infection cushions and deformed sclerotiorum. In addition, SsSm1 silencing caused weak tolerance to NaCl, sorbitol and SDS, and the sensitivity of mutants to carbendazim was also significantly decreased. Based on the above results, we speculate that this protein may be related to the development of hyphae, infection cushions and sclerotiorum, but the specific molecular mechanism needs to be studied further.


Assuntos
Ascomicetos/genética , Proteínas Fúngicas/metabolismo , Glycine max/microbiologia , Nicotiana/microbiologia , Doenças das Plantas/microbiologia , Sequência de Aminoácidos , Ascomicetos/crescimento & desenvolvimento , Ascomicetos/patogenicidade , Proteínas Fúngicas/genética , Hifas/genética , Hifas/crescimento & desenvolvimento , Hifas/patogenicidade , Filogenia , Folhas de Planta/genética , Folhas de Planta/crescimento & desenvolvimento , Folhas de Planta/metabolismo , Folhas de Planta/microbiologia , Interferência de RNA , Alinhamento de Sequência , Glycine max/genética , Glycine max/crescimento & desenvolvimento , Glycine max/metabolismo , Nicotiana/genética , Nicotiana/crescimento & desenvolvimento , Nicotiana/metabolismo
10.
Microb Pathog ; 108: 40-48, 2017 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-28438637

RESUMO

Succinate dehydrogenase (SDH) is one of the key enzymes of the tricarboxylic acid cycle (TCA cycle) and a proven target of fungicides for true fungi. To explore the roles of the SDHA gene in Phytophthora sojae, we first cloned PsSDHA to construct the PsSDHA silenced expression vector pHAM34-PsSDHA, and then utilized PEG to mediate the P. sojae protoplast transformation experiment. Through transformation screening, we obtained the silenced mutants A1 and A3, which have significant suppressive effect. Further study showed that the hyphae of the silenced mutant strains were shorter and more bifurcated; the growth of the silenced mutants was clearly inhibited in 10% V8 agar medium containing sodium chloride (NaCl), hydrogen peroxide (H2O2) or Congo Red, respectively. The pathogenicity of the silenced mutants was significantly reduced compared with the wild-type strain and the mock. The results could help us better to understand the position and function of SDH in P. sojae and provide a proven target of fungicides for the oomycete.


Assuntos
Phytophthora/enzimologia , Phytophthora/genética , Succinato Desidrogenase/genética , Succinato Desidrogenase/fisiologia , Sequência de Bases , Clonagem Molecular , Meios de Cultura/química , Regulação Fúngica da Expressão Gênica , Inativação Gênica/fisiologia , Genes Fúngicos , Peróxido de Hidrogênio/metabolismo , Hifas/citologia , Hifas/crescimento & desenvolvimento , Mutação , Fenótipo , Phytophthora/crescimento & desenvolvimento , Phytophthora/patogenicidade , Doenças das Plantas/microbiologia , Interferência de RNA , Análise de Sequência , Cloreto de Sódio/metabolismo , Glycine max/microbiologia , Estresse Fisiológico , Virulência
11.
Plant Cell Physiol ; 57(12): 2472-2484, 2016 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-27649734

RESUMO

SsCut, which functions as an elicitor, can induce plant immunity. In this study, we utilized Nicotiana benthamiana and virus-induced gene silencing to decrease the expression of > 2,500 genes individually. Using this forward genetics approach, several genes were identified that, when silenced, compromised SsCut-triggered cell death based on a cell death assay. A C2H2-type zinc finger gene was isolated from N. benthamiana Sequence analysis indicated that the gene encodes a 27 kDa protein with 253 amino acids containing two typical C2H2-type zinc finger domains; this gene was named NbCZF1 We found that SsCut-induced cell death could be inhibited by virus-induced gene silencing of NbCZF1 in N. benthamiana In addition, SsCut induces stomatal closure, accompanied by reactive oxygen species (ROS) production by NADPH oxidases and nitric oxide (NO) production. NbCZF1-silenced plants showed impaired SsCut-induced stomatal closure, decreased SsCut-induced production of ROS and NO in guard cells and reduced SsCut-induced resistance against Phytophthora nicotianae Taken together, these results demonstrate that the NbCZF1-ROS-NO pathway mediates multiple SsCut-triggered responses, including stomatal closure, hypersensitive responses and defense-related gene expression. This is the first report describing the function of a C2H2-type zinc finger protein in N. benthamiana.


Assuntos
Nicotiana/genética , Phytophthora/fisiologia , Doenças das Plantas/imunologia , Imunidade Vegetal , Proteínas de Plantas/metabolismo , Transdução de Sinais , Sequência de Aminoácidos , Dedos de Zinco CYS2-HIS2 , Morte Celular , Regulação da Expressão Gênica de Plantas , Inativação Gênica , Óxido Nítrico/metabolismo , Filogenia , Folhas de Planta/citologia , Folhas de Planta/genética , Folhas de Planta/imunologia , Folhas de Planta/fisiologia , Proteínas de Plantas/genética , Estômatos de Plantas/citologia , Estômatos de Plantas/genética , Estômatos de Plantas/imunologia , Estômatos de Plantas/fisiologia , Espécies Reativas de Oxigênio/metabolismo , Alinhamento de Sequência , Análise de Sequência de DNA , Nicotiana/citologia , Nicotiana/imunologia , Nicotiana/fisiologia
12.
Front Microbiol ; 7: 630, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27199956

RESUMO

Protein O-mannosylation is a type of O-glycosylation that is characterized by the addition of mannose residues to target proteins, and is initially catalyzed by evolutionarily conserved protein O-mannosyltransferases (PMTs). In this study, three members of PMT were identified in Magnaporthe oryzae, and the pathogenic roles of MoPmt2, a member of PMT2 subfamily, were analyzed. We found that MoPmt2 is a homolog of Saccharomyces cerevisiae Pmt2 and could complement yeast Pmt2 function in resistance to CFW. Quantitative RT-PCR revealed that MoPmt2 is highly expressed during conidiation, and targeted disruption of MoPmt2 resulted in defects in conidiation and conidia morphology. The MoPmt2 mutants also showed a distinct reduction in fungal growth, which was associated with severe alterations in hyphal polarity. In addition, we found that the MoPmt2 mutants severely reduced virulence on both rice plants and barley leaves. The subsequent examination revealed that the fungal adhesion, conidial germination, CWI and invasive hyphae growth in host cells are responsible for defects on appressorium mediated penetration, and thus attenuated the pathogenicity of MoPmt2 mutants. Taken together, our results suggest that protein O-mannosyltransferase MoPmt2 plays essential roles in fungal growth and development, and is required for the full pathogenicity of M. oryzae.

13.
Plant Dis ; 100(4): 824-828, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30688612

RESUMO

Infection by the clubroot pathogen Plasmodiophora brassicae on resistant and susceptible canola cultivars was investigated at various times following inoculation. Primary infection occurred on more than 90% of root hairs in both cultivars at 7 days after inoculation (dai), and thereafter declined to less than 20% at 14 to 35 dai. The amount of primary infection on the two cultivars was similar at each time point. Secondary infections were rare in both cultivars at 5 and 7 dai but became common after 14 dai. At 14 to 28 dai, the level of secondary infection was greater in the resistant cultivar than in the susceptible one. The in planta expression of 12 selected P. brassicae genes was investigated by reverse-transcription quantitative polymerase chain reaction. All genes were upregulated at 5 or 7 dai in the resistant cultivar. In the susceptible cultivar, the 12 genes could be classified into three groups according to their expression patterns: 2 genes showed an expression peak at 14 dai, 3 showed two expression peaks at 14 and 35 dai, and the others showed an expression peak at 35 dai. Results from this study will be useful in breeding for resistance and in selecting candidate pathogenicity genes for further studies.

14.
Appl Microbiol Biotechnol ; 99(19): 8075-88, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-26227409

RESUMO

The production of asexual spores plays a critical role in rice blast disease. However, the mechanisms of the genes involved in the conidiogenesis pathway are not well understood. F-box proteins are specific adaptors to E3 ubiquitin ligases that determine the fate of different substrates in ubiquitin-mediated protein degradation and play diverse roles in fungal growth regulation. Here, we identify a Saccharomyces cerevisiae Grr1 homolog, MoGrr1, in Magnaporthe oryzae. Targeted disruption of Mogrr1 resulted in defects in vegetative growth, melanin pigmentation, conidial production, and resistance to oxidative stress, and these mutants consequently exhibited attenuated virulence to host plants. Microscopy studies revealed that the inability to form conidiophores is responsible for the defect in conidiation. Although the Mogrr1 mutants could develop melanized appressoria from hyphal tips, the appressoria were unable to penetrate into plant tissues due to insufficient turgor pressure within the appressorium, thereby attenuating the virulence of the mutants. Quantitative RT-PCR results revealed significantly decreased expression of chitin synthase-encoding genes, which are involved in fungal cell wall integrity, in the Mogrr1 mutants. The Mogrr1 mutants also displayed reduced expression of central components of the MAP kinase and cAMP signaling pathways, which are required for appressorium differentiation. Furthermore, domain complementation analysis indicated that two putative protein-interacting domains in MoGrr1 play essential roles during fungal development and pathogenicity. Taken together, our results suggest that MoGrr1 plays essential roles in fungal development and is required for the full virulence of M. oryzae.


Assuntos
Parede Celular/metabolismo , Proteínas F-Box/metabolismo , Proteínas Fúngicas/metabolismo , Magnaporthe/metabolismo , Magnaporthe/patogenicidade , Oryza/microbiologia , Doenças das Plantas/microbiologia , Esporos Fúngicos/crescimento & desenvolvimento , Parede Celular/genética , Proteínas F-Box/genética , Proteínas Fúngicas/genética , Regulação Fúngica da Expressão Gênica , Hordeum/microbiologia , Magnaporthe/genética , Magnaporthe/crescimento & desenvolvimento , Melaninas/biossíntese , Estresse Oxidativo , Esporos Fúngicos/genética , Esporos Fúngicos/metabolismo , Esporos Fúngicos/patogenicidade , Virulência
15.
J Pharm Biomed Anal ; 115: 368-74, 2015 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-26279368

RESUMO

Berberrubine is an isoquinoline alkaloid isolated from Berberis vulgaris L, and it is readily derived from berberine. In this study, a sensitive and selective ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for the determination of berberrubine in rat plasma and mouse tissue has been developed. Magnoflorine was employed as an internal standard (IS), and liquid-liquid extraction by ethyl acetate was used for sample preparation. Chromatographic separation was achieved on a UPLC BEH C18 column (2.1mm×100mm, 1.7µm) with 0.1% formic acid and acetonitrile as the mobile phase with gradient elution. An electrospray ionization source was applied and operated in positive ion mode; multiple reactions monitoring (MRM) mode was used for quantification using target fragment ions m/z 322.0→307.0 for berberrubine and m/z 342.8→298.2 for IS. Calibration plots were linear in the range of 2-2000ng/mL for berberrubine in rat plasma and mouse tissue. Mean recoveries of berberrubine in rat plasma ranged from 79.6% to 84.8%. Intra-day and inter-day precision were less than 11%. The accuracy ranged from 93.6% to 106.8%. The method has also been successfully applied in pharmacokinetics and tissue distribution study of berberrubine. The absolute bioavailability of berberrubine was determined to be 31.6%. The results also show that berberrubine is rapidly absorbed and widely distributed in various tissues. The level of berberrubine in liver is highest, and followed by kidney, spleen and heart. Furthermore, the concentration of berberrubine in various tissues could also be predicted by a BP-ANN model.


Assuntos
Berberina/análogos & derivados , Cromatografia Líquida/métodos , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem , Administração Intravenosa , Administração Oral , Animais , Berberina/administração & dosagem , Berberina/sangue , Berberina/farmacocinética , Disponibilidade Biológica , Calibragem , Cromatografia Líquida/normas , Estabilidade de Medicamentos , Limite de Detecção , Modelos Lineares , Masculino , Camundongos , Modelos Biológicos , Ratos Sprague-Dawley , Padrões de Referência , Reprodutibilidade dos Testes , Espectrometria de Massas por Ionização por Electrospray/normas , Espectrometria de Massas em Tandem/normas , Distribuição Tecidual
16.
Mycologia ; 107(6): 1130-7, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26297780

RESUMO

Sclerotinia sclerotiorum is a necrotrophic plant-pathogenic fungus that infects more than 400 species of plants. In this study the nascent polypeptide-associated complex α subunit gene of S. sclerotiorum (SsNACα; accession No. XP_001593856.1) was cloned and characterized. The relative transcript expression of SsNACα at different morphological stages of asexual development of S. sclerotiorum were analyzed by quantitative real time PCR (qRT-PCR). RNAi-mediated gene silencing was successful for SsNACα, and the mutated strains exhibited less than 15% of the relative expression of SsNACα were obtained and used for studying the biological functions of the gene. A delay in sclerotial maturation for S. sclerotiorum was observed in the SsNACα mutants. The significant elevations for both the activities of pectin-degrading enzymes and the expression of polygalacturonase genes also were associated with the mutated strains, indicating that SsNACα could negatively influence polygalacturonases expression and modulate the pathogenicity of S. sclerotiorum.


Assuntos
Ascomicetos/enzimologia , Ascomicetos/patogenicidade , Proteínas Fúngicas/genética , Regulação Enzimológica da Expressão Gênica , Chaperonas Moleculares/metabolismo , Doenças das Plantas/microbiologia , Poligalacturonase/genética , Ascomicetos/genética , Ascomicetos/metabolismo , Brassica rapa/microbiologia , Proteínas Fúngicas/metabolismo , Regulação Fúngica da Expressão Gênica , Chaperonas Moleculares/genética , Poligalacturonase/metabolismo , Nicotiana/microbiologia , Virulência
17.
Bull Environ Contam Toxicol ; 95(1): 25-30, 2015 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-25792106

RESUMO

Graphene oxide (GO) nanosheets have been extensively investigated to fabricate the graphene in recent years. The migration of GO nanosheets into the environment could lead to the instability of biological system. In this study, the GO nanosheets were synthesized and were characterized by SEM, high resolution TEM, XRD, Raman, FTIR and XPS techniques. Toxicology testing of GO nanosheets against Paecilomyces catenlannulatus (P. catenlannulatus) was performed by measuring the efflux of cytoplasmic materials of P. catenlannulatus. Approximate 35 % of the bacteria could survive on the surface of GO nanosheets compared to the control sample (~92 %) within 3 h, indicating that GO nanosheets presented significantly antibacterial activities. It was observed that the concentration of RNA in the solution was obviously higher than that of control sample, which could be due to direct contact of the bacterial cell. The results showed that the damage of cell membrane of P. catenlannulatus was attributed to the direct contact of the P. catenlannulatus with the extremely sharp edges of GO nanosheets, which resulted in the P. catenlannulatus inactivation. The less resistant to the damage of cell membrane was observed with increasing of GO concentration and contact time.


Assuntos
Antibacterianos/farmacologia , Grafite/farmacologia , Nanoestruturas/química , Paecilomyces/efeitos dos fármacos , Antibacterianos/química , Relação Dose-Resposta a Droga , Grafite/química , Estresse Oxidativo/efeitos dos fármacos , Óxidos , Paecilomyces/metabolismo , Propriedades de Superfície , Fatores de Tempo
18.
Plant Mol Biol ; 86(4-5): 495-511, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-25149470

RESUMO

In this study, we report the cloning of the SsCut gene encoding cutinase from Sclerotinia sclerotiorum. We isolated a 609-bp cDNA encoding a polypeptide of 202 amino acids with a molecular weight of 20.4 kDa. Heterologous expression of SsCut in Escherichia coli (His-SsCut) caused the formation of lesions in tobacco that closely resembled hypersensitive response lesions. Mutational analysis identified the C-terminal-half peptide and the same amino acids indispensable for both enzyme and elicitor activity. His-SsCut was caused cell death in Arabidopsis, soybean (Glycine max), oilseed rape (Brassica napus), rice (Oryza sativa), maize (Zea mays), and wheat (Triticum aestivum), indicating that both dicot and monocot species are responsive to the elicitor. Furthermore, the elicitation of tobacco was effective in the induction of the activities of hydrogen peroxide, phenylalanine ammonia-lyase, peroxides, and polyphenol oxidase. His-SsCut-treated plants exhibited enhanced resistance as indicated by a significant reduction in the number and size of S. sclerotiorum, Phytophthora sojae, and P. nicotianae lesions on leaves relative to controls. Real-time PCR results indicated that the expression of defense-related genes and genes involved in signal transduction were induced by His-SsCut. Our results demonstrate that SsCut is an elicitor that triggers defense responses in plants and will help to clarify its relationship to downstream signaling pathways that induce defense responses.


Assuntos
Ascomicetos/genética , Hidrolases de Éster Carboxílico/genética , Proteínas Fúngicas/genética , Doenças das Plantas/genética , Plantas/genética , Sequência de Aminoácidos , Ascomicetos/metabolismo , Ascomicetos/fisiologia , Hidrolases de Éster Carboxílico/classificação , Hidrolases de Éster Carboxílico/metabolismo , Catecol Oxidase/metabolismo , Resistência à Doença/genética , Eletroforese em Gel de Poliacrilamida , Proteínas Fúngicas/metabolismo , Regulação da Expressão Gênica de Plantas , Interações Hospedeiro-Patógeno , Dados de Sequência Molecular , Fenilalanina Amônia-Liase/metabolismo , Filogenia , Doenças das Plantas/microbiologia , Folhas de Planta/genética , Folhas de Planta/metabolismo , Folhas de Planta/microbiologia , Plantas/microbiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Homologia de Sequência de Aminoácidos , Fatores de Tempo , Nicotiana/genética , Nicotiana/metabolismo , Nicotiana/microbiologia
19.
J Environ Radioact ; 137: 31-36, 2014 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24998746

RESUMO

The effect of Paecilomyces catenlannulatus (P. catenlannulatus) on removal of U(VI) onto illite as a function of contact time, pH, ionic strength, and solution concentration was conducted by batch techniques. The adsorption kinetics indicated that the removal of U(VI) on illite and illite coated P. catenlannulatus can be fitted by pseudo-second order kinetic model very well. The removal of U(VI) on illite and illite coated P. catenlannulatus increased with increasing pH from 1.0 to 7.0, whereas the decrease of U(VI) adsorption on illite and illite coated P. catenlannulatus was observed at pH > 7.5. The adsorption behavior of U(VI) on illite and illite coated P. catenlannulatus can be simulated by the double diffuse model under various pH conditions. The ionic strength-dependent experiments showed that the removal of U(VI) on illite was outer-sphere surface complexation, whereas the inner-sphere surface complexation predominated the U(VI) adsorption onto illite coated P. catenlannulatus at pH 5.0-7.0. The maximum adsorption capacity of U(VI) on illite and illite coated P. catenlannulatus calculated from Langmuir model at pH 5.0 and T = 298 K was 46.729 and 54.347 mg/g, respectively, revealing enhanced adsorption of U(VI) on illite coated P. catenlannulatus. This paper highlights the effect of microorganism on the removal of radionuclides from aqueous solutions in environmental pollution management.


Assuntos
Minerais/metabolismo , Paecilomyces/metabolismo , Poluentes Radioativos do Solo/metabolismo , Urânio/metabolismo , Modelos Biológicos , Monitoramento de Radiação , Fatores de Tempo
20.
Plant Signal Behav ; 7(10): 1284-8, 2012 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-22895102

RESUMO

Heterotrimeric G-proteins play an important regulatory role in multiple physiological processes, including the plant immune response, and substantial progress has been made in elucidating the G-protein-mediated defense-signaling network. This mini-review discusses the importance of G-proteins in plant immunity. We also provide an overview of how G-proteins affect plant cell death and stomatal movement. Our recent studies demonstrated that G-proteins are involved in signal transduction and induction of stomatal closure and defense responses. We also discuss future directions for G-protein signaling studies involving plant immunity.


Assuntos
Proteínas de Ligação ao GTP/metabolismo , Imunidade Vegetal/imunologia , Proteínas de Plantas/metabolismo , Morte Celular , Estômatos de Plantas/fisiologia , Transdução de Sinais/imunologia
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