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1.
BMC Plant Biol ; 19(1): 371, 2019 Aug 22.
Artigo em Inglês | MEDLINE | ID: mdl-31438856

RESUMO

BACKGROUND: Propamocarb (PM) is one of the main pesticides used for controlling cucumber downy mildew. However, due to its volatility and internal absorption, PM can easily form pesticide residues on cucumber fruits that seriously endanger human health and pollute the environment. The breeding of new cucumber varieties with a low abundance of PM residues via genetic methods constitutes an effective strategy for reducing pesticide residues and improving cucumber safety and quality. To help elucidate the molecular mechanism resulting in a low PM residue abundance in cucumber, we used the cucumber cultivar 'D0351' (which has the lowest PM residue content) as the test material and identified genes related to low PM residue abundance through high-throughput tag-sequencing (Tag-Seq). RESULTS: CsMAPEG was constitutively expressed and showed both varietal and organizational differences. This gene was strongly expressed in 'D0351'. The expression levels of CsMAPEG in different cucumber tissues under PM stress were as follows: fruit>leaf>stem>root. CsMAPEG can respond to salicylic acid (SA), gibberellin (GA) and Corynespora cassiicola Wei (Cor) stress and thus plays an important regulatory role in plant responses to abiotic and biological stresses. The PM residue abundance in the fruits of CsMAPEG-overexpressing plants was lower than those found in antisense CsMAPEG plants and wild-type plants at all tested time points. The results revealed that CsMAPEG played a positive role in reducing the PM residue abundance. A CsMAPEG sense construct increased the contents of SOD, POD and GST in cucumber fruits, enhanced the degradation and metabolism of PM in cucumber, and thus effectively reduced the pesticide residue abundance in cucumber fruits. CONCLUSIONS: The expression patterns of CsMAPEG in cucumber cultivars with high and low pesticide residue abundances and a transgenic verification analysis showed that CsMAPEG can actively respond to PM stress and effectively reduce the PM residue abundance in cucumber fruits. The results of this study will help researchers further elucidate the mechanism responsible for a low PM residue abundance in cucumber and lay a foundation for the breeding of new agricultural cucumber varieties with low pesticide residue abundances.


Assuntos
Carbamatos/farmacologia , Cucumis sativus/genética , Fungicidas Industriais/farmacologia , Genes de Plantas , Resíduos de Praguicidas , Clonagem Molecular , Cucumis sativus/efeitos dos fármacos , Cucumis sativus/enzimologia , Cucumis sativus/fisiologia , Perfilação da Expressão Gênica , Vetores Genéticos , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Transformação Genética
2.
Nat Commun ; 10(1): 3611, 2019 Aug 09.
Artigo em Inglês | MEDLINE | ID: mdl-31399587

RESUMO

The formicamycin biosynthetic gene cluster encodes two groups of type 2 polyketide antibiotics: the formicamycins and their biosynthetic precursors the fasamycins, both of which have activity against methicillin-resistant Staphylococcus aureus. Here, we report the formicapyridines which are encoded by the same gene cluster and are structurally and biosynthetically related to the fasamycins and formicamycins but comprise a rare pyridine moiety. These compounds are trace-level metabolites formed by derailment of the major biosynthetic pathway. Inspired by evolutionary logic we show that rational mutation of a single gene in the biosynthetic gene cluster encoding an antibiotic biosynthesis monooxygenase (ABM) superfamily protein leads to a significant increase both in total formicapyridine production and their enrichment relative to the fasamycins/formicamycins. Our observations broaden the polyketide biosynthetic landscape and identify a non-catalytic role for ABM superfamily proteins in type II polyketide synthase assemblages for maintaining biosynthetic pathway fidelity.

3.
Cancer Med ; 8(12): 5651-5661, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31364281

RESUMO

Homeobox A10 (HOXA10) has been implicated critical for the promotion of carcinogenesis, but the underlying mechanism between HOXA10 and malignant gastric cancer (GC) phenotype remains elusive. In the present study, we analyzed and validated that HOXA10 and BCL2 expressions were elevated both at the mRNA and protein levels in GC tissues. Upregulated HOXA10 promoted GC cell proliferation with reduced apoptosis in vitro and accelerated GC tumor growth in vivo. Bioinformatics analysis and quantitative real-time polymerase chain reaction (qRT-PCR) experiment inferred that HOXA10 might upregulate the expression of BCL2. By performing western blot, chromatin immunoprecipitation and quantitative PCR (ChIP-qPCR), and rescue experiment, we found that HOXA10 might bind to BCL2 promoter region, induce its expression, and thus inhibit intrinsic apoptosis pathway. Moreover, higher expression of HOXA10 and BCL2 predicted poor overall survival (OS) in GC patients. In summary, our study indicated that HOXA10 was upregulated in GC, and that HOXA10 might promote cell proliferation by elevating BCL2 expression and inhibiting apoptosis.

4.
Sensors (Basel) ; 19(12)2019 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-31200536

RESUMO

Because there are different types of BeiDou constellations with participating geostationary orbit (GEO) and inclined geosynchronous orbit (IGSO) satellites, the maneuvering frequency of BeiDou satellites is higher than that of other navigation systems. The satellite orbital maneuvers lead to orbital parameter failure for several hours from broadcast ephemeris. Due to the missing initial orbit, the maneuvering thrust, and the period of orbital maneuvering, the orbit products of maneuvering satellites cannot be provided by the International Global Navigation Satellite System (GNSS) Service (IGS) and International GNSS Monitoring and Assessment System (iGMAS). In addition, the period of unhealthy status and the orbital parameters of maneuvering satellites in broadcast ephemeris are unreliable, making the detection of orbital maneuver periods more difficult. Here, we develop a method to detect orbital maneuver periods involving two key steps. The first step is orbit prediction of maneuvering satellites based on precise orbit products. The second step is time period detection of orbit maneuvering. The start time detection factor is calculated by backward prediction orbit and pseudo-range observations, and the end time detection factor is calculated by forward prediction orbit and pseudo-range observations. Data of stations from the Multi-GNSS Experiment (MGEX) and iGMAS were analyzed. The results show that the period of orbit maneuvering could be detected accurately for BeiDou GEO and IGSO satellites. In addition, the orbital maneuver period of other GNSS medium Earth orbit (MEO) satellites could also be determined by this method. The results of period detection for orbit maneuvering provide important reference information for precision orbit and clock offset determination during satellite maneuvers.

5.
Sensors (Basel) ; 18(3)2018 Feb 28.
Artigo em Inglês | MEDLINE | ID: mdl-29495638

RESUMO

The orbital maneuvers of Global Navigation Satellite System (GNSS) Constellations will decrease the performance and accuracy of positioning, navigation, and timing (PNT). Because satellites in the Chinese BeiDou Navigation Satellite System (BDS) are in Geostationary Orbit (GEO) and Inclined Geosynchronous Orbit (IGSO), maneuvers occur more frequently. Also, the precise start moment of the BDS satellites' orbit maneuvering cannot be obtained by common users. This paper presented an improved real-time detecting method for BDS satellites' orbit maneuvering and anomalies with higher timeliness and higher accuracy. The main contributions to this improvement are as follows: (1) instead of the previous two-steps method, a new one-step method with higher accuracy is proposed to determine the start moment and the pseudo random noise code (PRN) of the satellite orbit maneuvering in that time; (2) BDS Medium Earth Orbit (MEO) orbital maneuvers are firstly detected according to the proposed selection strategy for the stations; and (3) the classified non-maneuvering anomalies are detected by a new median robust method using the weak anomaly detection factor and the strong anomaly detection factor. The data from the Multi-GNSS Experiment (MGEX) in 2017 was used for experimental analysis. The experimental results and analysis showed that the start moment of orbital maneuvers and the period of non-maneuver anomalies can be determined more accurately in real-time. When orbital maneuvers and anomalies occur, the proposed method improved the data utilization for 91 and 95 min in 2017.

6.
BMC Plant Biol ; 18(1): 16, 2018 01 18.
Artigo em Inglês | MEDLINE | ID: mdl-29347906

RESUMO

BACKGROUND: Cucumber downy mildew is among the most important diseases that can disrupt cucumber production. Propamocarb, also known as propyl-[3-(dimethylamino)propyl]carbamate (PM), is a systemic carbamate fungicide pesticide that is widely applied in agricultural production because of its high efficiency of pathogens control, especially cucumber downy mildew. However, residual PM can remain in cucumbers after the disease has been controlled. To explore the molecular mechanisms of PM retention, cucumber cultivars 'D9320' (with the highest residual PM content) and 'D0351' (lowest residual PM content) were studied. High-throughput tag-sequencing (Tag-Seq) results showed that the CsDIR16 gene was related to PM residue, which was verified using transgenic technology. RESULTS: We investigated the activity of a dirigent cucumber protein encoded by the CsDIR16 in gene response to stress induced by PM treatment. Gene-expression levels of CsDIR16 were up-regulated in the fruits, leaves, and stems of 'D0351' plants in response to PM treatment. However, in cultivar 'D9320', CsDIR16 levels were down-regulated in the leaves and stems after PM treatment, with no statistically significant differences observed in the fruits. Induction by jasmonic acid, abscisic acid, polyethylene glycol 4000, NaCl, and Corynespora cassiicola Wei (Cor) resulted in CsDIR16 up-regulation in 'D0351' and 'D9320'. Expression after salicylic acid treatment was up-regulated in 'D0351', but was down-regulated in 'D9320'. CsDIR16 overexpression lowered PM residues, and these were more rapidly reduced in CsDIR16(+) transgenic 'D9320' plants than in wild-type 'D9320' and CsDIR16(-) transgenic plants. CONCLUSIONS: Analyses of the CsDIR16-expression patterns in the cucumber cultivars with the highest and lowest levels of PM residue, and transgenic validation indicated that CsDIR16 plays a positive role in reducing PM residues. The findings of this study help understand the regulatory mechanisms occurring in response to PM stress in cucumbers and in establishing the genetic basis for developing low-pesticide residue cucumber cultivars.


Assuntos
Carbamatos/metabolismo , Cucumis sativus/genética , Fungicidas Industriais/metabolismo , Oomicetos/fisiologia , Doenças das Plantas/genética , Proteínas de Plantas/genética , Sequência de Aminoácidos , Sequência de Bases , Cucumis sativus/metabolismo , Resistência à Doença/genética , Doenças das Plantas/microbiologia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo
7.
Cell Death Dis ; 9(2): 27, 2018 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-29348665

RESUMO

The results of a cDNA  array revealed that protocadherin gamma subfamily A, 9 (PCDHGA9) was significantly decreased in SGC-7901 gastric cancer (GC) cells compared with GES-1 normal gastric cells and was strongly associated with the Wnt/ß-catenin and transforming growth factor-ß (TGF-ß)/Smad2/3 signaling pathway. As a member of the cadherin family, PCDHGA9 functions in both cell-cell adhesion and nuclear signaling. However, its role in tumorigenicity or metastasis has not been reported. In the present study, we found that PCDHGA9 was decreased in GC tissues compared with corresponding normal mucosae and its expression was correlated with the GC TNM stage, the UICC stage, differentiation, relapse, and metastasis (p < 0.01). Multivariate Cox analysis revealed that PCDHGA9 was an independent prognostic indicator for overall survival (OS) and disease-free survival (DFS) (p < 0.01). The effects of PCDHGA9 on GC tumor growth and metastasis were examined both in vivo and in vitro. PCDHGA9 knockdown promoted GC cell proliferation, migration, and invasion, whereas PCDHGA9 overexpression inhibited GC tumor growth and metastasis but induced apoptosis, autophagy, and G1 cell cycle arrest. Furthermore, PCDHGA9 suppressed epithelial-mesenchymal transition (EMT) induced by TGF-ß, decreased the phosphorylation of Smad2/3, and inhibited the nuclear translocation of pSmad2/3. Our results suggest that PCDHGA9 might interact with ß-catenin to prevent ß-catenin from dissociating in the cytoplasm and translocating to the nucleus. Moreover, PCDHGA9 overexpression restrained cell proliferation and reduced the nuclear ß-catenin, an indicator of Wnt/ß-catenin pathway activation, suggesting that PCDHGA9 negatively regulates Wnt signaling. Together, these data indicate that PCDHGA9 acts as a tumor suppressor with anti-proliferative activity and anti-invasive ability, and the reduction of PCDHGA9 could serve as an independent prognostic biomarker in GC.

8.
J Biotechnol ; 265: 116-118, 2018 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-29191667

RESUMO

Here we report the complete genome of the new species Streptomyces formicae KY5 isolated from Tetraponera fungus growing ants. S. formicae was sequenced using the PacBio and 454 platforms to generate a single linear chromosome with terminal inverted repeats. Illumina MiSeq sequencing was used to correct base changes resulting from the high error rate associated with PacBio. The genome is 9.6 Mbps, has a GC content of 71.38% and contains 8162 protein coding sequences. Predictive analysis shows this strain encodes at least 45 gene clusters for the biosynthesis of secondary metabolites, including a type 2 polyketide synthase encoding cluster for the antibacterial formicamycins. Streptomyces formicae KY5 is a new, taxonomically distinct Streptomyces species and this complete genome sequence provides an important marker in the genus of Streptomyces.


Assuntos
Genoma Bacteriano , Streptomyces/genética , Antibacterianos/metabolismo , Proteínas de Bactérias/metabolismo , Policetídeo Sintases/metabolismo , Streptomyces/metabolismo
9.
Pak J Med Sci ; 33(5): 1166-1170, 2017 Sep-Oct.
Artigo em Inglês | MEDLINE | ID: mdl-29142558

RESUMO

Objective: To evaluate the clinical effect of transpedical interbody bone grafting in the treatment of senile osteoporotic vertebral fracture. Methods: Eighty-six elders with osteoporotic vertebral fracture were selected and divided into a control group and a test group using random double-blind method. Patients in the control group were treated by short-segment transpedicular screw system internal fixation, while patients in the test group were treated by short-segment transpedicular screw system internal fixation in combination with transpedical interbody bone grafting. Operation related indexes and fracture recovery condition were compared between the two groups. Results: The overall effective rate of the test group was 93.02%, much higher than the control group (76.74%) (P<0.05). The difference of operation duration, intraoperative bleeding volume, length of hospital stay, fracture healing time, preoperative vertebral height loss and preoperative Cobb's angle between the two groups had no statistical significance (P>0.05). The postoperative pain score of the test group was lower than that of the control group, and the difference was statistically significant (P<0.05). The vertebral height loss and Cobb's angle of the test group were superior to those of the control group at the last follow up, and the difference had statistical significance (P<0.05). The incidence of internal fixator loosening of the test group was much lower than that of the control group (P<0.05). Conclusion: Short-segment transpedicular screw system internal fixation in combination with transpedical interbody bone grafting shows favorable effects in the treatment senile osteoporotic vertebral fracture, resulting in mild pain and less loss of vertebral height and angle; hence it is worth promotion in clinic.

10.
Sensors (Basel) ; 17(12)2017 Nov 29.
Artigo em Inglês | MEDLINE | ID: mdl-29186058

RESUMO

The frequent maneuvering of BeiDou Geostationary Orbit (GEO) and Inclined Geosynchronous Orbit (IGSO) satellites affects the availability of real-time orbit, and decreases the accuracy and performance of positioning, navigation and time (PNT) services. BeiDou satellite maneuver information cannot be obtained by common users. BeiDou broadcast ephemeris is the only indicator of the health status of satellites, which are broadcast on an hourly basis, easily leading to ineffective observations. Sometimes, identification errors of satellite abnormity also appear in the broadcast ephemeris. This study presents a real-time robust detection method for a satellite orbital maneuver with high frequency and high reliability. By using the broadcast ephemeris and pseudo-range observations, the time discrimination factor and the satellite identification factor were defined and used for the real-time detection of start time and the pseudo-random noise code (PRN) of satellites was used for orbital maneuvers. Data from a Multi-GNSS Experiment (MGEX) was collected and analyzed. The results show that the start time and the PRN of the satellite orbital maneuver could be detected accurately in real time. In addition, abnormal start times and satellite abnormities caused by non-maneuver factors also could be detected using the proposed method. The new method not only improves the utilization of observations for users with the data effective for about 92 min, but also promotes the reliability of real-time PNT services.

11.
Plant Mol Biol ; 95(4-5): 519-531, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29052099

RESUMO

KEY MESSAGE: Overexpression of CsERF025 induces fruit bending by promoting the production of ethylene. Cucumber fruit bending critically affects cucumber quality, but the mechanism that causes fruit bending remains unclear. To better understand this mechanism, we performed transcriptome analyses on tissues from the convex (C1) and concave (C2) sides of bending and straight (S) fruit at 2 days post anthesis (DPA). We identified a total of 281 differentially expressed genes (DEGs) from both the convex and concave sides of bent fruit that showed significantly different expression profiles relative to straight fruits. Of these 281 DEGs, 196 were up-regulated (C1/S_C2/S) and 85 were down-regulated (C1/S_C2/S). Among the 196 up-regulated DEGs, the transcriptional levels of genes related to ethylene biosynthesis and signaling pathways were significantly higher in bending fruit compared with straight fruit. CsERF025 showed the largest difference in expression between bending and straight fruit. CsERF025 is an AP2/ERF gene encoding a protein that localizes to the nucleus. Overexpression of this gene increased the bending rate of cucumber fruits and increased the angle of bending. CsERF025 increased both the expression of ethylene biosynthesis-related genes and the production of ethylene. The application of exogenous 1-aminocyclopropane-l-carboxylic acid (ACC) to straight fruits from control plants promoted fruit bending. Thus, CsERF025 enhances the production of ethylene and thereby promotes fruit bending in cucumber.


Assuntos
Cucumis sativus/genética , Etilenos/metabolismo , Regulação da Expressão Gênica de Plantas , Reguladores de Crescimento de Planta/metabolismo , Proteínas de Plantas/metabolismo , Transcriptoma , Cucumis sativus/crescimento & desenvolvimento , Frutas/genética , Frutas/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Plantas/genética
12.
BMC Plant Biol ; 17(1): 96, 2017 06 05.
Artigo em Inglês | MEDLINE | ID: mdl-28583084

RESUMO

BACKGROUND: Cucumber downy mildew, caused by P. cubensis, is an important leaf disease that can severely affect cucumber production. In recent years, cucumber target spot, caused by C. cassiicola, has been reported in both Asia and Europe and is now considered as a major disease disrupting cucumber production. Single-disease-resistant cucumber varieties have been unable to satisfy production needs. To explore the molecular mechanisms of cucumber resistance to these two diseases, cucumber cultivars D9320 (resistant to downy mildew and target spot) and D0401 (susceptible to downy mildew and target spot) were used as experimental materials in this study. We used transcriptome sequencing technology to identify genes related to disease resistance and verified using transgenic technology. RESULTS: We screened out the cucumber resistance-related gene CsERF004 using transcriptome sequencing technology. Induction by pathogens, salicylic acid (SA), and ethylene (ET) resulted in the up-regulation of CsERF004. Three treatments, namely, inoculation with C. cassiicola alone, inoculation with P. cubensis alone, and simultaneous inoculation with both pathogens, all resulted in the significant and sustained up-regulation of CsERF004 in the resistant cultivar D9320, during the early stage of infection. In the susceptible cultivar D0401, CsERF004 expression was also significantly up-regulated at the later stage of infection but to a lesser extent and for a shorter duration than in the resistant cultivar D9320. The CsERF004 gene encodes a protein localizes to the nucleus. The over-expression of CsERF004 in the susceptible cultivar D0401 resulted in the significant up-regulation of the CsPR1 and CsPR4 genes and increased the levels of SA and ET, which enhanced the resistance of cucumber to downy mildew and target spot. CONCLUSIONS: Analyses of the CsERF004 expression pattern in disease-resistant and susceptible cucumber cultivars and transgenic validation indicate that CsERF004 confers resistance to P. cubensis and C. cassiicola. The findings of this study can help to better understanding of mechanisms of response to pathogens and in establishment the genetic basis for the development of cucumber broad-spectrum resistant cultivars.


Assuntos
Ascomicetos/fisiologia , Cucumis sativus/genética , Resistência à Doença/genética , Oomicetos/fisiologia , Fatores de Transcrição/genética , Sequência de Aminoácidos , Cucumis sativus/metabolismo , Cucumis sativus/microbiologia , Etilenos/metabolismo , Regulação da Expressão Gênica de Plantas , Interações Hospedeiro-Patógeno/genética , Proteínas de Plantas/genética , Ácido Salicílico/metabolismo
13.
Nat Commun ; 8: 15935, 2017 06 26.
Artigo em Inglês | MEDLINE | ID: mdl-28649989

RESUMO

ß-Lactone natural products occur infrequently in nature but possess a variety of potent and valuable biological activities. They are commonly derived from ß-hydroxy-α-amino acids, which are themselves valuable chiral building blocks for chemical synthesis and precursors to numerous important medicines. However, despite a number of excellent synthetic methods for their asymmetric synthesis, few effective enzymatic tools exist for their preparation. Here we report cloning of the biosynthetic gene cluster for the ß-lactone antibiotic obafluorin and delineate its biosynthetic pathway. We identify a nonribosomal peptide synthetase with an unusual domain architecture and an L-threonine:4-nitrophenylacetaldehyde transaldolase responsible for (2S,3R)-2-amino-3-hydroxy-4-(4-nitrophenyl)butanoate biosynthesis. Phylogenetic analysis sheds light on the evolutionary origin of this rare enzyme family and identifies further gene clusters encoding L-threonine transaldolases. We also present preliminary data suggesting that L-threonine transaldolases might be useful for the preparation of L-threo-ß-hydroxy-α-amino acids.

14.
Chem Sci ; 8(4): 3218-3227, 2017 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-28507698

RESUMO

We report a new Streptomyces species named S. formicae that was isolated from the African fungus-growing plant-ant Tetraponera penzigi and show that it produces novel pentacyclic polyketides that are active against MRSA and VRE. The chemical scaffold of these compounds, which we have called the formicamycins, is similar to the fasamycins identified from the heterologous expression of clones isolated from environmental DNA, but has significant differences that allow the scaffold to be decorated with up to four halogen atoms. We report the structures and bioactivities of 16 new molecules and show, using CRISPR/Cas9 genome editing, that biosynthesis of these compounds is encoded by a single type 2 polyketide synthase biosynthetic gene cluster in the S. formicae genome. Our work has identified the first antibiotic from the Tetraponera system and highlights the benefits of exploring unusual ecological niches for new actinomycete strains and novel natural products.

15.
Front Plant Sci ; 7: 573, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27200046

RESUMO

Salinity severely threatens land use capability and crop yields worldwide. Understanding the mechanisms that protect soybeans from salt stress will help in the development of salt-stress tolerant leguminous plants. Here we initially analyzed the changes in malondialdehyde levels, the activities of superoxide dismutase and peroxidases, chlorophyll content, and Na(+)/K(+) ratios in leaves and roots from soybean seedlings treated with 200 mM NaCl at different time points. We found that the 200 mM NaCl treated for 12 h was optimal for undertaking a proteomic analysis on soybean seedlings. An iTRAQ-based proteomic approach was used to investigate the proteomes of soybean leaves and roots under salt treatment. These data are available via ProteomeXchange with the identifier PXD002851. In total, 278 and 440 proteins with significantly altered abundances were identified in leaves and roots of soybean, respectively. From these data, a total of 50 proteins were identified in the both tissues. These differentially expressed proteins (DEPs) were from 13 biological processes. Moreover, protein-protein interaction analysis revealed that proteins involved in metabolism, carbohydrate and energy metabolism, protein synthesis and redox homeostasis could be assigned to four high salt stress response networks. Furthermore, semi-quantitative RT-PCR analysis revealed that some of the proteins, such as a 14-3-3, MMK2, PP1, TRX-h, were also regulated by salt stress at the level of transcription. These results indicated that effective regulatory protein expression related to signaling, membrane and transport, stress defense and metabolism all played important roles in the short-term salt response of soybean seedlings.

16.
Plant J ; 86(6): 514-29, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-27121031

RESUMO

Although research has extensively illustrated the molecular basis of plant responses to salt and high-pH stresses, knowledge on carbonate alkaline stress is poor and the specific responsive mechanism remains elusive. We have previously characterized a Glycine soja Ca(2+) /CAM-dependent kinase GsCBRLK that could increase salt tolerance. Here, we characterize a methionine sulfoxide reductase (MSR) B protein GsMSRB5a as a GsCBRLK interactor by using Y2H and BiFc assays. Further analyses showed that the N-terminal variable domain of GsCBRLK contributed to the GsMSRB5a interaction. Y2H assays also revealed the interaction specificity of GsCBRLK with the wild soybean MSRB subfamily proteins, and determined that the BoxI/BoxII-containing regions within GsMSRBs were responsible for their interaction. Furthermore, we also illustrated that the N-terminal basic regions in GsMSRBs functioned as transit peptides, which targeted themselves into chloroplasts and thereby prevented their interaction with GsCBRLK. Nevertheless, deletion of these regions allowed them to localize on the plasma membrane (PM) and interact with GsCBRLK. In addition, we also showed that GsMSRB5a and GsCBRLK displayed overlapping tissue expression specificity and coincident expression patterns under carbonate alkaline stress. Phenotypic experiments demonstrated that GsMSRB5a and GsCBRLK overexpression in Arabidopsis enhanced carbonate alkaline stress tolerance. Further investigations elucidated that GsMSRB5a and GsCBRLK inhibited reactive oxygen species (ROS) accumulation by modifying the expression of ROS signaling, biosynthesis and scavenging genes. Summarily, our results demonstrated that GsCBRLK and GsMSRB5a interacted with each other, and activated ROS signaling under carbonate alkaline stress.


Assuntos
Fabaceae/enzimologia , Fabaceae/metabolismo , Metionina Sulfóxido Redutases/metabolismo , Proteínas de Plantas/metabolismo , Arabidopsis/metabolismo , Proteínas Quinases Dependentes de Cálcio-Calmodulina/genética , Proteínas Quinases Dependentes de Cálcio-Calmodulina/metabolismo , Calmodulina/genética , Calmodulina/metabolismo , Proteínas de Ligação a Calmodulina/genética , Proteínas de Ligação a Calmodulina/metabolismo , Fabaceae/genética , Regulação da Expressão Gênica de Plantas , Metionina Sulfóxido Redutases/genética , Proteínas de Plantas/genética , Ligação Proteica , Espécies Reativas de Oxigênio/metabolismo
17.
PLoS One ; 10(12): e0146163, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26717241

RESUMO

It is widely accepted that the 14-3-3 family proteins are key regulators of multiple stress signal transduction cascades. By conducting genome-wide analysis, researchers have identified the soybean 14-3-3 family proteins; however, until now, there is still no direct genetic evidence showing the involvement of soybean 14-3-3s in ABA responses. Hence, in this study, based on the latest Glycine max genome on Phytozome v10.3, we initially analyzed the evolutionary relationship, genome organization, gene structure and duplication, and three-dimensional structure of soybean 14-3-3 family proteins systematically. Our results suggested that soybean 14-3-3 family was highly evolutionary conserved and possessed segmental duplication in evolution. Then, based on our previous functional characterization of a Glycine soja 14-3-3 protein GsGF14o in drought stress responses, we further investigated the expression characteristics of GsGF14o in detail, and demonstrated its positive roles in ABA sensitivity. Quantitative real-time PCR analyses in Glycine soja seedlings and GUS activity assays in PGsGF14O:GUS transgenic Arabidopsis showed that GsGF14o expression was moderately and rapidly induced by ABA treatment. As expected, GsGF14o overexpression in Arabidopsis augmented the ABA inhibition of seed germination and seedling growth, promoted the ABA induced stomata closure, and up-regulated the expression levels of ABA induced genes. Moreover, through yeast two hybrid analyses, we further demonstrated that GsGF14o physically interacted with the AREB/ABF transcription factors in yeast cells. Taken together, results presented in this study strongly suggested that GsGF14o played an important role in regulation of ABA sensitivity in Arabidopsis.


Assuntos
Proteínas 14-3-3/fisiologia , Ácido Abscísico/fisiologia , Arabidopsis/fisiologia , Reguladores de Crescimento de Planta/fisiologia , Soja/fisiologia , Proteínas 14-3-3/genética , Arabidopsis/genética , Mapeamento Cromossômico , Germinação/fisiologia , Oryza/genética , Filogenia , Estômatos de Plantas/fisiologia , Plântula/crescimento & desenvolvimento , Alinhamento de Sequência , Transdução de Sinais/genética , Transdução de Sinais/fisiologia , Soja/genética
18.
Mol Plant ; 8(9): 1385-95, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-25936676

RESUMO

Plant pathogens deliver effector proteins that alter host processes to create an environment conducive to colonization. Attention has focused on identifying the targets of effectors and how their manipulation facilitates disease. RXLR effector Pi04089 from the potato blight pathogen Phytophthora infestans accumulates in the host nucleus and enhances colonization when transiently expressed in planta. Its nuclear localization is required for enhanced P. infestans colonization. Pi04089 interacts in yeast and in planta with a putative potato K-homology (KH) RNA-binding protein, StKRBP1. Co-localization of Pi04089 and StKRBP1, and bimolecular fluorescence complementation between them, indicate they associate at nuclear speckles. StKRBP1 protein levels increased when it was co-expressed with Pi04089. Indeed, such accumulation of StKRBP1 was observed also on the first day of leaf colonization by the pathogen. Remarkably, overexpression of StKRBP1 significantly enhances P. infestans infection. Mutation of the nucleotide-binding motif GxxG to GDDG in all three KH domains of StKRBP1 abolishes its interaction with Pi04089, its localization to nuclear speckles, and its increased accumulation when co-expressed with the effector. Moreover, the mutant StKRBP1 protein no longer enhances leaf colonization by P. infestans, implying that nucleotide binding is likely required for this activity. We thus argue that StKRBP1 can be regarded as a susceptibility factor, as its activity is beneficial to the pathogen.


Assuntos
Phytophthora infestans/fisiologia , Doenças das Plantas/microbiologia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Proteínas de Ligação a RNA/metabolismo , Solanum tuberosum/metabolismo , Solanum tuberosum/microbiologia , Motivos de Aminoácidos , Sequência de Aminoácidos , Núcleo Celular/metabolismo , Dados de Sequência Molecular , Mutação , Phytophthora infestans/crescimento & desenvolvimento , Phytophthora infestans/patogenicidade , Ligação Proteica , Estrutura Terciária de Proteína , Proteínas de Ligação a RNA/química , Virulência
19.
Anticancer Agents Med Chem ; 15(3): 277-84, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25353334

RESUMO

In this study, a new dithiolopyrrolone biosynthetic pathway was identified in Saccharothrix algeriensis NRRL B-24137, which was reported to produce a variety of dithiolopyrrolone natural products including thiolutin, a potential drug candidate for tumor angiogenesis inhibition. Bioinformatics analysis of the cluster revealed that it contains all the essential genes for holothin core biosynthesis and several other auxiliary genes. Interestingly, heterologous expression of the gene cluster in Streptomyces albus only induced the production of holomycin, implying that the gene responsible for the N4-methylation and the gene(s) involved in the formation of various acylated chains on N7 position of the holothin may locate outside the gene cluster. Incubation of holomycin with S-adenosyl-L-methionine (SAM) in the cell-free extract of Sa. algeriensis resulted in the production of thiolutin, suggesting that the N4-methyl group of thiolutin is originated from SAM, and the N4-methylation could be in the late stage of biosynthesis of thiolutin type dithiolopyrrolones. An evolution-based model for biosynthesis of thiolutin and its analogs was further proposed based on these results.


Assuntos
Actinomycetales/genética , Inibidores da Angiogênese/biossíntese , Vias Biossintéticas/genética , Família Multigênica/genética , Inibidores da Angiogênese/química , Inibidores da Angiogênese/isolamento & purificação , Produtos Biológicos/química , Produtos Biológicos/isolamento & purificação , Produtos Biológicos/metabolismo , Biologia Computacional , Estrutura Molecular , Pirrolidinonas/química , Pirrolidinonas/isolamento & purificação , Pirrolidinonas/metabolismo
20.
Chem Sci ; 6(2): 1414-1419, 2015 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-29861965

RESUMO

(2R3S4S)-5-Fluoro-2,3,4-trihydroxypentanoic acid (5-FHPA) has been discovered as a new fluorometabolite in the soil bacterium Streptomyces sp. MA37. Exogenous addition of 5-fluoro-5-deoxy-d-ribose (5-FDR) into the cell free extract of MA37 demonstrated that 5-FDR was an intermediate to a range of unidentified fluorometabolites, distinct from fluoroacetate (FAc) and 4-fluorothreonine (4-FT). Bioinformatics analysis allowed identification of a gene cluster (fdr), encoding a pathway to the biosynthesis of 5-FHPA. Over-expression and in vitro assay of FdrC indicated that FdrC is a NAD+ dependent dehydrogenase responsible for oxidation of 5-FDR into 5-fluoro-5-deoxy-lactone, followed by hydrolysis to 5-FHPA. The identity of 5-FHPA in the fermentation broth was confirmed by synthesis of a reference compound and then co-correlation by 19F-NMR and GC-MS analysis. The occurrence of 5-FHPA proves the existence of a new fluorometabolite pathway.

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