Arabidopsis RAB8A, RAB8B, and RAB8D Proteins Interact with Several RTNLB Proteins and Are Involved in the Agrobacterium tumefaciens Infection Process

2021 ◽  
Author(s):  
Fan-Chen Huang ◽  
Shin-Fei Chi ◽  
Pei-Ru Chien ◽  
Yin-Tzu Liu ◽  
Hsin-Nung Chang ◽  
...  
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Pseudomonas Syringae ◽  
Infection Process ◽  
Bimolecular Fluorescence Complementation ◽  
Virulence Proteins ◽  
Crown Gall Disease ◽  
Type Iv ◽  
Virulence Protein ◽  
Infection Processes

Abstract Arabidopsis thaliana small GTP-binding proteins, AtRAB8s, associate with the endomembrane system and modulate tubulovesicular trafficking between compartments of the biosynthetic and endocytic pathways. There are 5 members in Arabidopsis, namely AtRAB8A-8E. Yeast two-hybrid assays, bimolecular fluorescence complementation (BiFC) assays, and glutathione-S-transferase (GST) pull-down assays showed that RAB8A, 8B, and 8D interacted with several membrane-associated reticulon-like (AtRTNLB) proteins in yeast, plant cells, and in vitro. Furthermore, RAB8A, 8B, and 8D proteins showed interactions with the Agrobacterium tumefaciens virulence protein, VirB2, a component of a Type IV secretion system (T4SS). A. tumefaciens uses a T4SS to transfer T-DNA and Virulence proteins to plants, which causes crown gall disease in plants. The Arabidopsis rab8A, rab8B, and rab8D single mutants showed decrease levels of Agrobacterium-mediated root and seedling transformation, while the RAB8A, 8B, and 8D overexpression (O/E) transgenic Arabidopsis plants were hypersusceptible to A. tumefaciens and Pseudomonas syringae infections. RAB8A-8E transcripts accumulated differently in roots, rosette leaves, cauline leaves, inflorescence, and flowers of wild-type plants. In summary, RAB8A, 8B, and 8D interacted with several RTNLB proteins and participated in A. tumefaciens and P. syringae infection processes.

scholarly journals EFFICACY OF BIOCHEMICAL PREPARATIONS AND EXTRACT FROM Hypericum perforatum AGAINST BACTERIAL DISEASES

2019 ◽  
Vol 18 (3) ◽  
Author(s):  
Małgorzata Schollenberger ◽  
Sylwia Pudło ◽  
Elżbieta Paduch-Cichal ◽  
Ewa Mirzwa-Mróz
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Pseudomonas Syringae ◽  
Pathogenic Bacteria ◽  
Hypericum Perforatum ◽  
Agar Plate ◽  
Nutrient Agar ◽  
Bacterial Strains ◽  
Bacterial Diseases ◽  
Nutrient Agar Medium

The biotechnical preparations: Biosept Active (based on a grapefruit extract) and BioZell (based on thyme oil) as well as Hypericum perforatum extract, streptomycin solution and fungicide Champion 50WP (active ingredient substance – e.i. 50% copper hydroxide) were investigated for antimicrobial effects against plant pathogenic bacteria: Agrobacterium tumefaciens, Pseudomonas syringae pv. syringae and Xanthomonas ar- boricola pv. corylina. The screening was carried out in vitro on three media: Nutrient Agar (NA Difco), Pseudomonas Agar F (Merck) – analogue of King B and 523. In the experiments, the agar plate method was applied. There were no statistically significant differences in the effect of streptomycin and Champion 50WP on the growth inhibition of three bacteria strains for medium 523 and Nutrient Agar and of P. syringae pv. syringae and X. arboricola pv. corylina for medium King B. It was determined that the antibacterial activity of Biosept Active and BioZell biopreparations and H. perforatum extract against Agrobacterium tumefaciens (strain C58), Pseudomonas syringae pv. syringae (strain 760) and Xanthomonas arboricola pv. corylina (strain RIPF-x13) were dependent on the strain of pathogen as well as the growth medium used. According to the research results obtained, the Biosept Active preparation and H. perforatum extract demonstrated high bacteriostatic activity against three bacterial strains grown on the Nutrient Agar medium.

scholarly journals Type IV Secretion System Is Not Involved in Infection Process in Citrus

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Tiago Rinaldi Jacob ◽  
Marcelo Luiz de Laia ◽  
Leandro Marcio Moreira ◽  
Janaína Fernandes Gonçalves ◽  
Flavia Maria de Souza Carvalho ◽  
...  
Keyword(s):  
Cell Envelope ◽  
Positive Control ◽  
Secretion System ◽  
Infection Process ◽  
Type Iv Secretion ◽  
Target Cells ◽  
Type Iv Secretion System ◽  
In Planta ◽  
Type Iv

The type IV secretion system (T4SS) is used by Gram-negative bacteria to translocate protein and DNA substrates across the cell envelope and into target cells.Xanthomonas citrisubsp.citricontains two copies of the T4SS, one in the chromosome and the other is plasmid-encoded. To understand the conditions that induce expression of the T4SS inXcc, we analyzed,in vitroandin planta, the expression of 18 ORFs from the T4SS and 7 hypothetical flanking genes by RT-qPCR. As a positive control, we also evaluated the expression of 29 ORFs from the type III secretion system (T3SS), since these genes are known to be expressed during plant infection condition, but not necessarily in standard culture medium. From the 29 T3SS genes analyzed by qPCR, onlyhrpAwas downregulated at 72 h after inoculation. All genes associated with the T4SS were downregulated onCitrusleaves 72 h after inoculation. Our results showed that unlike the T3SS, the T4SS is not induced during the infection process.

scholarly journals Small heat-shock protein HspL is induced by VirB protein(s) and promotes VirB/D4-mediated DNA transfer in Agrobacterium tumefaciens

Microbiology ◽  
2009 ◽  
Vol 155 (10) ◽  
pp. 3270-3280 ◽  
Author(s):  
Yun-Long Tsai ◽  
Ming-Hsuan Wang ◽  
Chan Gao ◽  
Sonja Klüsener ◽  
Christian Baron ◽  
...  
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Heat Shock ◽  
Heat Shock Protein ◽  
Response Regulator ◽  
Small Heat Shock Protein ◽  
Protein S ◽  
Dna Transfer ◽  
Protein Accumulation ◽  
Crown Gall Disease ◽  
Type Iv

Agrobacterium tumefaciens is a Gram-negative plant-pathogenic bacterium that causes crown gall disease by transferring and integrating its transferred DNA (T-DNA) into the host genome. We characterized the chromosomally encoded alpha-crystallin-type small heat-shock protein (α-Hsp) HspL, which was induced by the virulence (vir) gene inducer acetosyringone (AS). The transcription of hspL but not three other α-Hsp genes (hspC, hspAT1, hspAT2) was upregulated by AS. Further expression analysis in various vir mutants suggested that AS-induced hspL transcription is not directly activated by the VirG response regulator but rather depends on the expression of VirG-activated virB genes encoding components of the type IV secretion system (T4SS). Among the 11 virB genes encoded by the virB operon, HspL protein levels were reduced in strains with deletions of virB6, virB8 or virB11. VirB protein accumulation but not virB transcription levels were reduced in an hspL deletion mutant early after AS induction, implying that HspL may affect the stability of individual VirB proteins or of the T4S complex directly or indirectly. Tumorigenesis efficiency and the VirB/D4-mediated conjugal transfer of an IncQ plasmid RSF1010 derivative between A. tumefaciens strains were reduced in the absence of HspL. In conclusion, increased HspL abundance is triggered in response to certain VirB protein(s) and plays a role in optimal VirB protein accumulation, VirB/D4-mediated DNA transfer and tumorigenesis.

scholarly journals The Vibrio cholerae VarS/VarA two-component system controls the expression of virulence proteins through ToxT regulation

Microbiology ◽  
2011 ◽  
Vol 157 (5) ◽  
pp. 1466-1473 ◽  
Author(s):  
Jeyoun Jang ◽  
Kyung-Tae Jung ◽  
Jungchan Park ◽  
Cheon-Kwon Yoo ◽  
Gi-Eun Rhie
Keyword(s):  
Vibrio Cholerae ◽  
Environmental Changes ◽  
Transcription Activator ◽  
Two Component System ◽  
Component System ◽  
Virulence Proteins ◽  
Two Component ◽  
Virulence Protein ◽  
El Tor

Although the conditions for inducing virulence protein expression in vitro are different, both classical and El Tor biotypes of Vibrio cholerae have been reported to regulate the expression of virulence proteins such as cholera toxin (CT) and toxin-coregulated pili (Tcp) through the ToxR/S/T system. The transcription activator ToxR responds to environmental stimuli such as pH and temperature and activates the second transcriptional regulator ToxT, which upregulates expression of virulence proteins. In addition to the ToxR/S/T signalling system, V. cholerae has been proposed to utilize another two-component system VarS/VarA to modulate expression of virulence genes. Previous study has shown that VarA of the VarS/VarA system is involved in the regulation of virulence proteins in the classical V. cholerae O395 strain; however, no further analysis was performed concerning VarS. In this study, we constructed varS mutants derived from the classical O395 and El Tor C6706 strains and demonstrated that VarS is also involved in the expression of the virulence proteins CT and Tcp from the V. cholerae classical and El Tor strains. This expression is through regulation of ToxT expression in response to environmental changes due to different toxin-inducing conditions.

scholarly journals VirE1-Mediated Resistance to Crown Gall in Transgenic Arabidopsis thaliana

2006 ◽  
Vol 96 (1) ◽  
pp. 105-110 ◽  
Author(s):  
Jodi Humann ◽  
Sarah Andrews ◽  
Walt Ream
Keyword(s):  
Arabidopsis Thaliana ◽  
Agrobacterium Tumefaciens ◽  
Crown Gall ◽  
Plant Cells ◽  
Bacterial Cells ◽  
Root Explants ◽  
Single Stranded Dna ◽  
Virulence Proteins ◽  
Crown Gall Disease ◽  
Efficient Transmission

Crown gall disease, caused by Agrobacterium tumefaciens, remains a serious agricultural problem despite current biocontrol methods. Agrobacterium tumefaciens transfers single-stranded DNA (T-strands) into plant cells along with several virulence proteins, including a single-stranded DNA-binding protein (VirE2). In plant cells, T-strands are protected from nucleases and targeted to the nucleus by VirE2, which is essential for efficient transmission (transfer and integration) of T-strands. VirE1 is the secretory chaperone for VirE2; it prevents VirE2 from forming aggregates and from binding the T-strands in bacterial cells. Therefore, we hypothesized that sufficient quantities of VirE1 expressed in plant cells might block T-DNA transmission by preventing VirE2 from binding T-strands. Here we show that root explants from Arabidopsis thaliana plants that expressed virE1 formed 3.5-fold fewer tumors than roots from plants without virE1. Also, this resistance was specific for VirE2-mediated Agrobacterium transformation. Plants that have been genetically altered to resist crown gall may prove more effective than biological control.

scholarly journals Elevated Temperature Differentially Affects Virulence, VirB Protein Accumulation, and T-Pilus Formation in Different Agrobacterium tumefaciens andAgrobacterium vitis Strains

2001 ◽  
Vol 183 (23) ◽  
pp. 6852-6861 ◽  
Author(s):  
Christian Baron ◽  
Natalie Domke ◽  
Michael Beinhofer ◽  
Siegfried Hapfelmeier
Keyword(s):  
Elevated Temperature ◽  
Agrobacterium Tumefaciens ◽  
Gene Transfer ◽  
Protein Accumulation ◽  
Temperature Sensitive ◽  
Class Iii ◽  
Agrobacterium Vitis ◽  
Virulence Proteins ◽  
Virulence Protein ◽  
Pilus Assembly

ABSTRACT That gene transfer to plant cells is a temperature-sensitive process has been known for more than 50 years. Previous work indicated that this sensitivity results from the inability to assemble a functional T pilus required for T-DNA and protein transfer to recipient cells. The studies reported here extend these observations and more clearly define the molecular basis of this assembly and transfer defect. T-pilus assembly and virulence protein accumulation were monitored in Agrobacterium tumefaciens strain C58 at different temperatures ranging from 20°C to growth-inhibitory 37°C. Incubation at 28°C but not at 26°C strongly inhibited extracellular assembly of the major T-pilus component VirB2 as well as of pilus-associated protein VirB5, and the highest amounts of T pili were detected at 20°C. Analysis of temperature effects on the cell-bound virulence machinery revealed three classes of virulence proteins. Whereas class I proteins (VirB2, VirB7, VirB9, and VirB10) were readily detected at 28°C, class II proteins (VirB1, VirB4, VirB5, VirB6, VirB8, VirB11, VirD2, and VirE2) were only detected after cell growth below 26°C. Significant levels of class III proteins (VirB3 and VirD4) were only detected at 20°C and not at higher temperatures. Shift of virulence-induced agrobacteria from 20 to 28 or 37°C had no immediate effect on cell-bound T pili or on stability of most virulence proteins. However, the temperature shift caused a rapid decrease in the amount of cell-bound VirB3 and VirD4, and VirB4 and VirB11 levels decreased next. To assess whether destabilization of virulence proteins constitutes a general phenomenon, levels of virulence proteins and of extracellular T pili were monitored in different A. tumefaciens and Agrobacterium vitis strains grown at 20 and 28°C. Levels of many virulence proteins were strongly reduced at 28°C compared to 20°C, and T-pilus assembly did not occur in all strains except “temperature-resistant” Ach5 and Chry5. Virulence protein levels correlated well with bacterial virulence at elevated temperature, suggesting that degradation of a limited set of virulence proteins accounts for the temperature sensitivity of gene transfer to plants.

scholarly journals Antibacterial effect of Artemisia and ginger extracts in controlling Agrobacterium tumefaciens in roses

2021 ◽  
pp. 1-5
Author(s):  
Alfred Njagi ◽  
Methuselah Nyamwange Mang'erere ◽  
Ezekiel Mugendi Njeru ◽  
Jonah Kiprono Birgen
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Crown Gall ◽  
Antibacterial Effect ◽  
Inhibition Zone ◽  
Stem Length ◽  
Copper Hydroxide ◽  
Cut Flowers ◽  
Promising Alternative ◽  
Crown Gall Disease

Rose is the world’s most traded cut flowers with 74 % being produced in Kenya. Pests like spider mites, caterpillar, aphids, thrips, nematodes and diseases such as crown gall, downy mildew, powdery mildew and botrytis highly compromise rose production. Crown gall disease caused by Agrobacterium tumefaciens is the most problematic disease of roses in Kenya, causing a production loss of up to 60 % depending on the age and variety of rose. An experiment to determine the antibacterial effect of Artemisia and ginger extracts was carried out in vitro, where the inhibitory zones around filter discs soaked with extracts on Muller Hinton agar was established. The extracts were emulsified with dimethylsulfoxide (50 %) and minimum inhibitory concentrations of Artemisia (125 mg/ml), ginger (62.5 mg/ml) and mixture of Artemisia and ginger (31.25 mg/ml) were used to soak the filter discs whereas the commercial recommended rate of copper hydroxide of 6.25 mg/ml was used. Artemisia and copper hydroxide (commercial antibiotic) had highest inhibition zone of 12.80 mm compared to ginger 10.60 mm. A mixture of Artemisia and Ginger had a slightly lower inhibition zone (10.20 mm) though not significantly different from ginger (P> 0.001). An eight-month greenhouse experiment was also done to determine the efficacy of the extracts on inoculated rose plants. The results showed that crown gall incidence and gall weight were low but not significantly different from copper hydroxide and Artemisia. Crop vigor, which was indicated by stem length was highest for Artemisia treatment with an average of 73.54 cm followed by copper hydroxide (67.25 cm) while ginger and mixture of ginger and Artemisia had 53.44 cm and 64.70 cm respectively. From the results of this research, Artemisia and Ginger extracts are promising alternative to control crown gall and possibly other diseases in field crops. Artemisia performance compares well with copper hydroxide and therefore the best alternative to replace copper hydroxide.

THE ACTIVITY OF ESSENTIAL OILS OBTAINED FROM SPECIES AND INTERSPECIES HYBRIDS OF THE Mentha GENUS AGAINST SELECTED PLANT PATHOGENIC BACTERIA

2018 ◽  
Vol 17 (6) ◽  
pp. 167-174 ◽  
Author(s):  
Małgorzata Schollenberger ◽  
Tomasz M. Staniek ◽  
Elżbieta Paduch-Cichal ◽  
Beata Dasiewicz ◽  
Agnieszka Gadomska-Gajadhur ◽  
...  
Keyword(s):  
Essential Oils ◽  
Pseudomonas Syringae ◽  
Pathogenic Bacteria ◽  
Antimicrobial Effect ◽  
Mentha Piperita ◽  
Mentha Spicata ◽  
Plant Essential Oils ◽  
Plant Pathogenic Bacteria ◽  
Interspecies Hybrids

Plant essential oils of six aromatic herb species and interspecies hybrids of the family Lamiaceae – chocolate mint (Mentha piperita × ‘Chocolate’), pineapple mint (Mentha suaveolens ‘Variegata’), apple mint (Mentha × rotundifolia), spearmint (Mentha spicata), orange mint (Mentha × piperita ‘Granada’) and strawberry mint (Mentha × villosa ‘Strawberry’) – were investigated for antimicrobial effects against plant pathogenic bacteria: Agrobacterium tumefaciens, Pseudomonas syringae pv. syringae and Xanthomonas arboricola pv. corylina. The screening was carried out in vitro on agar plates filled with the target organism. All essential oils screened exhibited a higher level of antibacterial activity against A. tumefaciens and X. arboricola pv. corylina than streptomycin used as a standard in all tests. The antimicrobial effect of streptomycin and five mint oils was at the same level for P. syringae pv. syringae. There were no significant differences in the influence of the chocolate mint oil on the growth inhibition of all bacteria tested. Plant essential oils from pineapple mint, apple mint, spearmint and strawberry mint showed the weakest antimicrobial activity against P. syringae pv. syringae and the strongest towards A. tumefaciens and X. arboricola pv. corylina. The essential oils from strawberry mint, pineapple mint, spearmint and apple mint had the strongest effect on A. tumefaciens, and the lowest inhibitory activity was exhibited by the chocolate mint and orange mint essential oils. X. arboricola pv. corylina was the most sensitive to the strawberry mint, pineapple mint and spearmint oils. The chocolate mint oil showed the greatest activity against P. syringae pv. syringae.

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