scholarly journals Expression of the gum Operon Directing Xanthan Biosynthesis in Xanthomonas campestris and Its Regulation In Planta

2001 ◽  
Vol 14 (6) ◽  
pp. 768-774 ◽  
Author(s):  
Adrian A. Vojnov ◽  
Holly Slater ◽  
Michael J. Daniels ◽  
J. Maxwell Dow
Keyword(s):  
Xanthomonas Campestris ◽  
Carbon Sources ◽  
Regulatory System ◽  
Extracellular Polysaccharide ◽  
In Planta ◽  
Glucuronidase Activity ◽  
Gum Gene Cluster ◽  
Diffusible Signal Factor ◽  
Turnip Leaves

The gum gene cluster of Xanthomonas campestris pv. campestris comprises 12 genes whose products are involved in the biosynthesis of the extracellular polysaccharide xanthan. These genes are expressed primarily as an operon from a promoter upstream of the first gene, gumB. Although the regulation of xanthan synthesis in vitro has been well studied, nothing is known of its regulation in planta. A reporter plasmid was constructed in which the promoter region of the gum operon was fused to gusA. In liquid cultures, the expression of the gumgusA reporter was correlated closely with the production of xanthan, although a low basal level of β-glucuronidase activity was seen in the absence of added carbon sources when xanthan production was very low. The expression of the gumgusA fusion also was subject to positive regulation by rpfF, which is responsible for the synthesis of the diffusible signal factor (DSF). The expression of the gumgusA fusion in bacteria recovered from inoculated turnip leaves was maximal at the later phases of growth and was subject to regulation by rpfF. These results provide indirect support for the operation of the DSF regulatory system in bacteria in planta.

scholarly journals Identification and Characterization of Two Novel DSF-Controlled Virulence-Associated Genes Within the nodB-rhgB Locus of Xanthomonas oryzae pv. oryzicola Rs105

2015 ◽  
Vol 105 (5) ◽  
pp. 588-596 ◽  
Author(s):  
Zhiwei Song ◽  
Yancun Zhao ◽  
Xingyang Zhou ◽  
Guichun Wu ◽  
Yuqiang Zhang ◽  
...  
Keyword(s):  
Extracellular Polysaccharide ◽  
Xanthomonas Oryzae ◽  
In Planta ◽  
Inoculation Density ◽  
Xanthomonas Species ◽  
Diffusible Signal Factor ◽  
Visible Effect ◽  
Extracellular Protease Activity ◽  
Epiphytic Survival

Xanthomonas oryzae pv. oryzicola and X. oryzae pv. oryzae are two pathovars of X. oryzae that cause leaf streak and blight in rice, respectively. These two bacterial pathogens cause different disease symptoms by utilizing different infection sites on rice. Compared with X. oryzae pv. oryzae, the molecular virulence mechanism of X. oryzae pv. oryzicola remains largely unknown. Previously, we identified a unique diffusible signal factor (DSF)-controlled virulence-related gene (hshB) in X. oryzae pv. oryzicola Rs105 located in the nodB-rghB locus, which is absent in X. oryzae pv. oryzae PXO99A. In the present study, we identified two additional genes within this locus (hshA and hshC) that were unique to X. oryzae pv. oryzicola Rs105 compared with X. oryzae pv. oryzae PXO99A, and we found that the transcription of these genes was regulated by DSF signaling in X. oryzae pv. oryzicola. The mutation of these genes impaired the virulence of the wild-type Rs105 when using a low inoculation density of X. oryzae pv. oryzicola. In contrast to hshB, the mutation of these genes did not have any visible effect on characterized virulence-related functions, including in vitro growth, extracellular polysaccharide production, extracellular protease activity, and antioxidative ability. However, we found that mutation of hshA or hshC significantly reduced the in planta growth ability and epiphytic survival level of X. oryzae pv. oryzicola cells, which was the probable mechanisms of involvement of these two genes in virulence. Collectively, our studies of X. oryzae pv. oryzicola have identified two novel DSF-controlled virulence-associated genes (hshA and hshC), which will add to our understanding of the regulatory mechanisms of conserved DSF virulence signaling in Xanthomonas species.

Strain distribution and in planta production of an extracellular polysaccharide depolymerase from Bradyrhizobium japonicum

1992 ◽  
Vol 38 (8) ◽  
pp. 857-861 ◽  
Author(s):  
Michael F. Dunn ◽  
Arthur L. Karr
Keyword(s):  
Bradyrhizobium Japonicum ◽  
Root Nodules ◽  
Extracellular Polysaccharide ◽  
Extracellular Polysaccharides ◽  
Neutral Sugar ◽  
In Planta ◽  
In Vitro Production ◽  
Polysaccharide Composition ◽  
Vitro Production

Thirty-four strains of Bradyrhizobium japonicum were screened for the in vitro production of an extracellular polysaccharide depolymerase active against the B. japonicum acidic extracellular polysaccharide that contains mannose, glucose, galactose, and 4-O-methylgalactose as neutral sugar components. Over 90% of tested strains producing this type of extracellular polysaccharide also produced the extracellular polysaccharide depolymerase, whereas strains producing a compositionally different extracellular polysaccharide did not. In addition, representatives of species related to B. japonicum by extracellular polysaccharide composition or host range were also phenotypically depolymerase negative. Depolymerase was also present in soybean root nodules formed by B. japonicum strain 2143. In contrast to the cell-associated depolymerase activity found in free-living cells of this strain, most of the depolymerase activity present in nodules is free of the bacteroids. The widespread occurrence of the depolymerase among B. japonicum strains and the spatiotemporal distribution of its activity in planta are consistent with the enzyme playing a role in the removal of surface extracellular polysaccharide from the microorganism during the infection of nodulation process. Key words: Bradyrhizobium japonicum, soybean, extracellular polysaccharides, extracellular polysaccharide depolymerase, bacteroids.

scholarly journals Detection and Visualization of an Exopolysaccharide Produced by Xylella fastidiosa In Vitro and In Planta

2007 ◽  
Vol 73 (22) ◽  
pp. 7252-7258 ◽  
Author(s):  
M. Caroline Roper ◽  
L. Carl Greve ◽  
John M. Labavitch ◽  
Bruce C. Kirkpatrick
Keyword(s):  
Xanthan Gum ◽  
Xanthomonas Campestris ◽  
Polyclonal Antibodies ◽  
Xylella Fastidiosa ◽  
Enzyme Linked Immunosorbent Assay ◽  
In Planta ◽  
Mannose Residue ◽  
Pantoea Stewartii ◽  
Vascular Occlusions

ABSTRACT Many phytopathogenic bacteria, such as Ralstonia solanacearum, Pantoea stewartii, and Xanthomonas campestris, produce exopolysaccharides (EPSs) that aid in virulence, colonization, and survival. EPS can also contribute to host xylem vessel blockage. The genome of Xylella fastidiosa, the causal agent of Pierce's disease (PD) of grapevine, contains an operon that is strikingly similar to the X. campestris gum operon, which is responsible for the production of xanthan gum. Based on this information, it has been hypothesized that X. fastidiosa is capable of producing an EPS similar in structure and composition to xanthan gum but lacking the terminal mannose residue. In this study, we raised polyclonal antibodies against a modified xanthan gum polymer similar to the predicted X. fastidiosa EPS polymer. We used enzyme-linked immunosorbent assay to quantify production of EPS from X. fastidiosa cells grown in vitro and immunolocalization microscopy to examine the distribution of X. fastidiosa EPS in biofilms formed in vitro and in planta and assessed the contribution of X. fastidiosa EPS to the vascular occlusions seen in PD-infected grapevines.

scholarly journals Development and Use of a Reverse Transcription-PCR Assay To Study Expression of Tri5 byFusarium Species In Vitro and In Planta

1999 ◽  
Vol 65 (9) ◽  
pp. 3850-3854 ◽  
Author(s):  
F. M. Doohan ◽  
G. Weston ◽  
H. N. Rezanoor ◽  
D. W. Parry ◽  
P. Nicholson
Keyword(s):  
Reverse Transcription ◽  
Infected Plant ◽  
Fusarium Culmorum ◽  
Pcr Analysis ◽  
Growth Stages ◽  
In Planta ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Glucuronidase Activity

ABSTRACT The Tri5 gene encodes trichodiene synthase, which catalyzes the first reaction in the trichothecene biosynthetic pathway. In vitro, a direct relationship was observed between Tri5expression and the increase in deoxynivalenol production over time. We developed a reverse transcription (RT)-PCR assay to quantifyTri5 gene expression in trichothecene-producing strains ofFusarium species. We observed an increase inTri5 expression following treatment of Fusarium culmorum with fungicides, and we also observed an inverse relationship between Tri5 expression and biomass, as measured by β-d-glucuronidase activity, during colonization of wheat (cv. Avalon) seedlings by F. culmorum. RT-PCR analysis also showed that for ears of wheat cv. Avalon inoculated with F. culmorum, there were different levels of Tri5 expression in grain and chaff at later growth stages. We used the Tri5-specific primers to develop a PCR assay to detect trichothecene-producing Fusariumspecies in infected plant material.

scholarly journals BDSF Is the Predominant In-Planta Quorum-Sensing Signal Used During Xanthomonas campestris Infection and Pathogenesis in Chinese Cabbage

2019 ◽  
Vol 32 (2) ◽  
pp. 240-254 ◽  
Author(s):  
Abdelgader Abdeen Diab ◽  
Xue-Qiang Cao ◽  
Hui Chen ◽  
Kai Song ◽  
Lian Zhou ◽  
...  
Keyword(s):  
Quorum Sensing ◽  
Chinese Cabbage ◽  
Xanthomonas Campestris ◽  
Growth Conditions ◽  
Infection Model ◽  
In Planta ◽  
Flight Mass Spectrometry ◽  
Diffusible Signal Factor ◽  
Liquid Chromatographic ◽  
Diverse Plant

Xanthomonas campestris pv. campestris uses the diffusible signal factor (DSF) family of quorum-sensing (QS) signals to coordinate virulence and adaptation. DSF family signals have been well-characterized using laboratory-based cell cultures. The in-planta QS signal used during X. campestris pv. campestris infection remains unclear. To achieve this goal, we first mimic in-planta X. campestris pv. campestris growth conditions by supplementing the previously developed XYS medium with cabbage hydrolysate and found that the dominant signal produced in these conditions was BDSF. Secondly, by using XYS medium supplemented with diverse plant-derived compounds, we examined the effects of diverse plant-derived compounds on the biosynthesis of DSF family signals. Several compounds were found to promote biosynthesis of BDSF. Finally, using an X. campestris pv. campestris ΔrpfB–Chinese cabbage infection model and an ultra-performance liquid chromatographic-time of flight-mass spectrometry–based assay, BDSF was found to comprise >70% of the DSF family signals present in infected cabbage tissue. BDSF at a concentration of 2.0 μM induced both protease activity and engXCA expression. This is the first report to directly show that BDSF is the predominant in-planta QS signal used during X. campestris pv. campestris infection. It provides a better understanding of the molecular interactions between X. campestris pv. campestris and its cruciferous hosts and also provides the logical target for designing strategies to counteract BDSF signaling and, thus, infection. Further studies are needed to get an exact idea about the DSF production dynamics of the wild-type strain inside the plant.

scholarly journals The RpfB-Dependent Quorum Sensing Signal Turnover System Is Required for Adaptation and Virulence in Rice Bacterial Blight Pathogen Xanthomonas oryzae pv. oryzae

2016 ◽  
Vol 29 (3) ◽  
pp. 220-230 ◽  
Author(s):  
Xing-Yu Wang ◽  
Lian Zhou ◽  
Jun Yang ◽  
Guang-Hai Ji ◽  
Ya-Wen He
Keyword(s):  
Quorum Sensing ◽  
Bacterial Blight ◽  
Xanthomonas Campestris ◽  
Deletion Mutant ◽  
Extracellular Polysaccharide ◽  
Xanthomonas Oryzae ◽  
Rice Bacterial Blight ◽  
Double Deletion ◽  
Diffusible Signal Factor ◽  
Bacterial Blight Pathogen

Xanthomonas oryzae pv. oryzae, the bacterial blight pathogen of rice, produces diffusible signal factor (DSF) family quorum sensing signals to regulate virulence. The biosynthesis and perception of DSF family signals require components of the rpf (regulation of pathogenicity factors) cluster. In this study, we report that RpfB plays an essential role in DSF family signal turnover in X. oryzae pv. oryzae PXO99A. The production of DSF family signals was boosted by deletion of the rpfB gene and was abolished by its overexpression. The RpfC/RpfG-mediated DSF signaling system negatively regulates rpfB expression via the global transcription regulator Clp, whose activity is reversible in the presence of cyclic diguanylate monophosphate. These findings indicate that the DSF family signal turnover system in PXO99A is generally consistent with that in Xanthomonas campestris pv. campestris. Moreover, this study has revealed several specific roles of RpfB in PXO99A. First, the rpfB deletion mutant produced high levels of DSF family signals but reduced extracellular polysaccharide production, extracellular amylase activity, and attenuated pathogenicity. Second, the rpfB/rpfC double-deletion mutant was partially deficient in xanthomonadin production. Taken together, the RpfB-dependent DSF family signal turnover system is a conserved and naturally presenting signal turnover system in Xanthomonas spp., which plays unique roles in X. oryzae pv. oryzae adaptation and pathogenesis.

scholarly journals Two Homologues of the Global Regulator Csr/Rsm Redundantly Control Phaseolotoxin Biosynthesis and Virulence in the Plant Pathogen Pseudomonas amygdali pv. phaseolicola 1448A

2020 ◽  
Vol 8 (10) ◽  
pp. 1536
Author(s):  
Diana Ramírez-Zapata ◽  
Cayo Ramos ◽  
Selene Aguilera ◽  
Leire Bardaji ◽  
Marta Martínez-Gil ◽  
...  
Keyword(s):  
Carbon Sources ◽  
Regulatory System ◽  
Mrna Levels ◽  
In Planta ◽  
Bacterial Pathogenicity ◽  
Regulatory Molecule ◽  
Transcriptional Regulatory ◽  
Carbon Storage Regulator ◽  
Functional Analyses ◽  
Pseudomonas Amygdali

The widely conserved Csr/Rsm (carbon storage regulator/repressor of stationary-phase metabolites) post-transcriptional regulatory system controls diverse phenotypes involved in bacterial pathogenicity and virulence. Here we show that Pseudomonas amygdali pv. phaseolicola 1448A contains seven rsm genes, four of which are chromosomal. In RNAseq analyses, only rsmE was thermoregulated, with increased expression at 18 °C, whereas the antagonistic sRNAs rsmX1, rsmX4, rsmX5 and rsmZ showed increased levels at 28 °C. Only double rsmA-rsmE mutants showed significantly altered phenotypes in functional analyses, being impaired for symptom elicitation in bean, including in planta growth, and for induction of the hypersensitive response in tobacco. Double mutants were also non-motile and were compromised for the utilization of different carbon sources. These phenotypes were accompanied by reduced mRNA levels of the type III secretion system regulatory genes hrpL and hrpA, and the flagellin gene, fliC. Biosynthesis of the phytotoxin phaseolotoxin by mutants in rsmA and rsmE was delayed, occurring only in older cultures, indicating that these rsm homologues act as inductors of toxin synthesis. Therefore, genes rsmA and rsmE act redundantly, although with a degree of specialization, to positively regulate diverse phenotypes involved in niche colonization. Additionally, our results suggest the existence of a regulatory molecule different from the Rsm proteins and dependent on the GacS/GacA (global activator of antibiotic and cyanide production) system, which causes the repression of phaseolotoxin biosynthesis at high temperatures.

scholarly journals The C-terminal domain of the type III secretion chaperone HpaB contributes to dissociation of chaperone-effector complex in Xanthomonas campestris pv. campestris

PLoS ONE ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. e0246033
Author(s):  
Yong-Liang Gan ◽  
Li-Yan Yang ◽  
Li-Chao Yang ◽  
Wan-Lian Li ◽  
Xue-Lian Liang ◽  
...  
Keyword(s):  
Xanthomonas Campestris ◽  
Pathogenic Bacteria ◽  
Terminal Region ◽  
Effector Proteins ◽  
Host Cells ◽  
Dependent Manner ◽  
In Planta ◽  
Terminal Domain ◽  
Self Association

Many animal and plant pathogenic bacteria employ a type three secretion system (T3SS) to deliver type three effector proteins (T3Es) into host cells. Efficient secretion of many T3Es in the plant pathogen Xanthomonas campestris pv. campestris (Xcc) relies on the global chaperone HpaB. However, how the domain of HpaB itself affects effector translocation/secretion is poorly understood. Here, we used genetic and biochemical approaches to identify a novel domain at the C-terminal end of HpaB (amino acid residues 137–160) that contributes to virulence and hypersensitive response (HR). Both in vitro secretion assay and in planta translocation assay showed that the secretion and translocation of T3E proteins depend on the C-terminal region of HpaB. Deletion of the C-terminal region of HpaB did not affect binding to T3Es, self-association or interaction with T3SS components. However, the deletion of C-terminal region sharply reduced the mounts of free T3Es liberated from the complex of HpaB with the T3Es, a reaction catalyzed in an ATP-dependent manner by the T3SS-associated ATPase HrcN. Our findings demonstrate the C-terminal domain of HpaB contributes to disassembly of chaperone-effector complex and reveal a potential molecular mechanism underpinning the involvement of HpaB in secretion of T3Es in Xcc.

THYROID STIMULATORS AND THYROID STIMULATION

1971 ◽  
Vol 66 (3) ◽  
pp. 558-576 ◽  
Author(s):  
Gerald Burke
Keyword(s):  
Regulatory System ◽  
Control Site ◽  
Thyroid Cell ◽  
Primary Control ◽  
Physico Chemical ◽  
Site Of Action ◽  
Long Acting

ABSTRACT A long-acting thyroid stimulator (LATS), distinct from pituitary thyrotrophin (TSH), is found in the serum of some patients with Graves' disease. Despite the marked physico-chemical and immunologic differences between the two stimulators, both in vivo and in vitro studies indicate that LATS and TSH act on the same thyroidal site(s) and that such stimulation does not require penetration of the thyroid cell. Although resorption of colloid and secretion of thyroid hormone are early responses to both TSH and LATS, available evidence reveals no basic metabolic pathway which must be activated by these hormones in order for iodination reactions to occur. Cyclic 3′, 5′-AMP appears to mediate TSH and LATS effects on iodination reactions but the role of this compound in activating thyroidal intermediary metabolism is less clear. Based on the evidence reviewed herein, it is suggested that the primary site of action of thyroid stimulators is at the cell membrane and that beyond the(se) primary control site(s), there exists a multifaceted regulatory system for thyroid hormonogenesis and cell growth.

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