Introduction of Genetic Material in Ralstonia solanacearum Through Natural Transformation and Conjugation

Perrier, Anthony; Barberis, Patrick; Genin, Stéphane

doi:10.1007/978-1-4939-7604-1_16

Cited by 22 publications

(15 citation statements)

References 16 publications

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“…The protocol of Perrier et al (2018) was followed with minor modifications. A single colony of R. solanacearum was inoculated into 20 ml of one‐quarter strength M63 minimal medium (15 mM ammonium sulphate, 1.8 µM iron(II) sulphate, 100 mM monopotassium phosphate, and 1 mM magnesium sulphate adjusted to pH 7 with KOH) supplemented with 2% glycerol (Miller, 1992) and incubated overnight at 28 °C with shaking (200 rpm).…”

Section: Methodsmentioning

confidence: 99%

Identification of RipAZ1 as an avirulence determinant of Ralstonia solanacearum in Solanum americanum

Moon

Pandey

Yoon

et al. 2021

Molecular Plant Pathology

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Ralstonia solanacearum causes bacterial wilt disease in many plant species. Type III‐secreted effectors (T3Es) play crucial roles in bacterial pathogenesis. However, some T3Es are recognized by corresponding disease resistance proteins and activate plant immunity. In this study, we identified the R. solanacearum T3E protein RipAZ1 (Ralstonia injected protein AZ1) as an avirulence determinant in the black nightshade species Solanum americanum. Based on the S. americanum accession‐specific avirulence phenotype of R. solanacearum strain Pe_26, 12 candidate avirulence T3Es were selected for further analysis. Among these candidates, only RipAZ1 induced a cell death response when transiently expressed in a bacterial wilt‐resistant S. americanum accession. Furthermore, loss of ripAZ1 in the avirulent R. solanacearum strain Pe_26 resulted in acquired virulence. Our analysis of the natural sequence and functional variation of RipAZ1 demonstrated that the naturally occurring C‐terminal truncation results in loss of RipAZ1‐triggered cell death. We also show that the 213 amino acid central region of RipAZ1 is sufficient to induce cell death in S. americanum. Finally, we show that RipAZ1 may activate defence in host cell cytoplasm. Taken together, our data indicate that the nucleocytoplasmic T3E RipAZ1 confers R. solanacearum avirulence in S. americanum. Few avirulence genes are known in vascular bacterial phytopathogens and ripAZ1 is the first one in R. solanacearum that is recognized in black nightshades. This work thus opens the way for the identification of disease resistance genes responsible for the specific recognition of RipAZ1, which can be a source of resistance against the devastating bacterial wilt disease.

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Section: Methodsmentioning

confidence: 99%

Identification of RipAZ1 as an avirulence determinant of Ralstonia solanacearum in Solanum americanum

Moon

Pandey

Yoon

et al. 2021

Molecular Plant Pathology

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“…Briefly, upstream (RSp1632) and downstream (RSp1635) regions were PCR amplified using the primer pairs 1632A/1632B and 1635C/1635D, respectively, and cloned into the EcoRI-HindIII-digested pk18mobsacb to generate pK18-RSp1632/1635 (listed below). The circularized plasmid was recombined in strain GMI1000 through natural transformation (Perrier et al, 2018). Kanamycin-resistant and sucrose-sensitive recombinant clones were first selected and in a second step, after overnight culture in BG medium , kanamycin-sensitive and sucroseresistant clones were screened by PCR using the primer pair 1632A/1635D to identify a RSp1632-RSp1635 recombinant.…”

Section: Data Availability Statementmentioning

confidence: 99%

Unravelling physiological signatures of tomato bacterial wilt and xylem metabolites exploited by Ralstonia solanacearum

Gerlin

Escourrou

Cassan

et al. 2021

Environmental Microbiology

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The plant pathogen Ralstonia solanacearum uses plant resources to intensely proliferate in xylem vessels and provoke plant wilting. We combined automatic phenotyping and tissue/xylem quantitative metabolomics of infected tomato plants to decipher the dynamics of bacterial wilt. Daily acquisition of physiological parameters such as transpiration and growth were performed. Measurements allowed us to identify a tipping point in bacterial wilt dynamics. At this tipping point, the reached bacterial density brutally disrupts plant physiology and rapidly induces its death. We compared the metabolic and physiological signatures of the infection to drought stress, and found that similar changes occur.Quantitative dynamics of xylem content enabled us to identify glutamine (and asparagine) as primary resources R. solanacearum consumed during its colonization phase. An abundant production of putrescine was also observed during the infection process and was strongly correlated with in planta bacterial growth. Dynamic profiling of xylem metabolites confirmed that glutamine is the favored substrate of R. solanacearum. On the other hand, a triple mutant strain unable to metabolize glucose, sucrose and fructose appears to be only weakly reduced for in planta growth and pathogenicity.

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“…Then, Rsc3174, PSI07_1860 and CMR15v4_mp10184 were cloned into pDONR207 vector using a BP reaction and in pNP329 using a LR reaction following the instructions of the manufacturer (LifeTechnologies, Carlsbad, CA, USA). The final expression vectors were transformed into the R. pseudosolanacearum GMI1000 strain and in the hrcV mutant (type III secretion defective mutant, used as a negative control) as previously described (Perrier, Barberis & Genin, 2018). In-vitro secretion assays and western blot analysis were performed as previously described (Lonjon et al, 2018).…”

Section: Methodsmentioning

confidence: 99%

Pangenomic type III effector database of the plant pathogenicRalstoniaspp.

Sabbagh

Carrère

Lonjon

et al. 2019

PeerJ

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Background The bacterial plant pathogenic Ralstonia species belong to the beta-proteobacteria class and are soil-borne pathogens causing vascular bacterial wilt disease, affecting a wide range of plant hosts. These bacteria form a heterogeneous group considered as a “species complex” gathering three newly defined species. Like many other Gram negative plant pathogens, Ralstonia pathogenicity relies on a type III secretion system, enabling bacteria to secrete/inject a large repertoire of type III effectors into their plant host cells. Type III-secreted effectors (T3Es) are thought to participate in generating a favorable environment for the pathogen (countering plant immunity and modifying the host metabolism and physiology). Methods Expert genome annotation, followed by specific type III-dependent secretion, allowed us to improve our Hidden-Markov-Model and Blast profiles for the prediction of type III effectors. Results We curated the T3E repertoires of 12 plant pathogenic Ralstonia strains, representing a total of 12 strains spread over the different groups of the species complex. This generated a pangenome repertoire of 102 T3E genes and 16 hypothetical T3E genes. Using this database, we scanned for the presence of T3Es in the 155 available genomes representing 140 distinct plant pathogenic Ralstonia strains isolated from different host plants in different areas of the globe. All this information is presented in a searchable database. A presence/absence analysis, modulated by a strain sequence/gene annotation quality score, enabled us to redefine core and accessory T3E repertoires.

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Introduction of Genetic Material in Ralstonia solanacearum Through Natural Transformation and Conjugation

Cited by 22 publications

References 16 publications

Identification of RipAZ1 as an avirulence determinant of Ralstonia solanacearum in Solanum americanum

Identification of RipAZ1 as an avirulence determinant of Ralstonia solanacearum in Solanum americanum

Unravelling physiological signatures of tomato bacterial wilt and xylem metabolites exploited by Ralstonia solanacearum

Pangenomic type III effector database of the plant pathogenicRalstoniaspp.

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