Quorum Sensing-Dependent Invasion of Ralstonia solanacearum into Fusarium oxysporum Chlamydospores

Tsumori, Chiaki; Matsuo, Shoma; Murai, Yuta; Kai, Kenji

doi:10.1128/spectrum.00036-23

Cited by 6 publications

(6 citation statements)

References 71 publications

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“…The vector was introduced and expressed in P. protegens Cab57 . The recombinant was named Cab57- cayG .…”

Section: Resultsmentioning

confidence: 99%

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Structures and Biosynthesis of Caryoynencins, Unstable Bacterial Polyynes from Pseudomonas protegens Recombinant Expressing the cayG Gene

Suenaga,

Katayama,

Kitamura

et al. 2023

J. Org. Chem.

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Bacteria in certain genera can produce "bacterial polyynes" that contain a conjugated C�C bond starting from a terminal alkyne. Protegenin A is a derivative of octadecanoic acid that contains an ene−tetrayne moiety. It was discovered in Pseudomonas protegens Cab57 and exhibits strong antioomycete and moderate antifungal activity. By introducing cayG, a cytochrome P450 gene from Burkholderia caryophylli, into P. protegens Cab57, protegenin A was converted into more complex polyynes, caryoynencins A−E. A purification method that minimized the degradation and isomerization of caryoynencins was established. For the first time, as far as we know, the 1 H and 13 C{ 1 H} NMR signals of caryoynencins were completely assigned by analyzing the NMR data of the isolated compounds and protegenin A enriched with [1-13 C]-or [2-13 C]-acetate. Through the structural analysis of caryoynencins D/E and bioconversion experiments, we observed that CayG constructs the allyl alcohol moiety of caryoynencins A−C through sequential hydroxylation, dehydration, and hydroxylation. The recombinant strain exhibited a stronger antioomycete activity compared to the wild-type strain. This paper proposes a stable purification and structural determination method for various bacterial polyynes, and P. protegens Cab57 holds promise as an engineering host for the production of biologically active polyynes.

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“…The vector was introduced and expressed in P. protegens Cab57 . The recombinant was named Cab57- cayG .…”

Section: Resultsmentioning

confidence: 99%

“…CayG (1.2 kb, containing NdeI- and Sal I-digestion sites, no codon optimization) was synthesized by Eurofins Genomics and inserted into a modified pDSK519 to create pCcayG. This plasmid was electroporated into Cab57 or Δ proA cells, and the kanamycin-resistant (50 μg/mL) recombinant was selected.…”

Section: Methodsmentioning

confidence: 99%

Structures and Biosynthesis of Caryoynencins, Unstable Bacterial Polyynes from Pseudomonas protegens Recombinant Expressing the cayG Gene

Suenaga,

Katayama,

Kitamura

et al. 2023

J. Org. Chem.

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“…EPS I, the major extracellular polysaccharide in the RSSC, is a virulence factor in both Rs and Rps ; it is critical for colonization of the stem and vertical spread in the stem [33,35,36]. It has been assumed EPS I has a major role in Ralstonia biofilm development as it does in several well-studied bacteria and because it is a component of the biofilm ECM, but studies of static biofilm formation yielded mixed results [53,54]. To investigate this further in both static and flowing environments, we used an EPS I-deficient epsB mutant.…”

Section: Resultsmentioning

confidence: 99%

“…While RSSC strains produce copious amounts of EPS in vitro and in the stem at high cell densities, EPS has not been directly visualized in biofilms in planta [33,69]. A recent study on endoparasitism of Fusarium oxysporum showed EPS I is a core component of Rs biofilms on fungal hyphae [54]. In Rps OE1-1, EPS I contributes to biofilm formation on PVC plates but contrarily, an EPS I-deficient mutant formed a similar number of mature biofilms in ex vivo apoplastic fluid [53,54].…”

Section: Discussionmentioning

confidence: 99%

Lectins and polysaccharide EPS I have flow-responsive roles in the attachment and biofilm mechanics of plant pathogenicRalstonia

Carter,

Tran,

Cope-Arguello

et al. 2024

Preprint

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Bacterial biofilm formation and attachment to hosts are mediated by carbohydrate- binding lectins, exopolysaccharides, and their interactions in the extracellular matrix (ECM). During tomato infectionRalstonia pseudosolanacearum(Rps) GMI1000 highly expresses three lectins: LecM, LecF, and LecX. The latter two are uncharacterized. We evaluated the roles in bacterial wilt disease of LecF, a fucose-binding lectin, LecX, a xylose-binding lectin, and theRpsexopolysaccharide EPS I. Interestingly, single and double lectin mutants attached to tomato roots better and formed more biofilm under static conditionsin vitro. Consistent with this finding, static bacterial aggregation was suppressed by heterologous expression oflecFGMI1000andlecXGMI1000in otherRalstoniastrains that naturally lack these lectins. Crude ECM from a ΔlecF/Xdouble mutant was more adhesive than the wild-type ECM, and LecF and LecX increasedRpsattachment to ECM. The enhanced adhesiveness of the ΔlecF/XECM could explain the double mutant’s hyper-attachment in static conditions. Unexpectedly, mutating lectins decreasedRpsattachment and biofilm viscosity under shear stress, which this pathogen experiences in plant xylem. LecF, LecX, and EPS I were all essential for biofilm development in xylem fluid flowing through cellulose-coated microfluidic channels. These results suggest that under shear stress, LecF and LecX increaseRpsattachment by interacting with the ECM and plant cell wall components like cellulose. In static conditions such as on root surfaces and in clogged xylem vessels, the same lectins suppress attachment to facilitate pathogen dispersal. Thus,Rpslectins have a dual biological function that depends on the physical environment.Author SummaryBacterial wilt diseases caused byRalstoniaspecies inflict significant losses on diverse, globally important agricultural plants. The pathogen first colonizes roots and ultimately the water-transporting xylem. There it attaches to host cell walls and other bacterial cells to form biofilms that eventually block xylem vessels and disrupt sap flow. It is not well known howRalstoniaspp. modulate attachment, but precise control of both attachment and dispersal is critical for successful host colonization over the disease cycle. Excessive adhesion could trap bacteria in a toxic or nutrient-depleted environment. Conversely, insufficient adhesion in a flowing environment could displace bacteria from an optimal niche. We provide evidence of dual, environment-specific roles of carbohydrate-binding lectins and exopolysaccharide EPS I inRalstonia pseudosolanacearum(Rps) attachment. In static conditions, whichRpsexperiences on a host root, two lectins suppress bacterial aggregation and adhesion to roots. However, in flowing conditions, whichRpsexperiences in healthy xylem vessels, the same two lectins and EPS I are essential for biofilm development. The lectins increase the biofilm viscosity and support colony structural integrity, likely by interacting with polysaccharides in the biofilm matrix. This novel multifunctionality of bacterial lectins reveals how pathogens adapt to a physically dynamic host environment.

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“…1 × 1 mm 2 agar) or F. oxysporum NBRC 31213 , (2 μL spore suspension) was placed on WYA-GlcNAc (2 mL/well, in a 12-well assay plate). A paper disk (6 mm; Toyo Roshi Kaisha) containing the test sample was placed on the other side of the inoculation site.…”

Section: Experimental Sectionmentioning

confidence: 99%

Disproof of the Structures and Biosynthesis of Ergoynes, Gs-Polyyne–l-Ergothioneine Cycloadducts from Gynuella sunshinyii YC6258

Kawahara,

Kai

2024

J. Org. Chem.

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Some bacteria produce “bacterial polyynes” bearing a conjugated CC bond that starts with a terminal alkyne. Ergoynes A and B have been reported as sulfur-containing metabolites from Gynuella sunshinyii YC6258. These compounds were thought to be formed by cycloaddition between a bacterial polyyne (named Gs-polyyne) and l-ergothioneine. The biosynthetic gene clusters (BGCs), which may contribute to their synthesis, were present in the YC6258 genome. The biosynthetic origin of Gs-polyyne is interesting considering its rare 2-isopentyl fatty acyl skeleton. Here, the structures and biosynthesis of Gs-polyyne and ergoynes were verified by analytical, chemical, and genetic techniques. In the YC6258 extract, which was prepared considering their instability, Gs-polyyne was detected as a major LC peak, and ergoynes were not detected. The NMR data of the isolated Gs-polyyne contradicted the proposed structure and identified it as the previously reported protegenin A. The expression of Gs-polyyne BGC in Escherichia coli BL21(DE3) also yielded protegenin A. The cyclization between protegenin A and l-ergothioneine did not proceed during sample preparation; a base, such as potassium carbonate, was required. Overall, Gs-polyyne was identified as protegenin A, while ergoynes were determined to be artifacts. This cyclization may provide a derivatization to stabilize polyynes or create new chemical space.

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Quorum Sensing-Dependent Invasion of Ralstonia solanacearum into Fusarium oxysporum Chlamydospores

Cited by 6 publications

References 71 publications

Structures and Biosynthesis of Caryoynencins, Unstable Bacterial Polyynes from Pseudomonas protegens Recombinant Expressing the cayG Gene

Structures and Biosynthesis of Caryoynencins, Unstable Bacterial Polyynes from Pseudomonas protegens Recombinant Expressing the cayG Gene

Lectins and polysaccharide EPS I have flow-responsive roles in the attachment and biofilm mechanics of plant pathogenicRalstonia

Disproof of the Structures and Biosynthesis of Ergoynes, Gs-Polyyne–l-Ergothioneine Cycloadducts from Gynuella sunshinyii YC6258

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