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MBio Jun 2020is an alphaproteobacterium belonging to the Bacteria from this order elongate their cell wall at the new cell pole, generated by cell division. Screening for protein...
is an alphaproteobacterium belonging to the Bacteria from this order elongate their cell wall at the new cell pole, generated by cell division. Screening for protein interaction partners of the previously characterized polar growth factors RgsP and RgsM, we identified the inner membrane components of the Tol-Pal system (TolQ and TolR) and novel Rgs (rhizobial growth and septation) proteins with unknown functions. TolQ, Pal, and all Rgs proteins, except for RgsE, were indispensable for cell growth. Six of the Rgs proteins, TolQ, and Pal localized to the growing cell pole in the cell elongation phase and to the septum in predivisional cells, and three Rgs proteins localized to the growing cell pole only. The putative FtsN-like protein RgsS contains a conserved SPOR domain and is indispensable at the early stages of cell division. The components of the Tol-Pal system were required at the late stages of cell division. RgsE, a homolog of the growth pole ring protein GPR, has an important role in maintaining the normal growth rate and rod cell shape. RgsD is a periplasmic protein with the ability to bind peptidoglycan. Analysis of the phylogenetic distribution of the Rgs proteins showed that they are conserved in and mostly absent from other alphaproteobacterial orders, suggesting a conserved role of these proteins in polar growth. Bacterial cell proliferation involves cell growth and septum formation followed by cell division. For cell growth, bacteria have evolved different complex mechanisms. The most prevalent growth mode of rod-shaped bacteria is cell elongation by incorporating new peptidoglycans in a dispersed manner along the sidewall. A small share of rod-shaped bacteria, including the alphaproteobacterial , grow unipolarly. Here, we identified and initially characterized a set of Rgs (rhizobial growth and septation) proteins, which are involved in cell division and unipolar growth of and highly conserved in Our data expand the knowledge of components of the polarly localized machinery driving cell wall growth and suggest a complex of Rgs proteins with components of the divisome, differing in composition between the polar cell elongation zone and the septum.
Topics: Agrobacterium tumefaciens; Bacterial Proteins; Cell Cycle; Cell Division; Cell Polarity; Nucleotidases; Phylogeny; RGS Proteins; Rhizobiaceae; Schizosaccharomyces pombe Proteins; Sinorhizobium meliloti
PubMed: 32605980
DOI: 10.1128/mBio.00306-20 -
BMC Research Notes Apr 2022'Candidatus Liberibacter asiaticus' (CLas) is associated with the devastating citrus 'greening' disease. All attempts to achieve axenic growth and complete Koch's...
OBJECTIVE
'Candidatus Liberibacter asiaticus' (CLas) is associated with the devastating citrus 'greening' disease. All attempts to achieve axenic growth and complete Koch's postulates with CLas have failed to date, at best yielding complex cocultures with very low CLas titers detectable only by PCR. Reductive genome evolution has rendered all pathogenic 'Ca. Liberibacter' spp. deficient in multiple key biosynthetic, metabolic and structural pathways that are highly unlikely to be rescued in vitro by media supplementation alone. By contrast, Liberibacter crescens (Lcr) is axenically cultured and its genome is both syntenic and highly similar to CLas. Our objective is to achieve replicative axenic growth of CLas via addition of missing culturability-related Lcr genes.
RESULTS
Bioinformatic analyses identified 405 unique ORFs in Lcr but missing (or truncated) in all 24 sequenced CLas strains. Site-directed mutagenesis confirmed and extended published EZ-Tn5 mutagenesis data, allowing elimination of 310 of these 405 genes as nonessential, leaving 95 experimentally validated Lcr genes as essential for CLas growth in axenic culture. Experimental conditions for conjugation of large GFP-expressing plasmids from Escherichia coli to Lcr were successfully established for the first time, providing a practical method for transfer of large groups of 'essential' Lcr genes to CLas.
Topics: Axenic Culture; Citrus; Liberibacter; Plant Diseases; Rhizobiaceae
PubMed: 35365194
DOI: 10.1186/s13104-022-05986-5 -
Philosophical Transactions of the Royal... May 2014Trade-offs between individual fitness and the collective performance of crop and below-ground symbiont communities are common in agriculture. Plant competitiveness for...
Trade-offs between individual fitness and the collective performance of crop and below-ground symbiont communities are common in agriculture. Plant competitiveness for light and soil resources is key to individual fitness, but higher investments in stems and roots by a plant community to compete for those resources ultimately reduce crop yields. Similarly, rhizobia and mycorrhizal fungi may increase their individual fitness by diverting resources to their own reproduction, even if they could have benefited collectively by providing their shared crop host with more nitrogen and phosphorus, respectively. Past selection for inclusive fitness (benefits to others, weighted by their relatedness) is unlikely to have favoured community performance over individual fitness. The limited evidence for kin recognition in plants and microbes changes this conclusion only slightly. We therefore argue that there is still ample opportunity for human-imposed selection to improve cooperation among crop plants and their symbionts so that they use limited resources more efficiently. This evolutionarily informed approach will require a better understanding of how interactions among crops, and interactions with their symbionts, affected their inclusive fitness in the past and what that implies for current interactions.
Topics: Biological Evolution; Breeding; Crops, Agricultural; Genetic Fitness; Humans; Mycorrhizae; Rhizobiaceae; Selection, Genetic; Species Specificity; Symbiosis
PubMed: 24686938
DOI: 10.1098/rstb.2013.0367 -
Journal of Proteomics Jun 2022The Asian citrus psyllid, Diaphorina citri, is the vector of Candidatus Liberibacter asiaticus (CLas), the presumed causative agent of citrus greening disease. For...
The Asian citrus psyllid, Diaphorina citri, is the vector of Candidatus Liberibacter asiaticus (CLas), the presumed causative agent of citrus greening disease. For successful transmission, CLas must cross the gut barrier, requiring interaction with proteins on the midgut epithelium. We compared the relative abundance of gut surface proteins for both adult and nymph D. citri, as nymphs are particularly susceptible to CLas infection. To enrich for gut surface proteins, brush border membrane vesicles were prepared from dissected guts, and proteins identified from triplicate samples run on a timsTOF mass spectrometer. A total of 1516 and 1219 proteins were identified from D. citri adults and nymphs respectively. Based on bioinformatics analysis software and manual curation, 112 adult and 87 nymph proteins were predicted to localize to the surface of the microvilli and were further categorized into integral membrane and glycosylphosphatidylinositol (GPI)-anchored proteins. Proteins exploited by insect pathogens such as aminopeptidase, alkaline phosphatase, cadherin, ABC transporters, and carboxypeptidase were among the most abundant proteins on the gut surface. In addition to providing insights into hemipteran gut physiology, the D. citri gut surface proteome will inform novel approaches to interfere with CLas interaction with the psyllid gut to prevent the spread of citrus greening. BIOLOGICAL SIGNIFICANCE: The Asian citrus psyllid (ACP), D. citri is one of the most serious pests of citrus worldwide. ACP transmits the pathogenic bacterium that causes citrus greening or huanglongbing (HLB), which has resulted in severe economic losses in global citriculture. The putative causative agent of this disease, the gram-negative bacterium Candidatus Liberibacter asiaticus (CLas), is vectored by the Asian citrus psyllid, D. citri, in a persistent and circulative manner. CLas must interact with gut surface proteins in order to enter midgut epithelial cells. However, the specific proteins exploited by CLas have yet to be identified. The characterization of the most abundant proteins on the surface of the D. citri gut provides insight into candidate receptors for CLas and other pathogens of D. citri. We hypothesize that pathogens of D. citri exploit the most abundant proteins on the surface of the gut for entry into the host insect. Importantly, the abundant gut surface proteins will provide the basis for novel approaches to disrupt CLas-D. citri interactions, with the goal of preventing further economic loss to the citrus industry.
Topics: Animals; Citrus; Hemiptera; Membrane Proteins; Nymph; Plant Diseases; Rhizobiaceae
PubMed: 35427801
DOI: 10.1016/j.jprot.2022.104580 -
Genome Biology and Evolution Nov 2020Rhizobia are soil bacteria capable of forming symbiotic nitrogen-fixing nodules associated with leguminous plants. In fast-growing legume-nodulating rhizobia, such as... (Comparative Study)
Comparative Study
Rhizobia are soil bacteria capable of forming symbiotic nitrogen-fixing nodules associated with leguminous plants. In fast-growing legume-nodulating rhizobia, such as the species in the family Rhizobiaceae, the symbiotic plasmid is the main genetic basis for nitrogen-fixing symbiosis, and is susceptible to horizontal gene transfer. To further understand the symbioses evolution in Rhizobiaceae, we analyzed the pan-genome of this family based on 92 genomes of type/reference strains and reconstructed its phylogeny using a phylogenomics approach. Intriguingly, although the genetic expansion that occurred in chromosomal regions was the main reason for the high proportion of low-frequency flexible gene families in the pan-genome, gene gain events associated with accessory plasmids introduced more genes into the genomes of nitrogen-fixing species. For symbiotic plasmids, although horizontal gene transfer frequently occurred, transfer may be impeded by, such as, the host's physical isolation and soil conditions, even among phylogenetically close species. During coevolution with leguminous hosts, the plasmid system, including accessory and symbiotic plasmids, may have evolved over a time span, and provided rhizobial species with the ability to adapt to various environmental conditions and helped them achieve nitrogen fixation. These findings provide new insights into the phylogeny of Rhizobiaceae and advance our understanding of the evolution of symbiotic nitrogen fixation.
Topics: Evolution, Molecular; Genome, Bacterial; Nitrogen Fixation; Phylogeny; Plasmids; Rhizobiaceae; Symbiosis
PubMed: 32687170
DOI: 10.1093/gbe/evaa152 -
Microbiology Spectrum Aug 2022Liberibacter solanacearum (CLso) haplotype D, transmitted by the carrot psyllid , is a major constraint for carrot production in Israel. Unveiling the molecular...
Liberibacter solanacearum (CLso) haplotype D, transmitted by the carrot psyllid , is a major constraint for carrot production in Israel. Unveiling the molecular interactions between the psyllid vector and CLso can facilitate the development of nonchemical approaches for controlling the disease caused by CLso. Bacterial surface proteins are often known to be involved in adhesion and virulence; however, interactions of CLso with carrot psyllid proteins that have a role in the transmission process has remained unexplored. In this study, we used CLso outer membrane protein (OmpA) and flagellin as baits to screen for psyllid interacting proteins in a yeast two-hybrid system assay. We identified psyllid vitellogenin (Vg) to interact with both OmpA and flagellin of CLso. As Vg and autophagy are often tightly linked, we also studied the expression of autophagy-related genes to further elucidate this interaction. We used the juvenile hormone (JH-III) to induce the expression of Vg, thapsigargin for suppressing autophagy, and rapamycin for inducing autophagy. The results revealed that Vg negatively regulates autophagy. Induced Vg expression significantly suppressed autophagy-related gene expression and the levels of CLso significantly increased, resulting in a significant mortality of the insect. Although the specific role of Vg remains obscure, the findings presented here identify Vg as an important component in the insect immune responses against CLso and may help in understanding the initial molecular response in the vector against Liberibacter. Pathogen transmission by vectors involves multiple levels of interactions, and for the transmission of liberibacter species by psyllid vectors, much of these interactions are yet to be explored. Liberibacter solanacearum (CLso) haplotype D inflicts severe economic losses to the carrot industry. Understanding the specific interactions at different stages of infection is hence fundamental and could lead to the development of better management strategies to disrupt the transmission of the bacteria to new host plants. Here, we show that two liberibacter membrane proteins interact with psyllid vitellogenin and also induce autophagy. Altering vitellogenin expression directly influences autophagy and CLso abundance in the psyllid vector. Although the exact mechanism underlying this interaction remains unclear, this study highlights the importance of immune responses in the transmission of this disease agent.
Topics: Animals; Autophagy; Flagellin; Hemiptera; Liberibacter; Plant Diseases; Rhizobiaceae; Vitellogenins
PubMed: 35863005
DOI: 10.1128/spectrum.01577-22 -
Journal of Bacteriology Jan 1991The lack of high-resolution genetic or physical maps for the family Rhizobiaceae limits our understanding of this agronomically important bacterial family. On the basis...
The lack of high-resolution genetic or physical maps for the family Rhizobiaceae limits our understanding of this agronomically important bacterial family. On the basis of statistical analyses of DNA sequences of the Rhizobiaceae and direct evaluation by pulsed-field agarose gel electrophoresis (PFE), five restriction endonucleases with AT-rich target sites were identified as the most rarely cutting: AseI (5'-ATTAAT-3'), DraI (5'-TTTAAA-3'), SpeI (5'-ACTAGT-3'), SspI (5'-AATAAT-3'), and XbaI (5'-TCTAGA-3'). We computed the sizes of the genomes of Bradyrhizobium japonicum USDA 424 and Rhizobium meliloti 1021 by adding the sizes of DNA fragments generated by SpeI digests. The genome sizes of R. meliloti 1021 and B. japonicum USDA 424 were 5,379 +/- 282.5 kb and 6,195 +/- 192.4 kb, respectively. We also compared the organization of the genomes of free-living and bacteroid forms of B. japonicum. No differences between the PFE-resolved genomic fingerprints of free-living and mature (35 days after inoculation) bacteroids of B. japonicum USDA 123 and USDA 122 were observed. Also, B. japonicum USDA 123 genomic fingerprints were unchanged after passage through nodules and after maintenance on a rich growth medium for 100 generations. We conclude that large-scale DNA rearrangements are not seen in mature bacteroids or during free-living growth on rich growth media under laboratory conditions.
Topics: Base Sequence; DNA Restriction Enzymes; DNA, Bacterial; Electrophoresis, Agar Gel; Genes, Bacterial; Molecular Sequence Data; Rhizobiaceae; Species Specificity; Substrate Specificity
PubMed: 1846148
DOI: 10.1128/jb.173.2.704-709.1991 -
International Journal of Molecular... Dec 2022" Liberibacter asiaticus" (CLas) is a phloem-restricted α-proteobacterium that is associated with citrus huanglongbing (HLB), which is the most destructive disease that...
Integrated Transcriptome and Metabolome Analysis Reveals Phenylpropanoid Biosynthesis and Phytohormone Signaling Contribute to " Liberibacter asiaticus" Accumulation in Citrus Fruit Piths (Fluffy Albedo).
" Liberibacter asiaticus" (CLas) is a phloem-restricted α-proteobacterium that is associated with citrus huanglongbing (HLB), which is the most destructive disease that affects all varieties of citrus. Although midrib is usually used as a material for CLas detection, we recently found that the bacterium was enriched in fruits, especially in the fruit pith. However, no study has revealed the molecular basis of these two parts in responding to CLas infection. Therefore, we performed transcriptome and UHPLC-MS-based targeted and untargeted metabolomics analyses in order to organize the essential genes and metabolites that are involved. Transcriptome and metabolome characterized 4834 differentially expressed genes (DEGs) and 383 differentially accumulated metabolites (DAMs) between the two materials, wherein 179 DEGs and 44 DAMs were affected by HLB in both of the tissues, involving the pathways of phenylpropanoid biosynthesis, phytohormone signaling transduction, starch and sucrose metabolism, and photosynthesis. Notably, we discovered that the gene expression that is related to beta-glucosidase and endoglucanase was up-regulated in fruits. In addition, defense-related gene expression and metabolite accumulation were significantly down-regulated in infected fruits. Taken together, the decreased amount of jasmonic acid, coupled with the reduced accumulation of phenylpropanoid and the increased proliferation of indole-3-acetic acid, salicylic acid, and abscisic acid, compared to leaf midribs, may contribute largely to the enrichment of CLas in fruit piths, resulting in disorders of photosynthesis and starch and sucrose metabolism.
Topics: Citrus; Liberibacter; Transcriptome; Plant Growth Regulators; Rhizobiaceae; Metabolome; Sucrose; Plant Diseases
PubMed: 36555287
DOI: 10.3390/ijms232415648 -
Microbiological Research Feb 2020Induction of systemic tolerance in sorghum [Sorghum bicolor (L.) Moench] against drought stress was studied by screening a large collection of rhizobacterial isolates...
Induction of systemic tolerance in sorghum [Sorghum bicolor (L.) Moench] against drought stress was studied by screening a large collection of rhizobacterial isolates for their potential to exhibit this essential plant growth-promoting trait. This was done by means of a greenhouse assay that measured the relative change in both plant height and -biomass (roots and shoots) between rhizobacteria-primed versus non-primed (naïve) plants under drought stress conditions. In order to elucidate the metabolomic changes in S. bicolor that conferred the drought stress tolerance after treatment (priming) with selected isolates, untargeted ultra-high performance liquid chromatography-high definition mass spectrometry (UHPLC-HDMS)-based metabolomics was carried out. Intracellular metabolites were methanol-extracted from rhizobacteria-primed and naïve S. bicolor roots and shoots. Extracts were analysed on a UHPLC-HDMS system and the generated data were chemometrically mined to determine signatory metabolic profiles and bio-markers related to induced systemic tolerance. The metabolomic results showed significant treatment-related differential metabolic reprogramming between rhizobacteria-primed and naïve plants, correlating to the ability of the selected isolates to protect S. bicolor against drought stress. The selected isolates, identified by means of 16S rRNA gene sequencing as members of the genera Bacillus and Pseudomonas, were screened for 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity by means of an in vitro assay and the presence of the acdS gene was subsequently confirmed by PCR for strain N66 (Pseudomonas sp.). The underlying key metabolic changes in the enhanced drought stress tolerance observed in rhizobacteria-primed S. bicolor plants included (1) augmented antioxidant capacity; (2) growth promotion and root architecture modification as a result of the upregulation of the hormones gibberellic acid, indole acetic acid and cytokinin; (3) the early activation of induce systemic tolerance through the signalling hormones brassinolides, salicylic acid and jasmonic acid and signalling molecules sphingosine and psychosine; (4) the production of the osmolytes proline, glutamic acid and choline; (5) the production of the epicuticular wax docosanoic acid and (6) ACC deaminase activity resulting in lowered ethylene levels. These results unravelled key molecular details underlying the PGPR-induced systemic tolerance in sorghum plants, providing insights for the plant priming for abiotic stress.
Topics: Adaptation, Physiological; Bacillus; Biomass; Droughts; Plant Development; Plant Roots; Pseudomonas; RNA, Ribosomal, 16S; Rhizobiaceae; Rhizobium; Soil; Soil Microbiology; Sorghum; Stress, Physiological
PubMed: 31865223
DOI: 10.1016/j.micres.2019.126388 -
Phytopathology Jul 2015Zebra chip (ZC) of potato is putatively caused by the fastidious, phloem-limited bacterium 'Candidatus Liberibacter solanacearum' (Lso), which is transmitted by the... (Review)
Review
Zebra chip (ZC) of potato is putatively caused by the fastidious, phloem-limited bacterium 'Candidatus Liberibacter solanacearum' (Lso), which is transmitted by the potato psyllid (Bactericera cockerelli). The disease, which significantly impacts both crop yield and quality, was first identified in the United States from south Texas in 2000. It reached epidemic levels in north Texas and certain production areas in Colorado, Nebraska, and New Mexico from 2004 to 2007 and it caused severe losses in fields in Oregon, Washington, and Idaho in 2011. The potato plant is susceptible to infection at all developmental stages, but disease management programs have focused on vector control through early and repeated insecticide applications, in an effort to minimize early to midseason infections which are most damaging. Growers often terminate spray programs 2 to 3 weeks prior to crop harvest due to lack of visible treatment effects on crop yield or quality. However, recent studies on vector transmission and host-pathogen interactions have revealed that late-season infections pose a significant, previously unrecognized, threat to crop quality. The pathogen can move from an infected leaf to tubers within 2 days; however, tubers infected less than 1 week before harvest will remain asymptomatic and the pathogen will be undetectable. When these tubers are placed into storage they are assumed to be disease free. However, Lso can continue to multiply in respiring tubers during storage, resulting in reduced tuber quality. Likewise, if plants become infected a few days before vines are killed, ZC can continue to develop in infected tubers before they are harvested. Perspectives on the significance of late-season infections and some of the more important issues associated with those infections are discussed.
Topics: Animals; Hemiptera; Host-Pathogen Interactions; Insect Control; Insect Vectors; Plant Diseases; Rhizobiaceae; Seasons; Solanum tuberosum
PubMed: 25894320
DOI: 10.1094/PHYTO-12-14-0365-FI