Publications

4364

Microbial communities with the ability to convert long-chain fatty acids (LCFA) coupled to sulfate reduction can be important in the removal of these compounds from wastewater. In this work, an enrichment culture, able to oxidize the long-chain fatty acid palmitate (C16:0) coupled to sulfate reduction, was obtained from anaerobic granular sludge. Microscopic analysis of this culture, designated HP culture, revealed that it was mainly composed of one morphotype with a typical collar-like cell wall invagination, a distinct morphological feature of theDesulfomonilegenus. 16S rRNA gene amplicon and metagenome-assembled genome (MAG) indeed confirmed that the abundant phylotype in HP culture belong toDesulfomonilegenus [ca.92% 16S rRNA gene sequences closely related toDesulfomonilespp.; andca. 82% whole genome shotgun (WGS)]. Based on similar cell morphology and average nucleotide identity (ANI) (77%) between theDesulfomonilesp. in HP culture and the type strainDesulfomonile tiedjeistrain DCB-1T, we propose a novel species designated as “CandidatusDesulfomonile palmitatoxidans.” This bacterium shares 94.3 and 93.6% 16S rRNA gene identity withDesulfomonile limimarisstrain DCB-MTandD. tiedjeistrain DCB-1T, respectively. Based on sequence abundance ofDesulfomonile-morphotype in HP culture, its predominance in the microscopic observations, and presence of several genes coding for enzymes involved in LCFA degradation, the proposed species “Ca.Desulfomonile palmitatoxidans” most probably plays an important role in palmitate degradation in HP culture. Analysis of the growth of HP culture andD. tiedjeistrain DCB-1Twith short- (butyrate), medium- (caprylate) and long-chain fatty acids (palmitate, stearate, and oleate) showed that both cultures degraded all fatty acids coupled to sulfate reduction, except oleate that was only utilized by HP culture. In the absence of sulfate, neither HP culture, norD. tiedjeistrain DCB-1Tdegraded palmitate when incubated withMethanobacterium formicicumas a possible methanogenic syntrophic partner. UnlikeD. tiedjeistrain DCB-1T, “Ca.Desulfomonile palmitatoxidans” lacks reductive dehalogenase genes in its genome, and HP culture was not able to grow by organohalide respiration. An emended description of the genusDesulfomonileis proposed. Our study reveals an unrecognized LCFA degradation feature of theDesulfomonilegenus.

AbstractA key feature that differentiates prokaryotic cells from eukaryotes is the absence of an intracellular membrane surrounding the chromosomal DNA. Here, we isolate a member of the ubiquitous, yet-to-be-cultivated phylum ‘Candidatus Atribacteria’ (also known as OP9) that has an intracytoplasmic membrane apparently surrounding the nucleoid. The isolate, RT761, is a subsurface-derived anaerobic bacterium that appears to have three lipid membrane-like layers, as shown by cryo-electron tomography. Our observations are consistent with a classical gram-negative structure with an additional intracytoplasmic membrane. However, further studies are needed to provide conclusive evidence for this unique intracellular structure. The RT761 genome encodes proteins with features that might be related to the complex cellular structure, including: N-terminal extensions in proteins involved in important processes (such as cell-division protein FtsZ); one of the highest percentages of transmembrane proteins among gram-negative bacteria; and predicted Sec-secreted proteins with unique signal peptides. Physiologically, RT761 primarily produces hydrogen for electron disposal during sugar degradation, and co-cultivation with a hydrogen-scavenging methanogen improves growth. We propose RT761 as a new species, Atribacter laminatus gen. nov. sp. nov. and a new phylum, Atribacterota phy. nov.

AbstractA recent survey in Germany revealed the wide presence of ‘Candidatus Phytoplasma ulmi’ in native elm stands. Accessions were studied for their genetic variability and phylogenetic relationship based on the conserved groEL and the variable imp gene. While the groEL sequences revealed a high intraspecific homology of more than 99%, the homology of the imp gene dropped to 71% between distantly related sequences. Twenty-nine groEL and 74 imp genotypes were distinguished based on polymorphic sites. Phylogenetic analysis of the groEL gene clustered all ‘Ca. P. ulmi’ strains and separated them from related phytoplasmas of the 16SrV group. The inferred phylogeny of the imp gene resulted in a different tree topology and separated the ‘Ca. P. ulmi’ genotypes into two clusters, one closely related to the flavescence dorée phytoplasma strain FD-D (16SrV-D), the other affiliated with the flavescence dorée phytoplasma strains FD-C and FD70 and the alder yellows phytoplasma (16SrV-C). In both phylograms, ‘Ca. P. ulmi’ genotypes from Scots elm trees formed a coherent cluster, while genotypes from European white elms and field elms grouped less strictly. The regional distribution pattern was congruent for some of the groEL and imp genotypes, but a strict linkage for all genotypes was not apparent.

<p>Las bacterias causantes del huanglongbing (HLB), <em>Canditatus</em> Liberibacter spp., son bacterias obligadas al floema de los cítricos y a diferentes sistemas del insecto vector,<em> Diaphorina citri</em>, por lo tanto, el enfoque genómico ha sido útil para estudiar sus mecanismos de patogenicidad. Dicho enfoque ha permitido identificar una copia homóloga del gen codificante de la enzima ATP/ADP translocasa, la cual tiene la capacidad de importar ATP y nucleótidos desde el hospedante, causando un parasitismo de energía considerable. Esta enzima se ha relacionado con la actividad endoparasítica de patógenos animales y humanos más que con fitopatógenos. El presente trabajo analizó la relación evolutiva entre la secuencia de aminoácidos de la ATP/ADP translocasa entre diferentes especies de <em>Ca</em>. Liberibacter y grupos como <em>Ricketssia</em> sp. y <em>Chlamydia</em> sp. Análisis filogenéticos muestran que la variación en la secuencia del gen codificante de la enzima está delimitada en clados correspondientes a las especies de <em>Ca</em>. Liberibacter, sugiriendo que la variación en la enzima responde a un proceso coevolutivo con los hospederos. Asimismo, la filogenia muestra que el ancestro común más cercano a <em>Ca</em>. Liberibacter podría ser un endosimbionte no patogénico del género <em>Ca</em>. Midichloria. Análisis de conservación de la secuencia de aminoácidos muestran que existen varias posiciones en la secuencia que podrían estar relacionadas con la variación específica de esta enzima dentro de <em>Ca</em>. Liberibacter. Este trabajo presenta la hipótesis de que el origen evolutivo de la capacidad de parasitismo energético del género <em>Ca</em>. Liberibacter, causantes del HLB, es un endosimbionte no patogénico.</p>

Abstract The potato psyllid, Bactericera cockerelli (Šulc), is a major pest of potato (Solanum tuberosum L.; Solanales: Solanaceae) as a vector of ‘Candidatus Liberibacter solanacearum’, the pathogen that causes zebra chip. Management of zebra chip is challenging in part because the noncrop sources of Liberibacter-infected psyllids arriving in potato remain unknown. Adding to this challenge is the occurrence of distinct genetic haplotypes of both potato psyllid and Liberibacter that differ in host range. Longleaf groundcherry (Physalis longifolia Nutt.) has been substantially overlooked in prior research as a potential noncrop source of Liberibacter-infected B. cockerelli colonizing fields of potato. The objective of this study was to assess the suitability of P. longifolia to the three common haplotypes of B. cockerelli (central, western, and northwestern haplotypes), and to two haplotypes of ‘Ca. L. solanacearum’ (Liberibacter A and B haplotypes). Greenhouse bioassays indicated that B. cockerelli of all three haplotypes produced more offspring on P. longifolia than on potato and preferred P. longifolia over potato during settling and egg-laying activities. Greenhouse and field trials showed that P. longifolia was also highly susceptible to Liberibacter. Additionally, we discovered that infected rhizomes survived winter and produced infected plants in late spring that could then be available for psyllid colonization and pathogen acquisition. Results show that P. longifolia is susceptible to both B. cockerelli and ‘Ca. L. solanacearum’ and must be considered as a potentially important source of infective B. cockerelli colonizing potato fields in the western United States.

Citrus huanglongbing (HLB) is a devastating disease for the citrus industry. The previous studies demonstrated that oxytetracycline and penicillin are effective antibiotics against Candidatus Liberibacter asiaticus (CLas). However, since CLas is uncultured, the mechanisms of action of antibiotics against CLas are still unclear. It was recently reported that the endophytic microbial communities are associated with the progression of citrus HLB after oxytetracycline and penicillin treatment. Therefore, we hypothesize that penicillin has greater antibacterial activity against CLas than oxytetracycline, which may be associated with the alteration of the structure and function of endophytic microbial communities in HLB-affected citrus in response to these antibiotics. To test this hypothesis, the microbiome of HLB-affected citrus leaves treated with these two antibiotics was analyzed using a metagenomic method. Our results indicate that the microbial structure and function in HLB-affected citrus were altered by these two antibiotics. The relative abundance of beneficial bacterial species, including Streptomyces avermitilis and Bradyrhizobium, was higher in penicillin-treated plants compared to those treated with oxytetracycline, and the relative abundance of the bacterial species (such as Propionibacterium acnes and Synechocystis sp PCC 6803) associated with CLas survival was lower for penicillin-treated plants compared to oxytetracycline-treated plants. These results indicate that penicillin has greater antibacterial activity against CLas. Based on the metagenomic analysis, this study elucidated the mechanism for the observed increase in antibacterial activity of penicillin against CLas. The data presented here are not only invaluable for developing eco-friendly and effective biocontrol strategies to combat citrus HLB, but also provide a method for revealing mechanism of antimicrobial against uncultured bacteria in host.

Health, growth, fruit production, and fruit quality of citrus crops are severely affected by tree infection with Candidatus Liberibacter asiaticus (CLas) and subsequent development of the disease huanglongbing (HLB). The use of HLB-tolerant rootstocks is one strategy that is used to ameliorate the effects of HLB in commercial production. Although there is a clear long-term rootstock effect to improve tree performance, such field trials take many years for establishment and evaluation, and this long time-period is very limiting to expeditious evaluation of new rootstocks. In this study, we have conducted a 50-week greenhouse experiment to evaluate rootstock influences on Valencia sweet orange tree response to CLas infection. The infection of trees with CLas reduced scion and rootstock growth, increased leaf yellowing, and reduced the number of leaves per tree and leaf area, regardless of rootstock. There were clear rootstock influences on some traits during the 50-week study. In general, infected trees on US-942 rootstock had lower CLas root titers, less reduction of the number of leaves, less reduction of leaf area, and less leaf yellowing, as compared with some of the other rootstocks. The 50-week greenhouse evaluation method provided results that corresponded well with results from long-term field testing, indicating this may be a useful tool to accelerate evaluation and selection of new rootstocks, as well as in testing other HLB management strategies.