Publications

4365

Recently, a novel species of the genusBorreliawas identified inBothriocroton concolorandIxodes holocyclusticks from echidnas. Analyses of 16S rRNA andflaBgenes identified three closely related genotypes of this bacterium (Borreliasp. Aus A-C) that were unique and distinct from previously described borreliae. Phylogenetic analyses offlaB(763 bp),groEL(1537 bp),gyrB(1702 bp) andglpQ(874 bp) gene sequences and concatenated sequences (3585 bp) of three gene loci (16S rRNA,flaBandgyrB) were consistent with previous findings and confirm that this novel species of the genusBorreliais more closely related to, yet distinct from, the Reptile-associated (REP) and Relapsing Fever (RF) groups. At theflaBlocus, genotypes A, B and C shared the highest percentage sequence similarities (87.9, 88 and 87.9 %, respectively) withB.orrelia turcica(REP), whereas at thegroELandgyrBloci, these genotypes were most similar (88.2–89.4 %) toB.orrelia hermsii(RF). At theglpQlocus, genotypes A and B were most similar (85.7 and 85.4 % respectively) toBorreliasp. Tortoise14H1 (REP). The presence of theglpQgene, which is absent in the Lyme Borreliosis group spirochaetes, further emphasises that the novel species of the genusBorreliacharacterized in the present study does not belong to this group. Phylogenetic analyses at multiple loci produced consistent topographies revealing the monophyletic grouping of this bacterium, therefore providing strong support for its species status. We propose the name ‘CandidatusBorreliatachyglossi’, and hypothesize that this species of the genusBorreliamay be endemic to Australia. The pathogenic potential of this bacterium is not yet known.

A recently described symbiosis between the metabolically streamlined nitrogen‐fixing cyanobacterium UCYN‐A and a single‐celled eukaryote prymnesiophyte alga is widely distributed throughout tropical and subtropical marine waters, and is thought to contribute significantly to nitrogen fixation in these regions. Several UCYN‐A sublineages have been defined based on UCYN‐A nitrogenase (nifH) sequences. Due to the low abundances of UCYN‐A in the global oceans, currently existing molecular techniques are limited for detecting and quantifying these organisms. A targeted approach is needed to adequately characterize the diversity of this important marine cyanobacterium, and to advance understanding of its ecological importance. We present findings on the distribution of UCYN‐A sublineages based on high throughput sequencing of UCYN‐A nifH PCR amplicons from 78 samples distributed throughout many major oceanic provinces. These UCYN‐A nifH fragments were used to define oligotypes, alternative taxonomic units defined by nucleotide positions with high variability. The data set was dominated by a single oligotype associated with the UCYN‐A1 sublineage, consistent with previous observations of relatively high abundances in tropical and subtropical regions. However, this analysis also revealed for the first time the widespread distribution of the UCYN‐A3 sublineage in oligotrophic waters. Furthermore, distinct assemblages of UCYN‐A oligotypes were found in oligotrophic and coastally influenced waters. This unique data set provides a framework for determining the environmental controls on UCYN‐A distributions and the ecological importance of the different sublineages.

The major citrus area of Brazil occupies near 450,000 ha between the Triângulo Mineiro (TM) region of Minas Gerais State and the south of São Paulo State (SPS). Significant climatic variation occurs between regions which could affect huanglongbing (HLB) progress, which is lower in TM. To investigate this possibility, young sweet orange shoots were sampled periodically over 2 years to determine ‘Candidatus Liberibacter asiaticus’ titers in naturally infected trees in orchards in Analândia, central SPS, and Frutal and Comendador Gomes, within TM. Data-loggers recorded local temperature and relative humidity hourly. In the laboratory, five ‘Ca. L. asiaticus’-free Diaphorina citri adults were placed on each sampled shoot for 48 h to feed and acquire the pathogen. Shoots and insects were individually analyzed by quantitative polymerase chain reaction to determine ‘Ca. L. asiaticus’ titers. The incidence of ‘Ca. L. asiaticus’-positive shoots, ‘Ca. L. asiaticus’ titers, and acquisition rates were lower for shoots from Comendador Gomes than those from Frutal or Analândia. Stronger association was observed between ‘Ca. L. asiaticus’ titers and the number of hours below 15°C (h < 15°C) or above 30°C (h > 30°C), and cumulative rainfall registered during the 15 days prior to sampling of shoots on each occasion. ‘Ca. L. asiaticus’ titers associated positively with h < 15°C and rainfall and negatively with h > 30°C. The slower spread and lower incidence of HLB in TM may be related to lower incidences of ‘Ca. L. asiaticus’-positive young shoots and lower titers of ‘Ca. L. asiaticus’ in the same shoots as a consequence of the warmer and drier conditions.

AbstractThe global deep subsurface biosphere is one of the largest reservoirs for microbial life on our planet. This study takes advantage of new sampling technologies and couples them with improvements to DNA sequencing and associated informatics tools to reconstruct the genomes of uncultivated Bacteria and Archaea from fluids collected deep within the Juan de Fuca Ridge subseafloor. Here, we generated two metagenomes from borehole observatories located 311 meters apart and, using binning tools, retrieved 98 genomes from metagenomes (GFMs). Of the GFMs, 31 were estimated to be >90% complete, while an additional 17 were >70% complete. Phylogenomic analysis revealed 53 bacterial and 45 archaeal GFMs, of which nearly all were distantly related to known cultivated isolates. In the GFMs, abundant Bacteria included Chloroflexi, Nitrospirae, Acetothermia (OP1), EM3, Aminicenantes (OP8), Gammaproteobacteria, and Deltaproteobacteria, while abundant Archaea included Archaeoglobi, Bathyarchaeota (MCG), and Marine Benthic Group E (MBG-E). These data are the first GFMs reconstructed from the deep basaltic subseafloor biosphere, and provide a dataset available for further interrogation.

El objetivo fue determinar diferencias en la actividad enzimática antioxidante y la concentración de fenoles totales en plantas de limón mexicano (Citrus aurantifolia (Christm.) Swingle), limón persa (Citrus latifolia Tanaka); y naranja dulce Valencia (Citrus sinensis (L.) Osbeck), infectadas bajo condiciones naturales de campo con Candidatus Liberibacter asiaticus (CaLas), en Bustamante, Tamaulipas, México. Se determinó la presencia de CaLas por qPCR; la actividad enzimática de fenilalanina amonio liasa (PAL), que se activa ante el ataque de patógenos; peroxidasa que interviene en la lignificación de paredes como respuesta a la infección; α-amilasa que degrada el almidón y se acumula en el tejido vascular, y compuestos fenólicos que cumplen funciones de defensa. Se realizó un análisis de contrastes. La concentración de proteínas totales mostró diferencias significativas entre especies entre especies (P< 0.0001). Con una media de 6.1 y 6.37 mg de proteínas por g de tejido fresco para árboles infectados de limón mexicano; y naranja dulce negativa a CaLas. La actividad peroxidasa presentó diferencia significativa para limón persa (P=0.0341), con una media de 1.96 U.mg de proteína-<sup>1</sup>. En naranja dulce se observó mayor actividad de α amilasa en los árboles infectados por CaLas (1.19 U.mg de proteína-<sup>1</sup>). La concentración de PAL y fenoles totales no presentaron diferencias significativas entre especies. En el presente estudio se observó que CaLas influye en la actividad enzimática de cítricos.

Abstract Ni.tro.so.cal.da'les. N.L. masc. n. Nitrosocaldus type genus of the order; ‐ ales ending to denote order; N.L. fem. pl. n. Nitrosocaldales the Nitrosocaldus order. Coccoid cells reproduce by binary fission. Thermophilic and neutrophilic. Obligately aerobic. Chemolithoautotrophic growth by ammonia oxidation to nitrite using CO 2 as carbon source. Some members can also use urea as an alternative source of energy for growth. Cell membrane is composed of isoprenoid tetraether lipids, including large amounts of acyclic and cyclized glycerol trialkyl glycerol tetraethers (GTGTs) and crenarchaeol (a unique glycerol dibiphytanyl glycerol tetraether (GDGT) containing one cyclohexane ring and four cyclopentane rings). Fully saturated and monounsaturated menaquinones with six isoprenoid units are present. They are globally distributed in various geothermal environments. Phylogenetic analyses of 16S rRNA and amoA gene sequences place the monophyletic order Ca . Nitrosocaldales as a basal lineage within the phylum Thaumarchaeota . This order can be distinguished from the closest order Nitrososphaerales by a lower mol% G + C content, a higher growth temperature (>60°C), and a higher relative abundance of GTGT. The order Ca . Nitrosocaldales is currently represented by a single family Ca . Nitrosocaldaceae. Type genus : Ca . Nitrosocaldus de la Torre, Walker, Ingalls, Könneke and Stahl 2008, 815. Taxonomic and Nomenclature Notes According to the List of Prokaryotic names with Standing in Nomenclature (LPSN), the taxonomic status of the order Candidatus Nitrosocaldales is: preferred name (not correct name) (last update, February 2025) * . LPSN classification: Archaea / Thermoproteati / Thermoproteota / incertae sedis / Candidatus Nitrosocaldales The order Candidatus Nitrosocaldales can also be recovered in the Genome Taxonomy Database (GTDB) as o__Nitrososphaerales (version v220) ** . GTDB classification: d__Archaea / p__Thermoproteota / c__Nitrososphaeria / o__Nitrososphaerales * Meier‐Kolthoff et al. ( 2022 ). Nucleic Acids Res , 50 , D801 – D807 ; DOI: 10.1093/nar/gkab902 ** Parks et al. ( 2022 ). Nucleic Acids Res , 50 , D785 – D794 ; DOI: 10.1093/nar/gkab776

Abstract Cen.ar.chae'um., Gr. adj. kainos recent, and Gr. adj. koinos common; Gr. adj. archaios ancient; N.L. neut. n. Cenarchaeum = genus of relatively recent (derived nonthermophilic phenotype) and common (non‐“extremophilic”) Archaea. The species Candidatus Cenarchaeum symbiosum of the genus Candidatus Cenarchaeum constitutes the sole archaeal symbiont of the temperate marine sponge Axinella mexicana . It represented the first tractable organism of the so‐called mesophilic crenarchaeota, now Nitrososphaeria, until pure isolates and enrichment cultures became available. It was also the source of the first assembled full genome of the Nitrososphaeria . Physiological and genomic analyses suggest that representatives of this genus are aerobic, psychrophilic, and capable of a chemolithoautotrophic metabolism, by oxidizing ammonia to nitrite. The environmental distribution of the genus so far has been confined to sponge‐associated communities. DNA G + C content (mol%) : 57.74. Type species : Candidatus Cenarchaeum symbiosum Preston, Wu, Molinsky and DeLong 1996, 6246. Taxonomic and Nomenclature Notes According to the List of Prokaryotic names with Standing in Nomenclature (LPSN), the taxonomic status of the genus Candidatus Cenarchaeum is: preferred name (not correct name) (last update, February 2025) * . LPSN classification: Archaea / Thermoproteati / Thermoproteota / Nitrososphaeria / Nitrosopumilales / Nitrosopumilaceae / Candidatus Cenarchaeum The genus Candidatus Cenarchaeum can also be recovered in the Genome Taxonomy Database (GTDB) as g__Cenarchaeum (version v220) ** . GTDB classification: d__Archaea / p__Thermoproteota / c__Nitrososphaeria / o__Nitrososphaerales / f__Nitrosopumilaceae / g__Cenarchaeum * Meier‐Kolthoff et al. ( 2022 ). Nucleic Acids Res , 50 , D801 – D807 ; DOI: 10.1093/nar/gkab902 ** Parks et al. ( 2022 ). Nucleic Acids Res , 50 , D785 – D794 ; DOI: 10.1093/nar/gkab776

Abstract Ni.tro.so.cal.da.ce'ae. N.L. masc. n. Nitrosocaldus type genus of the family; ‐ aceae ending to denote family; N.L. fem. pl. n. Nitrosocaldaceae the Nitrosocaldus family. Cells are irregular cocci and divide by binary fission. Strictly aerobic. Thermophilic, neutrophilic, and lithoautotrophic archaea that are able to gain energy via ammonia oxidation to nitrite. Membrane lipids contain glycerol dibiphytanyl glycerol tetraethers (GDGTs) bound to monoglycosidic, diglycosidic, phosphohexose, and hexose‐phosphohexose headgroups. Like other described Thaumarchaeota , members of this family contain crenarchaeol as a major membrane core lipid. However, their membrane lipids are characterized by the presence of the high abundances of acyclic and cyclized glycerol trialkyl glycerol tetraethers (GTGTs). Respiratory lipoquinones are saturated and monounsaturated menaquinones with six isoprenoid units. Cells and molecular markers have been found in neutral or slightly alkaline geothermal environments, such as hot springs and subsurface gold mines. At present, the family Ca . Nitrosocaldaceae only harbors a single genus, which is so far represented by only one species. Therefore, future phylogenetic analysis of additional cultivated representatives may reveal further subdivisions within the family. DNA G+C content (mol%):36.8–41.7%. Type genus: Candidatus Nitrosocaldus de la Torre, Walker, Ingalls, Könneke and Stahl 2008, 815. Taxonomic and Nomenclature Notes According to the List of Prokaryotic names with Standing in Nomenclature (LPSN), the taxonomic status of the family Candidatus Nitrosocaldaceae is: preferred name (not correct name) (last update, February 2025) * . LPSN classification: Archaea / Thermoproteati / Thermoproteota / incertae sedis / Candidatus Nitrosocaldales / Candidatus Nitrosocaldaceae The family Candidatus Nitrosocaldaceae can also be recovered in the Genome Taxonomy Database (GTDB) as f__Nitrosocaldaceae (version v220) ** . GTDB classification: d__Archaea / p__Thermoproteota / c__Nitrososphaeria / o__Nitrososphaerales / f__Nitrosocaldaceae * Meier‐Kolthoff et al. ( 2022 ). Nucleic Acids Res , 50 , D801 – D807 ; DOI: 10.1093/nar/gkab902 ** Parks et al. ( 2022 ). Nucleic Acids Res , 50 , D785 – D794 ; DOI: 10.1093/nar/gkab776