Publications

4495

In 2017, potato tubers suspected of being infected with the bacterium ‘Candidatus Liberibacter solanacearum’ were received from the Animal and Plant Health Inspection Service in the United States. A total of 368 chipping tubers were observed for internal symptoms of zebra chip disease, which is associated with ‘Ca. L. solanacearum’ infection in the United States, Mexico, Central America, and New Zealand. A single tuber sliced at the stem end showed classic zebra chip symptoms of darkened medullary rays, with streaking and necrotic flecking. The symptomatic tuber was confirmed positive for the bacterium by polymerase chain reaction targeting three different ‘Ca. L. solanacearum’ genes. Sequence analysis of these three genes, and subsequent BLAST analysis, identified the pathogen with 99, 98, and 97% identity to ‘Ca. L. solanacearum’ for the 16S ribosomal RNA gene, 50S ribosomal proteins L10/L12 genes, and the outer membrane protein gene, respectively. Sequence analysis did not identify the sample as one of the six known haplotypes of ‘Ca. L. solanacearum,’ indicating that a seventh haplotype of the pathogen was identified. This new haplotype, designated haplotype F, is now the third haplotype of the bacterium that infects Solanum tuberosum in the United States.

“ Candidatus Accumulibacter phosphatis” is widely found in full-scale wastewater treatment plants, where it has been identified as the key organism for biological removal of phosphorus. Since aeration can account for 50% of the energy use during wastewater treatment, microaerobic conditions for wastewater treatment have emerged as a cost-effective alternative to conventional biological nutrient removal processes. Our report provides strong genomics-based evidence not only that “ Ca . Accumulibacter phosphatis” is the main organism contributing to phosphorus removal under microaerobic conditions but also that this organism simultaneously respires nitrate and oxygen in this environment, consequently removing nitrogen and phosphorus from the wastewater. Such activity could be harnessed in innovative designs for cost-effective and energy-efficient optimization of wastewater treatment systems.

The symbiotic N 2 -fixing cyanobacterium UCYN-A, which is closely related to Braarudosphaera bigelowii , and its eukaryotic algal host have been shown to be globally distributed and important in open-ocean N 2 fixation. These unique cyanobacteria have reduced metabolic capabilities, even lacking genes for oxygenic photosynthesis and carbon fixation. Cyanobacteria generally use energy from photosynthesis for nitrogen fixation but require mechanisms for avoiding inactivation of the oxygen-sensitive nitrogenase enzyme by ambient oxygen (O 2 ) or the O 2 evolved through photosynthesis. This study showed that symbiosis between the N 2 -fixing cyanobacterium UCYN-A and its eukaryotic algal host has led to adaptation of its daily gene expression pattern in order to enable daytime aerobic N 2 fixation, which is likely more energetically efficient than fixing N 2 at night, as found in other unicellular marine cyanobacteria.

The microbial rare biosphere represents the largest pool of biodiversity on Earth and constitutes, in sum of all its members, a considerable part of a habitat’s biomass. Dormancy or starvation is typically used to explain the persistence of low-abundance microorganisms in the environment. We show that a low-abundance microorganism can be highly transcriptionally active while remaining in a zero-growth state for at least 7 weeks. Our results provide evidence that this zero growth at a high cellular activity state is driven by maintenance requirements. We show that this is true for a microbial keystone species, in particular a cosmopolitan but permanently low-abundance sulfate-reducing microorganism in wetlands that is involved in counterbalancing greenhouse gas emissions. In summary, our results provide an important step forward in understanding time-resolved activities of rare biosphere members relevant for ecosystem functions.

Abstract‘Candidatus Phytoplasma solani’ is the causal agent of Bois noir (BN) in grapevine (Vitis vinifera). It is usually detected in leaves, where typical disease symptoms are seen. However, little information is available on the presence of this phytoplasma in grapevine roots. Here, we investigated ‘Ca. P. solani’ in roots collected from 28 symptomatic, 27 recovered and eight asymptomatic grapevine plants. Protocols based on high-resolution melting (HRM) combined with real-time quantitative PCR (qPCR-HRM) and nested-qPCR-HRM were developed to identify ‘Ca. P. solani’ tuf-type variants with single nucleotide polymorphisms. In all, 21.4% of roots from symptomatic plants were positive to ‘Ca. P. solani’ using qPCR-HRM, and 60.7% with nested-qPCR HRM. Also, 7.4% of roots from recovered plants were positive using qPCR-HRM, which reached 44.4% using nested-qPCR HRM. These analyses identified tuf-type b1 on 88.2% of the positive samples from symptomatic grapevines, and 66.6% from recovered grapevines, with all other samples identified as tuf-type a. This study reports the presence of ‘Ca. P. solani’ in the roots of both symptomatic and recovered grapevines. These qPCR-HRM and nested-qPCR-HRM protocols can be applied to increase the sensitivity of detection of, and to simplify and speed up the screening for, ‘Ca. P. solani’ tuf-types.

AbstractMembers of the order Rickettsiales are often found in association with ciliated protists. An interesting case is the bacterial endosymbiont “Candidatus Megaira”, which is phylogenetically closely related to the pathogen Rickettsia. “Candidatus Megaira” was first described as an intracellular bacterium in several ciliate species. Since then it has been found in association with diverse evolutionary distantly-related hosts, among them other unicellular eukaryotes, and also algae, and metazoa, such as cnidarians. We provide the characterization of several new strains of the type species “Candidatus Megaira polyxenophila”, and the multidisciplinary description of a novel species, “Candidatus Megaira venefica”, presenting peculiar features, which highlight the diversity and variability of these widespread bacterial endosymbionts. Screening of the 16S rRNA gene short amplicon database and phylogenetic analysis of 16S rRNA gene hypervariable regions revealed the presence of further hidden lineages, and provided hints on the possibility that these bacteria may be horizontally transmitted among aquatic protists and metazoa. The phylogenetic reconstruction supports the existence of at least five different separate species-level clades of “Candidatus Megaira”, and we designed a set of specific probes allowing easy recognition of the four major clades of the genus.

Here we report the detection of Candidatus Rickettsia tarasevichiae from tick collected from human patient through DNA amplification. The tick was identified to be Haemaphysalis longicornis. We amplified and sequenced rickettsial citrate synthase gene (gltA), 190_kDa outer protein gene (ompA), and 120_ kDa outer protein gene (ompB). Sequencing results showed that gltA gene and ompA (GenBank accession No. KT899085, KT899079, respectively) shared 100% nucleotide sequence similarities with Candidatus R. tarasevichiae. In addition, ompB gene shared 96% nucleotide sequence similarity with R. canadensis. However, DNA extracted from patient serum showed 98% sequence similarity with R. japonica (AF123713.1). This result implies that H. longicornis ticks harbor both Candidatus R. tarasevichiae and R. japonica.

‘Candidatus Ornithobacterium hominis’ represents a new member of the Flavobacteriaceae detected in 16S rRNA gene surveys of people from South-East Asia, Africa and Australia. It frequently colonizes the infant nasopharynx at high proportional abundance, and we demonstrate its presence in 42 % of nasopharyngeal swabs from 12-month-old children in the Maela refugee camp in Thailand. The species, a Gram-negative bacillus, has not yet been cultured, but the cells can be identified in mixed samples by fluorescent hybridization. Here, we report seven genomes assembled from metagenomic data, two to improved draft standard. The genomes are approximately 1.9 Mb, sharing 62 % average amino acid identity with the only other member of the genus, the bird pathogen Ornithobacterium rhinotracheale . The draft genomes encode multiple antibiotic-resistance genes, competition factors, Flavobacterium johnsoniae -like gliding motility genes and a homologue of the Pasteurella multocida mitogenic toxin. Intra- and inter-host genome comparison suggests that colonization with this bacterium is both persistent and strain exclusive.