Publications

4954

A novel hyperthermophilic, heterotrophic and anaerobic archaeon, designated strain SLHALR2 T , was isolated from the ‘Cooking Pots’ hot springs at Hell’s Gate, Tikitere, New Zealand. During the exponential growth phase, cells of strain SLHALR2 T appear as long, thin rods to filaments, measuring 0.25–0.4 µm in width and 3.0–7.0 µm in length. Strain SLHALR2 T grows at temperatures ranging from 60 to 97 °C (optimum 80–85 °C), at pH values between 5.0 and 7.3 (optimum pH 6.0) and with 0–8 g l −1 sodium chloride concentration (optimum 0–1.0 g l −1 ). Elemental sulphur is reduced to hydrogen sulphide. Strain SLHALR2 T is an obligate organotroph capable of utilizing a variety of proteinaceous substrates and a limited range of sugars. Archaeal cell extracts improve growth but are not mandatory for growth. Phylogenetic analysis based on 16S rRNA gene sequence comparisons indicates that strain SLHALR2 T belongs to the genus Infirmifilum within the family Thermofilaceae , order Thermofilales (phylum Thermoproteota ). The strain shares 98.4–99% 16S rRNA gene sequence similarity with described Infirmifilum species. Despite this high level of 16S rRNA gene similarity with recently described Infirmifilum species, both average nucleotide identity (71–74%), average amino acid identity (67–75.5%) values and digital DNA–DNA hybridization values support the designation of strain SLHALR2 T as a distinct species within Infirmifilum . Based on phylogenomic analyses and the physiological characteristics of the novel isolate, strain SLHALR2 T is proposed as the type strain of a new species, Infirmifilum tikiterense sp. nov. (=OCM2026=UBOCC M-3571), and an emended description of the genus Infirmifilum is proposed.

Abstract Phytoplasma diseases are threat to sesame and its wild relatives, but information on their occurrence in exotic Sesamum species is limited. During field evaluation, Sesamum angolense , S. angustifolium , and S. latifolium exhibited characteristic phyllody and witches' broom symptoms, including virescence, leafy floral organs, polysepalous calyx, polypetalous corolla, flattened stamens, shortened internodes, excessive shoot proliferation, and leaf twisting and curling. Molecular detection using universal phytoplasma primers P1/P7 and tuf gene-specific primers fTufAy/rTufAy amplified products of approximately 1.8 kb and 940 bp, respectively, from symptomatic plants. Sequence analysis of the 16S rRNA and tuf genes revealed 99–100% nucleotide identity with members of the 'Candidatus Phytoplasma asteris ' group. Phylogenetic reconstruction based on both loci consistently assigned the isolates to the 16SrI-B subgroup of 'Ca. Phytoplasma asteris'. To the best of our knowledge, this is the first report of phytoplasma infection in S. angolense , S. angustifolium , and S. latifolium , characterized at the strain level using both 16S rRNA and tuf gene sequences. The findings expand the known host range of 16SrI-B phytoplasmas and demonstrate that these wild Sesamum species, which are valuable components of the tertiary gene pool of cultivated sesame, can serve as natural reservoirs of phytoplasma inoculum.

During field surveys conducted in 2024 and 2025 in pepper production areas in Altunhisar distinct of Niğde province, Türkiye, plants showing phytoplasma-like symptoms were observed. A total of 25 symptomatic pepper samples were collected, and total nucleic acids were extracted using the CTAB method. The samples were analyzed for phytoplasma infection by PCR amplification of the 16S rRNA gene region. In addition, two asymptomatic pepper plants were included as negative controls. PCR analysis revealed that all samples showing symptoms produced amplicons of the expected size, while no amplification was obtained from the asymptomatic plants. Sequencing and phylogenetic analyses revealed that the detected phytoplasma belonged to the 16SrVI-A subgroup, corresponding to ‘Candidatus Phytoplasma trifolii’. The subgroup assignment was further confirmed by in silico (virtual) RFLP analysis of the 16S rRNA gene. The representative 16S rRNA gene sequence of one strain obtained in this study was deposited in the GenBank database (GenBank accession number PX885052). Based on the available literature, this study provides the first molecular evidence of ‘Candidatus Phytoplasma trifolii’ infection in pepper plants grown in Niğde province, Türkiye. The findings contribute to the current knowledge on the distribution of pepper-infecting phytoplasmas in Türkiye and emphasize the need for further epidemiological and vector-related studies.

Abstract Syndrome Basses Richesses (SBR) is a phloem-limited disease of sugar beet caused by ‘ Candidatus Arsenophonus phytopathogenicus’ (ARSEPH), a bacterium currently considered a facultative endosymbiont of its insect vector, Pentastiridius leporinus . Due to increasing disease incidence and the need for reliable vector-free resistance screening methods, this study evaluated alternative ARSEPH transmission routes using chip grafting and the parasitic plant dodder ( Cuscuta campestris ). Given that Arsenophonus species are unculturable and their insect vectors have complex life cycles so laboratory propagation is labor intensive, the development of vector-independent systems is desirable for controlled pathogen transmission and host–pathogen interaction studies. Chip grafting enabled successful ARSEPH transmission to sugar beet, though with low efficiency (~ 9.6%) and delayed pathogen detection. While C. campestris acquired and maintained high ARSEPH titers, it failed to transmit the bacterium to healthy plants, indicating limited systemic movement. In contrast, vector-mediated transmission via P. leporinus achieved 100% efficiency within 25 days, confirming a strong biological compatibility. These findings underscore the limitations of non-vector-based approaches for phloem-restricted pathogens and highlight the need for methodological optimization. The study provides key insights into ARSEPH’s complex transmission biology and supports a functionally specialized association with its insect vector.

Abstract Huanglongbing (HLB), primarily induced by the phloem-restricted bacterium Candidatus Liberibacter asiaticus (CLas), poses a significant threaten to citrus production worldwide. CLas deploys Sec-dependent effectors (SDEs) to manipulate host cellular processes, yet the underlying virulence mechanisms remain incompletely understood. Here, we identify SDE4580, a conserved CLas effector localized to the plant plasma membrane, as a key virulence factor that promotes pathogen proliferation and suppresses host immunity across citrus and Arabidopsis. Transcriptomic analysis reveals that SDE4580 reprograms membrane-associated biological processes and defense responses. Through yeast two-hybrid screening and biochemical validation, we demonstrate that SDE4580 physically interacts with the citrus vesicle trafficking regulator CsPRA1.E, a Prenylated Rab Acceptor 1 family protein. Functional assays establish CsPRA1.E as a positive regulator of immunity that promotes apoplastic secretion of defense-related proteins and enhances resistance to Xanthomonas citri subsp. citri. Crucially, SDE4580 antagonizes CsPRA1.E-mediated resistance by suppressing defense protein secretion without inducing target degradation. These findings uncover a novel pathogenic strategy whereby CLas subverts host endomembrane trafficking to disarm plant immunity. Targeting the SDE4580-CsPRA1.E interface may offer a promising avenue for developing durable HLB control strategies.

The spread of the reed planthopper Pentastiridius leporinus and its associated pathogens Candidatus Arsenophonus phytopathogenicus (ARSEPH) and Candidatus Phytoplasma solani (PHYPSO) increasingly threatens sugar beet production in Central Europe. Current surveillance systems rely mainly on vector monitoring, visual field assessments and molecular diagnostics of individual vectors and plants, all of which provide valuable but spatially limited information. This study evaluated sugar beet pulp samples generated during routine factory quality assessment as a large- scale surveillance resource for PHYPSO. During the 2025/26 campaign, 4887 pulp samples representing 1954 delivery field locations were collected, of which 1237 samples of individual field locations with consistent beet quality parameters were analyzed by direct PCR. PHYPSO was detected in 214 samples (17.3%). The molecular workflow was not subgroup-specific and did not include genotyping; therefore, PHYPSO-positive samples could not be assigned to individual 16SrXII subgroups. Comparison with visual field assessments showed that 85.7% of positive molecular detections originated from fields without visible symptoms. A carry-over experiment demonstrated that pathogen-derived DNA may persist across consecutive samples under worst-case conditions and must therefore be considered during data interpretation. In contrast, ARSEPH was detected by direct PCR in only six of 1237 samples (0.5%), all originating from the Euskirchen growing region. Comparison with qPCR, single-beet diagnostics and the carry-over experiment indicated insufficient sensitivity of the current ARSEPH direct-PCR assay; therefore, the observed detection rate should not be interpreted as true pathogen prevalence. Nevertheless, the observed spatial distribution patterns of monitoring sites indicate that industrial pulp sample monitoring substantially expands surveillance coverage beyond conventional symptom-based approaches and can help identify regions warranting further epidemiological investigation.