Publications

4478

Huanglongbing (HLB) is a highly destructive citrus disease and is associated with a nonculturable bacterium, ‘Candidatus Liberibacter asiaticus’. ‘Ca. L. asiaticus’ in the United States was first found in Florida in 2005 and is now endemic there. In California, ‘Ca. L. asiaticus’ was first detected in Hacienda Heights in Los Angeles County in 2012 and has now been detected in multiple urban locations in southern California. Knowledge of ‘Ca. L. asiaticus’ strain diversity in California is important for HLB management. In this study, genomic diversity among 10 ‘Ca. L. asiaticus’ strains from six California locations were analyzed using a next-generation sequencing (NGS) (Illumina MiSeq and HiSeq) approach. Draft genome sequences of ‘Ca. L. asiaticus’ strains were assembled. Sequences of the 16S ribosomal RNA gene and nrdB confirmed ‘Ca. L. asiaticus’ identity. Prophages were detected in all ‘Ca. L. asiaticus’ strains. The California ‘Ca. L. asiaticus’ strains formed four prophage typing groups (PTGs): PTG1, with type 1 prophage only (strains from Anaheim, San Gabriel, and Riverside); PTG2, with type 2 prophage only (strains from Hacienda Heights); PTG1-3, with both type 1 and 3 prophages (a strain from Cerritos); and PTG1-2, with both type 1 and type 2 prophages (a strain from La Habra). Analyses of the terL sequence showed that all California ‘Ca. L. asiaticus’ strains were not introduced from Florida but likely from locations in Asia. Miniature inverted-repeat transposable elements were found in all ‘Ca. L. asiaticus’ strains, yet, a jumping-out event was detected in the ‘Ca. L. asiaticus’ strain from Cerritos. Altogether, this study demonstrated that the NGS approach focusing on prophage variation was sensitive and effective in revealing diversity of ‘Ca. L. asiaticus’ strains in California.

AbstractAfrican citrus greening (ACGD) and huanglongbing (HLB) diseases are the most damaging diseases of citrus worldwide. Currently, the disease has no cure and has been attributed to the collapse of the citrus industry in several countries. In Africa, the causative agent “Candidatus” Liberibacter africanus is vectored by African citrus triozid (ACT) Trioza erytreae Del Guercio (Hemiptera: Triozidae). African citrus triozid is native to Africa but has been recently reported in Asia and Europe. Apart from citrus, Murraya koenigii (L.) and Clausena anisata (Willd) Hook. F. ex Benth. are also considered as preferred host plants. At present, there is scant information on host plant suitability and preference of T. erytreae. Also, there are contradictory reports on its reproduction and survival on rutaceous and non‐rutaceous host plants. In the present study, we tested the suitability and preference of rutaceous and non‐rutaceous trees and shrubs as potential ACT host plants in choice and no‐choice bioassays. The development from egg to the adult stage was longest on Calodendrum capense (Wright & Arn.) Engl. Host plants of superior quality accordingly to several ACT's biological parameters measured also revealed significantly higher morphometric characteristics. Our findings on the host status of the five rutaceous plants imply that these plants can greatly influence the population dynamics of ACT as well as the epidemiology of ACGD, and these can be a useful guide in the area‐wide management of the pest in Kenya.

A novel slow-growing, facultatively anaerobic, filamentous bacterium, strain MO-CFX2T, was isolated from a methanogenic microbial community in a continuous-flow bioreactor that was established from subseafloor sediment collected off the Shimokita Peninsula of Japan. Cells were multicellular filamentous, non-motile and Gram-stain-negative. The filaments were generally more than 20 µm (up to approximately 200 µm) long and 0.5–0.6 µm wide. Cells possessed pili-like structures on the cell surface and a multilayer structure in the cytoplasm. Growth of the strain was observed at 20–37 °C (optimum, 30 °C), pH 5.5–8.0 (pH 6.5–7.0), and 0–30 g l−1 NaCl (5 g l−1 NaCl). Under optimum growth conditions, doubling time and maximum cell density were estimated to be approximately 19 days and ~105 cells ml−1, respectively. Strain MO-CFX2T grew chemoorganotrophically on a limited range of organic substrates in anaerobic conditions. The major cellular fatty acids were saturated C16 : 0 (47.9 %) and C18 : 0 (36.9 %), and unsaturated C18 : 1ω9c (6.0 %) and C16 : 1ω7 (5.1 %). The G+C content of genomic DNA was 63.2 mol%. 16S rRNA gene-based phylogenetic analysis showed that strain MO-CFX2T shares a notably low sequence identity with its closest relatives, which were Thermanaerothrix daxensis GNS-1T and Thermomarinilinea lacunifontana SW7T (both 85.8 % sequence identity). Based on these phenotypic and genomic properties, we propose the name Aggregatilinea lenta gen. nov., sp. nov. for strain MO-CFX2T (=KCTC 15625T, =JCM 32065T). In addition, we also propose the associated family and order as Aggregatilineaceae fam. nov. and Aggregatilineales ord. nov., respectively.

Apple, pear and apricot trees showing phytoplasma associated symptoms from Ankara and Isparta provinces were sampled and investigated to verify the presence of phytoplasma-associated diseases. Totally 31 samples were tested with phytoplasma universal and group specific primers and the PCR products were restricted using Tru1I, RsaI and SspI endonucleases. Different RFLP profiles were obtained and selected samples were directly sequenced. The apple samples were found infected with 16SrX-A (‘Candidatus Phytoplasma mali’), while the majority of the pear samples were infected with 16SrX-A and 16SrX-C subgroup phytoplasmas in mixed infection. The 16SrX-B (‘Candidatus Phytoplasma prunorum’), 16SrX-C (‘Candidatus Phytoplasma pyri’) and mixed infection of 16SrX-A/16SrX-C and 16SrX-C/16SrI (aster yellows) were detected in the apricot samples. In this study the 16SrI phytoplasmas in apricot and the mixed phytoplasma infections in pear and apricot trees were detected in Turkey for the first time.

AbstractThe Liberibacter genus comprises insect endosymbiont bacterial species that cause destructive plant diseases, including Huanglongbing in citrus and zebra chip in potato. To date, pathogenic ‘Candidatus Liberibacter spp.’ (CLs) remain uncultured, therefore the plant-associated Liberibacter crescens (Lcr), only cultured species of the genus, has been used as a biological model for in vitro studies. Biofilm formation by CLs has been observed on the outer midgut surface of insect vectors, but not in planta. However, the role of biofilm formation in the life cycle of these pathogens remains unclear. Here, a model system for studying CLs biofilms was developed using Lcr. By culture media modifications, bovine serum albumin (BSA) was identified as blocking initial cell-surface adhesion. Removal of BSA allowed for the first time observation of Lcr biofilms. After media optimization for biofilm formation, we demonstrated that Lcr attaches to surfaces, and form cell aggregates embedded in a polysaccharide matrix both in batch cultures and under flow conditions in microfluidic chambers. Biofilm structures may represent excellent adaptive advantages for CLs during insect vector colonization helping with host retention, immune system evasion, and transmission. Future studies using the Lcr model established here will help in the understanding of the biology of CLs.

“ Candidatus Thioglobus sp.” strain NP1 is an open-ocean isolate from the SUP05 clade of Gammaproteobacteria . Whole-genome comparisons of strain NP1 to other sequenced isolates from the SUP05 clade indicate that it represents a new species of SUP05 that lacks the ability to fix inorganic carbon using the Calvin-Benson-Bassham cycle.

Methanogenic archaea are major contributors to the global carbon cycle and were long thought to belong exclusively to the euryarchaeal phylum. Discovery of the methanogenesis gene cluster methyl-coenzyme M reductase (Mcr) in the Bathyarchaeota, and thereafter the Verstraetearchaeota, led to a paradigm shift, pushing back the evolutionary origin of methanogenesis to predate that of the Euryarchaeota. The methylotrophic methanogenesis found in the non-Euryarchaota distinguished itself from the predominantly hydrogenotrophic methanogens found in euryarchaeal orders as the former do not couple methanogenesis to carbon fixation through the reductive acetyl-CoA [Wood–Ljungdahl pathway (WLP)], which was interpreted as evidence for independent evolution of the two methanogenesis pathways. Here, we report the discovery of a complete and divergent hydrogenotrophic methanogenesis pathway in a thermophilic order of the Verstraetearchaeota, which we have named Candidatus Methanohydrogenales, as well as the presence of the WLP in the crenarchaeal order Desulfurococcales. Our findings support the ancient origin of hydrogenotrophic methanogenesis, suggest that methylotrophic methanogenesis might be a later adaptation of specific orders, and provide insight into how the transition from hydrogenotrophic to methylotrophic methanogenesis might have occurred.

Very thin filamentous cyanobacteria are ubiquitous in a wide range of environments. For many years they were traditionally studied according to their morphological properties only. With the introduction of additional taxonomic methods (cytomorphological analyses, molecular sequencing, exact ecological studies, better data about phytogeographical distribution), traditional genera such as Leptolyngbya and Phormidium were found to be polyphyletic. Phormidesmis belonged to a newly formed genus that was supposed to explain the variability of such very thin simple filamentous cyanobacteria. However, even after definition of Phormidesmis based on distinct cytomorphological and phylogenetic traits, the variability within this genus remained unresolved. Here we analyzed 26 Phormidesmis strains to describe the variability within this genus, classified two new species (P. arctica and P. communis) and transferred Leptolyngbya nigrescens into P. nigrescens. A tabular review of Phormidesmis species is included. The diacritical features of all these species are: width up to 1–4 µm, barrel-shaped cells, which can be shorter or slightly longer than wide, with apparent constrictions at the cross-walls. Our study shows that Phormidesmis is a morphologically and genetically well-defined genus with a global distribution. A newly described genus Leptodesmis has significant morphological similarities both with Phormidesmis and Leptolyngbya, however with intermediate phylogenetic position with significant divergence in 16S rRNA gene. Leptodesmis is cryptic both to Phormidesmis and Leptolyngbya. In the initial part of the life cycle resembles Leptolyngbya, the appearance of older trichomes change to Phormidesmis like morphology.