Publications

4378

ABSTRACT Insect intestinal tracts harbor several novel, deep-rooting clades of as-yet-uncultivated bacteria whose biology is typically completely unknown. Here, we report the isolation of the first representative of the termite group 1 (TG1) phylum from sterile-filtered gut homogenates of a humivorous scarab beetle larva. Strain Pei191 T is a mesophilic, obligately anaerobic ultramicrobacterium with a gram-negative cell envelope. Cells are typically rod shaped, but cultures are pleomorphic in all growth phases (0.3 to 2.5 μm long and 0.17 to 0.3 μm wide). The isolate grows heterotrophically on sugars and ferments d -galactose, d -glucose, d -fructose, d -glucosamine, and N -acetyl- d -glucosamine to acetate, ethanol, hydrogen, and alanine as major products but only if amino acids are present in the medium. PCR-based screening and comparative 16S rRNA gene sequence analysis revealed that strain Pei191 T belongs to the “intestinal cluster,” a lineage of hitherto uncultivated bacteria present in arthropod and mammalian gut systems. It is only distantly related to the previously described so-called “endomicrobia” lineage, which comprises mainly uncultivated endosymbionts of termite gut flagellates. We propose the name “ Elusimicrobium minutum ” gen. nov., sp. nov. (type strain, Pei191 T = ATCC BAA-1559 T = JCM 14958 T ) for the first isolate of this deep-branching lineage and the name “ Elusimicrobia ” phyl. nov. for the former TG1 phylum.

ABSTRACT The cold-water coral Lophelia pertusa (Scleractinia, Caryophylliidae) is a key species in the formation of cold-water reefs, which are among the most diverse deep-sea ecosystems. It occurs in two color varieties: white and red. Bacterial communities associated with Lophelia have been investigated in recent years, but the role of the associated bacteria remains largely obscure. This study uses catalyzed reporter deposition fluorescence in situ hybridization to detect the in situ location of specific bacterial groups on coral specimens from the Trondheimsfjord (Norway). Two tissue-associated groups were identified: (i) bacteria on the host's tentacle ectoderm, “ Candidatus Mycoplasma corallicola,” are flasklike, pointed cells and (ii) endoderm-associated bona fide TM7 bacteria form long filaments in the gastral cavity. These tissue-bound bacteria were found in all coral specimens from the Trondheimsfjord, indicating a closer relationship with the coral compared to bacterial assemblages present in coral mucus and gastric fluid.

ABSTRACT. Rickettsia‐like organisms (RLO) are obligate, often highly fastidious, intracellular bacterial parasites associated with a variety of vertebrate and invertebrate hosts. Despite their importance as causative agents of severe mortality outbreaks in farmed aquatic species, little is known about their life cycle and their host range. The present work reports the characterization of “Candidatus Cryptoprodotis polytropus,” a novel Rickettsia‐like bacterium associated with the common ciliate species Pseudomicrothorax dubius by means of the “Full‐Cycle rRNA Approach” and ultrastructural observations. The morphological description by in vivo and scanning electron microscopy and the 18S rRNA gene sequence of the host species is provided as well. Phylogenetic analysis based on the 16S rRNA gene supports the inclusion of “Candidatus Cryptoprodotis polytropus” within the family Rickettsiaceae (cl. Alphaproteobacteria) together with the genera Rickettsia and Orientia. Observations on natural ciliate populations account for the occasional nature of this likely parasitic association. The presence of a previously unknown RLO in ciliates sheds a new light on the possible role of protists as transient hosts, vectors or natural reservoir for some economically important pathogens.

Citrus huanglongbing (HLB) or yellow shoot disease (i.e., greening disease) is highly destructive to citrus production worldwide. Understanding the etiology of HLB is critical for managing the disease. HLB is currently associated with infection by ‘Candidatus Liberibacter spp.’ around the world, including China. However, Koch's postulates have not been fulfilled. In addition, other plant pathogens also may be involved in HLB. In a survey performed in Guangdong Province, P. R. China in 2006 and 2007, 141 citrus samples showing typical symptoms of HLB from 11 different cities were collected. Polymerase chain reaction (PCR) using phytoplasma-specific primer sets fU5/rU3 nested with primer set P1/P7 identified 110 (78.0%) positive samples. A 1,785-bp amplicon was obtained with primer set P1/P7. Analysis showed a 100% identity of this sequence in the region of 16S rDNA and 16S-23S rRNA intergenic transcribed spacer to three strains of ‘Candidatus Phytoplasma asteris’ (onion yellows [Japan], aster yellows ‘watercress’ [Hawaii], and valeriana yellows [Lithuania]). Of the 141 samples, 89 (63.1%) samples were positive for ‘Ca. Liberibacter asiaticus’. When mixed infection was considered, 69 (48.9%) samples were positive for both ‘Ca. P. asteris’ and ‘Ca. L. asiaticus’. Transmission electron microscopy (TEM) showed low titers of both walled and wall-less bodies in the phloem sieve tubes of HLB citrus. When transmission from symptomatic citrus to periwinkle (Catharanthus roseus) via dodder (Cuscuta campestris) was conducted, both phytoplasma and ‘Ca. L. asiaticus’ were detected from the affected periwinkle. In addition to yellowing/mottling, the infected periwinkle showed symptoms of virescence and phyllody which are commonly associated with phytoplasmal diseases. TEM analysis of affected periwinkle revealed pleomorphic and wall-less organisms, characteristic of phytoplasmas, filling some phloem sieve tubes. In contrast, walled bacteria were at low titer. This study showed that in addition to ‘Ca. L. asiaticus’, a phytoplasma related to ‘Ca. P. asteris’ could also be detected in citrus showing HLB symptoms in Guangdong.