Nutritional and functional biology of exudate-feeding ants

Feeding extensively on plant exudates and honeydews, many tropical arboreal ant species exhibit δ¹⁵N values characteristic of herbivores. Consistent with hypothesized herbivory, these taxa behave in feeding assays as though more N-deprived than are strictly carnivorous ants. However, to an as yet uncertain degree, relationships with N-upgrading and/or recycling microsymbionts may lower isotopic ratios, making ants appear to be more herbivorous than they actually are. Nutritional (N) contributions from microsymbionts have been inferred for a variety of ant taxa based on intracellular or extracellular associations between ants and bacteria. However, stronger and more specific inferences are possible when variability in microsymbiont locations within the digestive system is considered in the context of taxonomic variability in ant diets and digestive anatomy. Diets of exudate feeders may vary predictably in ratios of usable carbohydrates (CHOs) to N, depending on the extent to which they tend melezitose-producing Homoptera. Status of the peritrophic membrane, proventricular structure, and number and placement of Malpighian tubules can be interpreted as traits contributing to supply of N and/or CHOs to microsymbionts. In general, a more integrative understanding of ant diets, digestive anatomy, and associated microsymbionts helps to set out specific hypotheses to be tested experimentally and (where possible) in a phylogenetic context.     

GALEIDINIIUM RUGATUM GEN. ET SP. NOV. (DINOPHYCEAE), A NEW COCCOID DINOFLAGELLATE WITH A DIATOM ENDOSYMBIONT1

A new sand‐dwelling dinoflagellate from Palau, Galeidinium rugatum Tamura et Horiguchi gen. et sp. nov., is described. The life cycle of this new alga consists of a dominant nonmotile phase and a brief motile phase. The motile cell transforms itself directly into the nonmotile cell after swimming for a short period, and cell division takes place in the nonmotile phase. The nonmotile cell possesses a dome‐like cell covering, which is wrinkled and equipped with a transverse groove on the surface. The cell has 10–20 chloroplasts and a distinct eyespot. The motile cell is Gymnodinium‐like in shape. The dinoflagellate possesses an endosymbiotic alga to which the chloroplasts belong and which is separated from the host (dinoflagellate) cytoplasm by a unit membrane. The endosymbiont cytoplasm also possesses its own eukaryotic nucleus and mitochondria. The eyespot is surrounded by triple membranes and is located in the host cytoplasm. Photosynthetic pigment analysis, using HPLC, revealed that G. rugatum possesses fucoxanthin as the principal accessory pigment instead of peridinin. The rbcL tree showed that G. rugatum is monophyletic with Durinskia baltica (Levander) Carty et Cox and Kryptoperidinium foliaceum (Stein) Lindemann and that this clade is closely related to the pennate diatom, Cylindrotheca sp. The endosymbiont of G. rugatum is therefore shown to be a diatom. Phylogenetic analysis based on small subunit rDNA sequences demonstrated that G. rugatum, D. baltica, and K. foliaceum, all of which are known to harbor an endosymbiont of diatom origin, are closely related.     

EVOLUTION OF ASEXUALITY VIA DIFFERENT MECHANISMS IN GRASS THRIPS (THYSANOPTERA: Aptinothrips)

Asexual lineages can derive from sexual ancestors via different mechanisms and at variable rates, which affects the diversity of the asexual population and thereby its ecological success. We investigated the variation and evolution of reproductive systems in Aptinothrips, a genus of grass thrips comprising four species. Extensive population surveys and breeding experiments indicated sexual reproduction in A. elegans, asexuality in A. stylifer and A. karnyi, and both sexual and asexual lineages in A. rufus. Asexuality in A. stylifer and A. rufus coincides with a worldwide distribution, with sexual A. rufus lineages confined to a limited area. Inference of molecular phylogenies and antibiotic treatment revealed different causes of asexuality in different species. Asexuality in A. stylifer and A. karnyi has most likely genetic causes, while it is induced by endosymbionts in A. rufus. Endosymbiont‐community characterization revealed presence of Wolbachia, and lack of other bacteria known to manipulate host reproduction. However, only 69% asexual A. rufus females are Wolbachia‐infected, indicating that either an undescribed endosymbiont causes asexuality in this species or that Wolbachia was lost in several lineages that remained asexual. These results open new perspectives for studies on the maintenance of mixed sexual and asexual reproduction in natural populations.     

Facultative bacterial endosymbionts benefit pea aphids Acyrthosiphon pisum under heat stress

Abstract 1. Natural populations of pea aphids in California contain at least two facultative bacterial secondary symbionts (pea aphid secondary symbiont, PASS, or pea aphid rickettsia, PAR) in a range of frequencies throughout the state.
2. Two pea aphid clones without either of these facultative associates failed to reproduce in the first 8 days after the final moult if they had been heat-stressed for a period of about 4 h at 39 °C as 1-day-old larvae in the laboratory.
3. Aphids infected artificially with PASS, however, were able to produce up to 48% of the normal complement of offspring produced by PASS-positive aphids that had not been heat-stressed. Clones infected artificially with PAR did not have the same advantage as those with PASS after heat stress.
4. In aphids without PASS or PAR, heat stress reduced the number of bacteriocytes (in which the obligate primary symbiont, Buchnera , resides) to 7% of non-heat-stressed aphids, while aphids with only PASS retained 70% of their bacteriocytes. Bacteriocytes in aphids with PAR but not PASS were reduced to 42% of controls.
5. When larvae were heat-stressed as older instars (5 days old), a similar pattern emerged, though the effect of heat stress was less extreme. Clones containing PASS produced the most offspring, three to 14 times as many as aphids without PASS or PAR. Aphids with PAR only, or PASS and PAR together, had reduced or no advantage over aphids without facultative symbionts.
6. Aphids of all clones that had been heat-stressed as later instars gave birth to a variable number of stillborn offspring. Aphids without facultative symbionts produced the most stillborn larvae.
7. Field studies showed a higher incidence of PASS in aphids collected in California in summer compared with aphids from the same sites collected 2–4 months earlier. The difference was significant in two of three widely dispersed locations.     

Microbial manipulation of host sex determination. Endosymbiotic bacteria can directly manipulate their host's sex determination towards the production of female offspring

A recent study in the lepidopteran Ostrinia scapulalis shows that endosymbionts can actively manipulate the sex determination mechanism of their host. Wolbachia bacteria alter the sex-specific splicing of the doublesex master switch gene. In ZZ males of this female heterogametic system, the female isoform of doublesex is produced in the presence of the bacteria. The effect is a lethal feminization of genotypic males. Curing of ZW females leads to males that die, indicating that the bacteria have an obligate role in proper sex determination and development of their host. Microbial intervention with host sex determination may be a driving force behind the evolutionary turnover of sex determination mechanisms.     

柑橘木虱两地理种群的内共生菌群落组成及传菌能力

柑橘黄龙病（Huanglongbing, HLB）是经柑橘木虱Diaphorina citri传播的最主要柑橘病害之一, 危害严重时能对柑橘产业造成毁灭性的破坏。为了鉴定福建和海南2个地理种群柑橘木虱的内共生菌群落组成, 本研究对16S rRNA部分保守序列进行PCR扩增, 并利用特异性引物对不同内共生菌进行了感染率检测; 另外, 还通过人工接虫的方法, 探索柑橘木虱成虫在带黄龙病菌蕉柑Citrus reticulata cv. Tankan上的获菌能力, 以及带菌柑橘木虱成虫对黄岩蜜橘C. reticulata cv. Subcompressa的传菌能力。研究发现, 这2个地理种群的柑橘木虱含有相同的内共生菌组成, 包括α-Proteobacteria, Wolbachia spp., γ-Proteobacteria, mycetocyte symbionts, β-Proteobacteria, Oxalobacter和β-Proteobacteria, Herbaspirillum, 而且这2个地理种群柑橘木虱的4种内共生菌的携带率均在95%以上。柑橘木虱成虫在带菌蕉柑上饲菌28 d后, 带菌率可达到82%, 而带菌柑橘木虱成虫在黄岩蜜橘上传菌75 d后, 可导致橘树整体带菌。本研究为柑橘木虱的进一步研究和防虫治病途径提供了一些理论依据。     

A CARBON BUDGET FOR THE AUTOTROPHIC CILIATE MESODINIUM RUBRUM1

Extensive blooms of the autotrophic ciliate Mesodinium rubrum (Lohmann) occurred in the Peru coastal upwelling region at 15°S latitude in March through May 1977 and contributed significantly to the organic productivity of the region. From observations made during the JOINT-II oceanographic expedition, a budget of the carbon flux of these unusual photosynthetic organisms can be constructed. The light dependent C fixation was determined with short (1 h) incubations because of the organisms' sensitivity to confinement and rapid nutrient exhaustion. Maximum photosynthesis occurred at 50% of incident light with a maximum rate of particulate C synthesis of 2187 mg C · m^?3· h^?1. The specific carbon uptake rates were also high with a maximum light saturated value of 16.8 mg C ·(mg chl a)^?1· h^?1. The rate of excretion of dissolved organic C at the productivity maximum ranged from 16.1 to 181.1 mg C · m^?3· h^?1. The range of percent excretion was 1.8–12.5% the total C fixed, similar to the range found in both motile and nonmotile phytoplankton assemblages. Respiration, determined by the decrease in particulate C in the dark, averaged 4.6% of the previously fixed photosynthetic C · h^?1. M. rubrum actively took up amino acids and naturally occurring dissolved organic carbon. The C budget for this ciliate indicates that the daily contribution to the particulate food chain is large, although not as great as is indicated by short incubations. The contribution of M. rubrum to the productivity and elemental fluxes of upwelling and coastal ecosystems has been seriously underestimated.     

The candidate phylum 'Termite Group 1' of bacteria: phylogenetic diversity, distribution, and endosymbiont members of various gut flagellated protists

The candidate phylum 'Termite Group 1' (TG1) of bacteria, which is abundant in termite guts but has no culturable representative, was investigated with respect to the in situ localization, distribution, and diversity. Based on the 16S rRNA gene sequence analyses and FISH in termite guts, a number of lineages of TG1 members were identified as endosymbionts of a variety of gut flagellated protists from the orders Trichonymphida, Cristamonadida, and Oxymonadida that are mostly unique to termites. However, the survey in various environments using specific PCR primers revealed that TG1 members were also present in termites, a cockroach, and the bovine rumen that typically lack these protist orders. Most of the TG1 members from gut flagellates, termites, cockroaches, and the rumen formed a monophyletic subcluster that showed a shallow branching pattern in the phylogenetic tree, suggesting their recent diversification. Although endosymbionts of the same protist genera tended to be closely related, the endosymbiont lineages were often independent of the higher level classifications of their host protist and were dispersed in the phylogenetic tree. It appears that their cospeciation is not the sole rule for the diversification of TG1 members of endosymbionts.     

Natural occurrence of Mycobacterium as an endosymbiont of Acanthamoeba isolated from a contact lens storage case

Recent in vitro studies have revealed that a certain Mycobacterium can survive and multiply within free-living amoebae. It is believed that protozoans function as host cells for the intracellular replication and evasion of Mycobacterium spp. under harmful conditions. In this study, we describe the isolation and characterization of a bacterium naturally observed within an amoeba isolate acquired from a contact lens storage case. The bacterium multiplied within Acanthamoeba, but exerted no cytopathic effects on the amoeba during a 6-year amoebic culture. Transmission electron microscopy showed that the bacteria were randomly distributed within the cytoplasm of trophozoites and cysts of Acanthamoeba. On the basis of the results of 18S rRNA gene analysis, the amoeba was identified as A. lugdunensis. A 16S rRNA gene analysis placed this bacterium within the genus Mycobacterium. The bacterium evidenced positive reactivity for acid-fast and fluorescent acid-fast stains. The bacterium was capable of growth on the Middlebrook 7H11-Mycobacterium-specific agar. The identification and characterization of bacterial endosymbionts of free-living protozoa bears significant implications for our understanding of the ecology and the identification of other atypical mycobacterial pathogens.