Revised Systematics of Holospora-Like Bacteria and Characterization of “Candidatus Gortzia infectiva”, a Novel Macronuclear Symbiont of Paramecium jenningsi

The genus Holospora (Rickettsiales) includes highly infectious nuclear symbionts of the ciliate Paramecium with unique morphology and life cycle. To date, nine species have been described, but a molecular characterization is lacking for most of them. In this study, we have characterized a novel Holospora-like bacterium (HLB) living in the macronuclei of a Paramecium jenningsi population. This bacterium was morphologically and ultrastructurally investigated in detail, and its life cycle and infection capabilities were described. We also obtained its 16S rRNA gene sequence and developed a specific probe for fluorescence in situ hybridization experiments. A new taxon, “Candidatus Gortzia infectiva”, was established for this HLB according to its unique characteristics and the relatively low DNA sequence similarities shared with other bacteria. The phylogeny of the order Rickettsiales based on 16S rRNA gene sequences has been inferred, adding to the available data the sequence of the novel bacterium and those of two Holospora species (Holospora obtusa and Holospora undulata) characterized for the purpose. Our phylogenetic analysis provided molecular support for the monophyly of HLBs and showed a possible pattern of evolution for some of their features. We suggested to classify inside the family Holosporaceae only HLBs, excluding other more distantly related and phenotypically different Paramecium endosymbionts.     

“Candidatus Sonnebornia yantaiensis”, a member of candidate division OD1, as intracellular bacteria of the ciliated protist Paramecium bursaria (Ciliophora,Oligohymenophorea)

An intracellular bacterium was discovered in an isolate of Paramecium bursaria from a freshwater pond in Yantai, China. The bacteria were abundant and exclusively found in the cytoplasm of the host which, along with the green alga Chlorella, formed a three-partner consortium that could survive in pure water for at least one week. Cloning, sequencing and phylogenetic analysis of the bacterial 16S rRNA gene showed that the bacterium belonged to the uncultured candidate division OD1, which usually forms part of the rare biosphere. Transmission electron microscopy and fluorescence in situ hybridization (FISH) with specific probes showed that the bacteria were usually located close to the perialgal membranes of endosymbiotic Chlorella cells, and occasionally irregularly distributed throughout the host cytoplasm. The name “Candidatus Sonnebornia yantaiensis” gen. nov., sp. nov. is proposed for the new bacterium. A strongly supported monophyletic subclade, OD1-p, which included the new species, was recognized and this study highlights that protists can be important hosts for rare bacterial taxa.     

Paramecium calkinsi andP. putrinum (Ciliophora, Protista) harboring alpha-subgroup bacteria in the cytoplasm

Summary New intracellular bacteria were detected in the cytoplasm ofParamecium calkinsi andP. putrinum. Some of the bacteria were not evenly distributed in the cytoplasm of the host but were found in the center of the cell, eventually near the nuclei, but not in the cortex area, whereas another species was found in the cortex area. These peculiarities of intracellular bacteria localization in the host suggest that the conditions in various parts of the cytoplasm favor bacterial maintenance to different extent. Due to the results obtained by transmission electron microscopy and in situ hybridization using appropriate oligonucleotide probes, the bacteria, three or possibly four species, are Gram-negative and belong to the alpha-subgroup of proteobacteria. Bacteria from one stock ofP. calkinsi were found to be infectious for bacteria-free cells ofP. calkinsi andP. nephridiatum.     

Morphological and molecular investigations of Paramecium schewiakoffi sp. nov. (Ciliophora, Oligohymenophorea) and current status of distribution and taxonomy of Paramecium spp.

Paramecium schewiakoffi sp. nov. is described from a pond in Shanghai, China. It is a freshwater species belonging to the “aurelia” subgroup of the genus. It is of similar size and shape to P. jenningsi, but has a single large micronucleus of the “chromosomal” morphological type, while P. jenningsi has two smaller micronuclei. The general morphology, morphometric characteristics and nuclear reorganization pattern, a random amplified polymorphic DNA (RAPD) fingerprint pattern, and the small subunit rRNA gene sequence are presented for the species. Comparison of P. schewiakoffi with the other species of Paramecium indicates that it is a valid new species of the genus. Geographical locations reported for many Paramecium species do not support the theory that all ciliates have a cosmopolitan distribution. It is proposed that, in an extension of Jankowski's earlier suggestion, the genus Paramecium should be subdivided into four subgenera: Chloroparamecium, Helianter, Cypriostomum and Paramecium, on the basis of morphometric, biological and molecular differences.     

Geographical Distribution of Phacellaria Benth. (Santalaceae) and its Hosts

Based on the geographical distribution of the species of Phacellaria and its host plants in the world, we speculated on the possible time, sites, and migration of the origin of Phacellaria. The host plants of Phacellaria mainly belong to Loranthaceae. Plants of Phacellaria and their hosts are mainly distributed in tropical and subtropical areas. The plants of Phacellariamight have originated from a tropical area in the south of China before the Tertiary. Their ancestors were parasitic on the ancestors of some plants of Loranthaceae by chance during the Tertiary. It possibly took them millions of years to form a sturdy relationship with their hosts. Translated from Guihaia, 2005, 25(2) (in Chinese)     

In situ observation of the <Emphasis Type="Italic">Cardinium</Emphasis> symbionts of <Emphasis Type="Italic">Brevipalpus</Emphasis> (Acari: Tenuipalpidae) by electron microscopy

Brevipalpus (Acari: Tenuipalpidae) mites are important pests on a variety of host plant species. The mites damage their hosts directly by feeding and some species also serve as vectors of plant viruses. Among more than 200 described Brevipalpus species, three are recognized as vectors of plant viruses: B. phoenicis, B. californicus and B. obovatus. These species occur worldwide in subtropical and tropical regions. Brevipalpus mites reproduce mostly by thelytokous parthenogenesis and this condition was attributed to a bacterial endosymbiont, recently characterized as a member of the genus Cardinium. The same symbiont infects many other arthropods and is capable of manipulating their host reproduction in various ways. Generally the presence of Cardinium is determined by molecular, PCR based, techniques. In the current work we present visual evidence for the presence of these bacteria by transmission electron microscopy as a complement of previous detection by PCR. Cardinium is easily identified by the presence of a unique array of microtubule-like structures (ML) in the cell. Symbionts have been observed in several organs and eggs from different populations of all three Brevipalpus species known as vector of plant viruses. Cardinium cells were always immersed directly within the cytoplasm of infected cells. Bacteria were observed in all females of all instars, but were absent from all males examined. Females from some Brevipalpus populations were observed to be uninfected by Cardinium. This observation confirmed previous PCR-based results that these populations were aposymbiotic. The observed distribution of the bacteria suggests that these bacteria could have other functions in the mite biology beside feminization.     

鳗弧菌毒力相关基因的研究进展

鳗弧菌是引起多种海水鱼类出血性败血症的病原菌。其致病机理与各个毒力基因的协同作用密切相关。文中综述了鳗弧菌的主要毒力基因,包括编码外毒素、粘附因子、侵袭因子、细胞表面成分以及铁吸收系统的基因和部分检测方法。     

<Emphasis Type="Italic">Nramp1</Emphasis> deletion does not confer susceptibility to<Emphasis Type="Italic"> Rhodococcus equi</Emphasis> infection in mice

Rhodococcus equi is an intracellular bacterium that causes pneumonia in immunocompromised people and foals. The Nramp1 gene influences susceptibility to a variety of intracellular bacteria (including mycobacterial species), but not to Mycobacterium tuberculosis. In this study, we demonstrate that mice functionally deleted of the Nramp1 gene were not more susceptible to infection with virulent R. equi (ATCC 33701) than wild-type mice. Susceptibility of mice to infection with the intracellular bacterium R. equi is more similar to that of M. tuberculosis than to other intracellular bacteria, including other mycobacteria.     

Cystobasidiopsis nirenbergiae, a new agaricostilbomycete (Pucciniomycotina)

A new genus, Cystobasidiopsis, and a new species, Cystobasidiopsis nirenbergiae, are described for a fungus isolated from an arable loess soil in Ahlum near Braunschweig, Niedersachsen, Germany. An integrated analysis of morphological, ecological, ultrastructural and molecular data indicates that the new species belongs to the Chionosphaeraceae within the Agaricostilbales. Relevant characteristics of the new species are discussed and compared with those of related taxa.     

江苏省花椒属(芸香科)植物地理分布新记录

报道江苏省花椒属地理分布新记录植物1种——椿叶花椒(Zanthoxylum ailanthoides Sieb.et Zucc.),并对该省花椒属植物进行了补充记述。椿叶花椒分布于中国东南部(福建、浙江、江西、广东、广西、贵州、四川、云南、台湾)至东亚和东南亚的热带与亚热带地区,在江苏为新记录,且是该种分布的最北界。凭证标本存放于南京师范大学生命科学学院生物标本馆(NJNUH)。     

Vigna mungo, V. radiata and V. unguiculata plants sampled in different agronomical–ecological–climatic regions of India are nodulated by Bradyrhizobium yuanmingense

Vigna mungo, Vigna radiata and Vigna unguiculata are important legume crops cultivated in India, but little is known about the genetic resources in native rhizobia that nodulate these species. To identify these bacteria, a core collection of 76 slow-growing isolates was built from root nodules of V. mungo, V. radiata and V. unguiculata plants grown at different sites within three agro-ecological-climatic regions of India. The genetic diversity of the bacterial collection was assessed by restriction fragment length polymorphism (RFLP) analysis of PCR-amplified DNA fragments of the 16S–23S rDNA intergenic spacer (IGS) region, and the symbiotic genes nifH and nodC. One rDNA IGS type grouped 91% of isolates, but more diversity was found at the symbiotic loci (17 symbiotic genotypes). Overall, no host plant specificity was shown, the three host plant species sharing common bradyrhizobial genotypes that represented 62% of the collection. Similarly, the predominant genotypes were found at most sampling sites and in all agro-ecological-climatic regions. Phylogenies inferred from IGS sequencing and multi-locus sequence analysis of the dnaK, glnII and recA genes indicated that all isolates but one were clustered with the Bradyrhizobium yuanmingense species. The nifH phylogeny also grouped the different nif haplotypes within a cluster including B. yuanmingense, except for one infrequent nif haplotype which formed a new lineage within the Bradyrhizobium genus. These results may reflect a long history of co-evolution between B. yuanmingense and Vigna spp. in India, while intra-species polymorphism detected in the symbiotic loci may be linked with the long history of diversification of B. yuanmingense coinciding with that of its host legumes.     

A newHalophytophthora species,H. porrigovesica, from subtropical and tropical mangroves

A new oomycete was found from intertidal fallen leaves of mangroves in Japan and Thailand and is described here asHalophytophthora porrigovesica. This species is characterized by having an epapillate, ovate zoosporangium with a lens-shaped dehiscence plug-like material at the apex, and by forming an expanding long cylindrical vesicle prior to zoospore release. A key to 14 species and 2 varieties ofHalophytophthora including the new species is proposed. The subtropical (Iriomote is., Japan) strains and tropical (Thailand) strains were different in physiological properties and especially in the asexual reproduction. The subtropical strains showed a lower optimal temperature and wider range of suitable temperature and salinity for zoosporangium formation, whereas the tropical strains showed a higher optimal temperature and narrower range of temperature and salinity. These differences are explained as adaptations of the strains to the environmental conditions of their respective habitats. From the subtropical mangroves, six strains of the new species have been isolated only from submerged leaves ofSonneratia alba, while several strains have been isolated from tropical mangroves from the leaves of three species of mangrove trees,S. alba, Bruguiera gymnorrhiza andAvicennia alba. This indicates a change of taxon selectivity (host specificity) with the geographical distribution.     

The relationship between temperature,competition and the potential for colonization of a subtropical pond by Daphnia magna

The role that temperature plays in excluding large daphnid species from subtropical and tropical ponds, and competitive relationships between Daphnia laevis and Daphnia magna, are explored. D. magna, a large temperate species is probably not excluded from subtropical or tropical systems by elevated temperatures. However, D. magna was a poorer competitor, under a restricted set of laboratory conditions, than D. laevis, the only pond dwelling daphnid in subtropical Florida. Competition is proposed as one mechanism that may limit the number of daphnid species in subtropical and tropical ponds and lakes. Reduced environmental fluctuations in subtropical and tropical systems (compared with temperate systems) may allow zooplankton populations to reach an environmental carrying capacity where competition limits the number of similar, coexisting species in a habitat.     

黄翅大白蚁后肠优势菌大白蚁营发酵菌的全基因组序列分析

【目的】营发酵单胞菌属Dysgonomonas是黄翅大白蚁后肠的第二优势微生物。前期研究中,我们从黄翅大白蚁后肠分离出一种命名为大白蚁营发酵菌的新菌。为深入了解大白蚁营发酵菌在宿主白蚁体内发挥的作用和功能,有必要解析大白蚁营发酵菌的基因组序列信息。【方法】使用Illumina Miseq测序平台获取该菌的全基因组序列,将其全基因组序列经过注释的基因蛋白质序列提交COG和KEGG数据库进行BLASTp比对分析,确定该菌潜在的重要酶类和代谢途径,并对个别纤维素酶活进行检测。【结果】大白蚁营发酵菌整个基因组大小为4655756 bp,GC含量为38.54%,DDBJ数据库登录号为BBXL01000001–BBXL01000078。生物信息学分析结果表明菌株大白蚁营发酵菌具有多个木质纤维素降解酶基因,且具备完整的木质纤维素降解和乙酸、乳酸生成通路。此外发现该菌株中存在与氮源代谢和抵御病原体相关的基因。【结论】本研究首次解析大白蚁营发酵菌的全基因组序列,了解其基因组基本特征,初步探讨了该菌降解木质纤维素的过程,为细菌协助宿主白蚁降解木质纤维素提供了理论基础,同时为该菌可能参与宿主白蚁氮源代谢和抵御病原体入侵提供了依据。     

Mycotrophy of Annona cherimola and the morphology of its mycorrhizae

The mycotrophic character of Annona cherimola (Magnoliales), a tropical/subtropical plantation crop of interest, is described for the first time. This crop seems to depend on mycorrhizae (arbuscular) for optimal growth, with Glomus deserticola being the most effective endophyte tested. Study of the morphology of the arbuscular mycorrhizae in Annona roots showed exclusively intracellular hyphal development, with cell-to-cell fungal passage and an abundance of arbuscules and coiled hyphae within cells. Intercellular distributive hyphae were not observed. The morphology and the pattern of spread of the mycorrhizal colonization were similar for the different endophytes involved and appeared to be dependent on the host root. Such features of mycorrhizal colonization are characteristic of host species lacking intercellular air channels and have been described for some species of ecological interest, but they are not commonly noted in the mycorrhizal literature, especially that dealing with crop species. Some ecophysiological consequences of this pattern of colonization are discussed.     

Establishment,Maintenance, and Integration Mechanisms of Endosymbionts in Protozoa1

ABSTRACT. Important progress has been made in recent years towards better understanding of the establishment and maintenance of endosymbiosis in protozoa and of the eventual integrative mechanisms involved. Still, many problems remain to be investigated more thoroughly. In this paper, while treating and reviewing the subject broadly, particular and more detailed attention is given to three selected systems: endonuclear symbiosis by Holospora bacteria in Paramecium; algal (Chlorella) relationships with the “green”Paramecium (P. bursaria) as host; and the rod-shaped bacteria found in the cytoplasm of Amoeba proteus. Data concerning the physiology of food vacuoles, membrane transport, photobehavior, recognition specificity, enzyme activity, and the like are presented and reviewed and discussed in light of the growing literature on the overall subject of “endocytobiology.” Emphasized is the complicated network of interactions between symbiotic partners and the importance of the development of integrative mechanisms in the evolution of the many intimate associations known at the cellular level today.     

《广西植物志》秋海棠属(Begonia L.)增订

秋海棠属(Begonia L.)隶属于葫芦目(Cucurbitales)秋海棠科(Begoniaceae),有1 800余种,是世界第六大属,主要分布于美洲、非洲和亚洲的热带和亚热带地区,具有极高的观赏价值,亦可作为药用、食用、饮料和饲料等。我国有秋海棠属植物近300种,主要分布于我国的云南和广西等长江以南地区,其中云南有秋海棠属植物100余种。近年来,随着广西地区秋海棠属植物野外调查的不断深入,陆续发现并发表了大量秋海棠属新类群,使得广西产秋海棠属植物数目已超出1991年《广西植物志》(第一卷)收录的19种,因此亟需对发表于不同期刊中的类群进行系统地整理和汇总。鉴于此,该文通过查阅文献资料和考证标本信息,整理并汇总了广西产秋海棠属植物共84种,包括2个变种、11个亚种和46个特有种,新增的65种为《广西植物志》(第一卷)中未收录的类群,并补充了这些类群的特征集要、凭证标本信息和属下分组概况,为今后开展秋海棠属植物的分类修订以及系统学、进化生物学和保护生物学等研究提供了参考依据。     

Independent Origins of Vectored Plant Pathogenic Bacteria from Arthropod-Associated <Emphasis Type="Italic">Arsenophonus</Emphasis> Endosymbionts

The genus Arsenophonus (Gammaproteobacteria) is comprised of intracellular symbiotic bacteria that are widespread across the arthropods. These bacteria can significantly influence the ecology and life history of their hosts. For instance, Arsenophonus nasoniae causes an excess of females in the progeny of parasitoid wasps by selectively killing the male embryos. Other Arsenophonus bacteria have been suspected to protect insect hosts from parasitoid wasps or to expand the host plant range of phytophagous sap-sucking insects. In addition, a few reports have also documented some Arsenophonus bacteria as plant pathogens. The adaptation to a plant pathogenic lifestyle seems to be promoted by the infection of sap-sucking insects in the family Cixiidae, which then transmit these bacteria to plants during the feeding process. In this study, we define the specific localization of an Arsenophonus bacterium pathogenic to sugar beet and strawberry plants within the plant hosts and the insect vector, Pentastiridius leporinus (Hemiptera: Cixiidae), using fluorescence in situ hybridization assays. Phylogenetic analysis on 16S rRNA and nucleotide coding sequences, using both maximum likelihood and Bayesian criteria, revealed that this bacterium is not a sister taxon to “Candidatus Phlomobacter fragariae,” a previously characterized Arsenophonus bacterium pathogenic to strawberry plants in France and Japan. Ancestral state reconstruction analysis indicated that the adaptation to a plant pathogenic lifestyle likely evolved from an arthropod-associated lifestyle and showed that within the genus Arsenophonus, the plant pathogenic lifestyle arose independently at least twice. We also propose a novel Candidatus status, “Candidatus Arsenophonus phytopathogenicus” novel species, for the bacterium associated with sugar beet and strawberry diseases and transmitted by the planthopper P. leporinus.     

鸭疫里默氏杆菌铁离子代谢机制研究进展

铁离子是大多数细菌生存所必需的一种营养物质,但过多的铁离子会通过芬顿反应产生的活性氧对细菌造成损伤。因此,细菌通过摄取、调控、螯合、外排等机制维持体内铁离子的稳态。鸭疫里默氏杆菌(Riemerella anatipestifer)是一种最新被归类于威克斯菌科里氏杆菌属的革兰氏阴性菌。该菌主要感染禽类,参与该菌的铁离子代谢基因具有特别之处。本文对鸭疫里默氏杆菌铁离子代谢机制研究进展进行了系统总结和阐述,包括该菌的TonB系统、TonB依赖性受体、Fur蛋白及Dps蛋白等在铁离子转运、调控、螯合中的功能,以及以上蛋白在鸭疫里默氏杆菌致病中的作用,以期更全面地理解鸭疫里默氏杆菌铁代谢机制,并为进一步深入研究该菌铁离子代谢提供理论依据和参考。     

Interspecific competition between the braconid endoparasitoids Glyptapanteles porthetriae and Glyptapanteles liparidis in Lymantria dispar larvae

The effect of interspecific competition between the solitary endoparasitoid Glyptapanteles porthetriae Muesebeck (Hymenoptera: Braconidae) and the gregarious Glyptapanteles liparidis Bouché (Hymenoptera: Braconidae), was investigated in larvae of Lymantria dispar L. (Lepidoptera: Lymantriidae). Host larvae were parasitized by both wasp species simultaneously in premolt to the 2^nd or the 3^rd host instar or in an additional approach with a 4-day delay in parasitization by the second wasp species. Host acceptance experiments revealed that both wasp species do not discriminate between unparasitized host larvae and larvae parasitized previously by the same or the other species. In more than 90% female wasps parasitized the larva they encountered first. During the period of endoparasitic development, larvae of the competing parasitoid species never attacked the egg stage of the other species. When host larvae were parasitized simultaneously by both wasp species, the rate of successful development of both species depended on the age of the host larva at the time of its parasitization; G. liparidis emerged successfully from 44% of host larvae parasitized during the premolt to 2^nd instar, G. porthetriae from 28%, and in 20% of the hosts both parasitoid species were able to develop in one gypsy moth larva. However, when host larvae were parasitized simultaneously during premolt to the 3^rd instar, G. liparidis was successful in 90% of the hosts, compared to 8% from which only G. porthetriae emerged. In the experiments with delayed oviposition, generally the species that oviposited first succeeded in completing its larval development. Larvae of the species ovipositing with four days delay were frequently attacked and killed by larvae of the first parasitizing species or suffered reduced growth. As the secondary parasitoid species, G. porthetriae-larvae were never able to complete their development, whereas G. liparidis developed successfully in at least 12,5% of the multiparasitized host larvae. Thus, multiparasitism of gypsy moth larvae by both Glyptapanteles species corresponds to the contest type; however, G. porthetriae is only able to develop successfully as the primary parasitoid of young host larvae.