褶皱臂尾轮虫同胞物种复合类群分类的研究进展

褶皱臂尾轮虫包含复杂的同胞物种,对其分类研究经过了一个长期发展的过程。综述了轮虫分类手段的发展进程,介绍和分析了轮虫特别是褶皱臂尾轮虫同胞物种复合类群分类的研究进展。褶皱臂尾轮虫的L-型和S-型属于B.plicatilis和B.rotundiformis两个物种。这个复合类群中还包括更多的种类,如B.plicatilis sensu stricto、B.ibericus和B.rotundiformis,其中B.plicatilis sensu stricto属于L型,B.ibericus属于SM-型,B.rotundiformis属于SS-型。此类研究丰富了轮虫系统分类学,提高了水域生态调查的准确性。     

The association between the caprellid Pariambus typicus Krøyer (Crustacea,Amphipoda) and ophiuroids

The work of Kazutsugu Hirayama over the past 25 yearspromoted the wide use of the marine rotifer Brachionusplicatilis as an experimental model in zooplanktonecology. His reports about the nature of geneticvariation in the B. plicatilis complex stimulated usto investigate how mate recognition maintains speciesboundaries. For the past several years, we haveexamined chemical communication between female andmale B. plicatilis. Here we report on the comparativebinding of polyclonal antibody against the materecognition pheromone (MRP) to three B. plicatilisstrains and three B. rotundiformis strains.Quantification of anti-MRP binding permitsinvestigation of how the female mating signal differsamong closely related Brachionus species and strains.Antibody binding reflects differentiation independentof the male receptor which has been describedelsewhere. Anti-MRP bound to females of all sixstrains and was localized in the corona. Antibodybinding greatly reduced mating in all three B. plicatilis strains. However, antibody bindingsignificantly reduced mating in only one of the B. rotundiformis strains. The MRP of both species has asimilar molecular weight, but the differential bindingsuggests that the mate recognition pheromone onfemales has differentiated in B. plicatilis and B. rotundiformis.     

GABA, 5-HT and amino acids in the rotifers Brachionus plicatilis and Brachionus rotundiformis

gamma-Aminobutyric acid (GABA) and 5-hydroxytryptamine (5-HT) have been shown to increase the reproduction of the Brachionus plicatilis (NH3L strain). In the present study, the endogenous presence of GABA and 5-HT in the rotifers B. plicatilis (NH3L and Kamiura strains) and Brachionus rotundiformis (Langkawi strain) were confirmed by dot blot immunoassay and high-performance liquid chromatography (HPLC). HPLC showed that GABA and 5-HT concentrations in the three rotifer strains range from 71 to 188 pmol/mg and from 12 to 64 pmol/mg, respectively. A total of 33 amino acids were also detected in B. plicatilis and B. rotundiformis, with glutamic acid, serine, glycine, taurine, threonine, alanine, arginine, proline, valine and isoleucine in high concentrations relative to other amino acids.     

Interspecific interactions in the marine rotifer microcosm

Copepods and protozoans often co-exist in marinerotifer mass cultures. Interspecific interactionbetween the rotifer Brachionus rotundiformisTschugunoff and eight other zooplankton species,namely Brachionus plicatilis O. F. Müller (rotifer),Diaphanosoma celebensis Stingelin (cladoceran),Tigriopus japonicus Mori, Acartia sp. (copepod),Euplotes sp., Vorticella sp., an unidentifiedprotozoan species (P1 strain) (protozoan) and Artemiasp. (anostracan) at two developmental stages (nauplii –0.95 mm, 0 days old; adults – 3.3 mm, 19 days old) wereinvestigated in the laboratory.There was no contaminating species that contributedto an increase in rotifer population growth during theexperiments. Four types of interspecific interactionswere seen between B. rotundiformis and otherco-existing zooplankton species. These include effectson population growth: (1) both species declined, (2) onespecies is promoted while the other is not influenced,(3) one species is declined while the other is notinfluenced and (4) one species is promoted while theother declined. The first type was exhibited by B. rotundiformis vs B. plicatilis, B. rotundiformis vsD. celebensis and B. rotundiformis vs Artemia sp. The second type was exhibited by B. rotundiformis vsVorticella sp. and the third type by B. rotundiformisvs Euplotes sp. and B. rotundiformis vs T. japonicus. The fourth type was exhibited by B. rotundiformis vs Acartia sp. and B. rotundiformis vs P1 strain.     

Predatory interactions between a cyclopoid copepod and three sibling rotifer species

SUMMARY 1. Cyclopoid copepod predation on rotifers affects the dynamics and structure of zooplankton communities. We address the differential vulnerability of three sympatric rotifer sibling species belonging to the Brachionus plicatilis species complex. These co-occur with their cyclopoid predator, Diacyclops bicuspidatus odessanus .
2. Using video recording and tracking, we analysed the steps in predation including attack distance, attack angle, and rotifer species swimming in the presence and absence of the predator. Our results show the greater vulnerability of B. rotundiformis (the smallest species) to D. b. odessanus predation, which is associated with a high percentage of attacks after contact. Brachionus plicatilis (the biggest species) is the less vulnerable prey, with low percentage of attacks after contact and captures after attacks. Branchionus ibericus , the intermediate sized species, had also intermediate vulnerability.
3. The differential vulnerability provides insight into the coexistence and seasonal succession of these competing rotifer species. Our results show that the competitive superiority of B. rotundiformis may be balanced by its greater vulnerability to copepod predation.     

The mate recognition protein gene mediates reproductive isolation and speciation in the Brachionus plicatilis cryptic species complex

ABSTRACT: BACKGROUND: Chemically mediated prezygotic barriers to reproduction likely play an important role in speciation. In facultatively sexual monogonont rotifers from the Brachionus plicatilis cryptic species complex, mate recognition of females by males is mediated by the Mate Recognition Protein (MRP), a globular glycoprotein on the surface of females, encoded by the mmr-b gene family. In this study, we sequenced mmr-b copies from 27 isolates representing 11 phylotypes of the B. plicatilis species complex, examined the mode of evolution and selection of mmr-b, and determined the relationship between mmr-b genetic distance and mate recognition among isolates. RESULTS: Isolates of the B. plicatilis species complex have 1-4 copies of mmr-b, each composed of 2-9 nearly identical tandem repeats. The repeats within a gene copy are generally more similar than are gene copies among phylotypes, suggesting concerted evolution. Compared to housekeeping genes from the same isolates, mmr-b has accumulated only half as many synonymous differences but twice as many non-synonymous differences. Most of the amino acid differences between repeats appear to occur on the outer face of the protein, and these often result in changes in predicted patterns of phosphorylation. However, we found no evidence of positive selection driving these differences. Isolates with the most divergent copies were unable to mate with other isolates and rarely self-crossed. Overall the degree of mate recognition was significantly correlated with the genetic distance of mmr-b. CONCLUSIONS: Discrimination of compatible mates in the B. plicatilis species complex is determined by proteins encoded by closely related copies of a single gene, mmr-b. While concerted evolution of the tandem repeats in mmr-b may function to maintain identity, it can also lead to the rapid spread of a mutation through all copies in the genome and thus to reproductive isolation. The mmr-b gene is evolving rapidly, and novel alleles may be maintained and increase in frequency via asexual reproduction. Our analyses indicate that mate recognition, controlled by MMR-B, may drive reproductive isolation and allow saltational sympatric speciation within the B. plicatilis cryptic species complex, and that this process may be largely neutral.     

Speciation in ancient cryptic species complexes: evidence from the molecular phylogeny of Brachionus plicatilis (Rotifera)

Continental lake-dwelling zooplanktonic organisms have long been considered cosmopolitan species with little geographic variation in spite of the isolation of their habitats. Evidence of morphological cohesiveness and high dispersal capabilities support this interpretation. However, this view has been challenged recently as many such species have been shown either to comprise cryptic species complexes or to exhibit marked population genetic differentiation and strong phylogeographic structuring at a regional scale. Here we investigate the molecular phylogeny of the cosmopolitan passively dispersing rotifer Brachionus plicatilis (Rotifera: Monogononta) species complex using nucleotide sequence variation from both nuclear (ribosomal internal transcribed spacer 1, ITS1) and mitochondrial (cytochrome c oxidase subunit I, COI) genes. Analysis of rotifer resting eggs from 27 salt lakes in the Iberian Peninsula plus lakes from four continents revealed nine genetically divergent lineages. The high level of sequence divergence, absence of hybridization, and extensive sympatry observed support the specific status of these lineages. Sequence divergence estimates indicate that the B. plicatilis complex began diversifying many millions of years ago, yet has showed relatively high levels of morphological stasis. We discuss these results in relation to the ecology and genetics of aquatic invertebrates possessing dispersive resting propagules and address the apparent contradiction between zooplanktonic population structure and their morphological stasis.     

The use of rotifers for the maintenance of monoalgal mass cultures of Spirulina

Zooplankton was successfully used for the biological control of unicellular algal contaminants in Spirulina mass cultures even under conditions adverse to the growth of Spirulina (maximal winter daily temperature of approximately 10 degrees C and very low bicarbonate concentration). Brachionus plicatilis (Rotifera) was the most successful species of zooplankton used. The interrelationships between Spirulina, green unicellular contaminant, and B. plicatilis were studied under various conditions. Two species of unicellular contaminant were used; Monoraphidium minutum was isolated from local cultures and Chlorella vulgaris, obtained from contaminated Spirulina cultures in Israel. The rotifer B. plicatilis successfully controlled the population size of both contaminants whether they were introduced in a single addition or as a daily dose. The biological control of the unicellular contaminants allows Spirulina to be cultured in a medium low in bicarbonate, thereby reducing the cost of the medium and increasing the quantity of CO(2) that may be freely absorbed from the atmosphere at the optimal pH for Spirulina cultivation.     

Phylogenetic study of two truffles, Tuber formosanum and Tuber furfuraceum, identified from Taiwan

Truffles are one of the most valuable edible fungi and have drawn extensive research interests worldwide. In Taiwan, two species of truffle, Tuber formosanum and Tuber furfuraceum , have been identified and reported. Although the morphological features of these two truffles have been described, lack of molecular identification has led to difficulties with firmly establishing their relatedness to other truffles. In this study, we utilized the internal transcribed spacer (ITS) and β-tubulin gene sequences to generate the phylogenetic relationship of T. formosanum and T. furfuraceum with other taxonomic relatives. Our analysis revealed five/three major phylogenetic clades according to the 5.8S-ITS2/β-tubulin gene sequences and corroborated with their morphological characterization. Tuber formosanum highly resembles the Tuber indicum B complex, while T. furfuraceum is most similar to Tuber huidongense . Based on a molecular clock, we estimated that T. furfuraceum and T. formosanum would have diverged from their close relatives in mainland China between 10.2 and 4.1 Ma, respectively. Based on the results, we propose that these two Tuber species found in Taiwan might originate from the common ancestors with some truffle species in China. However, due to a long divergence time and geographical separation, they have evolved into indigenous species of Taiwan.     

A quorum-quenching approach to investigate the conservation of quorum-sensing-regulated functions within the Burkholderia cepacia complex

Taxonomic studies of the past few years have shown that the Burkholderia cepacia complex, a heterogeneous group of B. cepacia-like organisms, consists of at least nine species. B. cepacia complex strains are ubiquitously distributed in nature and have been used for biocontrol, bioremediation, and plant growth promotion purposes. At the same time, B. cepacia complex strains have emerged as important opportunistic pathogens of humans, particularly those with cystic fibrosis. All B. cepacia complex species investigated thus far use quorum-sensing (QS) systems that rely on N-acylhomoserine lactone (AHL) signal molecules to express certain functions, including the production of extracellular proteases, swarming motility, biofilm formation, and pathogenicity, in a population-density-dependent manner. In this study we constructed a broad-host-range plasmid that allowed the heterologous expression of the Bacillus sp. strain 240B1 AiiA lactonase, which hydrolyzes the lactone ring of various AHL signal molecules, in all described B. cepacia complex species. We show that expression of AiiA abolished or greatly reduced the accumulation of AHL molecules in the culture supernatants of all tested B. cepacia complex strains. Phenotypic characterization of wild-type and transgenic strains revealed that protease production, swarming motility, biofilm formation, and Caenorhabditis elegans killing efficiency was regulated by AHL in the large majority of strains investigated.     

Persistent genetic signatures of colonization in Brachionus manjavacas rotifers in the Iberian Peninsula

Recent phylogeographical assessments have consistently shown that continental zooplankton display high levels of population subdivision, despite the high dispersal capacity of their diapausing propagules. As such, there is an apparent paradox between observed cosmopolitanism in the zooplankton that is associated with long-distance dispersal, and strong phylogeographical structures at a regional scale. Such population dynamics, far from migration-drift equilibrium, have been shown in the rotifer species complex Brachionus plicatilis, a group of over a dozen species inhabiting salt lakes and coastal lagoons worldwide. Here we present the mitochondrial DNA phylogeography of one of these species, Brachionus manjavacas, in the Iberian Peninsula, where it often co-occurs with the morphologically similar species B. plicatilis sensu stricto. We obtained sequences from 233 individuals from diapausing eggs and clonal cultures from 16 lakes in the Iberian Peninsula, and a Tunisian lake. Two strongly supported deep mitochondrial DNA clades were found (A and B). Phylogenetic and nested clade analysis showed that clade A has a strong phylogeographical structure, with a strong similarity of phylogeographical patterns between B. manjavacas clade A and B. plicatilis s.s. These include (i) signatures of allopatric fragmentation between central and southern populations, and (ii) range expansions in the Iberian Peninsula, both likely to have occurred during the Pleistocene. We find evidence for a glacial refugium in the Guadiana basin. Clades A and B co-occurred in several of these lakes because of range expansion and secondary contact between both clades. The co-occurrence between B. plicatilis s.s. and B. manjavacas is not recent, and both species might have experienced similar environmental challenges during the Pleistocene. The strong correlation of genetic and geographical distance found suggests that historical events can lead to such correlation, mirroring the effects of 'isolation by distance' in equilibrium populations.     

营养强化时褶皱臂尾轮虫对饵料微藻的摄食

选取适宜浓度利用单种微藻和混合微藻对轮虫进行营养强化,采用实验生态学方法研究了轮虫滤水率和摄食率的动态变化.结果表明:微藻浓度、微藻种类和培养时间均对轮虫的滤水率和摄食率有显著影响;轮虫对几种单种微藻的滤水率和摄食率均随培养时间的延长而下降,在实验条件下,6h内轮虫对3种微藻的滤水率大小顺序为小球藻＞球等鞭金藻＞牟氏角毛藻,12h内轮虫对3种和,微藻的滤水率大小顺序则为球等鞭金藻＞小球藻＞牟氏角毛藻;轮虫在混合微藻中的选择顺序为球等鞭金藻＞小球藻＞牟氏角毛藻.     

Application of unicellular algae Chlorella vulgaris for the mass-culture of marine rotifer Brachionus

Condensed suspension of Chlorellavulgaris was used for the food of the rotifer Brachionus plicatilis and B. rotundiformis inplace of Nannochloropsis oculata. Thisreport describes the characteristics of C. vulgaris as arotifer food in comparison with N. oculata and thepresent status of this field.The cell components of C. vulgarissuch as protein content, amino acids, minerals andvitamins are generally similar to those of N. oculata. However, the taxonomic status of thesealgal species are different. Based on thesimilarity of cell components, the dietary value ofC. vulgaris is equal in value to that of N. oculata for rotifer growth. Dietary value ofC. vulgaris can be improved by addition ofvitamin B₁₂. This improved C. vulgaris is currently widely used as an indispensable food organism for rotifer culture. Recent investigationshave shown that the use of the condensed suspensionof C. vulgaris makes it possible tosignificantly increase the rotifer density atharvest. Application of condensed C. vulgaris has made rotifer culture quite easy because theculture of N. oculata is no longer required,and intensive rotifer production in aquaculture cannow be realized.     

In silico analysis of Burkholderia pseudomallei genome sequence for potential drug targets

Recent advances in DNA sequencing technology have enabled elucidation of whole genome information from a plethora of organisms. In parallel with this technology, various bioinformatics tools have driven the comparative analysis of the genome sequences between species and within isolates. While drawing meaningful conclusions from a large amount of raw material, computer-aided identification of suitable targets for further experimental analysis and characterization, has also led to the prediction of non-human homologous essential genes in bacteria as promising candidates for novel drug discovery. Here, we present a comparative genomic analysis to identify essential genes in Burkholderia pseudomallei. Our in silico prediction has identified 312 essential genes which could also be potential drug candidates. These genes encode essential proteins to support the survival of B. pseudomallei including outer-inner membrane and surface structures, regulators, proteins involved in pathogenenicity, adaptation, chaperones as well as degradation of small and macromolecules, energy metabolism, information transfer, central/intermediate/miscellaneous metabolism pathways and some conserved hypothetical proteins of unknown function. Therefore, our in silico approach has enabled rapid screening and identification of potential drug targets for further characterization in the laboratory.     

Genetic variation among marine Brachionus strains and function of mate recognition pheromone (MRP)

Recent studies on morphology and genetics of marinerotifer populations have demonstrated the existence ofconsiderable variation. Differences in theglycoprotein structure of a mate recognition pheromone(MRP) probably have a primary role in maintainingspecies boundaries among Brachionus species.This study examined factors involved in mating toclarify their relation with genetic variability. Threeexperiments were performed using 3 Brachionusplicatilis strains (Russia, Germany and Tokyo) and 4B. rotundiformis strains (Hamana, Fiji, Thai andSpain).Selfing and cross mating of 7 rotifer strains wasconducted in the first experiment. Russian malesmated with B. plicatilis females in thefollowing increasing order, based on mating attempts:German, Tokyo and Russia. There was little matingattempts with females of B. rotundiformisstrains. In a second set of experiments, the binding of anantibody (anti-MRP) derived from the MRP of Russianstrain was tested. A fluorescent label was attached toanti-MRP and the antibody reacted with the MRP onfemales of seven strains. The fluorescence intensity,indicating the degree of antibody binding, wasmeasured with epifluorescence microscopy and imageanalysis. The binding intensity of the anti-MRP to theMRP was in the following increasing order: Hamana,Fiji, Spain, German, Tokyo, Thai and Russia.It was expected that when the anti-MRP binds tothe MRP of a female, the males recognition of thefemale would be inhibited. In the third experiment,male mating with females exposed to the anti-MRP wascompared with unexposed females. Russia Tokyo, Thaiand Spain females exposed to anti-MRP, elicited fewermale mating attempts than unexposed females. Matingfrequency and anti-MRP binding significantlycorrelated with genetic distance obtained from isozymeanalysis.     

蒙古裸腹溞与褶皱臂尾轮虫的种间关系

在实验室内将蒙古裸腹溞按0．06、0．10、0．30、0．60个·ml^-1的密度与褶皱臂尾轮虫(密度为0．3个·ml。)进行}昆合培养,同时按上述密度分别单种培养了蒙古裸腹溞和褶皱臂尾轮虫作为对照．结果表明,蒙古裸腹溞与褶皱臂尾轮虫之间存在种间竞争,当二者共存时,褶皱臂尾轮虫对蒙古裸腹溞种群产生压制作用,使蒙古裸腹溞在混合培养体系中以很低的密度存在,而褶皱臂尾轮虫种群受蒙古裸腹溞影响不大．通过进一步的饥饿实验发现90％的褶皱臂尾轮虫经过144h饥饿后仍然存活,而经过120h饥饿后的蒙古裸腹溞100％死亡,褶皱臂尾轮虫耐饥饿能力较强是其在与蒙古裸腹溞竞争中获胜的原因之一。     

A Quorum-Quenching Approach To Investigate the Conservation of Quorum-Sensing-Regulated Functions within the Burkholderia cepacia Complex

Taxonomic studies of the past few years have shown that the Burkholderia cepacia complex, a heterogeneous group of B. cepacia-like organisms, consists of at least nine species. B. cepacia complex strains are ubiquitously distributed in nature and have been used for biocontrol, bioremediation, and plant growth promotion purposes. At the same time, B. cepacia complex strains have emerged as important opportunistic pathogens of humans, particularly those with cystic fibrosis. All B. cepacia complex species investigated thus far use quorum-sensing (QS) systems that rely on N-acylhomoserine lactone (AHL) signal molecules to express certain functions, including the production of extracellular proteases, swarming motility, biofilm formation, and pathogenicity, in a population-density-dependent manner. In this study we constructed a broad-host-range plasmid that allowed the heterologous expression of the Bacillus sp. strain 240B1 AiiA lactonase, which hydrolyzes the lactone ring of various AHL signal molecules, in all described B. cepacia complex species. We show that expression of AiiA abolished or greatly reduced the accumulation of AHL molecules in the culture supernatants of all tested B. cepacia complex strains. Phenotypic characterization of wild-type and transgenic strains revealed that protease production, swarming motility, biofilm formation, and Caenorhabditis elegans killing efficiency was regulated by AHL in the large majority of strains investigated.     

Long-term coexistence of rotifer cryptic species

Despite their high morphological similarity, cryptic species often coexist in aquatic habitats presenting a challenge in the framework of niche differentiation theory and coexistence mechanisms. Here we use a rotifer species complex inhabiting highly unpredictable and fluctuating salt lakes to gain insights into the mechanisms involved in stable coexistence in cryptic species. We combined molecular barcoding surveys of planktonic populations and paleogenetic analysis of diapausing eggs to reconstruct the current and historical coexistence dynamics of two highly morphologically similar rotifer species, B. plicatilis and B. manjavacas. In addition, we carried out laboratory experiments using clones isolated from eight lakes where both species coexist to explore their clonal growth responses to salinity, a challenging, highly variable and unpredictable condition in Mediterranean salt lakes. We show that both species have co-occurred in a stable way in one lake, with population fluctuations in which no species was permanently excluded. The seasonal occurrence patterns of the plankton in two lakes agree with laboratory experiments showing that both species differ in their optimal salinity. These results suggest that stable species coexistence is mediated by differential responses to salinity and its fluctuating regime. We discuss the role of fluctuating salinity and a persistent diapausing egg banks as a mechanism for species coexistence in accordance with the 'storage effect'.     

Playing Darwin. Part A. Experimental Evolution in Drosophila

In 2009 we celebrate Charles Darwin’s second centenary, and 150 years since the publication of ‘The Origin of Species’. After so many years, what has changed in the way we understand Evolution? Obviously we have now a full understanding of the mechanisms underlying heritability. Many molecular tools are available, allowing among other things to reconstruct more accurately the evolutionary history of species and use a comparative approach to infer evolutionary processes. But we can also study evolution in action. Such studies—Experimental Evolution—help us to characterize in detail the evolutionary processes and patterns as a function of environmental challenges, the previous history and present state of populations, and the interactions between such factors. We have now a wide variety of organisms that have been studied with this approach, exploring a diversity of potentialities, in biological characteristics and genetic tools, and covering a variety of evolutionary questions. In this short article I will illustrate the potentialities of Experimental Evolution, focusing in three studies in Drosophila. These and other studies of Experimental Evolution illustrate that Evolution is often local, involving complex patterns and processes, which lead both to specific adaptations and to biological diversity, as Darwin already stated clearly in ‘The Origin of Species’.     

Isolation and characterization of soluble boron complexes in higher plants. The mechanism of phloem mobility of boron.

Boron (B) polyol complexes have been isolated and characterized from the phloem sap of celery (Apium graveolens L.) and the extrafloral nectar of peach (Prunus persica L.). In celery the direct analysis of untreated phloem sap by matrix-assisted laser desorption-Fourier transform mass spectrometry, with verification by high-performance liquid chromagraphy and gas chromatography-mass spectrometry, revealed that B is present in the phloem as the mannitol-B-mannitol complex. Molecular modeling further predicted that this complex is present in the 3,4 3',4' bis-mannitol configuration. In the extrafloral nectar of peach, B was present as a mixture of sorbitol-B-sorbitol, fructose-B-fructose, or sorbitol-B-fructose. To our knowledge, these findings represent the first successful isolation and characterization of soluble B complexes from higher plants and provide a mechanistic explanation for the observed phloem B mobility in these species.