Species Structure in Euplotes crassus (Ciliophora, Hypotrichida)

ABSTRACT. We observed different low salinity tolerances between two groups of populations of Euplotes crassus. After mating analysis, the less tolerant populations could be assigned to three mating groups. Two of them were separated by preconjugative barriers but both conjugated with a third one. A breeding test showed evidence of (potential) introgression mediated by this "bridging" group. On the other hand, the more tolerant populations formed a fourth mating group unable to conjugate with the others and characterized by distinct zymogram patterns. We conclude that natural introgression among the studied populations is not complete and that the species structure of E. crassus comprises intrafertile sets of populations among which gene flow may be difficult or virtually absent.     

Time-course analysis of nuclear events during conjugation in the marine ciliate Euplotes vannus and comparison with other ciliates (Protozoa,Ciliophora)

Ciliates represent a morphologically and genetically distinct group of single-celled eukaryotes that segregate germline and somatic functions into two types of nuclei and exhibit complex cytogenetic events during the sexual process of conjugation, which is under the control of the so-called “mating type systems”. Studying conjugation in ciliates may provide insight into our understanding of the origins and evolution of sex and fertilization. In the present work, we studied in detail the sexual process of conjugation using the model species Euplotes vannus, and compared these nuclear events with those occurring in other ciliates. Our results indicate that in E. vannus: 1) conjugation requires about 75 hours to complete: the longest step is the development of the new macronucleus (ca. 64h), followed by the nuclear division of meiosis I (5h); the mitotic divisions usually take only 2h; 2) there are three prezygotic divisions (mitosis and meiosis I and II), and two of the eight resulting nuclei become pronuclei; 3) after the exchange and fusion of the pronuclei, two postzygotic divisions occur; two of the four products differentiate into the new micronucleus and macronucleus, respectively, and the parental macronucleus degenerates completely; 4) comparison of the nuclear events during conjugation in different ciliates reveals that there are generally three prezygotic divisions while the number of postzygotic divisions is highly variable. These results can serve as reference to investigate the mating type system operating in this species and to analyze genes involved in the different steps of the sexual process.     

A Morphometric Study of Euryhalinity in Marine Populations of the Ciliate Genus Euplotes

ABSTRACT. Twenty different clonal strains of marine and brackish Euplotes , representing four morphotypes, were tested for hyposalinity tolerance by a method which gradually acclimated the cells to lower salinity medium. The lowest salinities in which the strains could thrive ranged from 60% of normal seawater to complete freshwater. The morphological effects of culture medium salinity were also examined for two strains of a small "Euplotes charon" morphotype, as well as for two mating compatible "Euplotes vannus" strains and several of their exconjugates. There were no differences between the euryhaline strains grown in fresh or saltwater, except for a slight increase in overall cell size in one strain when cultured in freshwater medium. E. vannus strains increased in overall cell size with decreased salinity; also, the dorsal surface of the cells can become disorganized when the cells are cultured in 30% normal seawater.     

农业生态系统中新物种引进的初步探讨

在现有的农业生态系统中,引进某些新物种,往往可以使整个系统的功能发生变化。如果新物种引进得当,则可产生良好的效果,系统功能得以改善,效益提高;反之,则会造成人力、物力、财力的损失,削弱系统的功能,甚至可能遗患无穷。因此,对农业生态系统中     

Rare, Fragile Species, Small Populations, and the Dilemma of Collections

Considerable sums of money are aimed at biodiversity conservation programs at present, and although collection of specimens is necessary for the advancement of taxonomy and systematics, we detected a dilemma between conservation and collection of rare species with small populations. Collecting may act synergistically with other factors to increase the risk of, or even drive species to extinction. We present some examples we believe show the conflict between collecting rare species with a small population and the conservation programs for those species. We also stress the need to review the Minimum Viable Population Size concept as it can be used as a justification for collecting small, rare, and declining population species.     

Evidence for Gene Duplication and Allelic Codominance (not Hierarchical Dominance) at the Mating‐Type Locus of the Ciliate,Euplotes crassus

The high‐multiple mating system of Euplotes crassus is known to be controlled by multiple alleles segregating at a single locus and manifesting relationships of hierarchical dominance, so that heterozygous cells would produce a single mating‐type substance (pheromone). In strain L‐2D, now known to be homozygous at the mating‐type locus, we previously identified two pheromones (Ec‐α and Ec‐1) characterized by significant variations in their amino acid sequences and structure of their macronuclear coding genes. In this study, pheromones and macronuclear coding genes have been analyzed in strain POR‐73 characterized by a heterozygous genotype and strong mating compatibility with L‐2D strain. It was found that POR‐73 cells contain three distinct pheromone coding genes and, accordingly, secrete three distinct pheromones. One pheromone revealed structural identity in amino acid sequence and macronuclear coding gene to the Ec‐α pheromone of L‐2D cells. The other two pheromones were shown to be new and were designated Ec‐2 and Ec‐3 to denote their structural homology with the Ec‐1 pheromone of L‐2D cells. We interpreted these results as evidence of a phenomenon of gene duplication at the E. crassus mating‐type locus, and lack of hierarchical dominance in the expression of the macronuclear pheromone genes in cells with heterozygous genotypes.     

Factors that Control the Species Composition of Freshwater Phytoplankton,with Special Attention to Nutrient Concentrations

Variations of species composition and population size of planktonic algae were studied in relation to the different nutrient levels in a eutrophic pond and an oligotrophic lake. The results obtained were discussed in comparison with the changes in photosynthesis and chlorophyll of several algal populations cultured under different nutrient conditions. As based on unialgal culture experiments two types of algae (the Chlorella-type and the Melosira-type) could be distinguished with regard to variations of photosynthesis and chlorophyll in response to different nutrient levels. Distribution of the Chlorella-type algae may be confined to eutrophic waters, while the Melosira-type algae can be distributed widely in waters from oligotrophic to eutrophic.     

多次交配对松褐天牛精子数量消耗、产卵量和孵化率的影响

通过解剖松褐天牛雌虫受精囊和室内饲养观察,研究了多次交配对松褐天牛雌虫受精囊内精子数量消耗、产卵量、卵孵化率、产卵历期和寿命的影响,并对松褐天牛雌性生殖系统、精子形态及精子在雌性生殖系统中的分布动态进行了详细的描述,其结果表明:多次交配的松褐天牛雌虫受精囊内的精子数量长期保持在12万个左右,而只交配一次的松褐天牛雌虫受精囊内精子数量不断消耗,交配16天后降到5万个以下。多次交配的雌虫产卵量(167·0870粒)和卵孵化率(94·38%)都显著大于单次交配雌虫的产卵量(113·5217粒)和卵孵化率(83·79%);但是多次交配的雌虫与单次交配的雌虫相比较,产卵历期和寿命差异都不显著。     

Genetics and Morphology Characterize the Dinoflagellate Symbiodinium voratum,n. sp., (Dinophyceae) as the Sole Representative of Symbiodinium Clade E

Dinoflagellates in the genus Symbiodinium are ubiquitous in shallow marine habitats where they commonly exist in symbiosis with cnidarians. Attempts to culture them often retrieve isolates that may not be symbiotic, but instead exist as free‐living species. In particular, cultures of Symbiodinium clade E obtained from temperate environments were recently shown to feed phagotrophically on bacteria and microalgae. Genetic, behavioral, and morphological evidence indicate that strains of clade E obtained from the northwestern, southwestern, and northeastern temperate Pacific Ocean as well as the Mediterranean Sea constitute a single species: Symbiodinium voratum n. sp. Chloroplast ribosomal 23S and mitochondrial cytochrome b nucleotide sequences were the same for all isolates. The D1/D2 domains of nuclear ribosomal DNA were identical among Western Pacific strains, but single nucleotide substitutions differentiated isolates from California (USA) and Spain. Phylogenetic analyses demonstrated that S. voratum is well‐separated evolutionarily from other Symbiodinium spp. The motile, or mastigote, cells from different cultures were morphologically similar when observed using light, scanning, and transmission electron microscopy; and the first complete Kofoidian plate formula for a Symbiodinium sp. was characterized. As the largest of known Symbiodinium spp., the average coccoid cell diameters measured among cultured isolates ranged between 12.2 (± 0.2 SE) and 13.3 (± 0.2 SE) μm. Unique among species in the genus, a high proportion (approximately 10–20%) of cells remain motile in culture during the dark cycle. Although S. voratum occurs on surfaces of various substrates and is potentially common in the plankton of coastal areas, it may be incapable of forming stable mutualistic symbioses.     

Molecular Phylogeny and Genetic Divergence of the Introduced Populations of Japanese Guppies, Poecilia reticulata

A phylogenetic tree of guppies from seven introduced Japanese populations was constructed using a mitochondrial gene, and the relationship between these Japanese populations and indigenous South American populations was examined. Phylogenetic trees constructed by maximum parsimony and neighbor joining methods indicated four major groups: Group A: Trinidadian populations; Group B: the northernmost Okinawa populations; Group C: the northern Okinawa populations; Group D: Shimoda populations and the remainder of Trinidadian, Venezuelan, and southern Okinawa populations. Considerable genetic differences were observed among the haplotypes within each Okinawa population similar to that found among haplotypes of different native populations, but not within the Shimoda population, because each Okinawa population included haplotypes belonging to different groups. Genetic differences between Groups B, C, and D might reflect those of the introduced lineages; however, these differences might not result from the divergence after introduction events. These results suggested that multiple introductions of different breeding lineages might cause considerable divergence within and between Okinawa populations.     

Genetic analysis of mating type differentiation in Paramecium tetraurelia. III. A mutation restricted to mating type E and affecting the determination of mating type

In P. tetraurelia each cell is determined to express only one of the two complementary mating types, O and E. This determination is under cytoplasmic control and seems to be achieved only by the commitment or noncommitment to the expression of mating type E. All the previously known mutations affecting the differentiation of mating type prevent the expression of the E mating type (O-restricted mutations) without affecting the determination process. An E-restricted mutation was obtained: mtF^E. Its phenotypic properties indicate that the mutation affects the determination process itself. When an O cell becomes mtF^E/mtF^E it acquires the E mating type and an E-determining cytoplasm. We propose that this constitutive determination for the E mating type is due to the inefficiency of a factor which is normally active in an O cell. This factor would act like a repressor and stabilize the E functions under an inactive state.     

Improved description of the bipolar ciliate,Euplotes petzi,and definition of its basal position in the Euplotes phylogenetic tree

Data improving the characterization of the marine Euplotes species, E. petzi Wilbert and Song, 2008, were obtained from morphological, ecological and genetic analyses of Antarctic and Arctic wild-type strains. This species is identified by a minute (mean size, 46 μm × 32 μm) and ellipsoidal cell body which is dorsally decorated with an argyrome of the double-patella type, five dorsal kineties (of which the median one contains 8–10 dikinetids), five sharp-edged longitudinal ridges, and a right anterior spur. Ventrally, it bears 10 fronto-ventral, five transverse, two caudal and two marginal cirri, 30–35 adoral membranelles, and three inconspicuous ridges. Euplotes petzi grows well at 4 °C on green algae, does not produce cysts, undergoes mating under the genetic control of a multiple mating-type system, constitutively secretes water-borne pheromones, and behaves as a psychrophilic microorganism unable to survive at >15 °C. While the α-tubulin gene sequence determination did not provide useful information on the E. petzi molecular phylogeny, the small subunit rRNA (SSU rRNA) gene sequence determination provided solid evidence that E. petzi clusters with E. sinicus Jiang et al., 2010a, into a clade which represents the deepest branch at the base of the Euplotes phylogentic tree.     

Mating Behavior and Species Status of Host-Associated Populations of the Polyphagous Thrips, <Emphasis Type="Italic">Frankliniella schultzei</Emphasis>

The thrips taxon, Frankliniella schultzei Trybom, could possibly comprise a complex of unrecognised cryptic species. We therefore made a range of reproductive behavioral observations on F. schultzei individuals from different host-associated populations to investigate their species status. We first described the mating behavior of F. schultzei taken from flowers of a primary host species, Malvaviscus arboreus. The pattern of male-female interaction was complex and protracted, with several behavioral steps being performed. These steps were similar for all mating pairs. Most females responded to male presence before physical contact, suggesting that male pheromones may be involved. We then compared mating behaviors, brood sizes and sex ratios of F. schultzei pairs derived from different host plant species. All pairs in crosses regardless of their original host plant species produced female offspring, indicating successful mating. Frequencies of behaviors were similar for all pairs. Only those crosses involving F. schultzei individuals from Erythrina crista-galli flowers differed from the general pattern in pre-copulation and copulation times. The females from E. crista-galli also produced significantly fewer offspring than females from other crosses. Finally we used traps baited with males from two populations, but set among flowers of one host plant species, to test for differential attraction of females. We found no evidence that the different host-associated populations investigated differ from one another functionally in their sex pheromone system. However, these results do not yet discount the possibility of cryptic species within the taxon F. schultzei. Populations of F. schultzei on many other host species need to be investigated from this perspective.     

Genetic diversity and population structure in the outcrossing populations of Equisetum arvense and E. hyemale (Equisetaceae)

An allozyme examination was conducted to study the mating systems and genetic differentiation of populations of Equisetum arvense and E. hyemale. The study revealed that the rate of intragametophytic selfing in these homosporous pteridophytes is very low, i.e., on average 0.020 and 0.019, respectively, despite the potential hermaproditism and selfing of the gametophytes. Most populations consisted of numerous genotypes, and the average heterozygosities of E. arvense and E. hyemale equalled 0.092 and 0.134, respectively. The commonly observed excess of the heterozygote genotypes indicates that there are interclonal differences in the frequency of vegetative reproduction. The level of genetic divergence among populations was considerable even within a limited geographic area. It is suggested that the life history of Equisetum, characterized by the inefficiency of spore germination and gametophyte reproduction in noncolonizing situations, limits the level of gene flow and leads to a great genetic divergence between populations.     

Species concepts and the determination of historic gene flow patterns in the Eulemur fulvus (Brown Lemur) complex

Eulmur fulvus , a complex comprising six subspecies, is a classic example of species status conferred through evolutionary taxonomy. We used the phylogcnctic species concept as an alternative method to the biological species concept for determining historic patterns of gene flow between the various E. julvus subspecies and for conferring species status. In this paper, we used population aggregation analysis to determine the proper species partitions and cladistic analysis to reconstruct the evolutionary relationships of the different populations in the Eulemur fulvus complex. We sequenced three mtDNA gene regions (d-loop, 12S, and cyt-b) and one nuclear region, casein kinase, for a total of 1247 bases. Through population aggregation analysis, we determined that the E. fulvus complex should be split into three units; one unit supported by six diagnostic sites comprising E.f. albocollans , one unit supported by three diagnostic sites comprising E.f. collaris , and one unit supported by two diagnostic sites comprising the four other subspecies. Although all six subspecies in the E. julvus complex share a common ancestor, we found in our cladistic analysis that E. J. collaris and E. j. albocollans share a common ancestor that more recently split off from the common ancestor of the four other E. fulvus subspecies.     

Use of AFLP Analyses to Assess Genetic Variation in Morone and Thunnus Species

The amplified fragment length polymorphism (AFLP) technique was used to assess genetic variation across the genera of 2 Morone and 3 Thunnus species. The AFLP profiles from 23 primers revealed higher levels of polymorphism in each of 3 Thunnus species than in either of the 2 Morone species. However, extensive variation (20 of 23 primers) was observed between the 2 Morone species, but much less variation was observed among the 3 Thunnus species. In addition, comparisons of banding patterns indicated that albacore is divergent from both Atlantic northern bluefin tuna and yellowfin tuna. This result is consistent with the findings of several previous studies employing either allozymes or mitochondrial DNA. Overall, these results demonstrate that AFLP is a useful technique for the assessment of both intraspecies and interspecies variation of fish. Furthermore, the species-specific patterns produced by AFLP can be used for the identification of closely related species.     

游仆虫Rab蛋白基因的克隆表达与纯化

为对单细胞原生动物纤毛虫中Rab蛋白的功能进行研究 ,进而探讨以胞吞和胞吐为主要物质交换途径的纤毛虫中囊泡定向运输的机理 .利用PCR技术从游仆虫大核DNA及cDNA中扩增出rab基因 ,并进行了序列分析 ,该基因全长为 783bp ,两端为端粒序列 ,编码框为 6 2 4bp ,编码 2 0 7个氨基酸 ,开放读框中有 3个TGA ,在此编码半胱氨酸 .利用定点突变将rab基因中 3个TGA突变为通用半胱氨酸密码子TGC .将游仆虫Rab蛋白基因构建于原核表达载体pGEX 4T 2中 ,得到的重组质粒pGEX Eorab1转化至大肠杆菌BL2 1(DE3)中 ,IPTG诱导表达 .表达产物与抗GST抗体在 4 9kD处有很强的交叉反应 .融合蛋白GST EoRab1通过亲和层析柱纯化和凝血酶的切割 ,再经两步纯化得到电泳纯的游仆虫Rab蛋白 .