西藏湖泊长刺溞复合种的系统进化关系

本文通过对西藏湖泊长刺溞复合种(Daphnia longispina complex)中分布最广的3个物种, 即长刺溞(D. longispina)、盔形溞(D. galeata)和颈齿溞(D. dentifera)线粒体COI基因序列以及GenBank中欧洲的长刺溞、加拿大的颈齿溞和我国东部低海拔地区的盔型溞COI基因序列的比较分析, 研究了西藏湖泊长刺溞复合种的系统进化关系, 发现西藏地区的盔型溞、颈齿溞和长刺溞均已出现较大分化。颈齿溞种群内遗传差异度为0.33-2.32%, 盔型溞为0.33-2.74%, 长刺溞的遗传差异度最高, 为1.31-5.50%。基于COI基因序列构建的最大似然树和贝叶斯系统树均表明, 长刺溞复合种由3个进化分支组成, 分别对应长刺溞、盔型溞和颈齿溞, 三者之间的遗传差异度为9.40-16.98%(Kimura 2-parameter双参数模型)。基于COI基因单倍型(haplotype)所构建的网络关系也支持上述3个分支的存在。早期记录虽然显示长刺溞在我国分布较广, 但本次调查只在班公错有发现, 相比之下, 盔形溞和颈齿溞则分布更广。我们的研究表明, 由于形态学鉴定上的局限性, 早期的长刺溞记录很可能混杂了容易引起混淆的盔型溞或颈齿溞。     

松毛虫属（Dendrolimus) 部分种类亲缘关系与遗传分化的等位酶分析

采用等位酶聚丙烯酰胺凝胶电泳技术对松毛虫属5个种和亚种的野生种群进行了亲缘关系和遗传变异的研究.8种等位酶系统(乳酸脱氢酶LDH、苹果酸脱氢酶MDH、苹果酸酶ME、乙醇脱氢酶ADH、甲酸脱氢酶FDH、谷氨酸脱氢酶GDH、过氧化物酶POD、过氧化氢酶CAT)共检测到12个基因位点,其中6个位点为多态位点,检测到15个等位基因.松毛虫属5个种和亚种的总体水平多态位点比率P＝50%,平均有效基因数A = 1.917,平均期望杂合度He =0.267,平均遗传距离为0.0730～0.5701.遗传参数表明松毛虫属昆虫种间存在较高程度的遗传变异,聚类图和遗传距离数据表明赤松毛虫与马尾松毛虫亲缘关系最近,落叶松毛虫与思茅松毛虫亲缘关系最远.     

Intra-specific rDNA-ITS restriction site variation and an improved protocol to distinguish species and hybrids in the Daphnia longispina complex

A collaborative research effort was undertaken to evaluate the robustness of a recently developed genetic tool for species identification of members in the morphologically variable Daphnia longispina species complex. This genetic method, based on restriction fragment length polymorphism (RFLP) of the internal transcribed spacer region (ITS) of nuclear ribosomal DNA (rDNA) with restriction enzymes Mwo I and Sau96 I [Billiones et al., 2004. Hydrobiologia 526: 43–53], was applied to many different European populations. Results were compared with two or more independently obtained characters (morphology, allozymes, mitochondrial DNA (mtDNA), or cloned rDNA-ITS sequences). Individuals of most taxa were readily identified, but unexpected ITS-RFLP patterns were found in many individuals indicated by other markers to be D. galeata or one of its hybrids. Among 43 investigated D. galeata populations (902 specimen analysed by ITS-RFLP), deviant RFLP fragment patterns occurred in 26 (i.e., more than half) of the populations. The deviant patterns could be attributed to the loss of one single restriction site in the ITS2 region. This loss made the distinction of D. galeata from other species unreliable, and F1 hybrids could not be identified. Future users should be aware of this shortcoming of the Billions et al. [2004. Hydrobiologia 526: 43–53] protocol. As a solution to this problem, we present an improved genetic identification protocol based on a simple double digestion of the rDNA-ITS region with the restriction enzymes BsrB I and EagI. Sequence analyses of rDNA-ITS clones and preliminary testing indicate that the new protocol is unaffected by the rDNA variation which troubled the Mwo I/Sau96 I protocol. Further, the new protocol identifies all European species of the D. longispina complex, as well as their F1 hybrids. However, a wider screening is required to verify its general utility for all species, since yet unknown variation may occur. Guest editor: Piet Spaak Cladocera: Proceedings of the 7th International Symposium on Cladocera     

Allozyme variation and genetic relationships among species in the Carex willdenowii complex (Cyperaceae)

A taxonomic study by Naczi, Reznicek, and Ford (American Journal of Botany, 85, 434-447, 1998) has determined that three species (Carex willdenowii, C. basiantha, and C. superata) can be recognized within the C. willdenowii complex. To determine the amount of genetic divergence within and between these species, allozyme analyses were conducted on 14 populations distributed from Pennsylvania to eastern Texas. Seventeen loci were surveyed, 13 of which were polymorphic, with all populations being polymorphic at one or more loci. Interspecific genetic identities ranged from 0.560 (C. willdenowii and C. basiantha) to 0.807 (C. basiantha and C. superata). Alleles for the isozymes Aat-1, Dia-1, Idh-2, Mdh-2, Per-1, Pgm-1, and Pgm-2 served to distinguish C. willdenowii from C. basiantha and C. superata. Carex basiantha and C. superata were recognized by alleles of Mdh-2, Pgm-1, and Tpi-2. The genetic identities of populations within species were high and exceeded 0.957. A caespitose growth habit and perigynia in close proximity to the staminate flowers suggest adaptations for selfing and therefore low levels of heterozygosity. Paradoxically, the values for expected heterozygosities (Hexp) were always lower than those obtained by direct count (Hobs): F values were highly negative, indicating heterozygous excess. Disassortative mating and selection are discussed as possible mechanisms for maintaining heterozygous excess within populations.     

Exploring the reticulate evolution in the Asplenium pekinense complex and the A. varians complex (Aspleniaceae)

The Asplenium pekinense complex mainly comprises one diploid, A. sarelii Hook. (rare), one autotetraploid, A. pekinense Hance (best known and very common), and shares two allotetraploids, A. anogrammoides Christ (common but often misidentified) and A. altajense (Komarov) Grubov (rare and endemic) with the A. varians complex. The latter is further constituted by two diploids, A. tenuicaule Hayata (widespread) and A. semivarians Viane & Reichstein (rare), as well as other three tetraploids, A. kansuense Ching (barely known), A. varians Wallich ex Hooker & Greville (well‐known, relatively common, and morphologically variable), and A. kukkonenii Viane & Reichstein (rare and often misidentified). These two species complexes are notorious for their taxonomic difficulty based on general morphology, which is mainly caused by their history of reticulate evolution. Here, we collected most species within the two complexes, and obtained ploidy information by spore size measurement and flow cytometry investigation. Phylogenetic analyses using DNA markers representing maternally inherited chloroplast and biparentally inherited nuclear genomes helped to reconstruct the reticulate evolution history. The present results support previous hypotheses that A. sarelii is the ancestor of both A. pekinense and A. anogrammoides, as well as that A. tenuicaule is the common progenitor of A. anogrammoides, A. varians, and A. kukkonenii. We also unraveled the autotetraploid origin of A. kansuense from A. tenuicaule for the first time, and found that A. altajense shares essentially identical genomes with A. anogrammoides.     

Clonal diversity,clonal persistence and rapid taxon replacement in natural populations of species and hybrids of the Daphnia longispina complex

Hybridization is common among cyclical parthenogens, especially in zooplankton species assemblages of the genus Daphnia. To explore hybridization dynamics and the extent of clonal diversity in the Daphnia longispina complex, we analysed population structure in eight permanent lakes. Based on 15 microsatellite loci, three major taxonomic units emerged: two species, D. galeata and D. longispina and their F1 hybrids, supported by factorial correspondence analysis and two Bayesian methods. At the same time, the detection of backcross classes differed between methods. Mean clonal diversity was lowest in the F1 hybrids, as expected from the high rate of asexual reproduction. Within taxa, replicated genotypes were of clonal origin, but clonal lineages persisted in subsequent years in only one of three resampled lakes. In another lake, the taxon composition changed from being dominated by hybrids to complete dominance by one parental taxon. Such a year‐to‐year taxon replacement has not been reported for the D. longispina complex before. Our data on this hybrid complex illustrate that high‐resolution genotyping is essential for the understanding of ecological and evolutionary outcomes of hybridization in partially clonal taxa.     

Population structure, gene flow and evolutionary relationships in four species of the genera Tomocerus and Pogonognathellus (Collembola, Tomoceridae)

Genetic diversity and gene flow at 10 allozyme loci were investigated in 13 natural populations of four species of the collembolan genera Tomocerus and Pogonognathellus. Levels of observed heterozygosity were found to vary from 0.033 in P. longicornis to 0.120 in T. vulgaris. Average Nei's genetic distances (D) ranged from D = 0.222 between populations of P. ftavescens to D = 0.647 between populations of T. vulgaris. Genetic distances between species were always 1.000 and the highest value (D = 4.321) was between P. longicomis and one population of T. vulgaris. Values of F_sr were very high in all species (from 0.567 to 0.696) and levels of gene flow (Nm) derived from F_ST and the private allele method were low (Nm <1). Gene flow was significandy higher only in two subsets of populations of T. vulgaris. The Plio-Pleistocenic geological rearrangements and the effect of stochastic events, such as genetic drift, are invoked to explain the different levels of genetic divergence between and within species. Distance- and character-based approaches were used to reconstruct evolutionary relationships between and within species. While the monophyly of all species was confirmed, the results did not univocally support the monophyly of the two genera, leaving the question of their generic or subgeneric status unresolved.     

Coding of intraspecific nucleotide polymorphisms: A tool to resolve reticulate evolutionary relationships in the ITS of beech trees (Fagus L., Fagaceae)

The internal transcribed spacers ITS1 and ITS2 of the nuclear ribosomal DNA (rDNA) have recently been found to display remarkable intraspecific polymorphism, a feature suggested as limiting their value for phylogenetic reconstructions. A comparative study of oligonucleotide motives and intraindividual nucleotide variability across all species of the tree genus Fagus (beech) shows, however, that this intraspecific ITS polymorphism follows a particular pattern, which can be used to detect reticulation and ancient polymorphism within the genus. Coding ITS polymorphisms as phylogenetically informative characters, moreover, resulted in better‐resolved phylogenies than traditional ‘base‐per‐base’ maximum parsimony and maximum likelihood analyses.     

Apomixis and reticulate evolution in the Asplenium monanthes fern complex

Background and Aims

Asexual reproduction is a prominent evolutionary process within land plant lineages and especially in ferns. Up to 10 % of the approx. 10 000 fern species are assumed to be obligate asexuals. In the Asplenium monanthes species complex, previous studies identified two triploid, apomictic species. The purpose of this study was to elucidate the phylogenetic relationships in the A. monanthes complex and to investigate the occurrence and evolution of apomixis within this group.

Methods

DNA sequences of three plastid markers and one nuclear single copy gene were used for phylogenetic analyses. Reproductive modes were assessed by examining gametophytic and sporophyte development, while polyploidy was inferred from spore measurements.

Key Results

Asplenium monanthes and A. resiliens are confirmed to be apomictic. Asplenium palmeri, A. hallbergii and specimens that are morphologically similar to A. heterochroum are also found to be apomictic. Apomixis is confined to two main clades of taxa related to A. monanthes and A. resiliens, respectively, and is associated with reticulate evolution. Two apomictic A. monanthes lineages, and two putative diploid sexual progenitor species are identified in the A. monanthes clade.

Conclusions

Multiple origins of apomixis are inferred, in both alloploid and autoploid forms, within the A. resiliens and A. monanthes clades.     

Parental and hybrid Daphnia from the D. longispina complex: long‐term dynamics in genetic structure and significance of overwintering modes

In recent decades, hybridization has become a focus of attention because of its role in evolutionary processes. However, little is known about changes in genetic structure within and between parental species and hybrids over time. Here, we studied processes of genetic change in parental species and hybrids from the Daphnia longispina complex (Crustacea, Cladocera) over a period of six years across ten habitats. These cyclical parthenogens respond to fluctuating environments by switching from asexual to sexual reproduction. Importantly, sexually produced diapausing eggs, which resist extreme conditions such as low temperatures and serve as dispersal stages, are produced to a lower extent by hybrids. Long‐term microsatellite data revealed clear differences between hybrids and parental species. In hybrids, clonal diversity values were lower, whereas heterozygosity and linkage disequilibrium values were higher compared to parental species. Clonal diversity of hybrids responded to the strength of the winter, with cold winters resulting in few genotypes in the following spring. In time windows when only asexual hybrid females survive, priority effects will favour the establishment of the hybrid offspring before hatchlings from parental diapause eggs can enter the community. The constant high levels of heterozygosity maintained by clonal reproduction in hybrids might lead to their successful establishment over time, when they are able to escape competition from both parental species. Although we found evidence that hybrids diversity depends on fluctuating environments, a direct link between hybrid abundance and the strength of winter was missing. Because of reduced adaptability in clonally reproducing hybrids, multiple factors must contribute to promoting their long‐term success in fluctuating environments.     

Ecological genetics of Norwegian Daphnia. II. Distribution of Daphnia longispina genotypes in relation to short-wave radiation and water colour

In the mountain range of South Norway, transparent and melanic members of the Daphnia longispina complex occur. Melanic populations inhabit clearwater lakes and ponds, while transparent populations are found in ponds with slightly humic water. Mixed populations have not been detected. The distribution patterns of the two morphs are related to ambient levels of short-wave radiation, and the light absorptive properties of the inhabited waters. The pigment present is probably melanin, which is deposited in the dorsally directed parts of the carapace, head shield and antennae. Allozyme studies indicate that these distinct morphs are only remotely related, the alpine transparent group being closer to lowland, likewise transparent, populations. A melanic population found at Svalbard is closely related to the melanic mainland populations. Clonal diversity in ponds and shallow lakes is very low, in contrast to populations of deeper lakes.Deceased.     

Molecular and morphological evidence for a new grass genus,Dupontiopsis (Poaceae tribe Poeae subtribe Poinae s.l.), endemic to alpine Japan,and implications for the reticulate origin of Dupontia and Arctophila within Poinae s.l.

Phylogenetic analyses within Poaceae tribe Poeae subtribes Puccinellinae (=Coleanthinae), Phleinae, Poinae s.l. (including Alopecurinae), and Miliinae (PPAM clade), revealed that one species formerly placed in Poa represents a new monotypic genus belonging to subtribe Poinae s.l., Dupontiopsis gen. nov., D. hayachinensis comb. nov. (based on Poa hayachinensis), endemic to wet, gravelly, serpentine, alpine habitats in northern Japan. This genus forms a strongly supported clade (DAD) with two circumarctic Poinae genera, Arctophila and Dupontia, in phylogenetic analyses of plastid and nuclear ribosomal DNA sequence data. Both morphology and DNA sequence analyses provide support for D. hayachinensis as a lineage distinct from either Arctophila or Dupontia, with moderate DNA support for a position as sister to these two genera. Dupontiopsis resembles these other monotypic genera in its several-flowered spikelets, lemmas usually 3-nerved, with frequently awned attenuate scarious apices (as in Dupontia) and calluses with a crown of hairs around the base of the lemma, but differs in its keeled lemmas, scabrous palea keels, glumes shorter than the first lemma. Our investigation suggests that the most recent shared ancestor of the DAD clade evolved from a single hybridization event, as a hexaploid, probably in western Beringia. The probable parentage of the ancestor is considered to be within the Poinae–Alopecurinae clade excluding Poa. We provide evidence for possible secondary hybridization and introgression of duodecaploid Dupontia fisheri with Puccinellia. A key to perennial genera of PPAM with hairy calluses, and a supplemental table of morphological characters in the genera accepted in PPAM are provided.     

Allozyme variation and population differentiation of the Aconitum delavayi complex (Ranunculaceae) in the Hengduan Mountains of China

The Aconitum delavayi complex is a group of four climbing species with trisect-leaves occurring in the Hengduan Mountains. The species of this complex are highly localized on very narrow regions with quite small population sizes. Because of rapid environmental changes recently in the Hengduan Mountains, this complex shows complicated morphological variability, which makes it difficult to delimit species. In the present study, 10 enzyme systems coding for 14 putative loci were employed to detect the interspecific and intraspecific genetic variation of the complex. In addition to low genetic diversity within all eight populations surveyed, the results indicate that A. episcopale is a distinct species because of high genetic identities among its three populations. Very low genetic divergence among populations of A. stapfianum and A. delavayi suggests that the two species should be treated as a single one.     

Allozyme variation and evolutionary relationships of grain amaranths (Amaranthus spp.)

Summary Allozyme studies in amaranth provided useful assays of genetic variation in order to verify the patterns inferred from morphological traits, for elucidating the genetic structure of landraces, and for the studies of evolutionary relationships among wild, weedy and crop species. Thirty-four populations of cultivated New World amaranths were surveyed along with 21 weedy New World populations for allozyme variation at nine electrophoretic enzyme loci. Eleven populations of cultivated amaranths from the Indian State of Uttar Pradesh and six from Nepal were also surveyed for a comparison. In the New World populations, heterozygosity was low, and different populations ranged from 0 to 44% polymorphic loci. Adjacent populations were often fixed for different alleles or had very different allele frequencies at certain loci, with no apparent geographical patterns. Diversity index H was partitioned into the intra- and interpopulation as well as the interspecific components of variability. The crop versus weed genetic distances were the largest, whereas the intra- and interpopulation components of H were about equal. Genetic structure of all three species of the New World amaranths together can be described as a collection of distinct populations, each more or less a heterogeneous collection of highly homozygous individuals. The North Indian populations showed relatively less allozyme variability with the most common alleles same as those of Mexican landraces. Alleles at several loci proved to be diagnostic of the crop and weed groups, and of the three individual crop species. Genetic distances based on pooled gene frequencies showed the three crop species to be generally more closely related inter se than they were to their putative weedy progenitor species, respectively (with the exception of the weed-crop pair A. quitensis and A. caudatus). This implies a single domestication event involving A. hybridus as the common ancestor rather than three separate domestication events. Close similarity between A. caudatus and A. quitensis might have resulted from transdomestication based on a weedy or semi-domesticated species having migrated from Meso-America to South America. This preliminary report must now be expanded by further ecogeographical, cytogenetic and population studies on new extensive collections from the areas of early domestication. Some evidence of recent introgression and/or segregation of crop-weed hybrids between A. caudatus and A. retroflexus is available in the form of rare individuals in crop populations with crop allozyme genotypes except for a single homozygous weedy allele.     

中国沿海中部珊瑚菜居群等位酶变异及其遗传多样性

利用聚丙烯酰胺凝胶电泳方法,分析了我国沿海中部（江苏、山东和浙江）海滨沙滩珊瑚菜（Glehnia littoralis Fr.Schmidt ex Miq.)7个居群8种酶系统19个位点的等位基因遗传变异特征,结果表明居群内多态位点比率平均为82．4％,每一位点平均等位基因数为2．77,有效等位基因数为2．24,固定指数F的平均值为－0．091。珊瑚菜居群基因多样性80．9％产生于居群内,19．1％产生于居群间。居群间的遗传距离平均为0．317,遗传一致性为0．728,居群内维持着较高水平的遗传多样性。据此,对珊瑚菜的渐危机理进行了讨论。     

Genetic assessment of species boundaries in the North American Daphnia longispina complex (Crustacea: Daphniidae)

Species boundaries in the North American Daphnia longispina group have proved difficult to establish on the basis of morphology alone. This confusion may be due to hydridization, phenotypic plasticity or the existence of sibling species. We therefore used genetic analysis to delineate species boundaries by examining 27 North American populations belonging to the longispina complex for variation at 15–26 allozyme loci. The populations consisted of Daphnia thorata from two western sites and two eastern sites, Daphnia galeata mendotae from its type location and seven sites across its range, and Daphnia rosea from eight temperate and seven arctic sites. Two populations from the Eurasian longispina complex were also included for reference. Populations assigned to D. galeata mendotae formed a genetically cohesive group, whereas a genetic dichotomy was found between temperate and arctic D. rosea , suggesting that this taxon includes two species. Genetic analysis also confirmed the distinctness of western D. thorata from other members of the longispina group. Unexpectedly, eastern populations resembling D. thorata were genetically more similar to temperate D. rosea than to any helmeted species ( D. galeata, Daphnia hyalina or D. thorata ). Our results suggest that the helmet character is a poor indicator of phylogenetic relationships, as the genetic ability to produce this feature has been lost or acquired several times in the evolution of the longispina group.     

Taxonomic and evolutionary relationships in the genus Vella L. (Cruciferae)

Thirty-two differential characters are numerically analysed for the six existing taxa of the genus Vella L. The closely related monotypic genus Boleum Desv. is also included for comparison. Most of the characters used are morphological, but cariological, ecological and biogeographical aspects have also been studied. The results indicate that four distinct species, one ( V. pseudocytisus L.) with three sub-species, should be distinguished. The new combination V. anremerica (Lit. et Maire) Gómez-Campo (basionym: V. pseudocytisus L. subsp. anremerica Lit. et Maire) and the new name V. pseudocytisus L. subsp. paui Gómez-Campo for V. pseudocytisus L. var. glabrescens Willk., nornen nudum are presented. A key is given to aid identification of these taxa. Their conservation status is also discussed.     

Growth of Daphnia longispina L. in a polyhumic lake under various availabilities of algal,bacterial and detrital food

The availability and importance of food sources for growth of Daphnia longispina L. from a highly coloured fishless lake with anoxic hypolimnion were assessed by combining in situ and laboratory experiments. In in situ experiments populations were enclosed in tubes with natural temperature stratification and with or without anoxic hypolimnion. In the laboratory experiments the importance of food source (littoral zone vs pelagic epilimnion) was assessed by enclosing moss thalli and a natural zooplankton population in a large-scale flow-through system supplying food for experimental Daphnia. Growth of juveniles of Daphnia in epilimnetic water was determined in batch culture experiments and the importance of increasing concentrations of bacteria and algae for their growth and development was investigated with a small-scale flow-through system. Access to the anoxic hypolimnion enhanced the growth of Daphnia by 23–24%. Growth rates in the tubes with anoxic hypolimnion were 0.36 and 0.16 d^–1 in July and August respectively. In tubes without anoxia the corresponding values were 0.29 and 0.13. In batch-cultures the highest growth rate determined was 0.16 and the overall rates were lower than in in situ experiments. In batch culture Daphnia was able to grow in darkness for 10 days with a rate of 0.16. In the large-scale flow-through system Daphnia population fed with littoral water reproduced well despite the low concentration of algae and increased its number by a factor of c. 32 in 10 days. However, the animals were small and net production of Daphnia population thus lower under the littoral influence than in the control treatment. Population could survive and grew slowly on pelagial water processed by a natural zooplankton community and with very little algae left. It is thus possible that bacteria serve as a life-support system enabling the population survival over periods of algal shortage. Small-scale flow-through experiments revealed that Daphnia longispina is able to mature and reproduce on a bacterial diet if the food concentration is high enough and Daphnia on bacterial food could achieve growth rates similar to those on an algal diet. The threshold food level for Daphnia longispina was estimated to be c. 18.5 g C 1^–1. Detrital material is of limited value in nutrition of Daphnia even in a lake where more than 75% of carbon is bound in particulate detritus.