Genetic divergence in Northamerican freshwater planarians of the Dugesia dorotocephala group (Turbellaria, Tricladida, Paludicola)

The genetic differentiation between the members of the Dugesia (Girardia) dorotocephala group was analyzed by means of multilocus electrophoresis, and comared to that of another planarian secies, D. tahitiensis, also belonging to the subgenus Girardia. The species examined were: D. dorotocephala s.s (2n = 16), D. arizonensis (2n = 8), D. jenkinsae (2n = 8), and the above mentioned D. tahitiensis (2n = 16). The former three species inhabit North America, and show different proportion of fissiparous and sexual individuals; the latter species inhabits Polynesia and is fully asexual. A total of 11 enzyme loci were genetically analyzed: Mdh-1, Mdh-2, Zdh-1, Idh-2, G3pdh, Got-1, Ck, Pgm-2, Ada, Mpi, and Gpi. Low values of observed mean heterozygosity per locus (Ho) were found in the populations studied, ranging from 0 to 0.18 (average 0.08. In asexual populations (except that of D. tahitiensis) fixed heterozygosity was observered in all the individuals for 1 or 2 loci. The genetic divergence between the species examined is very high, with many loci showing discriminating alleles in different taxa (Nei's genetic distance varies from 0.871 to 1.759). The populations of D. dorotocehala s.s., on the contrary, appear to be genetically quite homogenous average D= 0.019), and the genetic distance values are apparently unrelated to their geographic location and to their way of reproduction. The genetic distance between D. tahitiensis, a species not included in the D. dorotocephala group and D. dorotocephala s.s. is 1.314 and hence similar to the D value between two members of;he dorotocephala group: D. dorotocephala and D. jenkinsae (D = 1.303). The genetic relationships among the populations studied were established by UPGMA cluster analysis and multidimensional scaling. The descendence of the North American species with 2n = 8 from a dorotocephala-like ancestor with 2n = 16 is considered. It is suggested that the latter, as well as a tahitiensis-like line, also having 2n = 16, have originated from a common ancestor by geographic isolation.     

Taxonomic revision of Dictyochales (Dictyochophyceae) based on morphological,ultrastructural, biochemical and molecular data

The taxonomy of the siliceous members of Dictyochales has been based exclusively on the structure of their silica skeletons. In this study, other morphological, ultrastructural, pigment and molecular characters, in addition to the silica skeletons, were used in the systematics of the siliceous members of this group. As very little is known about the ultrastructure of Dictyocha octonaria, cells of both the skeleton‐bearing and naked forms were also studied. A cladistic analysis based on morphological data and a molecular phylogeny based on nuclear coded small subunit ribosomal DNA retrieved a well‐supported monophyletic Dictyochales. D. octonaria and D. speculum were resolved together with strong support. There was no support for a clade for the three species currently placed in Dictyocha; D. fibula is clearly distinct from the other two Dictyocha taxa, D. speculum and D. octonaria. It is highly likely there are two or three undescribed species within D. octonaria/D. speculum clade and two undescribed genera within the Dictyochales, based on the positions of sequences from uncultured eukaryotes present in GenBank. These findings necessitate a taxonomic revision of the three siliceous, skeleton‐bearing species. Because D. fibula is the type species of Dictyocha, we propose that D. octonaria and D. speculum be reassigned respectively to the genus Octactis as O. octonaria Hovasse and O. speculum (Ehrenberg) F. H. Chang, J. M. Grieve & J. E. Sutherland, comb. nov.     

Evolution,phylogeography and taxonomy of allopolyploid Dactylorhiza (Orchidaceae) and its implications for conservation

This review is based on recent molecular studies of Dactylorhiza (Orchidaceae). Most of the studies have focused on the allotetraploid members of the genus in general and on D. majalis ssp. lapponica in particular. It was concluded that most of the allotetraploid taxa have derived from hybridizations between the parental lineages D. maculata s.l. and D. incarnata s.l., with D. maculata s.l. serving as the seed parent. Evidence of multiple origins was found both among northern European allotetraploids as well as among Greek allotetraploids. Introgression from both parental lineages and hybridizations between independently derived polyploid lineages was also detected. The three morphologically similar taxa D. majalis ssp. traunsteineri, ssp. lapponica and ssp. russowii should be treated as one and most of the Greek allotetraploids should be regarded as regional variants of the southeastern European D. majalis ssp. cordigera. The Balkans and the Alps most probably served as refugia for the genus during the last glaciations and at least two waves of immigration reached Scandinavia. Finally, we suggest that the conservation of allotetraploid Dactylorhiza should emphasize important geographic areas and habitats and that the allopolyploids should have the same conservation status as the diploids.     

Polyphyly of intraspecific groups of <Emphasis Type="Italic">Umbelopsis ramanniana</Emphasis> and their genetic and morphological variation

The taxonomic positions of three intraspecific groups of Umbelopsis ramanniana in the genus Umbelopsis were analyzed based on the nucleotide sequences of their nuclear large subunit ribosomal DNA (nLSU rDNA) D1/D2 region. The examined members of the genus Umbelopsis were resolved into two major clades, Clades I and II. The intraspecific groups of U. ramanniana were nested within Clade II together with U. westeae, U. swartii, U. autotrophica, U. gibberisopra, U. angularis, and U. fusiformis. In this major clade, the intraspecific groups of U. ramanniana were split into three polyphyletic subclades. This suggests that U. ramanniana is an assemblage of several genetically distinct species. Interestingly, in spite of the diverse sporangiospore shapes of the members of Clade II, the genetic variation among them was small. It is considered that their flexible sporangia membranes make it possible for them to develop various sporangiospore shapes.     

Interrelationships among three derivative forms of the immigrans-Hirtodrosophila radiation: evidence from heterochromatin and polytene chromosomes

The phylogenetic relationships among three derivative forms of the immigrans-Hirtodrosophila radiation viz. Chaetodrosophilella, Zaprionus and the immigrans species group are examined, by comparing the banding patterns of their polytene chromosomes and by analysing the nature of their heterochromatin. Based on the results of these studies it is concluded that D. quadrilineata has a very strong affinity with the members of the immigrans species group, while the genus Zaprionus represents a very distinct evolutionary lineage. This study further indicates that D. quadrilineata is a karyotypically primitive species having five pairs of rods and one pair of dots, while the karyotype of other members of the immigrans species group appears to have undergone modifications through fusions, fissions, inversions and the addition or deletion of heterochromatin to dots and other chromosomes.     

Cytogenetics of natural populations of Drosophila

D. bipectinata and D. malerkotliana differ from each other by three overlapping inversions in IIL, two included inversions in IIIL and two overlapping inversions in IIIR. These inversions were analysed on the basis of the salivary chromosome maps of D. malerkotliana. Bock's (1971) data revealed that the four members of the bipectinata species complex differ from each other with respect to overlapping inversions. The reason why the ancestral population which may have been heterozygous for common inversions split into at least four groups, each leading to the formation of a new species, and the possible mechanism of the origin of sexual isolation between the groups is discussed.     

Induction by alkaloids and phenobarbital of Family 4 Cytochrome P450s in Drosophila : evidence for involvement in host plant utilization

In vertebrates, cytochrome P450s of the CYP2 and CYP3 families play a dominant role in drug metabolism, while in insects members of the CYP6 and CYP28 families have been implicated in metabolism of insecticides and toxic natural plant compounds. A degenerate 3^′ RACE strategy resulted in the identification of fifteen novel P450s from an alkaloid-resistant species of Drosophila. The strong (17.4-fold) and highly specific induction of a single gene (CYP4D10) by the toxic isoquinoline alkaloids of a commonly utilized host-plant (saguaro cactus) provides the first indication that members of the CYP4 family in insects may play an important role in the maintenance of specific insect-host plant relationships. Strong barbiturate inducibility of CYP4D10 and two other D. mettleri P450 sequences of the CYP4 family was also observed, suggesting a pattern of xenobiotic responsiveness more similar to those of several vertebrate drug-metabolizing enzymes than to putative vertebrate CYP4 homologs. Received: 14 August 1997 / Accepted: 24 March 1998     

The evolutionary genetics of thehobo transposable element in theDrosophila melanogaster complex

Hobo elements are a family of transposable elements found inDrosophila melanogaster and its three sibling species:D. simulans, D. mauritiana andD. sechellia. Studies inD. melanogaster have shown thathobo may be mobilized, and that the genetic effects of such mobilizations included the general features of hybrid dysgenesis: mutations, chromosomal rearrangements and gonadal dysgenis in F1 individuals. At the evolutionary level somehobo-hybridizing sequences have also been found in the other members of themelanogaster subgroup and in many members of the relatedmontium subgroup. Surveys of older collected strains ofD. melanogaster suggest that completehobo elements were absent prior to 50 years ago and that they have recently been introduced into this species by horizontal transfer. In this paper we review our findings and those of others, in order to precisely describe the geographical distribution and the evolutionary history ofhobo in theD. melanogaster complex. Studies of the DNA sequences reveal a different level of divergence between the groupD. melanogaster, D. simulans andD. mauritiana and the fourth speciesD. sechellia. The hypothesis of multiple transfers in the recent past into theD. melanogaster complex from a common outside source is discussed.     

Differences in male courtship song among the species of thequinaria group ofDrosophila

Courtship song is known to vary among several groups of closely related species ofDrosophila. The present study investigates differences in song among four members of thequinaria group of fungal-breedingDrosophila which are known to coexist in Britain and the near-continent. Results show that the species can be distinguished by several song parameters including interpulse interval.D. phalerata is very different from the other three species, while there is considerable overlap betweenD. kuntzei andD. limbata in several characters.D. transversa appears to be more similar to the latter species than it is toD. phalerata. The question of mate choice and the potential for reproductive character displacement are discussed.     

Taxonomic and molecular studies on Drosophila sinobscura and D. hubeiensis,two sibling species of the D. obscura group*

Allozyme electrophoresis and three different DNA sequences (ATOC180 satellite DNA, 5SrDNA repeats, and parts of the Adh gene) were used to compare the two closely related East Asian sibling species Drosophila sinobscura and D. hubeiensis producing fertile hybrids in the laboratory. The data were also applied to establish their phylogenetic relationships to the other species of the D. obscura group. Genetic divergence in 5SrDNA repeats and specifically in the Adh gene separate the two species clearly from each other and justify their species status. Both species are related to the European species of the D. obscura group but the presence of members of the ATOC 180 satellite DNA family, specific and common to the species triad D. ambigua, D. tristis and D. obscura, in the genomic DNA of D. sinobscura and D. hubeiensis put the two sibling species in their close neighbourhood.     

Systematic analysis of Dendrobium Swartz section Dendrocoryne in the Australian region

Dendrobium sect. Dendrocoryne is a difficult taxonomic group of allied species and species complexes, occurring on the eastern Australian coastline, Lord Howe Island and New Caledonia. Significant morphological characters were assessed from all key areas of the distribution range. Patterns of variation between species, varieties and hybrids were investigated by principal coordinate analysis. The analyses affirm 12 species in the sect. Dendrocoryne described by Schlechter (1912, 1982) and Dockrill (1969, 1992), including a cluster of small statured D. gracilicaule, D. adae, D. fleckeri, and D. finniganense, and three variable species complexes – D. speciosum, D. kingianum and D. tetragonum. Recently described D. finniganense and D. callitrophilum are interpreted as members of the section. Phylogenetic relationships are presented based on parsimony analysis of 31 morphological characters. Analyses do not support proposals to recognize new genera for D. tetragonum, D. callitrophilum and D. aemulum, or to create new species within D. tetragonum, D. jonesii, D. speciosum and D. kingianum. The phylogeny indicates that rainforest taxa are earlier lineages and xerophytic taxa are more derived.     

Heterochromatin in mitotic chromosomes of theVirilis species group ofDrosophila

The amount of heterochromatin in the genome of ten members of thevirilis species group was determined as the length of C-band chromosome material relative to the total karyotype length. Thevirilis phylad (Drosophila virilis, D. novamexicana, D. americana americana, andD. americana texana) has significantly greater amounts of heterochromatin in the genome than do members of the montana phylad (D. montana, D. lacicola, D. flavomontana, D. borealis, D. ezoana, D. littoralis). Thus, the significant karyotypic change accompanying diversification of these species has involved reduction in their total constitutive heterochromatin. These changes have apparently involved reductions in the amount of centromeric heterochromatin in the autosomes.     

The autosomal recessive nonsyndromic deafness locus DFNB72 is located on chromosome 19p13.3

We ascertained three consanguineous Pakistani families (PKDF291, PKDF335 and PKDF793) segregating nonsyndromic recessive hearing loss. The hearing loss segregating in PKDF335 and PKDF793 is moderate to severe, whereas it is profound in PKDF291. The maximum two-point LOD scores are 3.01 (D19S1034), 3.85 (D19S894) and 3.71 (D19S894) for PKDF291, PKDF335 and PKDF793, respectively. Haplotype analyses of the three families define a 1.16 Mb region of overlap of the homozygous linkage intervals bounded by markers D19S216 (20.01 cM) and D19S1034 (20.75 cM). These results define a novel locus, DFNB72, on chromosome 19p13.3. There are at least 22 genes in the 1.16 Mb interval, including PTPRS, ZNRF4 and CAPS. We identified no pathogenic variants in the exons and flanking intronic sequences of these three genes in affected members of the DFNB72 families. DFNB72 is telomeric to DFNB68, the only other known deafness locus with statistically significant support for linkage to chromosome 19p.     

Comparative mapping of cosmids and gene clones from a 1.6 Mb chromosomal region of Drosophila melanogaster in three species of the distantly related subgenus Drosophila

The successful hybridization of cosmid clones from Drosophila melanogaster (Sophophora subgenus) to the salivary gland chromosomes of other species as distantly related as those in the Drosophila subgenus attests their great potential for unravelling genome evolution. We have carried out, using 28 cosmids and 13 gene clones, a study of the organization of the D. melanogaster 95A-96A chromosomal region in three Drosophila subgenus species: D. repleta, D. buzzattii and D. virilis. These clones were first used to built an accurate map of this 1.6 Mb region of D. melanogaster chromosome 3R (Muller’s element E). Then, they were hybridized and mapped to the homologous chromosome 2 of the other three distantly related species. The studied region is disseminated over 13 different sites of chromosome 2 in the Drosophila subgenus species, which implies a minimum of 12 inversion breakpoints fixed between the two subgenera. Extrapolation to the entire chromosome gives 90 fixed inversions. The D. melanogaster Pp1-96A-Acr96Aa segment conserved in D. repleta and D. buzzatii is longer than previously thought and is also conserved in D. virilis. In addition, three other D. melanogaster segments conserved in the three Drosophila subgenus species were found. Finally, our data indicate significant statistical differences in the evolution rate of Muller’s element E among lineages, a result that agrees well with the previous cytogenetic data. Received: 22 July 1998; in revised form: 11 November 1998 / Accepted: 12 November 1998     

小麦苗期盐胁迫下DREB基因的表达

为了从全基因组和转录组水平鉴定响应盐胁迫的小麦DREB (dehydration responsive element binding,DREB) 基因,该研究对小麦耐盐材料CH7034苗期施加盐胁迫后的根部样本进行Illumina转录组测序,从中分离TaDREB家族成员的表达数据和可变剪接信息,并对其下游靶基因进行预测;利用 qRT PCR对盐胁迫响应TaDREB成员和预测靶基因进行验证。结果显示：(1)从小麦中共鉴定出48个DREB成员(204个拷贝序列),命名为TaDREB1~TaDREB48,分布于21条染色体。(2)TaDREB家族分为14组(G1~G14),位于G2、G5、G10和G14的TaDREB成员受NaCl胁迫后转录水平均无显著变化,其余组中共有25个(52%) TaDREB成员表现出对盐胁迫不同程度的响应;其中有9个成员在盐胁迫后持续上调(含5个新报道基因),有2个成员表现为持续下调;蛋白互作预测结果显示,下调成员TaDREB35的编码蛋白可能会受到1个小麦RING型E3泛素连接酶作用而降解。(3)盐胁迫后有9个成员TaDREB3、TaDREB6、TaDREB16、TaDREB19、TaDREB21、TaDREB24、TaDREB25.12、TaDREB43和TaDREB47发生了可变剪切变化。(4)从转录组差异表达基因中进一步鉴定出3个起始密码子上游2 000 bp序列,包含DRE/CRT元件且在A/B/D组间表达趋势一致的候选靶基因TaRD29、TaGLOS和TaCKX。(5)qRT PCR验证结果显示,上调成员中,除TaDREB19外,其余成员以及TaDREB16均表现出持续上升的趋势;下调成员中只有TaDREB25和TaDREB35的表达量呈持续下降的趋势;3个预测靶基因的表达量均持续上升,验证结果与转录组测序结果一致。该研究鉴定出的11个盐胁迫响应TaDREB成员以及预测的3个下游靶基因为小麦耐盐机制解析和分子育种奠定了基础。     

Morphological and chromosomal variation of the Dryopteris varia (L.) Kuntze complex (Dryopteridaceae) in Korea

The Dryopteris varia complex is highly variable in morphology, resulting in taxonomic confusion in delimiting taxon boundaries and determining relationships. We have examined the variation in morphology, chromosome number and mode of reproduction of the Korean members of the D. varia complex to clarify their taxonomic identities. Landmark analysis of the leaf blades and pinnae and the principal components analysis of 31 morphological characters revealed seven entities within the D. varia complex in Korea; these comprise D. varia s. str., D. pacifica, D. sacrosancta, D. bissetiana, D. saxifraga, D. saxifragivaria, and the Suak population which is considered to be a new taxon. Mitotic chromosome counts and examination of reproduction modes indicated that D. bissetiana in Korea appears to be agamosporous with diploid (2n = 82) or triploid (2n = 123) chromosome numbers. However, D. saxifraga is sexual diploid or agamosporous triploid, and the other taxa are agamosporous triploid. Dryopteris bissetiana, D. saxifraga and D. saxifragivaria are similar in major morphological characteristics, but show differences in attachment and shape of rachis scales. The results also suggest that agamosporous triploid D. saxifragivaria was probably derived from hybridization between sexual diploid D. saxifraga and D. bissetiana.     

Many corals host thermally resistant symbionts in high-temperature habitat

Physiologically distinct lines of dinoflagellate symbionts, Symbiodinium spp., may confer distinct thermal tolerance thresholds on their host corals. Therefore, if a coral can alternately host distinct symbionts, changes in their Symbiodinium communities might allow corals to better tolerate increasing environmental temperatures. However, researchers are currently debating how commonly coral species can host different symbiont types. We sequenced chloroplast 23 s rDNA from the Symbiodinium communities of nine reef-building coral species across two thermally distinct lagoon pools separated by ~500 m. The hotter of these pools reaches 35°C in the summer months, while the other pool’s maximum temperature is 1.5°C cooler. Across 217 samples from nine species, we found a single haplotype in both Symbiodinium clades A and D, but four haplotypes in Symbiodinium clade C. Eight of nine species hosted a putatively thermally resistant member of clade D Symbiodinium at least once, one of which hosted this clade D symbiont exclusively. Of the remaining seven that hosted multiple Symbiodinium types, six species showed higher proportions of the clade D symbiont in the hotter pool. Average percentage rise in the frequency of the clade D symbiont from the hotter to cooler pool was 52% across these six species. Even though corals hosted members of both the genetically divergent clades D and C Symbiodinium, some showed patterns of host–symbiont specificity within clade C. Both Acropora species that hosted clade C exclusively hosted a member of sub-clade C2, while all three Pocillopora species hosted a member of sub-clade C1 (sensu van Oppen et al. 2001). Our results suggest that coral–algal symbioses often conform to particular temperature environments through changes in the identity of the algal symbiont.     

Colonization of North America by Drosophila subobscura: Ecological analysis of three communities of drosophilids in California

We have carried out bimonthly collections of Drosophila species during a 16 month period in three localities of California (Eureka, Davis, and Gilroy). D. subobscura, which has colonized North America only a few years ago, is well established and has an annual abundance cycle similar to that observed in the Old World. The differences between the three localities account for 26.4% of the species diversity. Davis and Gilroy exhibit similarities attributable to their similar environmental conditions. Seasonality accounts for 46% and 50.4 % of the species diversity in Davis and Gilroy respectively, but only for 39.4% in Eureka. Differences between years explain only a small percentage of the total species diversity. A correspondence analysis of the association between species and months of collection shows ecological similarities between D. subobscura and the Nearctic members of the obscura species group.     

Differences in genetic structure and ecological diversity between parental forms and hybrids in a Daphnia species complex

Hybridization is a common phenomenon in Daphnia species complexes. Hybrids often dominate in Daphnia populations; therefore it is worthwhile to look for principal differences between parental and hybrid populations with respect to their genetic structure and clonal differentiation. We studied natural populations of members of the Daphnia galeata/hyalina/cucullata complex in three lakes. In one of these lakes, one parental species (D. galeata) and one hybrid (D. galeata × cucullata) were investigated more intensively. The frequency of sexual reproduction was higher in parental populations, whereas clonal diversity was higher in hybrid populations. Ecological differentiation among clonal groups was more pronounced in the D. galeata × cucullata hybrid compared to D. galeata, whereas selection intensity was weaker. These results are discussed with respect to stability of clonal groups, multiple hybridizations and selective constraints.     

The neo-X neo-Y sex pair in Acrididae,its structure and association

In most of the fifty described cases of neo-X neo-Y sex determining systems in Acrididae the pairing regions during meiosis are limited to distal regions. A comparative study on the structure and pairing mechanisms of Dichroplus silveiraguidoi (2n=8); Dichroplus bergi (2n=22) and Dichroplus vittatus (2n=20) has been undertaken. — The sex bivalents of these three grasshoppers are different: the neo-X centromere is associated with the neo-Y telomere in D. silveiraguidoi; in D. bergi the neo-X is related through the short arm telomere to the centromere of neo-Y and both members of the sex pair are associated by the telomeres in D. vittatus. Centromeric and telomeric C-band positive blocks are present in both members of the pair in the three species. D. silveiraguidoi also presents an interstitial block in the neo-X. These blocks are brightly fluorescent with quinacrine mustard and Hoechst 33258 at low concentration (0.05 g/ml). The region of neo-X corresponding to the primitive X takes an intermediate staining during the early meiotic prophase with C-banding and Hoechst 33258. — The structure of the sex bivalent and the particular staining of the X region are discussed in relation to the available information on the presence of different types of DNA in this segment. The possibility that the neo-X interstitial block of D. silveiraguidoi plays a role in preventing the spreading of heterochromatinization along the chromosome is also discussed. The classical interpretation of the neo-X neo-Y association during meiotic prophase as the result of a terminalized chiasma is considered in the light of optic and electronmicroscopic data. Other possible mechanisms of relationship between both chromosomes are also presented by these three orthopteran species.