Genetic variation in sympatric populations of diploid and polyploid brine shrimp (Artemia parthenogenetica)

We examined genetic variation in sympatric diploid and polyploid brine shrimp Artemia parthenogenetica from each of three populations (China, Italy and Spain). Italian and Spanish tetraploids are closely related (I=0.964). Diploids and tetraploids within each of the two European populations are also closely related (mean I=0.905). Most alleles found in diploids also exist in sympatric polyploids. In contrast, the asexual Artemia (2N, 4N and 5N) in our study share few alleles with their close sexual relative, A. tunisiana (mean I=0.002). These results, as well as the work of other authors, strongly suggest that at least the tetraploid Artemia in our study have an autopolyploid origin.Clonal diversity of polyploid Artemia can be very high at least in some population. Both diploids and polyploids had low clonal diversities in the populations dominated by polyploids and high clonal diversities in the population dominated by diploids.The most common genotypes of sympatric diploid and polyploid Artemia frequently differed. Some alleles occurred only in diploids, while others were restricted to polyploids. These results suggest that polyploidy in Artemia has led to genetic divergence from diploid progenitors, and that ploidy-level variation must also be considered in developing an understanding of spatial and temporal allozyme polymorphism in asexual populations.     

Characterization of brine shrimp Artemia from Cam Ranh Bay in Central Vietnam

Artemia cysts collected from inoculation experiments in Cam Ranh salterns are evaluated for their potential use in aquaculture. Cyst biometrics, hatching quality, naupliar fatty acid profile and naupliar growth were measured and compared to reference Artemia strains. Cyst characteristics reveal that the parthenogenetic strain (PR China) used in inoculations, was eliminated from the environment and that the remaining brine shrimp are likely to be composed of Macau and Great Salt Lake Artemia strains, and of their cross-breds. Differences in cyst diapause deactivation characteristics between Macau and Great Salt Lake Artemia may have resulted in the disappearance of Macau Artemia during the rainy season and the persistence of Great Salt Lake Artemia during the following dry season.     

Molecular phylogenetics and asexuality in the brine shrimp Artemia

Explaining cases of long-term persistence of parthenogenesis has proven an arduous task for evolutionary biologists. Interpreting sexual-asexual interactions though has recently advanced owing to methodological design, increased taxon sampling and choice of model organisms. We inferred the phylogeny of Artemia, a halophilic branchiopod genus of sexual and parthenogenetic forms with cosmopolitan distribution, marked geographic patterns and ecological partitioning. Joint analysis of newly derived ITS1 sequences and 16S RFLP markers from global isolates indicates significant interspecific divergence as well as pronounced diversity for parthenogens, matching that of sexual ancestors. Maximum parsimony, maximum likelihood, and Bayesian methods were largely congruent in reconstructing the phylogeny of the genus. Given the current sampling, at least four independent origins of parthenogenesis are deduced. Molecular clock calibrations based on biogeographic landmarks indicate that the lineage leading to A. persimilis diverged from the common ancestor of all Artemia species between 80 and 90 MYA at the time of separation of Africa from South America, whereas parthenogenesis first appeared at least 3 MYA. Common mitochondrial DNA haplotypes delineate A. urmiana and A. tibetiana as possible maternal parents of several clonal lineages. A novel topological placement of A. franciscana as a sister clade to all Asian Artemia and parthenogenetic forms is proposed and also supported by ITS1 length and other existing data.     

Guanylate requirement for patterning the postcephalic body region of the brine shrimp Artemia

Mycophenolic acid (MA) reduces growth and survival rates of Artemia larvae fed hypoxanthine. The inhibitory effect of MA is suppressed by dietary guanosine as expected for inosinate dehydrogenase inhibition. Pulse MA treatments lasting 24 h imposed on larvae from 24 to 96 h posthatch result in the production of abnormal adults with thoracic, genital or abdominal defects. The MA-induced anomalies are interpreted as a disruption of pattern formation. Since dietary guanosine restores normal pattern in adults developing from larvae, MA-treated during early stages, it is concluded that guanylate production is required at critical developmental times to establish normal developmental fates.     

Reproductive strategies of diploid and polyploid populations of arcticDraba (Brassicaceae)

132 cultivated populations (2x–16x) of 15 arctic-alpine species ofDraba were investigated to clarify a possible relationship between reproductive strategies and polyploid evolution in the genus. The populations were exclusively sexual and produced viable seed after spontaneous self-pollination, but showed large variation both in traits promoting cross-pollination and in autogamous fruit and seed set. Traits promoting cross-pollination, e.g., floral display, protogyny, and delayed selfing, were positively correlated, and these traits were negatively correlated with autogamous fruit and seed set. All diploid and many polyploid populations had high autogamous seed set and small, unscented, non-protogynous, and rapidly selfing flowers. In contrast, all populations with low autogamous seed set and large, scented, and strongly protogynous flowers with distinctly delayed selfing were polyploid. These results are consistent with those previously obtained from enzyme electrophoresis, suggesting that the genetically depauperate diploids are extreme inbreeders and that the highly fixed-heterozygous polyploids vary from extreme inbreeders to mixed maters. The reproductive data lend additional support to the hypothesis that allopolyploidy in arcticDraba serves as an escape from genetic depauperation caused by uniparental inbreeding at the diploid level.     

Characterizing bisexual Artemia populations by isoelectric focusing

Isoelectric focusing (IEF) of protein extracts, -Esterase and Phosphoglucose isomerase, from groups of Artemia adults from different bisexual populations were studied. Both gave clear separation between the Old and New World species. The second was more polymorphic, allowing a discrimination among Mediterranean populations.     

Comparative demography of sympatric populations of Lacerta vivipara and Lacerta agilis

Summary A mark-recapture study was carried out in sympatric populations of Lacerta agilis and Lacerta vivipara in the Netherlands from 1976 to 1982. In most years the age structure of both populations was pyramidal. For both species life expectation of females was higher and on average they did live longer. Hence the sex ratio for adults deviated significantly from 1.0 in favour of females. Maximum age for Lacerta vivipara was 8 years (female) and for Lacerta agilis 12 years (male). The density of both species fluctuated around 100/ha. The biomass of Lacerta agilis was twice that of Lacerta vivipara. In Lacerta vivipara the 3rd and 4th calendar year class supplied 78% of total net reproduction; in Lacerta agilis the 4th, 5th, and 6th calendar year classes supplied 68%. In both populations the population replacement rate was 2. Population turnover time was 4.83 years for Lacerta agilis and 2.81 for Lacerta vivipara. The life history strategy of the Lacerta vivipara population is compared with six other European Lacertavivipara populations.     

Further characterization of the cathepsin L-associated protein and its gene in two species of the brine shrimp, Artemia

The major cysteine protease in embryos and larvae of the brine shrimp Artemia franciscana is a heterodimer composed of a cathepsin L-like polypeptide of 28.5 kDa and a 31.5 kDa polypeptide called the cathepsin L-associated protein or CLAP. In a previous study, CLAP was shown to be a cell adhesion protein containing two Fas I domains and two GTP/ATP binding sites known as Walker A and B motifs. Here, we have characterized CLAP and its genes to better understand the role of this protein in Artemia development. The polymerase chain reaction was used to investigate the structure of the CLAP gene in two species of Artemia, the New World bisexual diploid A. franciscana and the Old World parthenogenetic tetraploid Artemia parthenogenetica. The protein coding region of the CLAP gene from each species was 99.5% identical for a protein of 332 amino acids, while the 3' non-coding region, representing nearly 45% of the gene, was only 86% identical between the two related species. However, while the CLAP gene is intronless in A. franciscana, in A. parthenogenetica the gene contained a mini-intron of 30 base pairs in the 3' non-coding region. The sequences representing the CLAP gene in A. franciscana and A. parthenogenetica have been entered into the NCBI database as AY757920 and DQ100385, respectively. Northern blot analysis showed that while the cathepsin L gene is expressed constitutively in Artemia franciscana embryos and young larvae, the CLAP gene is not expressed in late embryos and young larvae. In contrast, Western blots indicated that CLAP is present in developing embryos and young larvae, at least to the first larval molt, supporting results obtained previously showing CLAP's resistance to degradation by its dimeric partner, cathepsin L. At the protein level we showed that the GTP/ATP binding sites in CLAP are functional with rate constants of 0.024 and 0.022 for GTP and ATP hydrolase activity, respectively. GTP but not ATP also had a slight stimulatory effect on cathepsin L activity of the heterodimeric protease containing CLAP. Our results support the hypothesis that CLAP plays an important role in targeting and expression regulation of cathepsin L activity during early development of Artemia.     

Stress tolerance during diapause and quiescence of the brine shrimp,Artemia

Oviparously developing embryos of the brine shrimp, Artemia, arrest at gastrulation and are released from females as cysts before entering diapause, a state of dormancy and stress tolerance. Diapause is terminated by an external signal, and growth resumes if conditions are permissible. However, if circumstances are unfavorable, cysts enter quiescence, a dormant stage that continues as long as adverse conditions persist. Artemia embryos in diapause and quiescence are remarkably resistant to environmental and physiological stressors, withstanding desiccation, cold, heat, oxidation, ultraviolet radiation, and years of anoxia at ambient temperature when fully hydrated. Cysts have adapted to stress in several ways; they are surrounded by a rigid cell wall impermeable to most chemical compounds and which functions as a shield against ultraviolet radiation. Artemia cysts contain large amounts of trehalose, a non-reducing sugar thought to preserve membranes and proteins during desiccation by replacing water molecules and/or contributing to vitrification. Late embryogenesis abundant proteins similar to those in seeds and other anhydrobiotic organisms are found in cysts, and they safeguard cell organelles and proteins during desiccation. Artemia cysts contain abundant amounts of p26, a small heat shock protein, and artemin, a ferritin homologue, both ATP-independent molecular chaperones important in stress tolerance. The evidence provided in this review supports the conclusion that it is the interplay of these protective elements that make Artemia one of the most stress tolerant of all metazoan organisms.     

The American brine shrimp as an exotic invasive species in the western Mediterranean

The hypersaline environments and salterns present in the western Mediterranean region (including Italy, southern France, the Iberian Peninsula and Morocco) contain autochthonous forms of the brine shrimp Artemia, with parthenogenetic diploid and tetraploid strains coexisting with the bisexual species A. salina. Introduced populations of the American brine shrimp A. franciscana have also been recorded in these Mediterranean environments since the 1980s. Based on brine shrimp cyst samples collected in these countries from 1980 until 2002, we were able to establish the present distribution of autochthonous brine shrimps and of A. franciscana, which is shown to be an expanding invasive species. The results obtained show that A. franciscana is now the dominant Artemia species in Portuguese salterns, along the French Mediterranean coast and in Cadiz bay (Spain). Co-occurrence of autochthonous (parthenogenetic) and American brine shrimp populations was observed in Morocco (Mar Chica) and France (Aigues Mortes), whereas A. franciscana was not found in Italian cyst samples. The results suggest these exotic A. franciscana populations originate as intentional or non-intentional inoculations through aquacultural (hatchery effluents) or pet market activities, and suggest that the native species can be rapidly replaced by the exotic species. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of water-borne copper on respiratory and cardiac function during the early ontogeny of the brine shrimp,Artemia franciscana Kellogg 1908 (Branchiopoda: Anostraca)

There was no direct effect of copper on the ontogeny or function of the heart of the brine shrimp Artemia franciscana in sea water (salinity= 36 mg·ml^-1, 25°C). There was, however, an indirect effect as an increase in copper concentration resulted in a reduced growth rate. There was no difference between the critical O₂ tensions of newly hatched (stage 0/1) nauplii of control and treated (<0.32 and 10.11 mol·l^-1 copper, respectively) individuals. However by developmental stages 4–6, when both the heart and thoracic gills are in the process of differentiating, respiratory performance had improved (i.e. critical O₂ tension decreased from 6.27±0.45 to 4.69±0.24 kPa) in control but not in copper-treated individuals. It is suggested that respiratory impairment of stages 4–6 individuals is unlikely to be related to differences in cardiac performance or cellular respiration. Instead it may be related to metal-related damage to newly differentiating gill tissue and/or by copper in some way compromising the normal ontogenic shift in haemoglobin O₂ affinity. Copper-related respiratory impairment develops at a critical point in brine shrimp organogenesis when a good supply of O₂ is essential for normal development and if compromised may reduce the ability of this species to survive copper exposure.Abbreviations BL body length - BW body weight - HR heart rate - HM heavy metals - SW sea water - P _c critical oxygen tension     

Life history characteristics of the clam<Emphasis Type="Italic"> Mya arenaria</Emphasis> in the White Sea

Dynamics of Mya arenaria beds in two bights of the Chupa Inlet (Kandalaksha Bay, White Sea) were studied on a long-term basis. Observations were carried out at 1– to 3-year intervals from 1979 up to 1999. The studied soft-shell clam beds were characterised by a substantial instability of age structure. Since 1988, only one year-class has dominated in the beds while other generations have been scarce and recruitment was not observed. This pattern of Mya bed dynamics was related neither to interannual environmental changes nor to differential reproduction success or predation effects in the benthic assemblages. Favourable conditions for spat formation in 1988 (low abundance of other M. arenaria generations), as well as for juvenile survival during the following winter, resulted in high abundance of juveniles in both investigated locations in 1989. The mortality rate (μ) in this 1988 generation varied throughout the period of investigation and was related to age. The mortality level decreased for the first 2–4 years of the life cycle, then stabilised for the next 3–4 years, and eventually increased in subsequent years. Overall μ values ranged from 0 to 1.68 year^–1. The oldest specimens observed were 17 years old and had a maximum shell length of 79 mm. Significant differences in average growth rates were observed between molluscs of different locations. Communicated by H.-D. Franke     

Interdomain linkage in the polymeric hemoglobin molecule of Artemia

Artemia has evolved the longest known concatenation of hemoglobin domains, the subunit containing nine domains and the subunit having a similar size. Translation of the cDNA sequence of the subunit reveals eight regions of inter-domain polypeptide linking together the nine heme-binding domains, together with partially analogous sequences preceding the first domain and following the last. Analysis of the structural possibilities of the linker sequences suggests how the domains may be organized in the subunit.The interdomain linker sequences were 14%–64% identical (62%–91% similar by Dayhoff substitution matrix) and approximately 14 residues in length including a consensus -Val-Asp-Pro-Val-Thr-Gly-Leu-. The linker composition resembled that of the 11 amino acid pre-A leader sequence of Petromyzon marinus (lamprey) hemoglobin V, the structure of which is known. Prediction of structure from the Artemia linker sequences indicated a nonhelical, turn-associated linker which could be modeled to the Petromyzon leader. Measurements confirmed that such a structure could support the packing of nine Artemia domains into a polymeric subunit of annular shape, two of which subunits (which can be similar or dissimilar) comprise the physiological molecule.The position of interdomain introns and the character of a variable residue early in the linker are compatible with the nine-domain polymer having evolved through gene duplication reflected in globin domain fusion incorporating an extension specifically of the N-terminus. The multiplication of an original single-domain globin gene to give the present nine is estimated from sequence differences, allowing for multiple mutations at individual sites, to have occurred in a period at least 500–700 million years ago.Correspondence to: C.N.A. Trotman 1444     

Characterisation of high-altitude Artemia populations from the Qinghai-Tibet Plateau,PR China

The brine shrimp Artemia was discovered in a number of saline lakes on the Qinghai-Tibet Plateau, widely diverging in chemical composition. Several lakes were athalassohaline, with relatively high amounts of trace elements. Common environmental factors are their high altitude (exceeding 4500 m) and the low average annual temperatures. A number of Artemia populations in this area were analysed to assess their preference for low temperatures and an athalassohaline medium. Furthermore, their characteristics were compared with Artemia tibetiana, the species recently described for one lake in this area. All samples contained a variable mixture of parthenogenetic and bisexual individuals. A cross-breeding test of the sample from Jingyu Lake showed cross-fertility both with A. tibetiana and A. sinica. All populations showed similarities to A. tibetiana: a large cyst diameter and naupliar length, high HUFA content and a high tolerance to low temperatures, as compared to the control A. franciscana samples. These can thus be considered as recurrent characteristics of the populations from the high-altitude low-temperature environment on the Qinghai-Tibet Plateau, although further research is needed to identify their exact species status.     

Life history variation among low-arctic clones of obligately parthenogenetic Daphnia pulex: a diploid-polyploid complex

Summary Laboratory life table experiments were conducted using nine clones of obligately parthenogenetic Daphnia pulex that were collected from a site in the Canadian low-arctic. Two of the nine clones were diploids, while the other seven clones were polyploids. Significant clonal differences in age at first reproduction, size at first reproduction, number of offspring in each of the first three broods, offsrring sizes for the first two broods, and intrinsic rates of natural increase were detected. Differences in life histories were evident between polyploids and diploids. Generally, polyploid clones reached maturity at later ages, matured at larger sizes, produced smaller broods, and larger offspring than the diploid clones. The data are discussed in reference to potential biotic (i.e. invertebrate predation) and abiotic factors (i.e. physicochemical gradients) that may influence life history variation in this clonal assemblage.     

International study on Artemia. LVI. Characterization of two Artemia populations from Namibia and Madagascar: cytogenetics, biometry, hatching characteristics and fatty acid profiles

Two parthenogenetic Artemia populations from southern Africa, one from Swakopmund saltworks (Namibia) and another from Ankiembe saltworks (Madagascar) have been studied. The population from Namibia is mainly diploid (2n=42) with few tetraploid individuals (4n=84), while the one from Madagascar was found to be triploid (3n=63). No chromocenters have been observed in either of the two populations. The Namibian population has smaller cysts and nauplii compared to those of the Madagascar population. Discriminant analysis revealed significant differences in the biometry of the adults from the two populations. The two populations exhibited very good hatching characteristics. The study of fatty acid methyl esters revealed that the Namibian population belongs to the fresh water type of Artemia showing low levels of eicosapentaenoic acid, whereas the population from Madagascar displayed exceptionally high levels of eicosapentaenoic acid, belonging to the marine water type.     

Life history and population dynamics of Atriplex triangularis

The dynamics of a population of the stoloniferous Potentilla anserina was studied during four years. The mortality of established ramets followed a log-linear pattern superimposed on a seasonal fluctuation with an increased mortality during the period of vegetative spread. The major external mortality factors were unfavourable weather conditions such as drought and freezing. The turnover rate of the population was slow (31.6 yr). Vegetative spread dominated the recruitment to the population. Seedlings occurred but were exposed to a much higher mortality than vegetative daughter ramets. The juvenile period of seedlings is at least 4 yr in the field. This contrast to vegetative daughter ramets which are potentially flowering and stoloniferous the year after they are established, even if age affects both reproduction and vegetative spread. The density of most of the population is below the level where intraspecific regulative effects operate. However, in very dense stands both stolon and flower production were negatively density dependent.Nomenclature follows Lid (1974), Norsk og Svensk Flora, 2nd ed. Oslo.I am indebted to Lenn Jerling, Peter Torstensson, Anders Telenius and Gudrún Jónsdóttir for their help, and inspiring criticism throughout this study.     

Heterochromatin differentiation and phylogenetic relationship of the A genomes in diploid and polyploid wheats

Summary Heterochromatin differentiation, including band size, sites, and Giemsa staining intensity, was analyzed by the HKG (HCl-KOH-Giemsa) banding technique in the A genomes of 21 diploid (Triticum urartu, T. boeoticum and T. monococcum), 13 tetraploid (T. araraticum, T. timopheevi, T. dicoccoides and T. turgidum var. Dicoccon, Polonicum), and 7 cultivars of hexaploid (T. aestivum) wheats from different germplasm collections. Among wild and cultivated diploid taxa, heterochromatin was located mainly at centromeric regions, but the size and staining intensity were distinct and some accessions' genomes had interstitial and telomeric bands. Among wild and cultivated polyploid wheats, heterochromatin exhibited bifurcated differentiation. Heterochromatinization occurred in chromosomes 4A^t and 7A^t and in smaller amounts in 2A^t, 3A^t, 5A^t, and 6A^t within the genomes of the tetraploid Timopheevi group (T. araraticum, and T. timopheevi) and vice versa within those of the Emmer group (T. dicoccoides and T. turgidum). Similar divergence patterns occurred among chromosome 4A^a and 7A^a of cultivars of hexaploid wheat (T. aestivum). These dynamic processes could be related to geographic distribution and to natural and artifical selection. Comparison of the A genomes of diploid wheats with those of polyploid wheats shows that the A genomes in existing diploid wheats could not be the direct donors of those in polyploid wheats, but that the extant taxa of diploids and polyploids probably have a common origin and share a common A-genomelike ancestor.Contribution of the College of Agricultural Sciences, Texas Tech Univ. Journal No. T-4-233.