Further studies with interspecific hybridization among mediterranean/African lupin species

Summary Interspecific hybridization has a role to play in the domestication of wild species through the introgression of desirable genes from related domesticated species. Almost complete genetic isolation among the Mediterranean-African rough-seeded lupin species has hitherto ruled out introgression within that group. Recent work in Western Australia with L. atlanticus, L. cosentinii, L. pilosus and L. digitatus has nevertheless suggested that hybrid sterility may be overcome if specially selected lines of each are used in crossing. We have now selected F₂-F₅ plants and backcrosses (Bc₁; Bc₂) from L. atlanticus/L. cosentinii, some with improvement in seed fertility combined with domestication genes from either species. Relatively better F₁-F₂ plant fertility in subsequent crosses of L. atlanticus/L. digitatus than in L. atlanticus/L. cosentinii or L. digitatus/L. cosentinii indicate closer relationship between L. atlanticus and L. digitatus than in the other cross combinations. Use of embryo culture may help to overcome L. pilosus/L. atlanticus F₁ plant sterility. Some of the interspecific selections could act as genetic bridges between L. cosentinii and L. pilosus.     

Isolation of seven polymorphic microsatellites in Ophioblennius atlanticus atlanticus (Perciformes,Blenniidae)

We isolated and characterized seven polymorphic microsatellite loci of Ophioblennius atlanticus atlanticus (Valenciennes, 1836) using an optimized protocol to construct and screen a microsatellite‐enriched genomic library. The analysis of variability was performed in 16 specimens from Faial Island (Azores, Portugal). The mean number of alleles was 8.71 ± 2.43 and the level of expected heterozygosity ranged from 0.764 to 0.903. The total exclusionary probabilities using these loci for the first and the second parent were 0.985 and 0.998, respectively, suggesting that these microsatellites are a useful tool for large‐scale parentage analysis.     

Evolution of the third eye: a phylogenetic comparative study of parietal‐eye size as an ecophysiological adaptation in Liolaemus lizards

The parietal, or third, eye is a photosensory organ situated in the middle of the skull of many lizards. Despite many hypotheses, its exact ecological functions are still unclear. Studies have compared the presence and absence of a functioning parietal eye, although there are no quantitative studies of parietal‐eye traits in relation to ecology, physiology or behaviour. In the present study, we report the first comparative study of relative parietal‐eye size in relation to climatic and thermophysiological variables. We studied thirty species of Liolaemus, a genus of South‐American lizards inhabiting a range of climatic conditions, but found little evidence for adaptation to thermal environment, in that parietal‐eye size did not vary meaningfully with latitude, altitude or any measures of environmental temperature. Neither did it relate to thermophysiology; there was a weak relation to thermal tolerance, although this was partially confounded with body size, which explained 23% of the among‐species variance after controlling for within‐species variation. The negative results obtained could not be explained by phylogenetic constraints because we found no evidence of phylogenetic inertia. We also observed high intraspecific variation indicating that parietal‐eye size may not be under strong selection for accuracy. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101 , 870–883.     

Resurrected Ceriodaphnia quadrangula highlight differences between pheno‐ and genotypic expressions

The hatching of cladoceran ephippia from a 15‐cm long sediment core was investigated, and Ceriodaphnia quadrangula clones were isolated from different sediment layers. Bosmina microfossil data were also analyzed, and compared with the corresponding data from a Pb210 dated core, which allowed us to infer the age of the sediment layers. Using changes in Bosmina microfossil morphologies, we were, furthermore, able to infer the presence of different regimes of fish predation. C. quadrangula was found to hatch in layers with an inferred age of approximately a century. Newly hatched individuals had smaller eye‐size in sediment layers corresponding to high predation by young‐of‐the‐year perch. Newly hatched individuals also generally had a marked neck‐spine. In contrast, morphological characters of C. quadrangula clones reared in the laboratory over several generations showed no variation in relation to predation regime, indicating the absence of fixed genotype level changes. Furthermore, the laboratory grown clones only rarely produced a neck‐spine. The results suggest phenotypic variation in response to the regime under which ephippia were produced.     

A new cottid species, Icelus sekii (Perciformes: Cottoidei), from Hokkaido, Japan

A new cottid species, Icelus sekii, is described on the basis of six specimens collected from off Rausu and Urakawa, Hokkaido Island, Japan. This species is distinguished from its congeners by the following combination of characters: supraocular and parietal spines absent; nuchal spine obscure; uppermost preopercular spine unbranched; no scales between dorsal scale row and lateral line scale row, and no scales below lateral line scale row; supraocular, parietal, and nuchal cirri present; five dark brown saddles dorsolaterally; anal fin rays 13; pectoral fin rays 15; vertebrae 12 + 24–25 = 36–37. Icelus sekii can be mature at the smallest size among the species of Icelus. As a secondary sexual character, the male holotype has unique ensiform flaps on the distal tips of the first dorsal fin.     

The Pacific Notocotylus atlanticus (Digenea: Notocotylidae)

Recently two unidentified Notocotylus species have been recovered from a mallard in Hokkaido, Japan. Preliminary data indicated that one of them may belong to N. atlanticus—a species found in Europe and on the Atlantic coast of North America. Now we have sequenced partial cox1 for several European N. atlanticus isolates and performed a barcode gap analysis. It has shown the conspecificity of European and Japanese worms, bringing up discussion on distribution, transmission and host specificity in this species.     

Taxonomic status of southern South American Conepatus (Carnivora: Mephitidae)

Despite recent taxonomic evaluations of Mephitidae and North American hog‐nosed skunks, southern South American species of Conepatus have not been thoroughly examined in a systematic context. Conepatus chinga and Conepatus humboldtii were described more than 150 years ago, based on external characters such as hair coloration and size. Although historically recognized as valid species, to date no detailed systematic analysis has been performed for either of these taxa. Herein, we evaluated the taxonomic status of C. chinga and C. humboldtii within the southern part of South America using geometric morphometrics of the skull and mandible, mitochondrial DNA analysis using the cytochrome b and cytochrome oxidase c subunit I genes, and also control region and pelage pattern variation. We failed to find morphological (skull shape and pelage coloration patterns) or molecular differences between these two species; thus, we considered that the specimens assigned to C. chinga and C. humboldtii belong to the same species. Our results indicate that environmental variation seems to be responsible for shape and size variation in Conepatus skulls from southern South America. © 2013 The Linnean Society of London     

Variation in the growth of lupin species and genotypes on alkaline soil

Commercial lupins grow poorly on alkaline and neutral fine-textured soils. Genotypic variation exists among lupins. The present study compared the growth of 13 lupin genotypes, including introduced cultivars and wild types, in an alkaline loamy soil and an acid loamy soil.Plants grown in the alkaline and acid soils did not show obvious symptoms of iron deficiency at any stage. There was however a large variation of shoot fresh weight among genotypes in response to the alkaline soil with L. atlanticus and L. pilosus being more tolerant than L. luteus, L. cosentinii, L. albus and L. angustifolius. Some variation also existed among genotypes of L. angustifolius. In addition, root growth was retarded on the alkaline soil except for L. atlanticus, L. pilosus P20955 and L. albus Kiev mutant. In the alkaline soil, root growth at week 2 correlated well with the shoot fresh weight at week 12. The results suggest that early root elongation may be useful for screening tolerant genotypes for alkaline soils.     

Analysis of asymmetries in the African fruit bats Eidolon helvum and Rousettus egyptiacus (Mammalia: Megachiroptera) from the islands of the Gulf of Guinea. I. Variance and size components of bilateral variation

A set of cranial characters was examined in the fruit bats Rousettus egyptiacus and Eidolon helvum to compare trends and relative importance of major components of bilateral morphometric variation, and their relationship with character size. Using two‐way, sides‐by‐individuals ANOVA , four components of variation were estimated for each bilateral variable: individual variation (I), directional asymmetry (DA), non‐directional asymmetry (NDA) and measurement error (E). Both species exhibit similar major trends of variation in asymmetry across characters, as shown by principal component analysis, using variance components as variables. Degree of interspecific congruence among characters was confirmed by a two‐way ANOVA with species and variance components as fixed factors. Congruence of asymmetry patterns between species suggests that the concept of population asymmetry parameter (PAP) could be extended to higher hierarchies. PAPs above the species level may result from common mechanisms or similar developmental constraints acting on species’ buffering capacities and morphological integration processes.     

Occurrence and abundance of three larval codlet species (Bregmacerotidae, Teleostei) in the Southwest Atlantic Ocean (12–22°S)

There are still only a few studies related to Bregmacerotidae larvae in Brazil, although these larvae are frequently collected in oceanic ichthyoplankton samples. Thus this study analyzes the occurrence and abundance of Bregmaceros larvae along the Brazilian central coast and describes larval characteristics. During three oceanographic cruises samples were collected between the Real River (12°S) and the São Tomé Cape (22°S). Oblique hauls were conducted from a maximum depth of 200 m, using bongo nets of 330‐ and 500‐μm mesh size. Among the 951 bregmacerotids collected, three species were identified corresponding to 89%Bregmaceros atlanticus, 8%Bregmaceros cantori and 3%Bregmaceros n. sp. B. atlanticus was widely distributed along the study area and was more frequent in stations situated beyond the 200‐m isobath; this species was more abundant during the winter, showing a peak of density between Vitória and the São Tomé Cape (355 larvae per 10 m²). Bregmaceros cantori densities were very similar during the three cruises; the highest value (25.6 larvae per 10 m²) was recorded during the winter and higher concentrations were observed in neritic stations along the 200‐m isobath. Bregmaceros n. sp. was rare in collections made during the three cruises; it was concentrated along the Bahia coast (12–18°S), and was more frequent in stations situated in the oceanic region. Bregmaceros atlanticus varying from 2.0 to 6.0 mm and Bregmaceros n. sp. larvae varying from 3.0 to 6.0 mm were present in all cruises, indicating that they spawn year‐round. Bregmaceros cantori did not show a distribution pattern among the different length classes, probably associated with the fact that this species has a coastal distribution.     

Comparative morphology of the pectoral fin spine of the Persian sturgeon Acipenser persicus,the Russian sturgeon Acipenser gueldenstaedtii,and the Starry sturgeon Acipenser stellatus in Iranian waters of the Caspian Sea

The morphological characteristics of the pectoral fin spine were compared in three species of sturgeon, the Persian sturgeon (Acipenser persicus), the Russian sturgeon (Acipenser gueldenstaedtii), and the Starry sturgeon (Acipenser stellatus), all sampled from the Caspian Sea. On the basis of morphological characters of the pectoral fin spine, 62.2% of the individuals were correctly classified into separate groups. The cluster analysis also divided the three species into two major subgroups. Acipenser persicus and A. gueldenstaedtii were grouped together, suggesting a similar evolutionary basis. Significant morphological heterogeneity in pectoral fin spine characteristics was observed among the three sturgeon species. Principal component analysis identified the largest differences were in the pectoral fin spine size and the angle between distal pectoral fin spine and the horizontal line (A°). The first and second principal components (PC1 and PC2) of all observations accounted for 64.19% and 14.33% of the total variation, respectively. The combination of all analyses showed the relevance of applying pectoral fin spine shape for interspecific distinction of the three species of sturgeons.     

Genetic connectivity and phenotypic plasticity in the cyprinodont Aphanius farsicus from the Maharlu Basin,south‐western Iran

Meristic and morphometric characteristics, including otolith data, of the Farsi tooth‐carp Aphanius farsicus, which is endemic to the endorheic Maharlu Basin in south‐western Iran, were analysed for a sample of 92 individuals from four spring‐streams; DNA sequence data (cytochrome b gene) are presented for 29 specimens. Some phenotypic variation was detected but the genetic data clearly indicate connectivity between the populations. Possible links between phenotypic variation and environmental variables such as water temperature, habitat size and absence or presence of predators and competitors are discussed. Based on a literature survey and the new data, it is concluded that population connectivity is maintained during times of droughts via large aquifers that formed during the late Pliocene to early Pleistocene, when the extant endorheic Maharlu Basin was created. Based on new data presented here and previous work, it is apparent that plastic and constant characteristics are present in Aphanius species, and that, if a population becomes isolated, a given trend of evolution may give rise to a taxonomically useful characteristic.     

Variation in seed packaging of a fleshy‐fruited conifer provides insights into the ecology and evolution of multi‐seeded fruits

• The study of intraspecific seed packaging (i.e. seed size/number strategy) variation across different populations may allow better understanding of the ecological forces that drive seed evolution in plants. Juniperus thurifera (Cupressaceae) provides a good model to study this due to the existence of two subspecies differentiated by phenotypic traits, such as seed size and cone seediness (number of seeds inside a cone), across its range.
• The aim of this study was to analyse seed packaging (seed mass and cone seediness) variation at different scales (subspecies, populations and individuals) and the relationship between cone and seed traits in European and African J. thurifera populations.
• After opening more than 5300 cones and measuring 3600 seeds, we found that seed packaging traits followed different patterns of variation. Large‐scale effects (region and population) significantly contributed to cone seediness variance, while most of the seed mass variance occurred within individuals. Seed packaging differed between the two sides of the Mediterranean Sea, with African cones bearing fewer but larger seeds than the European ones. However, no differences in seed mass were found between populations when taking into account cone seediness. Larger cones contained more pulp and seeds and displayed a larger variation in individual seed mass.
• We validated previous reports on the intraspecific differences in J. thurifera seed packaging, although both subspecies followed the same seed size/number trade‐off. The higher seediness and variation in seed mass found in larger cones reveals that the positive relationship between seed and cone sizes may not be straightforward.We hypothesise that the large variation of seed size found within cones and individuals in J. thurifera, but also in other fleshy‐fruited species, could represent a bet‐hedging strategy for dispersal.

     

Reproduction and larval ecology of the Arctic fish speciesArtediellus atlanticus (Cottidae)

 State of maturity, egg numbers, as well as gonadosomatic index (GSI), and relative fecundity were determined in 209 females of the cottid species Artediellus atlanticus. Specimens were caught by Agassiz and bottom trawls during two expeditions of RV “Polarstern” to the shelf off northeast Greenland (ARK VII/2; July/ August 1990) as well as to the Barents Sea and the shelf areas surrounding Svalbard (ARK VIII/2 – Arctic EPOS; June/July 1991). Analysis of the state of maturity showed that A. atlanticus spawns from late summer to autumn. Individuals from northern populations spawn later (November) than those from more southern locations (July to September). The total number of eggs (40–216) was small due to their large diameter and the small size of the fish. The mean GSI of females from the central Barents Sea (36.9±6.5%; n=24) was significantly higher (P<0.001) than the GSI of females caught off eastern Svalbard (18.7±4.0%; n=14), compared at stage IV of maturity. The mean relative fecundity was significantly higher (P<0.001) for females of the central Barents Sea (17.9±2.3; n=19) than for females caught on more northern stations east of Svalbard (6.9±1.8; n=17) and north of Svalbard (6.4±0.5; n=7). Individuals of A. atlanticus that were maintained at 0°C spawned in the aquaria, allowing observation of the development of their eggs and larvae for 1 year. The eggs needed more than 200 days to develop. The larvae hatched at a very advanced stage and were similar to the adults in appearance and behaviour. The larvae grew at a rate of 0.02 mm per day and showed a benthic mode of life. Received: 7 April 1995/Accepted: 30 July 1995     

Thermal,trophic and metabolic life histories of inaccessible fishes revealed from stable‐isotope analyses: a case study using orange roughy Hoplostethus atlanticus

A time‐resolved record of inhabited water depth, metabolic rate and trophic behaviour of the orange roughy Hoplostethus atlanticus was recovered from combined stable‐isotope analyses of otolith and muscle tissue. The results demonstrate that H. atlanticus from the north‐east Atlantic Ocean have a complex life history with three distinct depth‐stratified life stages. Early juvenile H. atlanticus occupy relatively shallow habitats, juvenile H. atlanticus show a deep‐demersal phase, rising at sexual maturity, and adult H. atlanticus exploit increasingly deep habitats with increasing age. At all sampled sizes, H. atlanticus muscle tissues have an isotopic composition suggesting a benthic rather than benthopelagic or pelagic diet. Isotopic measures of relative metabolic rate provide an insight into energy partitioning throughout ontogeny. Hoplostethus atlanticus have relatively low metabolic rates compared to coexisting deep‐water benthic fishes, consistent with their unusually high longevity. Surprisingly, lifetime fastest growth rates are achieved during juvenile stages when otolith isotopes imply deep‐water residency and relatively low metabolic rates. Fast growth may be sustained during a period of high efficiency associated with reduced metabolic costs of prey capture or predator evasion. The stable‐isotope approach can be applied to any teleost and provides a rapid, cost‐effective technique for studying deep‐water fish communities.     

Maternal age and spine development in the rotifer Brachionus calyciflorus: increase of spine length with birth orders1

1. Maternal effects have long been known to influence phenotypic plasticity in rotifers. Females in Brachionus calyciflorus and several other species produce long‐spined offspring when the predatory rotifer Asplanchna is present; B. calyciflorus also develops short spines when food concentrations are low. These spines protect against predation and decrease food threshold concentrations. 2. Some strains of B. calyciflorus develop long spines even in the absence of Asplanchna and other environmental stimuli. We demonstrate in this study that spine length in such cases is dependent on the age of the mother. 3. In strains from Florida and Georgia, offspring spine length increased significantly with birth order, sometimes to lengths formerly observed only in the presence of Asplanchna. Significant variation in this trait was found among and within clones of a strain. Offspring body size also increased with maternal age. This is the first time maternal age has been shown to affect rotifer morphology. 4. These birth‐order effects may have important ecological implications and explain phenotypic plasticity and polymorphism in body size and spine length in populations when predators are absent and food concentrations are high. They may be a bet‐hedging mechanism to assure adaptation to rapid changes in predation pressure or food conditions.     

Population systematics of the snake genus Naja (Reptilia: Serpentes: Elapidae) in Indochina: Multivariate morphometrics and comparative mitochondrial DNA sequencing (cytochrome oxidase I)

We analyze the population systematics of Asiatic cobras in Indochina, China and the Andaman Islands by means of comparative sequencing of the cytochrome oxidase subunit I gene of the mitochondrial DNA molecule and multivariate analysis of morphological characters. Canonical variate analysis and mtDNA sequence information reveal that the cobras of this region comprise four distinct species: Naja atra from China and northern Vietnam, Naja kaouthia from Burma, central Thailand, Cambodia and southern Vietnam, Naja siamensis from Thailand, Cambodia and southern Vietnam, and Naja sagittifera from the Andaman Islands. The subspecies N. kaouthia suphanensis Nutaphand 1986 shows no mtDNA sequence difference from typical N. kaouthia from central Thailand, and multivariate analysis does not reveal differences in general phenotypic profile; the subspecies is therefore synonymised with Naja kaouthia. The cytochrome oxidase subunit I gene, little used in molecular taxonomy, is shown to be well suited for studies at the species level, as it shows taxonomically useful levels of interspecific divergence but low levels of intraspecific variation; this is particularly relevant for studies of rare species, where sample size is a problem. The combination of multivariate morphometrics and molecular systematics can be particularly powerful in resolving systematic problems in such cases.     

Mitochondrial DNA analysis reveals hidden genetic diversity in captive populations of the threatened American crocodile (Crocodylus acutus) in Colombia

Identification of units within species worthy of separate management consideration is an important area within conservation. Mitochondrial DNA (mtDNA) surveys can potentially contribute to this by identifying phylogenetic and population structure below the species level. The American crocodile (Crocodylus acutus) is broadly distributed throughout the Neotropics. Its numbers have been reduced severely with the species threatened throughout much of its distribution. In Colombia, the release of individuals from commercial captive populations has emerged as a possible conservation strategy that could contribute to species recovery. However, no studies have addressed levels of genetic differentiation or diversity within C. acutus in Colombia, thus complicating conservation and management decisions. Here, sequence variation was studied in mtDNA cytochrome b and cytochrome oxidase I gene sequences in three Colombian captive populations of C. acutus. Two distinct lineages were identified: C. acutus‐I, corresponding to haplotypes from Colombia and closely related Central American haplotypes; and C. acutus‐II, corresponding to all remaining haplotypes from Colombia. Comparison with findings from other studies indicates the presence of a single “northern” lineage (corresponding to C. acutus‐I) distributed from North America (southern Florida), through Central America and into northern South America. The absence of C. acutus‐II haplotypes from North and Central America indicates that the C. acutus‐II lineage probably represents a separate South American lineage. There appears to be sufficient divergence between lineages to suggest that they could represent two distinct evolutionary units. We suggest that this differentiation needs to be recognized for conservation purposes because it clearly contributes to the overall genetic diversity of the species. All Colombian captive populations included in this study contained a mixture of representatives of both lineages. As such, we recommend against the use of captive‐bred individuals for conservation strategies until further genetic information is available.     

A clinally varying promoter polymorphism associated with adaptive variation in wing size in Drosophila

Body size often shows adaptive clines in many ectotherms across altitude and latitude, but little is known about the genetic basis of these adaptive clines. Here we identify a polymorphism in the Dca (Drosophila cold acclimation) gene in Drosophila melanogaster that influences wing size, affects wing:thorax allometry and also controls a substantial proportion of the clinal wing‐size variation. A polymorphism in the promoter region of Dca had two common alleles showing strong reciprocal clinal variation in frequency with latitude along the east coast of Australia. The Dca‐237 allele increased towards the tropics where wing size is smaller. A within‐population association study highlighted that an increase in the frequency of this allele decreased wing size but did not influence thorax size. A manipulated increase in the level of expression of Dca achieved through UAS‐GAL4 was associated with a decrease in wing size but had no effect on thorax size. This was consistent with higher Dca expression levels in family lines with higher frequency of the Dca‐237 allele. Genetic variation in the promoter region of the Dca gene appears to influence adaptive size variation in the eastern Australian cline of Drosophila melanogaster and accounts for more than 10% of the genetic variation in size within and between populations.