Behavioural effects of cadmium (Cd) in free-living willow ptarmigan (<Emphasis Type=Italic>Lagopus lagopus</Emphasis>)

The objective of this study was to examine effects of sublethal cadmium (Cd) contamination on parental behaviour and chick survival in willow ptarmigan (Lagopus lagopus) in an area that does not have high environmental levels of Cd due to geology but have high environmental levels of Cd due to anthropogenic deposition and acidification. One group of willow ptarmigan hens were exposed to Cd (83.2 mg Cd/L) after egg-laying using osmotic minipumps; whereas, another group given saline served as a control. No statistically significant difference in Cd concentration between the two groups was found. There was a significant correlation between the Cd concentrations in the liver and kidney of the birds and between Cd and metallothionein (MT) concentration in the kidney. No significant differences were found between the control hens and the Cd-contaminated hens with respect to any of the behaviour parameters. Age seemed to be more important than Cd treatment as adult hens showed more distraction display, had lower flushing distance, and higher brood attendance than juveniles. There were no difference in chick survival between Cd-contaminated and control hens, and there was no correlation between Cd concentrations and chick survival. The non-significant effects of Cd concentrations on parental behaviour may be due high environmental levels of Cd caused by anthropogenic deposition and acidification. We suggest that willow ptarmigan in the area have gradually evolved abilities to cope with high environmental levels of Cd, for instance, by binding Cd to MT in kidney and liver. This may suggest that Cd exerts a strong selection pressure on birds causing a relatively rapid evolution of protection mechanisms.     

Fluctuating asymmetry of <Emphasis Type="Italic">Amphibalanus</Emphasis> (<Emphasis Type="Italic">Balanus</Emphasis>) <Emphasis Type="Italic">amphitrite</Emphasis> (Cirripedia: Thoracica) in association with shore height and metal pollution

The level of fluctuating asymmetry (FA), which is defined as random deviations from perfect bilateral symmetry in the morphological traits of an organism, increases with increasing developmental instability, and it may be used as an indicator of environmental and/or genetic stresses. This study attempted to relate FA levels in the opercular plates of the barnacle Amphibalanus (Balanus) amphitrite with shore height and body trace metal concentrations. Barnacles were collected from both low and mid shores at six coastal locations with various degrees of marine pollution in Hong Kong. Four opercular traits, namely scutum length, tergum length, scutum width, tergum width were measured in the specimen while concentrations of five common trace metals (Cd, Cu, Cr, Mn and Zn) were determined in their body tissues using inductively coupled plasma-atomic emission spectrophotometry. Among the four traits, only tergum length and scutum width fulfilled the assumption of FA while their measurement errors were low. Across all sites, mid-shore A. amphitrite consistently exhibited a significantly higher FA level in scutum width than that in low-shore barnacles, but FA levels in tergum length were similar between the two shore heights. These results suggested that FA was trait-dependent, and mid-shore barnacles were possibly under high physical stress, such as desiccation and high temperature, leading to high developmental instability. Although no positive association was observed between FA and metal contamination in low-shore A. amphitrite, FA level in tergum length significantly increased with decreasing body concentration of manganese (Mn) in these barnacles. Such a negative relationship may be explained by the mechanism of bioaccumulation and physiological role of Mn in A. amphitrite with respect to the formation of barnacle shell plates. Handling editor: T. P. Crowe     

Metal accumulation and arbuscular mycorrhizal status in metallicolous and nonmetallicolous populations of <Emphasis Type="Italic">Pteris vittata</Emphasis> L. and <Emphasis Type="Italic">Sedum alfredii</Emphasis> Hance

Although Pteris vittata L. and Sedum alfredii Hance have been identified as an As hyperaccumulator and a Zn/Cd hyperaccumulator, respectively, for a few years, variations in metal accumulation among populations and their arbuscular mycorrhizal (AM) status have not been fully explored. Six populations of P. vittata and four populations of S. alfredii from southeast China were investigated. Up to 1,373 As, 680 Pb, 376 Zn, 4.8 Cd, 169 Cu mg kg⁻¹ in fronds of P. vittata and 358 As, 2,290 Pb, 23,403 Zn, 708 Cd, 342 Cu mg kg⁻¹ in shoots of S. alfredii were detected. Constitutive properties of As and Zn hyperaccumulation in metallicolous populations of P. vittata and S. alfredii, respectively, were confirmed. However, Cd hyperaccumulation in S. alfredii varied among populations. The two hyperaccumulators varied in efficiency in taking up other heavy metals. Different metal tolerance strategies adopted by the two hyperaccumulators varied among plant species and metal species. Low to moderate levels of AM colonization in P. vittata (4.2–12.8%) and S. alfredii (8.5–45.8%) were observed at uncontaminated and metal-contaminated sites. The relationship between metal concentrations and AM colonization in the two hyperacumulators was also examined. The abundance of AM fungal spores ranged from 16 to 190 spores per 25 g soil. Glomus microaggregatum, Glomus mosseae, Glomus brohultii and Glomus geosporum were the most common species associated with both P. vittata and S. alfredii. To our knowledge, this is the first report of AM fungal status in rhizosphere of P. vittata and S. alfredii.     

Transgenic Indian mustard (<Emphasis Type="Italic">Brassica juncea</Emphasis>) plants expressing an <Emphasis Type="Italic">Arabidopsis</Emphasis> phytochelatin synthase (<Emphasis Type="Italic">AtPCS1</Emphasis>) exhibit enhanced As and Cd tolerance

Phytochelatins (PCs) are post-translationally synthesized thiol reactive peptides that play important roles in detoxification of heavy metal and metalloids in plants and other living organisms. The overall goal of this study is to develop transgenic plants with increased tolerance for and accumulation of heavy metals and metalloids from soil by expressing an Arabidopsis thaliana AtPCS1 gene, encoding phytochelatin synthase (PCS), in Indian mustard (Brassica juncea L.). A FLAG-tagged AtPCS1 gDNA, under its native promoter, is expressed in Indian mustard, and transgenic pcs lines have been compared with wild-type plants for tolerance to and accumulation of cadmium (Cd) and arsenic (As). Compared to wild type plants, transgenic plants exhibit significantly higher tolerance to Cd and As. Shoots of Cd-treated pcs plants have significantly higher concentrations of PCs and thiols than those of wild-type plants. Shoots of wild-type plants accumulated significantly more Cd than those of transgenic plants, while accumulation of As in transgenic plants was similar to that in wild type plants. Although phytochelatin synthase improves the ability of Indian mustard to tolerate higher levels of the heavy metal Cd and the metalloid As, it does not increase the accumulation potential of these metals in the above ground tissues of Indian mustard plants.     

<Emphasis Type="Italic">Ty</Emphasis>1<Emphasis Type="Italic">/copia</Emphasis>- and <Emphasis Type="Italic">Ty</Emphasis>3<Emphasis Type="Italic">/gypsy</Emphasis>-like DNA sequences in <Emphasis Type="Italic">Helianthus </Emphasis>species

Two repeated DNA sequences isolated from a partial genomic DNA library of Helianthus annuus, p HaS13 and p HaS211, were shown to represent portions of the int gene of a Ty3 /gypsy retroelement and of the RNase-Hgene of a Ty1 /copia retroelement, respectively. Southern blotting patterns obtained by hybridizing the two probes to BglII- or DraI-digested genomic DNA from different Helianthus species showed p HaS13 and p HaS211 were parts of dispersed repeats at least 8 and 7 kb in length, respectively, that were conserved in all species studied. Comparable hybridization patterns were obtained in all species with p HaS13. By contrast, the patterns obtained by hybridizing p HaS211 clearly differentiated annual species from perennials. The frequencies of p HaS13- and p HaS211-related sequences in different species were 4.3x10(4)-1.3x10(5) copies and 9.9x10(2)-8.1x10(3) copies per picogram of DNA, respectively. The frequency of p HaS13-related sequences varied widely within annual species, while no significant difference was observed among perennial species. Conversely, the frequency variation of p HaS211-related sequences was as large within annual species as within perennials. Sequences of both families were found to be dispersed along the length of all chromosomes in all species studied. However, Ty3 /gypsy-like sequences were localized preferentially at the centromeric regions, whereas Ty1/ copia-like sequences were less represented or absent around the centromeres and plentiful at the chromosome ends. These findings suggest that the two sequence families played a role in Helianthusgenome evolution and species divergence, evolved independently in the same genomic backgrounds and in annual or perennial species, and acquired different possible functions in the host genomes.     

New combinations and typifications in <Emphasis Type="Italic">Bistorta</Emphasis>, <Emphasis Type="Italic">Persicaria</Emphasis>, <Emphasis Type="Italic">Polygonum,</Emphasis> and <Emphasis Type="Italic">Rumex</Emphasis> (Polygonaceae)

New combinations are proposed in anticipation of the Polygonaceae treatment in the forthcoming volume of Intermountain Flora: Polygonum kelloggii var. esotericum, P. kelloggii var. watsonii , Rumex densiflorus var. pycnanthus , R. salicifolius var. utahensis, and R. occidentalis var. tomentellus. Typifications are proposed to facilitate ongoing studies in Polygonaceae and to maintain current usage.     

Predation of arctic fox (<Emphasis Type="Italic">Vulpes lagopus</Emphasis>) pups by common ravens (<Emphasis Type="Italic">Corvus corax</Emphasis>)

Identifying correctly trophic interactions is important for understanding population dynamics and ecosystem functioning. However, some predator–prey relationships may still remain undetected, due to the difficulty of observing rare predation events. We report the first observation of predation of arctic fox (Vulpes lagopus) pups by common ravens (Corvus corax). The predation event was witnessed on Bylot Island, Nunavut, Canada, through an automatic camera placed on a den. On June 8, 2013, the day following pup emergence from the den, the complete litter of four was killed and taken away by a pair of ravens despite the intermittent presence of the mother. This event lasted 2.5 h, occurred during a low lemming year, and resulted in the fox pair failing their reproduction. Only two other fox litters were present that summer in our 600 km² study area, so this litter predated by ravens accounted for 25 % (4/16) of the pups produced. Our report shows how continuous monitoring of dens using automatic cameras can allow documentation of rare events. In addition to food competition and cache raiding, pup predation contributes to the antagonistic interactions between arctic foxes and ravens in the High Arctic, which may intensify during low lemming years. This observation allows a better understanding of species interactions within the Arctic predator guild.     

Pedogamy in <Emphasis Type="Italic">Neidium</Emphasis> (<Emphasis Type="Italic">Bacillariophyceae</Emphasis>)

Single (unpaired) vegetative cells of freshwater pennate diatom Neidium cf. ampliatum differentiated into gametangia and produced a single zygote (auxospore) via a pedogamic process. The gametic nuclei fused after auxospore expansion had begun. The auxospore expanded in parallel to the apical axis of the gametangium.     

<Emphasis Type="Italic">In silico</Emphasis> and <Emphasis Type="Italic">in situ</Emphasis> characterization of the zebrafish (<Emphasis Type="Italic">Danio rerio</Emphasis>) <Emphasis Type="Italic">gnrh3</Emphasis> (sGnRH) gene

Background

Gonadotropin releasing hormone (GnRH) is responsible for stimulation of gonadotropic hormone (GtH) in the hypothalamus-pituitary-gonadal axis (HPG). The regulatory mechanisms responsible for brain specificity make the promoter attractive for in silico analysis and reporter gene studies in zebrafish (Danio rerio).

Results

We have characterized a zebrafish [Trp⁷, Leu⁸] or salmon (s) GnRH variant, gnrh 3. The gene includes a 1.6 Kb upstream regulatory region and displays the conserved structure of 4 exons and 3 introns, as seen in other species. An in silico defined enhancer at -976 in the zebrafish promoter, containing adjacent binding sites for Oct-1, CREB and Sp1, was predicted in 2 mammalian and 5 teleost GnRH promoters. Reporter gene studies confirmed the importance of this enhancer for cell specific expression in zebrafish. Interestingly the promoter of human GnRH-I, known as mammalian GnRH (mGnRH), was shown capable of driving cell specific reporter gene expression in transgenic zebrafish.

Conclusions

The characterized zebrafish Gnrh3 decapeptide exhibits complete homology to the Atlantic salmon (Salmo salar) GnRH-III variant. In silico analysis of mammalian and teleost GnRH promoters revealed a conserved enhancer possessing binding sites for Oct-1, CREB and Sp1. Transgenic and transient reporter gene expression in zebrafish larvae, confirmed the importance of the in silico defined zebrafish enhancer at -976. The capability of the human GnRH-I promoter of directing cell specific reporter gene expression in zebrafish supports orthology between GnRH-I and GnRH-III.

     

<Emphasis Type="Italic">Quinua</Emphasis> and Relatives (<Emphasis Type="Italic">Chenopodium</Emphasis> sect.<Emphasis Type="Italic">Chenopodium</Emphasis> subsect.<Emphasis Type="Italic">Celluloid</Emphasis>)

Traditionally viewed as an Andean grain crop,Chenopodium quinoa Willd. includes domesticated populations that are not Andean, and Andean populations that are not domesticated. Comparative analysis of leaf morphology and allozyme frequencies have demonstrated that Andean populations, both domesticated(quinua) and free-living(ajara), represent an exceptionally homogeneous unit that is well differentiated from allied domesticates of coastal Chile(quingua) and freeliving populations of the Argentine lowlands(C. hircinum). This pattern of relationships indicates that Andean populations represent a monophyletic crop/weed system that has possibly developed through cyclic differentiation (natural vs. human selection) and introgressive hybridization. Relative levels of variation suggest that this complex originated in the southern Andes, possibly from wild types allied withC. hircinum, with subsequent dispersal north to Colombia and south to the Chilean coast. Coastal populations were apparently isolated from post-dispersal differentiation and homogenization that occurred in the Andes. Other data point toward a center of origin in the northern Andes with secondary centers of genetic diversity subsequently developing in the southern Andes and the plains of Argentina. Comparative linkage of South American taxa, all tetraploid, with North American tetraploids of the subsection will eventually clarify this problem. While the possibility of a direct phyletic connection betweenC. quinoa and the Mexican domesticate(C. berlandieri subsp. nuttalliae,) cannot be excluded, available evidence indicates that the latter represents an autonomous lineage that is associated with the basal tetraploid, C. b. subsp.berlandieri, through var.sinuatum, whereas South American taxa show possible affinities to either var. zschackei or var.berlandieri. An extinct domesticate of eastern North America,C. b. subsp.jonesianum, represents either another instance of independent domestication, possibly from subsp. b. var.zschackei, or a northeastern outlier of subsp.nuttalliae.     

Presence of moose (<Emphasis Type="Italic">Alces alces</Emphasis>) in Southeastern Germany

Germany is situated on the western border of the distribution area of moose (Alces alces) in Europe. In the past, the number of moose sightings has been very limited and irregular in the state of Bavaria (SE Germany). Nearby populations of moose are located in neighboring countries to the east of Germany, in the Czech Republic, and Poland. A monitoring system for moose was established following a considerable increase in moose observations in Bavaria in 2007. As no reproduction has yet been recorded, moose observation has so far been attributed only to migrating individuals. Cows are more often registered than bulls (27 of 35 observations between 2004 and 2008). The reasons for the increase in migratory activities of moose have not yet been identified.     

Polymorphism of <Emphasis Type="Italic">yellow</Emphasis> locus in <Emphasis Type="Italic">Drosophila melanogaster</Emphasis> mutants from natural populations

We have studied the molecular characteristics of the yellow locus (y; 1–0.0), which determines the body color of phenotypically wild-type and mutant alleles isolated in different years from geographically distant populations of Drosophila melanogaster. According to the Southern blot, data restriction maps of the yellow locus of all examined strains differ from one another, as well as from Oregon stock. FISH analysis shows that, in the neighborhood of the yellow locus in the X chromosome, neither P nor hobo elements are found in y^1–775 stock, while only hobo is found in these region in y^1–859 and y^1–866 stocks, only the P element is found in y⁺sn⁸⁴⁹ stock, and both elements are found in y^1–719 stock. Thus, all yellow mutants studied are of independent origin. Locus yellow located on the end of X chromosome (region 1A5–8 on the cytologic map) carries significantly more transposon than retrotransposon induced mutations compared to the white locus (region 3C2). It is possible that, at the ends of Drosophila melanogaster chromosomes, transposons are more active than retrotransposons.     

Floral development and systematic position of <Emphasis Type="Italic">Arillastrum</Emphasis>, <Emphasis Type="Italic">Allosyncarpia</Emphasis>, <Emphasis Type="Italic">Stockwellia</Emphasis> and <Emphasis Type="Italic">Eucalyptopsis</Emphasis> (Myrtaceae)

We have investigated the floral ontogeny of Arillastrum, Allosyncarpia, Stockwellia and Eucalyptopsis (of the eucalypt group, Myrtaceae) using scanning electron microscopy and light microscopy. Several critical characters for establishing relationships between these genera and to the eucalypts have been determined. The absence of compound petaline primordia in Arillastrum, Allosyncarpia, Stockwellia and Eucalyptopsis excludes these taxa from the eucalypt clade. Post-anthesis circumscissile abscission of the hypanthium above the ovary in Stockwellia, Eucalyptopsis and Allosyncarpia is evidence that these three taxa form a monophyletic group; undifferentiated perianth parts and elongated fusiform buds are characters that unite Stockwellia and Eucalyptopsis as sister taxa. No floral characters clearly associate Arillastrum with either the eucalypt clade or the clade of Stockwellia, Eucalyptopsis and Allosyncarpia.We gratefully acknowledge Clyde Dunlop and Bob Harwood (Northern Territory Herbarium) for collecting specimens of Allosyncarpia, and Bruce Gray (Atherton) for collecting specimens of Stockwellia. The Australian National Herbarium (CANB) kindly lent herbarium specimens of Eucalyptopsis for examination. This research was supported by a University of Melbourne Research Development Grant to Andrew Drinnan.     

<Emphasis Type="Italic">Agrobacterium</Emphasis><Emphasis Type="Italic">tumefaciens</Emphasis>-mediated transformation of callus cells of <Emphasis Type="Italic">Crataegus</Emphasis><Emphasis Type="Italic">aronia</Emphasis>

A genetic transformation system has been developed for callus cells of Crataegus aronia using Agrobacterium tumefaciens. Callus culture was established from internodal stem segments incubated on Murashige and Skoog (MS) medium supplemented with 5 mg l⁻¹ Indole-3-butyric acid (IBA) and 0.5 mg l⁻¹ 6-benzyladenine (BA). In order to optimize the callus culture system with respect to callus growth and coloration, different types and concentrations of plant growth regulators were tested. Results indicated that the best average fresh weight of red colored callus was obtained on MS medium supplemented with 2 mg l⁻¹ 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.5 mg l⁻¹ kinetin (Kin) (callus maintenance medium). Callus cells were co-cultivated with Agrobacterium harboring the binary plasmid pCAMBIA1302 carrying the mgfp5 and hygromycin phosphotransferase (hptII) genes conferring green fluorescent protein (GFP) activity and hygromycin resistance, respectively. Putative transgenic calli were obtained 4 weeks after incubation of the co-cultivated explants onto maintenance medium supplemented with 50 mg l⁻¹ hygromycin. Molecular analysis confirmed the integration of the transgenes in transformed callus. To our knowledge, this is the first time to report an Agrobacterium-mediated transformation system in Crataegus aronia.     

<Emphasis Type="Italic">Chenopodium</Emphasis>: a prospective plant for phytoextraction

Leaf samples were collected from 40 accessions of Chenopodium spp. and assessed for six heavy metals (Fe, Zn, Cu, Ni, Cr and Cd) accumulation to explore the use of Chenopodium for phytoextraction of heavy metals. The results suggest that Chenopodium spp. have the ability to accumulate large quantities of heavy metals in the leaf tissues even when they are present in low concentrations in the soil. C. quinoa is a better accumulator of Ni, Cr and Cd than the rest of the species, while C. album accessions are good copper accumulators. Bioconcentration factor for chromium ranged from 0.36 (C. album “Chandanbathua”) to 6.57 (C. quinoa Ames 13719) with 13 accessions of C. quinoa scoring above the mean value. High heritability coupled with high genetic advance was recorded for Ni, Cr and Cd, which indicated a major role of additive gene action in the inheritance of these characters. Zinc showed significant positive association with iron (0.351**), nickel (0.659**), chromium (0.743**) and cadmium (0.288**). Nickel was significantly and negatively associated with copper (−0.663**), while it was positively and significantly correlated with chromium (0.682**) and cadmium (0.461**). Considering the accumulation efficiency of Chenopodium spp. with respect to heavy metals, this genus should be further explored for decontamination of metal polluted soils, with plant breeding playing an important role in evolving new plant types with higher capacity of heavy metal accumulation.     

The influence of cadmium on life-history parameters and gut microflora of <Emphasis Type="Italic">Archegozetes longisetosus</Emphasis> (Acari: Oribatida) under laboratory conditions

We tested the effect of cadmium (25, 130 μg Cd g⁻¹), administered via Chinese cabbage (Brassica chinensis L.) as food on life-history parameters and gut microflora of tritonymphs and adults of the oribatid mite, Archegozetes longisetosus Aoki. Both concentrations of Cd had an adverse effect on offspring mortality, and the higher concentration also reduced female fecundity, as well as the number of bacteria, fungi and actinomycetes, and it changed the community structure of bacteria; the proportion of gram-negative bacteria increased while that of gram-positive bacteria declined. Interestingly, at the lower Cd concentration microflora was more abundant and diverse than in the control group, especially in the tritonymphs, although the mean activity of gut microflora was reduced. The higher Cd concentration reduced microflora activity both in the tritonymphs and adults.     

Biosorption of heavy metals by the exopolysaccharide produced by <Emphasis Type="Italic">Paenibacillus jamilae</Emphasis>

The biosorption of several toxic heavy metals (Pb, Cd, Co, Ni, Zn and Cu) by the exopolysaccharide (EPS) produced by Paenibacillus jamilae, a potential biosorbent for metal remediation and recovery was studied. Firstly, the biochemical composition of this bacterial polymer was determined. Glucose was the most abundant neutral sugar, followed by galactose, rhamnose, fucose and mannose. The polymer presented a high content of uronic acids (28.29%), which may serve as binding sites for divalent cations. The presence of carboxylic groups was also detected by infrared spectroscopy. The EPS presented an interesting affinity for Pb in comparison with the other five metals. Lead biosorption (303.03 mg g⁻¹) was tenfold higher (in terms of mg of metal adsorbed per gram of EPS) than the biosorption of the rest of metals. Biosorption kinetics, the effect of pH and the effect of competitive biosorption were determined. Finally, we found that the EPS was able to precipitate Fe(III), but the EPS-metal precipitate did not form with Fe(II), Pb(II), Cd(II), Co(II), Ni(II), Cu(II) and Zn(II).     

<Emphasis Type="Italic">Galleria</Emphasis><Emphasis Type="Italic">mellonella</Emphasis> are Resistant to <Emphasis Type="Italic">Pneumocystis</Emphasis><Emphasis Type="Italic">murina</Emphasis> Infection

Studying Pneumocystis has proven to be a challenge from the perspective of propagating a significant amount of the pathogen in a facile manner. The study of several fungal pathogens has been aided by the use of invertebrate model hosts. Our efforts to infect the invertebrate larvae Galleria mellonella with Pneumocystis proved futile since P. murina neither caused disease nor was able to proliferate within G. mellonella. It did, however, show that the pathogen could be rapidly cleared from the host.