FOOD-INDUCED BODY PIGMENTATION QUESTIONS THE TAXONOMIC VALUE OF COLOUR IN THE SELF-FERTILIZING SLUG CARINARION SPP

Body pigmentation is a popular taxonomic marker in slugs to discriminateclosely related species. However, the genetic background ofbody pigmentation is known only for a few species, while inmany others body pigmentation is influenced by age, food and/orclimate. In this study, we investigated the effects of differentfood items on body pigmentation expression in two selfing pulmonategastropods, Arion (Carinarion) silvaticus and Arion (Carinarion) fasciatus.Both species mainly differ in the distribution of yellow-orangegranules on the body, which in A. fasciatus are concentratedin lateral bands, and in A. silvaticus are evenly scattered.Animals were raised individually under the same conditions, whilethey laid eggs as a consequence of selfing. This F₁ generation wasafterwards divided into two groups, which were fed with different fooditems. A diet of carrot, lettuce or paper had no effect on the distributionof the yellow-orange granules in A. silvaticus, but provokeda loss of the yellow-orange lateral bands in A. fasciatusso that externally these F₁ specimens became similar to A. silvaticus.In both species, a diet of nettle resulted in a strong yellow-orangepigmentation, which often formed yellow-orange lateral bands.These results indicate that food can probably influence the'species-specific' body pigmentation in Carinarion, and thusquestion the reliability of colour traits to distinguish A.silvaticus and A. fasciatus. (Received 5 May 2000; accepted 29 September 2000)     

An integrative approach to characterize cryptic species in the Thoracostoma trachygaster Hope, 1967 complex (Nematoda: Leptosomatidae)

Nematode diversity may seriously be underestimated when taking into account cryptic speciation. Thoracostoma trachygaster is commonly found in kelp holdfasts along the California coastline and was recently shown to consist of at least two distinct molecular clades (I and II). Here, we provide detailed morphological analysis of both clades, based on measurements taken from video vouchers of respectively eight and 16 individuals from the previous study, as well as 80 newly collected specimens from four Californian beaches. The latter were vouchered, measured, and then subjected to molecular analyses of the mitochondrial cytochrome oxidase c subunit I (COI) gene, and the ribosomal D2D3 and internal transcribed spacer (ITS) regions. This integrative approach shows that the three molecular clades are phylogenetically and morphologically distinct species, but a combination of morphological characters is needed to distinguish them. Two new species, Thoracostoma fatimae sp. nov. and Thoracostoma igniferum sp. nov. , are identified and described. The spicule length of T. fatimae sp. nov. is significantly shorter than that of T. trachygaster. Thoracostoma igniferum sp. nov. can be distinguished by the irregular posterior edge of the cephalic capsule and the two internal subdorsal tropis‐like projections in the wall of the cephalic capsule, which are lacking in T. fatimae sp. nov. and T. trachygaster. © 2012 The Linnean Society of London, Zoological Journal of the Linnean Society, 2012, 164 , 18–35.     

Spring feeding by pink-footed geese reduces carbon stocks and sink strength in tundra ecosystems

Tundra ecosystems are widely recognized as precious areas and globally important carbon (C) sinks, yet our understanding of potential threats to these habitats and their large soil C store is limited. Land‐use changes and conservation measures in temperate regions have led to a dramatic expansion of arctic‐breeding geese, making them important herbivores of high‐latitude systems. In field experiments conducted in high‐Arctic Spitsbergen, Svalbard, we demonstrate that a brief period of early season belowground foraging by pink‐footed geese is sufficient to strongly reduce C sink strength and soil C stocks of arctic tundra. Mechanisms are suggested whereby vegetation disruption due to repeated use of grubbed areas opens the soil organic layer to erosion and will thus lead to progressive C loss. Our study shows, for the first time, that increases in goose abundance through land‐use change and conservation measures in temperate climes can dramatically affect the C balance of arctic tundra.     

Contribution of subsurface peat to CO2 and CH4 fluxes in a neotropical peatland

Tropical peatlands play an important role in the global carbon cycling but little is known about factors regulating carbon dioxide (CO₂) and methane (CH₄) fluxes from these ecosystems. Here, we test the hypotheses that (i) CO₂ and CH₄ are produced mainly from surface peat and (ii) that the contribution of subsurface peat to net C emissions is governed by substrate availability. To achieve this, in situ and ex situ CO₂ and CH₄ fluxes were determined throughout the peat profiles under three vegetation types along a nutrient gradient in a tropical ombrotrophic peatland in Panama. The peat was also characterized with respect to its organic composition using ¹³C solid state cross‐polarization magic‐angle spinning nuclear magnetic resonance spectroscopy. Deep peat contributed substantially to CO₂ effluxes both with respect to actual in situ and potential ex situ fluxes. CH₄ was produced throughout the peat profile with distinct subsurface peaks, but net emission was limited by oxidation in the surface layers. CO₂ and CH₄ production were strongly substrate‐limited and a large proportion of the variance in their production (30% and 63%, respectively) was related to the quantity of carbohydrates in the peat. Furthermore, CO₂ and CH₄ production differed between vegetation types, suggesting that the quality of plant‐derived carbon inputs is an important driver of trace gas production throughout the peat profile. We conclude that the production of both CO₂ and CH₄ from subsurface peat is a substantial component of the net efflux of these gases, but that gas production through the peat profile is regulated in part by the degree of decomposition of the peat.     

A new role for glutathione in the regulation of root architecture linked to strigolactones

Reduced glutathione (GSH) is required for root development, but its functions are not characterized. The effects of GSH depletion on root development were therefore studied in relation to auxin and strigolactone (SL) signalling using a combination of molecular genetic approaches and pharmacological techniques. Lateral root (LR) density was significantly decreased in GSH synthesis mutants (cad2‐1, pad2‐, rax1‐), but not by the GSH synthesis inhibitor, buthionine sulfoximine (BSO). BSO‐induced GSH depletion therefore did not influence root architecture in the same way as genetic impairment. Root glutathione contents were similar in the wild‐type seedlings and max3‐9 and max4‐1 mutants that are deficient in SL synthesis and in the SL‐signalling mutant, max2‐1. BSO‐dependent inhibition of GSH synthesis depleted the tissue GSH pool to a similar extent in the wild‐type and SL synthesis mutants, with no effect on LR density. The application of the SL analogue GR24 increased root glutathione in the wild‐type, max3‐9 and max4‐1 seedlings, but this increase was absent from max2‐1. Taken together, these data establish a link between SLs and the GSH pool that occurs in a MAX2‐dependent manner.     

Integrative taxonomy in two free-living nematode species complexes

Integrative taxonomy considers species boundaries from multiple, complementary perspectives, with the main objective being to compare the observed data against the predictions of the methodologies used. In the present study we used three methods for delineating species boundaries within the cosmopolitan nematode species Rhabditis ( Pellioditis ) marina and Halomonhystera disjuncta . First, phylogenetic relationships among molecular sequences from the mitochondrial cytochrome oxidase c subunit 1 gene (COI), and from two nuclear regions, internal transcribed spacer (ITS) and D2D3, were analysed. Subsequently, multivariate morphometric analysis was used to investigate whether concordant molecular lineages were also morphologically distinct. When morphological differences were found, typological taxonomy was performed to identify fixed or non-overlapping characters between lineages. Interbreeding experiments were conducted between the two closest related lineages of R . ( P. ) marina to investigate potential reproductive isolation. This integrative approach confirmed the presence of several species within each nominal species: molecular lineages were concordant across two independent loci (COI and ITS), and were characterized by significant morphological divergence. Most lineages were also detectable in the D2D3 region, but were less resolved. The two lineages investigated in our study did not produce offspring. Our results highlight that classical taxonomy grossly underestimates species diversity within the phylum Nematoda.  © 2008 The Linnean Society of London, Biological Journal of the Linnean Society , 2008, 94 , 737–753.     

Development of polymorphic nuclear microsatellite markers for polyploid and diploid members of the orchid genus Dactylorhiza

Dactylorhiza traunsteineri is an allotetraploid species that belongs to the large Dactylorhiza incarnata/maculata polyploid complex of the Eurasian genus Dactylorhiza (Orchidaceae). Here, eight polymorphic microsatellite loci were isolated using selective hybridization according to the FIASCO protocol (fast isolation by AFLP of sequences containing repeats) with slight modifications. The number of alleles per locus ranged from two to seven. All loci were possible to amplify in several other species of Dactylorhiza, using the same primers.     

Polyploid evolution and plastid DNA variation in the Dactylorhiza incarnata/maculata complex (Orchidaceae) in Scandinavia

The Dactylorhiza incarnata/maculata complex (Orchidaceae) was used as a model system to understand genetic differentiation processes in a naturally occurring polyploid complex with much of ongoing diversification and wide distribution in recently glaciated areas in northern Europe. Data were obtained for 12 hypervariable regions in the plastid DNA genome. A total of 166 haplotypes were found in a sample of 1099 plants. Allopolyploid taxa have inherited their plastid genomes from D. maculata s.l. Overall haplotype diversity of the combined group of allopolyploid taxa was comparable to that of maternal D. maculata s.l., but populations of allopolyploids were also more strongly differentiated from each other and contained lower numbers of haplotypes than populations of D. maculata s.l. In addition to haplotypes found in extant D. maculata s.l., the allopolyploids also contained several distinct and widespread haplotypes that were not found in any of the parental lineages. Some of these haplotypes were shared between widespread allopolyploids. Divergent allopolyploids with small distributions did not seem to originate from local polyploidization events, but rather as segregates of already existing allopolyploids. Genetic diversification of allopolyploid Dactylorhiza is the result of repeated polyploid formation, secondary hybridization and introgression between already existing polyploids and extant representatives of parental lineages, hybridization between independently derived polyploid lineages, and phyletic diversification in the group of allopolyploids. Although some polyploid taxa must have evolved after the last glaciation, genetic material from the parental lineages has been transferred continuously for longer periods of time. This combination of processes may explain the taxonomic complexity encountered in Dactylorhiza and other polyploid complexes distributed in previously glaciated parts of Europe.