Isolation of highly polymorphic microsatellite markers from the intertidal barnacle Chthamalus montagui Southward

This study reports the isolation and characterization of seven highly polymorphic microsatellite loci in Chthamalus montagui (Crustacea, Cirripedia). The loci were isolated from a library constructed from genomic DNA enriched for CA repeats. The markers yielded three to 43 alleles per locus (mean 16.7) in samples averaging 49 individuals. Observed heterozygosity ranged from 0.08 to 0.58 (mean 0.39). These microsatellite loci will be valuable tools for population genetic studies of this species.     

Effect of rock type on the recruitment and early mortality of the barnacle Chthamalus montagui

Discontinuities in distribution of sessile species due to changes in available habitat have rarely been investigated. In the English Channel, soft sedimentary rocks including chalk could potentially present a barrier to eastward range extension of the southern barnacle Chthamalus montagui. To test the effect of rock type on recruitment and mortality, prepared settlement tiles of four calcareous rocks that form important platforms close to existing limits of distribution were fixed to the shore at a site in south-west England, known to have consistently high larval settlement. Prior to fixation, the roughness parameter “R_a” and Potential Settlement Sites (PSS) index was measured. Recruitment was monitored photographically on four occasions during the settlement season and subsequent mortality for 7 months after the settlement season. By the end of the settlement season, recruitment on the hard Kimmeridge Cementstone was significantly less than on the other three rock types and attributed to low surface roughness and fewest Potential Settlement Sites (PSS), yet only 28% of variance could be attributed to PSS. Post-recruitment mortality was more variable, however after 7 months there were significant differences between rock types. Simultaneous surveys of post-recruitment mortality in natural populations showed that although recruitment on chalk can potentially be high, survivorship was relatively low. Thus rock type could be influential in setting species geographical limits.     

The vertical distribution of Chthamalus montagui and Chthamalus stellatus (Crustacea,Cirripedia) in two areas of the NW Mediterranean Sea

The Mediterranean Sea is characterised by a small tidal range (0.3–1 m). Despite this, intertidal communities are well established and their upper limits often extend above mean high water level. Organisms living in the intertidal region and in the supralittoral zone rely on both tides and wave action to perform their biological functions. Lack of food, desiccation and predation are common stresses in such a harsh environment. The present study deals with the vertical distribution of two species of intertidal barnacles, Chthamalus montagui Southward and Chthamalus stellatus (Poli), which are the main constituents of the barnacle belt along Mediterranean rocky shores. Previous work, carried out in the Atlantic, showed that where the distribution ranges of the two Chthamalus species overlap, C. montagui is more common in the upper barnacle zone while C. stellatus is dominant lower down. The main aims of our study are: (1) to establish if there is a relationship between position and extension of the barnacle belt on the shore and tidal range and/or wave exposure, (2) to test the hypothesis that in the study areas C. montagui is more abundant than C. stellatus high on the shore, and that the pattern is reversed lower down. Barnacle populations were monitored in summer 1998 in the Gulf of Genoa (Ligurian Sea) and in the Gulf of Trieste (North-Adriatic Sea). The two areas differ in tidal range and hydrodynamism, the former presenting quite strong wave action and a tidal range of 30 cm, the latter having limited wave action and 1 m tidal range. Three shores were randomly selected in each gulf and two transects on each shore. Counts of barnacles in 10 ^* 10 cm quadrats were done at different shore heights along each transect. The data was subjected to analysis of variance. Results showed that a more pronounced hydrodynamic regime corresponded to a shift of the barnacle belt towards the higher shore (Gulf of Genoa), while in more sheltered areas (Gulf of Trieste), the barnacle distribution was confined to the intertidal region. The relative spatial distribution of C. montagui and C. stellatus within the barnacle belt varied locally, even between transects on the same shore, and this obscured the distribution pattern along the vertical gradient. Nevertheless, it was still possible to conclude that at mid and high shore in Genoa, C. stellatus was more abundant than C. montagui, while in Trieste the pattern was reversed.     

Climate change and historical biogeography of the barnacle Semibalanus balanoides

Aim Marine sessile inter‐tidal invertebrates are expected to undergo shifts in distribution due to climate change. Using a combination of survey and transplant data with thermal modelling, we investigated the role of climate on the poleward contraction of the southern range edge of the north temperate barnacle Semibalanus balanoides. Location Western Atlantic of the United States. Methods Barnacle surveys were conducted along the east coast of the United States in 1963 and 2007. Presence, absence and abundance data were collected and the time periods were compared. Transplant experiments monitoring survival with relation to temperature were conducted upon S. balanoides along the more southerly portion of their range, and modelling predicting barnacle survival with relation to biogeography was completed. Results The southern limit of S. balanoides has contracted approximately 350 km to the north. Main conclusions The changes thus far observed in climate along the east coast of the United States have contributed to the southern limit range contraction of S. balanoides. Further changes in the biogeography of S. balanoides are expected with continued climate warming.     

Barnacle species as biomonitors of metal contamination in the northwest coast of Portugal: Ecological quality classification approach

This work discusses the simultaneous use of barnacles C. montagui and P. pollicipes as biomonitors of metal (Cd, Cr, Cu, Fe, Mn, and Zn) contamination in the northwest (NW) coast of Portugal during the four seasons of 2011. The metal concentrations in soft tissues of both barnacles species, metal bioaccumulation factors (BAFs), and ecological quality classifications of coastal waters are presented. The most and the least metal-contaminated locations were, respectively, location 5 (“Cabo do Mundo”) and location 10 (“Moledo”). The most and the least metal-contaminated seasons of 2011 were, respectively, autumn and winter. Temporal comparisons with previous works of 2009–2011 showed that C. montagui bioaccumulated Cd, Cr, and Fe at higher rates and P. pollicipes accumulated more Zn besides Cd and Fe. In addition, global mean BAFs showed that the best season to collect both barnacle species for metal contamination assessment/monitoring purposes in future works would be spring and that P. pollicipes would be the best barnacle species to monitor metals. In terms of ecological quality classification, all NW coast of Portugal should be classified as “Class III – Remarkably Polluted” due the high concentrations of Zn found in barnacles species during the four seasons of 2011. This work suggested that both barnacle species are suitable species to be included in the Portuguese Environmental Specimen Banks under European Marine Strategy Framework Directive 2008/56/EC.     

Spatial and temporal patterns in the recruitment of the intertidal barnacle Chthamalus malayensis Pilsbry (Crustacea: Cirripedia) on the equatorial shores of Peninsular Malaysia and Singapore

Spatial and temporal variation in the recruitment of the intertidal barnacle Chthamalus malayensis was examined over one year (September 2003-August 2004) on the equatorial shores of southern Peninsular Malaysia and Singapore. A nested sampling design was applied for the first time on these shores, over three spatial scales - hundreds of kilometres, kilometres to tens of kilometres and tens of metres - and temporal variation was determined through monthly sampling of recruits. Shores within 2° north of the equator on the East and West coasts of southern Peninsular Malaysia and the southern coast of Singapore were selected as study sites. Generally, all three coasts recruited throughout the year, with varying intensities. There was a clear regional pattern, where the largest number of recruits was found on the East Coast of Malaysia, and the least in Singapore. Analyses of variance (ANOVA) showed that variability occurred on the smallest scale, but only in nine of the twelve months examined, consequently resulting in significant temporal and spatial interaction. Calculated variance components indicated that small-scale variation accounted for most of the overall variability. The potential causes of the spatial and temporal patterns of C. malayensis recruitment, and implications on tropical-temperate comparisons will be discussed.     

Phylogeography of the cold‐water barnacle Chthamalus challengeri in the north‐western Pacific: effect of past population expansion and contemporary gene flow

Aim Phylogeographical patterns of marine organisms in the north‐western Pacific are shaped by the interaction of past sea‐level fluctuations during glacial maxima and present‐day gene flow. This study examines whether observed population differentiation in the barnacle Chthamalus challengeri, which is endemic to the north‐western Pacific, can be explained by the interactions between historical glacial events and patterns of contemporary gene flow. Location Eleven locations in the north‐western Pacific. Methods Partial sequences of mitochondrial cytochrome c oxidase subunit I (COI), 12S, 16S and nuclear internal transcribed spacer 1 (ITS1) were obtained from 312 individuals. Parsimony haplotype networks and analysis of molecular variance (AMOVA) were used to determine whether the observed genetic structure corresponds to marine provinces (Kuroshio Current and China Sea Coastal Provinces), zoogeographical zones (oriental and Japan warm‐temperate zones) and/or potential refugial areas (Sea of Japan and East China Sea) in the north‐western Pacific. Neutrality tests, mismatch distribution analysis and Bayesian skyline plots were used to infer the demographic history of C. challengeri. Results In total, 312, 117, 182 and 250 sequences were obtained for COI, 12S, 16S and ITS1, respectively. A panmictic population was revealed, which did not conform to the ‘isolation by distance’ model. None of the a priori population groupings based on marine provinces, zoogeographical zones or potential refugial areas was associated with observed genetic patterns. Significant negative values from neutrality tests and the unimodal mismatch distribution and expansion patterns in Bayesian skyline plots for the COI and 16S data sets indicate a population expansion in the mid‐Pleistocene (c. 200 ka). Information from the fossil record suggests that there has been a northward range shift of this species from the East China Sea or the Palaeo‐Pacific coast of Japan to the Sea of Japan since the mid‐Pleistocene. Main conclusions Chthamalus challengeri has experienced a population expansion and range shift since the mid‐Pleistocene. The observed lack of population differentiation can be explained by this past population expansion and present‐day wide‐scale larval dispersal (owing to the long planktonic larval duration) across marine provinces, which have led to the successful establishment of the species in different zoogeographical zones and habitats.     

三种植物生长调节剂对密叶绢藓(Entodon challengeri)孢子萌发、原丝体发育及芽体发生的影响

研究了3种植物生长调节剂苯基噻二唑基脲(TDZ)、6-苄基腺嘌呤(6-BA)、萘乙酸(NAA)对密叶绢藓[Entodon challengeri(Paris)Cardot]孢子萌发、原丝体发育及芽体发生的影响,并对整个发育过程进行了显微观察和照相,结果表明:(1)3种植物生长调节剂对密叶绢藓孢子萌发影响不显著;(2)在原丝体发育阶段,1.0 mg/LNAA对原丝体初期的发育促进效果显著,0.4 mg/L TDZ对原丝体发育中期分枝的形成促进效果显著,6-BA处理效果不显著;(3)3种植物生长调节剂单独处理均促进芽体的发生,但0.4 mg/L TDZ效果最佳。而1.5 mg/L 6-BA+TDZ组合处理效果更加显著;(4)芽体的发生数量与芽体的长势无正相关性。