Figure 4. Geochemistry on sediment core MD04-2879

Anaerobic ammonium oxidation (anammox) has been recognized as an important process converting fixed nitrogen to N2 in many marine environments, thereby having a major impact on the present-day marine nitrogen cycle. However, essentially nothing is known about the importance of anammox in past marine nitrogen cycles. In this study, we analyzed the distribution of fossil ladderane lipids, derived from bacteria performing anammox, in a sediment core from the northern Arabian Sea. Concentrations of ladderane lipids varied between 0.3 and 5.3 ng/g sediment during the past 140 ka, with high values observed during the Holocene, intervals during the last glacial, and during the penultimate interglacial. Maxima in ladderane lipid abundances correlate with high total organic carbon (4-6%) and elevated d15N (>8 per mil) values. Anammox activity, therefore, seems enhanced during periods characterized by an intense oxygen minimum zone (OMZ). Low concentrations of ladderanes (<0.5 ng/g sediment), indicating low-anammox activity, coincide with periods during which the OMZ was severely diminished. Since anammox activity covaried with OMZ intensity, it may play an important role in the loss of fixed inorganic nitrogen from the global ocean on glacial-interglacial timescales, which was so far attributed only to heterotrophic denitrification.

Microsatellite genotypic data of Euptelea pleiospermum in STRUCTURE format

We sampled leaves from 678 individuals in 21 natural populations (30-36 individuals per population), covering the entire distribution of Euptelea pleiospermum in China.Total DNA was isolated from about 50 mg powdered leaf tissue following the protocol of a DNA extraction kit (Tiangen Biotech Co., LTD., Beijing, China). We used seven fluorescence-labeled microsatellite loci (EP036, EP059, EP081, EP087, EP091, EP278 and EP294; Zhang et al., 2008) to genotype our 678 DNA samples.

Investigation of N. pachyderma from the North Atlantic

The shells of the planktonic foraminifer Neogloboquadrina pachyderma have become a classical tool for reconstructing glacial-interglacial climate conditions in the North Atlantic Ocean. Palaeoceanographers utilize its left- and right-coiling variants, which exhibit a distinctive reciprocal temperature and water mass related shift in faunal abundance both at present and in late Quaternary sediments. Recently discovered cryptic genetic diversity in planktonic foraminifers now poses significant questions for these studies. Here we report genetic evidence demonstrating that the apparent 'single species' shell-based records of right-coiling N. pachyderma used in palaeoceanographic reconstructions contain an alternation in species as environmental factors change. This is reflected in a species-dependent incremental shift in right-coiling N. pachyderma shell calcite d18O between the Last Glacial Maximum and full Holocene conditions. Guided by the percentage dextral coiling ratio, our findings enhance the use of d18O records of right-coiling N. pachyderma for future study. They also highlight the need to genetically investigate other important morphospecies to refine their accuracy and reliability as palaeoceanographic proxies.

Molecular differentiation by PCR-RFLP technique shows that Antarctic fishes of the genus Notothenia, claimed to be two species, are phenotypic varieties with the same genetic pattern (Table 1)

The taxonomy of Antarctic fishes has been predominantly based on morphological characteristics rather than on genetic criteria. A typical example is the Notothenia group, which includes N. coriiceps Richardson, 1844, N. neglecta Nybelin, 1951 and N. rossii Richardson, 1844. The Polymerase Chain Reaction and Restriction Fragment Length Polymorphism (PCR-RFLP) technique was used to determine whether N. coriiceps Richardson, 1844 and N. neglecta Nybelin, 1951 are different or whether they are the same species with morphological, physiological and behavioural variability. N. rossii was used as control. Mitochondrial DNA (mtDNA) was isolated from muscle specimens of N. coriiceps Richardson, 1844, N. neglecta Nybelin, 1951 and N. rossii, which were collected in Admiralty Bay, King George Island. The DNA was used to amplify a fragment (690 base pairs) of the mitochondrial gene coding region of NADH dehydrogenase subunit 2. Further, the amplicon was digested with the following restriction enzymes: DdeI, HindIII and RsaI. The results showed a variation of the digestion pattern of the fragment amplified between N. rossii, and N. coriiceps Richardson, 1844 or N. neglecta Nybelin, 1951. However, no differences were found between N. coriiceps Richardson, 1844 and N. neglecta Nybelin, 1951, on the grounds of the same genetic pattern shown by the two fish.

Evidence of cryptic and pseudocryptic speciation in the Paracalanus parvus species complex (Crustacea, Copepoda, Calanoida), supplementary material

Introduction Many marine planktonic crustaceans such as copepods have been considered as widespread organisms. However, the growing evidence for cryptic and pseudo-cryptic speciation has emphasized the need of re-evaluating the status of copepod species complexes in molecular and morphological studies to get a clearer picture about pelagic marine species as evolutionary units and their distributions. This study analyses the molecular diversity of the ecologically important Paracalanus parvus species complex. Its seven currently recognized species are abundant and also often dominant in marine coastal regions worldwide from temperate to tropical oceans. Results COI and Cytochrome b sequences of 160 specimens of the Paracalanus parvus complex from all oceans were obtained. Furthermore, 42 COI sequences from GenBank were added for the genetic analyses. Thirteen distinct molecular operational taxonomic units (MOTU) and two single sequences were revealed with cladistic analyses (Maximum Likelihood, Bayesian Inference), of which seven were identical with results from species delimitation methods (barcode gaps, ABDG, GMYC, Rosenberg's P(AB)). In total, 10 to 12 putative species were detected and could be placed in three categories: (1) temperate geographically isolated, (2) warm-temperate to tropical wider spread and (3) circumglobal warm-water species. Conclusions The present study provides evidence of cryptic or pseudocryptic speciation in the Paracalanus parvus complex. One major insight is that the species Paracalanus parvus s.s. is not panmictic, but may be restricted in its distribution to the northeastern Atlantic.