ALBERT

Description: The genus Ulva is has broadly negative connotations because of its ability to form harmful “green tides” and the problems it causes with precise species identification, due to its morphological plasticity. During recent years, tides of unattached Ulva compressa U. Linneus 1753 with an atypical sheet-like morphology were for the first time observed in the German Baltic. Here we report that this nuisance alga is conspecific with the type strain of U. mutabilis Föyn 1958 from Faro in Portugal, an important model organism to study morphogenesis, morphogenetics and mutualistic interactions. Different approaches were used to examine conspecificity: (1) Comparisons on vegetative and reproductive features of cultured material of Ulva mutabilis and German Ulva compressa resulted in congruent results proving that a certain morphogenetic mutation pattern is shared. Spontaneous mutations of “slender-like” thalli are appearing whilst the common form exhibits a “leaf-like” wildtype morphology. (2) Interbreeding experiments of gametes of Ulva compressa and Ulva mutabilis were successful and showed a fertile first-generation offspring exhibiting the typical wildtype morphology similar to the phenotype of the parental generation. (3) Phylogenetic and species delimitation analyses were carried out on 128 tufA sequences of Ulva compressa specimens sampled in 2014–2016 in Germany and on tufA sequences of two clones of the strains Ulva mutabilis (sl-G[mt+]) and Ulva mutabilis (wt-[mt-]) to identify Molecular Operational Taxonomic Units (MOTUs). The Generalized Mixed Yule-Coalescent (GMYC) method comprises one major MOTU containing all included sequences of Ulva compressa and Ulva mutabilis, while reference sequences included in the analysis clustered outside this MOTU. This highly supports the monophyly of Ulva compressa and Ulva mutabilis, which can be treated as the same species. As a consequence, U. mutabilis is also a suitable model for future studies of green tides and their molecular and morphogenetic basis in the Baltic Sea.

Description: Invasive species are one of the principal components of global change along with ocean warming at the global scale, or overfishing and deoxygenation at the regional scale. Seaweeds represent up to 40% of all introduced marine species and some seaweeds can significantly affect the composition and functioning of marine benthic communities. Within ten years of its first discovery in the Kiel Fjord in 2005 the East Asian red seaweed Gracilaria vermiculophylla has spread approximately 100 km eastward and 120 km northward along the German Baltic Sea coast, now inhabiting many lagoons and sheltered bays between the German-Danish border and Neustadt. During the first two years after its discovery Gracilaria vermiculophylla increased its biomass in the Kiel Fjord massively. However, this was followed by a sudden decline in late summer 2008, when the alga decayed in nearly all inhabited parts of the bay within few weeks. Co-cultivation of healthy Gracilaria from unaffected environments with small amounts of decaying material from the Kiel Fjord in laboratory assays demonstrated that the decay was apparently caused by an infectious disease. Thus, 59 different species of epibacteria isolated from Gracilaria were tested for their capacity to induce decay in a bleaching assay. Out of these, three were found to induce the disease, while 19 others significantly reduced the risk of decay and were thus protective. When protectors and pathogens were tested together, the protective strains fully prevented the negative impact of the bleachers, hinting at the presence of an associational defence offered by Gracilaria’s epibacteria. Presence of such an associational resistance was also supported in a follow- up bioassay where surface extract of Gracilaria and its associated microbiome attracted the beneficial strains, but deterred the detrimental ones. Thus, we suggest that the breakdown in 2008 was due to a collapse of such associational resistance provided by bacterial partners.

Description: Fouling is a stressor that might determine the fate of seaweeds, but reports of algal adaptation to epibiosis are scarce. Previous comparisons have shown resistance to epibionts can be higher in non-native than in resident seaweed species, but we do not know whether it is an intrinsic trait of the non-natives or it has been acquired during the invasion process. We here compared native and non-native populations of the same algal species to elucidate this question. Resistance against two groups of epiphytes was assessed in living thalli and in artificial substrata coated with surface extracts, both gained from four Asian (native) and four European (non-native) populations of the red alga Gracilaria vermiculophylla. Two diatom species and two filamentous macroalgae were used as micro- and macro-epiphytes, and one of each type was collected in Asia, while the other came from Europe. Laboratory assays were done in both distributional ranges of G. vermiculophylla and in different seasons. We used a fully crossed design with the factors (i) ‘Origin of Gracilaria’, (ii) ‘Origin of epiphytes’, (iii) ‘Season’ and (iv) ‘Solvent used for extraction’. Both groups of epiphytes, regardless of their origin, attached less to living thalli and to surface extracts from non-native G. vermiculophylla. Fewer diatoms attached to hexane-based extracts, while fewer Ceramium filaments settled on extracts gained with dichloromethane. Our results show for the first time that non-native individuals of a seaweed are better defended against epiphytes than native conspecifics. Furthermore, we found evidence that at least a part of the defence is based on extractable secondary metabolites. We suggest that an enhanced defence against epiphytes after introduction is one reason for G. vermiculophylla’s invasion success. Our observation may also apply to other basibiont–epibiont interactions and could be a key feature of seaweed bioinvasions.

Description: The recent introduction of Gracilaria vermiculophylla (Rhodophyta) to the Kiel Fjord area was a reason for concern, since this red macroalga performs best under mesohaline conditions and thus appears well adapted to thrive and spread in the Baltic Sea environment, A systematic survey on a coastal range of 500 km in 2006 and 2007 indicated considerable Multiplication and spreading of G. vermiculophylla within Kiel Fjord, but provided little evidence of long-distance transport. Nonetheless, flow-through growth experiments conducted at a range of salinities under ambient light showed that G. vermiculophylla should be able to grow in most of the Baltic Sea. Growth declined only below a salinity of 5.5. High water temperatures in summer seem to reduce resistance against low salinity. Growth of G, vermiculophylla in the SW Baltic is limited by light and is only possible during summer and above a depth of 3 m. Drifting fragments are dispersed by currents. Either they sink to deeper waters, where they degrade, or they accumulate in shallow and sheltered waters, where they form perennial mats. These overgrow not only soft bottom sediments, but also stones, which are an important habitat to Fucus vesiculosus, the main native perennial alga in the Baltic Sea. As compared to F. vesiculosus, G. vermiculophylla seems to represent a preferred refuge for mesograzers and other invertebrates, particularly in winter. Nonetheless, feeding trials showed that potential grazers avoided G. vermiculophylla relative to F vesiculosus. Daily biomass uptake by grazers associated with G. vermiculophylla in nature did not exceed 2 g kg(-1) and is 〈11% of average daily net growth (18.5 g kg(-1)) in the first 2 m below sea level. Consequently, feeding may not be sufficient to control the spread of G. vermiculophylla in the SW Baltic. Our study suggests that absence of feeding enemies and adaptation to brackish water may allow G. vermiculophylla to invade most shallow coastal waters of the inner Baltic Sea despite light limitation.

Description: We assessed the effects of light limitation and temperature shift on palatability and induced antiherbivore defense in the brown alga Fucus vesiculosus L. Incubation for 2 wk at light intensities above the compensation point of photosynthesis and in the absence of grazers increased the palatability of F. vesiculosus and its subsequent consumption by the omnivorous isopod Idotea baltica Pallas. This effect correlated with an increased C:N ratio and mannitol content in the algal tissue, presumably due to increased photosynthetic carbon fixation. Mannitol, the primary product of photosynthesis in F. vesiculosus, proved to be a feeding cue for I. baltica, and depletion of the mannitol pool may therefore account for the reduced palatability during light limitation. At light intensities above the compensation point of photosynthesis, F. vesiculosus responded with decreasing palatability when it was exposed to I. baltica grazing. Irrespective of the preceding light regime, such defense induction was prevented during incubation under light limitation. Thus, under low light, defense induction is not only inhibited, but also less necessary due to the relative absence of feeding cues. Upward or downward shifts in water temperature by approximately 10°C also inhibited inducible defense in F. vesiculosus. However, such shifts did not affect algal growth and were therefore the consequence of an impairment of specific defense-related components rather than of resource limitation, unless compensatory growth was given priority over defense.

Description: Differences with respect to anti-herbivore defense were investigated in invasive and native populations of the seaweed Gracilaria vermiculophylla. Specimens from 6 native populations in East Asia and from 8 populations invasive in Europe and the Mexican Pacific coast were maintained under identical conditions and offered to herbivorous snails from both the native range (Littorina brevicula) and Europe (L. littorea) in no-choice feeding assays. L. brevicula consumed in total significantly larger amounts of G. vermiculophylla tissue than did L. littorea. Further, both snail species least consumed the seaweed specimens originating from either non-native populations or from populations native to the Korean East Sea/Sea of Japan. The Korean East Sea/Sea of Japan had previously been identified as putative donor region of all the invasive populations of G. vermiculophylla. Thus, populations in the donor region as well as non-native populations in different invaded realms feature an increased capacity to resist feeding pressure. Differences in nutrient content did not account for the observed patterns of consumption, as palatability and carbon to nitrogen (C:N) ratio were not significantly correlated. Thus, mechanical or chemical defenses or the content of feeding cues influenced the behavior of the snails. We suggest that low palatability contributed to the invasion success of the species.

Description: The impact of moderate environmental stress may be modulated by stress-induced shifts of biotic interactions such as host – epibiont relationships. We studied the stress regime in shallow Western Baltic habitats, the variability of fouling at different temporal and spatial scales, and whether common stressors - low light, high temperature, grazing – affect the abundance and composition of the biofilm on a regionally important macroalga, the bladder wrack Fucus vesiculosus. We further explore the alga’s capacity to chemically modulate the recruitment of microfoulers and whether this ability is impacted by stress. In laboratory, mesocosm and field experiments fouling pressure and epibiotic cover on the algae varied strongly with changing environmental conditions such as temperature, irradiance, depth or grazing. The expectation that abiotic stress affects the fouling-modulating ability of the alga and, thus, indirectly produces the observed variability of epibiosis was not generally confirmed. Indeed, while the strength of chemical antifouling resistance varied seasonally, with a maximum in winter/spring and a minimum in late summer, this could not be related to temporal patterns of environmental stress, fouling pressure, or growth of Fucus. Only the seasonal variation in reproduction seemed to be in phase with antifouling activity. Controlled experiments confirmed that resistance strength was not affected by temperature or grazing, and only moderately by light. We conclude that the fouling modulation ability of Fucus vesiculosus may suffer from light-reduction (e.g. by eutrophication effects) while they are not sensitive to the predicted warming or enhanced grazing.

Description: To evaluate the importance of anti-herbivore resistance for algal invasion success we compared resistance traits among specimens of the red macroalga Gracilaria vermiculophylla from six native populations in Korea and China and eight invasive populations in Europe and Mexico that were maintained under identical conditions in the laboratory. Herbivorous snails both from the native range (Littorina brevicula) and from the invaded range (Littorina littorea) consumed significantly less of seaweed specimens originating from non-native populations. Metabolome profiling revealed that this preference was correlated with an increased woundactivated production of deterring prostaglandins and hydroxyeicosatetraenoic acids. Thus, invasive populations of G. vermiculophylla are more strongly defended against challenge by herbivores and other biological enemies that cause local tissue or cell disruption and activate oxylipin production. Anthropogenic distribution of genotypes adapted to resist elevated feeding pressure probably contributed to the invasion success of this species.

Description: Grazing by the isopod Idotea baltica induces chemical defenses in the brown seaweed Fucus vesiculosus. A combination of a 33 day induction experiment, feeding choice assays and functional genomic analyses was used to investigate temporal defense patterns and to correlate changes in palatability to changes in gene expression. Despite permanent grazing, seaweed palatability varied over time. Controls were significantly more consumed than grazed pieces only after 18 and 27 days of grazing. Relative to controls, 562/402 genes were up-/down-regulated in seaweed pieces that were grazed for 18 days, i.e. when defense induction was detected. Reprogramming of the regulative expression orchestra (translation, transcription), up-regulation of genes involved in lipid and carbohydrate metabolism, intracellular trafficking, defense and stress response, as well as downregulation of photosynthesis was found in grazed seaweed. These findings indicate short-term temporal variation in defenses and that modified gene expression patterns arise at the same time when grazed seaweed pieces show reduced palatability. Several genes with putative defensive functions and cellular processes potentially involved in defence, such as reallocation of resources from primary to secondary metabolism, were revealed

Description: Bacterial epiphytes of Gracilaria conferta were quantified. Saprophytic bacteria reached 350 times and agar degraders 25000 times higher numbers g-1 algal wet wt on tissues infected with the 'white tips disease', as compared to healthy tissues. A bacterial inducing agent of the 'white tips disease' was detected. Addition of 10(2) to 10(3) cells of this isolate ml-1 medium led to increased rates of infection. This effect did not occur if the isolate was autoclaved before addition. The virulent bacteria could always be isolated from infected tissues. It frequently, but not always, infected G. conferta and should be regarded as a facultative parasite. Several factors influenced the disease development. Temperatures above 20-degrees-C, in combination with photon flux densities of more than 200 muE m-2 s-1, increased the rate of infection. Relatively low amounts (more than 25 mug ml-1) of certain organic nutrients (peptone and yeast extract) led to strong manifestations of the disease. Addition of agar did not cause any symptoms, while 5 mg l-1 of the antibiotic rifampicin prevented the alga from being infected.