Transcriptome and functional analyses of phenotypic plasticity in sea grape Caulerpa okamurae.

Caulerpa is a marine green macroalga distinguished by a large single cell with multiple nuclei. It also exhibits remarkable morphological intraspecies variations, in response to diverse environmental types. However, the molecular mechanisms underlying this phenotypic plasticity remain poorly understood. In this work, we compare the transcriptomes of Caulerpa okamurae Weber Bosse, 1897 displaying altered phenotypes of cultivation and natural phenotypes and investigate significantly regulated genes and their biological functions using differential expression analyses. We observe light-harvesting complex upregulation and cellular framework stability downregulation in altered phenotypes compared to the natural phenotypes. Intertidal macrophytes reduce light capture to avoid photodamage and regulate their morphology to protect against wave damage. In contrast, the lower light conditions and the cultivation environment augment light capture and increase a morphology prioritizing light trapping. Moreover, the addition of simulated wave-sweeping stimuli induces a return to the natural morphology under high-light conditions, showing how mechanical stress affects morphological organization in C. okamurae. We provide detailed gene expression patterns in C. okamurae under varying light intensities and water conditions, suggesting a distinct influence on its morphological traits.

Marine macroalga Caulerpa: role of its metabolites in modulating cancer signaling.

Cancer, the leading causes of death worldwide, causes multiple metabolic and physiological alterations, leading to an unregulated proliferation of cells. The existing anticancer therapies are usually nonspecific with side effects and or are extremely expensive, thus hunt for better therapeutics is still on, specially efforts are made to look for naturally occurring molecules. Sea harbors several organisms which are unexplored for their biological potentials. Green macroalga genus, Caulerpa, is one such invaluable repository of bioactive metabolites like alkaloids, terpenoids, flavonoids, steroids and tannins with reported bioactivities against many diseases including cancer. Anti-cancerous metabolites of Caulerpa like caulerpenyne (Cyn), caulerpin, caulersin, and racemosin C, possess unique structural moieties and are known to exhibit distinct effects on cancer cells. Theses metabolites are reported to affect microtubule dynamics, unfolded protein response, mitochondrial health, cell cycle progression, metabolic and stress pathways by their cross-talk with signalling proteins like AMPK, GRP78, GADD153, Bid, Bax, AIF, Bcl2, P21, cyclin D, cyclin E, caspase 9, and PTP1B. Targeting of multiple cancer hallmarks by Caulerpa metabolites, with concomitant modulations of multiple signalling cascades, displays its multifactorial approach against cancer. Evaluation of anti-cancer properties of this genus is particularly important as Caulerpa species are widely edible and utilized in several delicacies in the coastal countries. This is the first review article providing a consolidated information about the role of Caulerpa in cancer with major contributing metabolites and plausible modulations in cancer signaling and prospects.

Habitat heterogeneity promotes the coexistence of exotic seaweeds.

Despite the progressive accumulation of exotic species in natural communities, little effort has been devoted to elucidating the mechanisms underpinning the coexistence of invaders in environmentally and biologically heterogeneous systems. The exotic seaweeds, Asparagopsis taxiformis and Caulerpa racemosa, exhibit a segregated distribution on Mediterranean rocky reefs. A. taxiformis dominates assemblages in topographically complex habitats, but is virtually absent on homogenous platforms. In contrast, C. racemosa achieves extensive cover in both types of habitat. We assessed whether differences in their distribution were generated by biotic interactions (between invaders and/or between invaders and natives) or by environmental constraints. Three models were proposed to explain seaweed distribution patterns: (1) invaders inhibit one another; (2) native assemblages, differing between complex and simple habitats, prevent the establishment/spread of one invader, but not that of the other; and (3) environmental conditions regulate the establishment/persistence of the seaweeds in different habitats. We removed the dominant invader and resident assemblages in each type of habitat. Moreover, A. taxiformis thalli were transplanted into the habitat dominated by C. racemosa to establish whether its failure to colonize the simple habitat was due to the lack of propagules or post-recruitment mortality. C. racemosa spread in the complex habitat was not influenced by the removal of resident assemblages, but it was slightly enhanced by A. taxiformis removal. Neither C. racemosa removal nor that of resident assemblages promoted A. taxiformis colonization and survival in simple habitats. Our results suggest that heterogeneity in environmental conditions can promote invader coexistence by mitigating the effects of negative biotic interactions. Therefore, the accumulation of introduced species in native communities does not necessarily imply established invaders fostering further invasion.

Insights into the bacterial community composition of farmed Caulerpa lentillifera: A comparison between contrasting health states.

The bacterial communities of Caulerpa lentillifera were studied during an outbreak of an unknown disease in a sea grape farm from Vietnam. Clear differences between healthy and diseased cases were observed at the order, genus, and Operational Taxonomic Unit (OTU) level. A richer diversity was detected in the diseased thalli of C. lentillifera, as well as the dominance of the orders Flavobacteriales (phylum Bacteroidetes) and Phycisphaerales (Planctomycetes). Aquibacter, Winogradskyella, and other OTUs of the family Flavobacteriaceae were hypothesized as detrimental bacteria, this family comprises some well-known seaweed pathogens. Phycisphaera together with other Planctomycetes and Woeseia were probably saprophytes of C. lentillifera. The Rhodobacteraceae and Rhodovulum dominated the bacterial community composition of healthy C. lentillifera. The likely beneficial role of Bradyrhizobium, Paracoccus, and Brevundimonas strains on nutrient cycling and phytohormone production was discussed. The bleaching of diseased C. lentillifera might not only be associated with pathogens but also with an oxidative response. This study offers pioneering insights on the co-occurrence of C. lentillifera-attached bacteria, potential detrimental or beneficial microbes, and a baseline for understanding the C. lentillifera holobiont. Further applied and basic research is urgently needed on C. lentillifera microbiome, shotgun metagenomic, metatranscriptomic, and metabolomic studies as well as bioactivity assays are recommended.

The seaweed Caulerpa racemosa on Mediterranean rocky reefs: from passenger to driver of ecological change.

Disentangling the ecological effects of biological invasions from those of other human disturbances is crucial to understanding the mechanisms underlying ongoing biotic homogenization. We evaluated whether the exotic seaweed, Caulerpa racemosa, is the primary cause of degradation (i.e., responsible for the loss of canopy-formers and dominance by algal turfs) on Mediterranean rocky reefs, by experimentally removing the invader alone or the entire invaded assemblage. In addition, we assessed the effects of enhanced sedimentation on the survival and recovery of canopy-forming macroalgae at a relatively pristine location and how their loss affects the ability of C. racemosa to conquer space. C. racemosa did not invade dense canopy stands or influence their recovery in cleared plots. Competition with C. racemosa could not explain the rarity of canopy-forming species at degraded sites. Removing the assemblages invaded by C. racemosa and preventing reinvasion did not trigger the transition from algal turfs to canopies, but it enhanced the cover of morphologically complex erect macroalgae under some circumstances. Once established, C. racemosa, enhancing sediment accumulation, favors algal turfs over erect algal forms and enables them to monopolize space. Our results show that introduced species that rely on disturbance to establish can subsequently become the main drivers of ecological change.

Variable direct and indirect effects of a habitat-modifying invasive species on mortality of native fauna.

Habitat-modifying invasive species can influence rates of predation on native prey either directly by providing protective structure or indirectly by modifying traits of prey species responding to the habitat. The alga Caulerpa taxifolia is one of the most successful invasive species of shallow-water marine systems globally, often provisioning habitat in areas previously lacking in vegetated structure. We experimentally evaluated the direct effect of Caulerpa to provide refuge for the native clam Anadara trapezia and how this balances with its influence on two trait-mediated indirect interactions that may increase Anadara's susceptibility to predators. Specifically, Caulerpa's alteration of physical and chemical properties of the surrounding water and sediment deteriorate Anadara's condition and predator resistance properties and also cause Anadara, though normally buried, to project from beneath the sediment, exposing it to predators. Our results show that Anadara are somewhat (but not consistently) protected from predators by living among Caulerpa. Shallow burial depth did not counteract this protective effect. However at times of year when predator activity diminishes and conducive environmental conditions develop, negative effects of Caulerpa habitat such as hypoxia and lowered flow may dominate. Under such situations, poor clam condition accentuates Anadara's susceptibility to mortality. Ultimately, a slight and inconsistent positive effect of Caulerpa to protect Anadara from predators is exceeded by the strong negative effect of Caulerpa on clam mortality, which is heightened by clams' weakened condition produced by chronic exposure to Caulerpa. Our results show that invasive habitat-modifying species can affect mortality of native species not simply through obvious positive direct effects of their protective structure, but indirectly through contrasting negative modification of the traits of prey species responding to the habitat.

Native species behaviour mitigates the impact of habitat-forming invasive seaweed.

Habitat-forming invasive species cause large, novel changes to the abiotic environment. These changes may elicit important behavioural responses in native fauna, yet little is known about mechanisms driving this behaviour and how such trait-mediated responses influence the fitness of native species. Low dissolved oxygen is a key abiotic change created by the habitat-forming invasive seaweed, Caulerpa taxifolia, which influences an important behavioural response (burrowing depth) in the native infaunal bivalve Anadara trapezia. In Caulerpa-colonised areas, Anadara often emerged completely from the sediment, and we experimentally demonstrate that water column hypoxia beneath the Caulerpa canopy is the mechanism instigating this "pop-up" behaviour. Importantly, pop-up in Caulerpa allowed similar survivorship to that in unvegetated sediment; however, when we prevented Anadara from popping-up, they suffered >50% mortality in just 1 month. Our findings not only highlight the substantial environmental alteration by Caulerpa, but also an important role for the behaviour of native species in mitigating the effects of habitat-forming invasive species.

Behavioural interactions between ecosystem engineers control community species richness.

Behavioural interactions between ecosystem engineers may strongly influence community structure. We tested whether an invasive ecosystem engineer, the alga Caulerpa taxifolia, indirectly facilitated community diversity by modifying the behaviour of a native ecosystem engineer, the clam Anadara trapezia, in southeastern Australia. In this study, clams in Caulerpa-invaded sediments partially unburied themselves, extending >30% of their shell surface above the sediment, providing rare, hard substrata for colonization. Consequently, clams in Caulerpa had significantly higher diversity and abundance of epibiota compared with clams in unvegetated sediments. To isolate the role of clam burial depth from direct habitat influences or differential predation by habitat, we manipulated clam burial depth, predator exposure and habitat (Caulerpa or unvegetated) in an orthogonal experiment. Burial depth overwhelmingly influenced epibiont species richness and abundance, resulting in a behaviourally mediated facilitation cascade. That Caulerpa controls epibiont communities by altering Anadara burial depths illustrates that even subtle behavioural responses of one ecosystem engineer to another can drive extensive community-wide facilitation.

Synergistic reduction of a native key herbivore performance by two non-indigenous invasive algae.

Native generalist grazers can control the populations of non-indigenous invasive algae (NIIA). Here, it was found that the simultaneous consumption of two co-occurring NIIA, Caulerpa cylindracea and C. taxifolia var. distichophylla, hinders the grazing ability of the main Mediterranean herbivorous, the native sea urchin Paracentrotus lividus. The ingestion of any of the two NIIA alone did not produce any difference in sea urchin righting time with respect to usual algal diet. In contrast, the simultaneous consumption of both NIIA, which grow intermingled in nature and are consumed by P. lividus, retarded its righting behavior. Such result reveals substantial physiological stress in the sea urchin, which resulted in reduced motility and coordination. The reported findings reveal the potential of NIIA co-occurrence to escape the supposed control exerted by the main native generalist grazer in Mediterranean sublittoral communities, which in turn can be locked in an "invaded" state.

Negative effects of warming on seagrass seedlings are not exacerbated by invasive algae.

The observed and projected rise in sea surface temperature challenges marine biodiversity worldwide, and particularly in temperate ecosystems dealing with the arrival of novel species of tropical provenance. When the impacted biota are early life stages of ecosystem engineers, the effects of those impacts are of major concern for ecologists and coastal managers. We experimentally examined the individual and potential additive effects of seawater warming and the presence of the invasive algae on the development of seedlings of the seagrass Posidonia oceanica in a three-month mesocosm experiment. Whereas the presence of the invasive algae (Caulerpa cylindracea and Lophocladia lallemandii) did not result in detrimental effects on seedlings, warming negatively affected seedling development. Interestingly, the presence of both invasive algae may ameliorate the negative effects of warming.

Changes in amphipod (Crustacea) assemblages associated with shallow-water algal habitats invaded by Caulerpa racemosa var. cylindracea in the western Mediterranean Sea.

The effects of the invasive species Caulerpa racemosa var. cylindracea (hereafter C. racemosa) on amphipod assemblages associated with shallow-water rocky habitats were studied. Two habitats located along the SE Iberian Peninsula were compared; invaded and non-invaded. The results showed that growth of C. racemosa affects habitat structure, influencing the species composition and biomass of macroalgae, and detritus accumulation. In turn, such changes in habitat features affected the associated amphipod assemblages with different ecological requirements. However, the species richness of amphipods was relatively high in both habitats, while the species composition of amphipods changed completely. For example, some species such as Ampithoe ramondi and Hyale schmidti did not colonize invaded habitats, while others such as Apocorophium acutum were favoured by the spread of C. racemosa. Habitat invasion by C. racemosa can have an important influence on biotic assemblages, modifying both habitat structure and the associated fauna, with unknown effects on the overall ecosystem.

The spread of Caulerpa racemosa var. cylindracea in the Mediterranean Sea: an example of how biological invasions can influence beta diversity.

The present study aims to evaluate if invasion of Caulerpa racemosa var. cylindracea can affect both alpha and beta diversity in Mediterranean coastal system. The following hypotheses were tested: (i) measures of alpha diversity differ between invaded and non-invaded assemblages (ii) this pattern is consistent between different habitats, (iii) assemblages from habitats which are naturally very different tend to be more similar when invaded compared to non-invaded situations. Results showed that Mediterranean benthic assemblages invaded by C. racemosa var. cylindracea were characterized by lower values of alpha diversity than non-invaded assemblages; moreover, differences between deep (25 m) and shallow water (5m) assemblages appeared lower in invaded than in non-invaded areas, with a decrease of beta diversity. This homogenisation of habitats was related to a lack of several characteristic species and to a higher abundance of few opportunistic species in invaded areas, particularly turf-forming algae. Moreover, variability in taxa composition was lower in invaded than in non-invaded assemblages in both habitats.

The Caulerpa racemosa invasion: a critical review.

Caulerpa racemosa var. cylindracea is a marine Chlorophyta introduced into the Mediterranean Sea from south-western Australia. Since 1990, it has been invading the Mediterranean Sea and the Canary Islands, raising ecological problems. Although this invasion event can be considered as one of the most serious in the history of species introduced into the Mediterranean Sea, C. racemosa has not triggered as much attention as the famous "killer alga"Caulerpa taxifolia. The aim of the present study was: (i) to summarize the current state of knowledge with regard to the distribution, the various biological and ecological characteristics of the introduced C. racemosa and its impact on the Mediterranean coastal environment; (ii) to discuss the various hypotheses regarding the explanation for its rapid and successful spread; (iii) to investigate the disparity in the treatment of C. racemosa and Caulerpa taxifolia invasions; and (iv) to outline future research needs.

Biotic resistance and vegetative propagule pressure co-regulate the invasion success of a marine clonal macrophyte.

Propagule pressure is considered a major driver of plant invasion success. Great propagule pressure would enable invasive species to colonize new areas overcoming the resistance of native species. Many highly invasive aquatic macrophytes regenerate from vegetative propagules, but few studies have experimentally investigated the importance of propagule pressure and biotic resistance, and their interaction, in determining invasion success. By manipulating both recipient habitat and the input of vegetative propagules of the invasive seaweed Caulerpa cylindracea in mesocosm, we examined whether higher propagule pressure would overcome the resistance of a native congeneric (Caulerpa prolifera) and influence its performance. With the native, C. cylindracea population frond number decreased irrespectively of pressure level. High propagule pressure did not increase stolon length and single plant size decreased due to the effects of intra- and interspecific competition. Native biomass decreased with increasing C. cylindracea propagule pressure. These results indicate that higher propagule pressure may fail in enhancing C. cylindracea invasion success in habitats colonized by the native species, and they suggest that biotic resistance and propagule pressure co-regulate the invasion process. These findings emphasize the need to preserve/restore native seaweed populations and may help to design effective management actions to prevent further C. cylindracea spread.

Field transplantation of seagrass (Posidonia oceanica) seedlings: Effects of invasive algae and nutrients.

Seedlings are a key life stage in seagrasses, providing genetic diversity and being a useful tool for restoration. We examined the influence of increased sediment nutrients and the presence of the invasive macroalga Caulerpa cylindracea on the success of in situ transplanting Posidonia oceanica seedlings in a six-month experiment. Our results indicate that one-year old seedlings successfully survive in the field and their survival and growth are positively affected by the presence of C. cylindracea. Furthermore, nutrient addition in the sediment had positive effects on both C. cylindracea (increasing its cover) and seedlings (increasing leaf development), and the increased C. cylindracea cover did not result in detrimental effects on seedlings. Therefore, biological invasions and nutrient addition do not reinforce each other in the short term to negatively impact transplanted seedlings, which highlights facilitative interactions between invasive algae and native seagrass and provides useful information for successful strategies of seagrass restoration.

Under the canopy: Community-wide effects of invasive algae in Marine Protected Areas revealed by metabarcoding.

We analysed with multigene (18S and COI) metabarcoding the effects of the proliferation of invasive seaweeds on rocky littoral communities in two Spanish Marine Protected Areas. The invasive algae studied were Caulerpa cylindracea, Lophocladia lallemandii and Asparagopsis armata. They are canopy-forming, landscape-dominant seaweeds, and we were interested in their effects on the underlying communities of meiobenthos and macrobenthos, separated in two size fractions through sieving. A new semiquantitative treatment of metabarcoding data is introduced. The results for both markers showed that the presence of the invasive seaweed had a significant effect on the understory communities for Lophocladia lallemandii and Asparagopsis armata but not for Caulerpa cylindracea. Likewise, changes in MOTU richness and diversity with invasion status varied in magnitude and direction depending on the alga considered. Our results showed that metabarcoding allows monitoring of the less conspicuous, but not least important, effects of the presence of dominant invasive seaweeds.

Urban seagrass: status of Posidonia oceanica facing the Genoa city waterfront (Italy) and implications for management.

A system of five adjacent Posidonia oceanica meadows facing the waterfront of Genoa city (Ligurian Sea, NW Mediterranean) was investigated over different spatial scales (meters-kilometers) using three environmental indices: conservation index (CI), substitution index (SI) and phase-shift index (PSI). CI revealed differences mostly at large spatial scale, distinguishing the poor condition of the meadows closest to Genoa centre and harbour from the comparatively healthy condition of the farthest meadows. SI showed differences mostly at small spatial scale (i.e., within meadows), suggesting the influence of local factors in the re-colonisation of regressed meadows by the seagrass Cymodocea nodosa and/or the invasive alga Caulerpa racemosa. Mapping of PSI showed that the meadows closest to Genoa centre and harbour have undergone a nearly total phase shift and have no real potential for recovery: attempts to re-establish P. oceanica there might be a waste of time and money. On the contrary, the meadows farthest from Genoa centre and harbour showed a comparatively low level of phase shift and could still fully recover given specific management actions.

Habitat associations of an expanding native alga.

There are many examples of native macrophytes becoming locally dominant and spreading outside their traditional distributions, but the causes and impacts are often not understood. In New South Wales, Australia, the green alga Caulerpa filiformis is undergoing a range expansion and has transitioned from a subdominant to a dominant alga on several rocky shores around the Sydney coastline. Here we investigated relationships between established patches of C. filiformis, the habitat it occupies and associated algal communities at multiple subtidal sites over the green alga's 700 km range. We tested the following predictions: 1) C. filiformis cover differs among substrata, being greatest on turf-forming algae; 2) C. filiformis cover is positively related to environmental variables linked to increased sedimentation (e.g. reduced reef width, surface slope, increased rugosity and distance from shore); 3) occurrence of C. filiformis is associated with a change in macrophyte community structure and a reduction of macrophyte richness; 4) intact native algal canopies inhibit C. filiformis spread, but turf-forming algae and bare sand are susceptible to invasion. Substratum associations were highly consistent among sites, but contrary to our prediction, C. filiformis was most commonly associated with rock or rock + sand substratum and less frequently associated with turf-forming algae substratum. C. filiformis cover was negatively correlated with reef width, which explained most of the variation observed, although local scale variables distance from shore, reef slope, and water depth were also correlated with C. filiformis cover. Algal diversity and community composition typically differed in the presence of C. filiformis, often with a reduction of algal abundances, in particular Sargassum spp., although results varied among substrata and sites. However, monitoring of borders suggested that C. filiformis does not invade and outcompete undisturbed adjacent canopy-forming algae over a 12 month period. Our results suggest that disturbance processes (possibly linked to sedimentation) acting at the site and quadrat scale are likely important determinants of C. filiformis cover and spread, and hence its potential ecological impacts.

Differential response of coral communities to Caulerpa spp. bloom in the reefs of Indian Ocean.

Coral reef ecosystems are disturbed in tandem by climatic and anthropogenic stressors. A number of factors act synergistically to reduce the live coral cover and threaten the existence of reefs. Continuous monitoring of the coral communities during 2012-2014 captured an unprecedented growth of macroalgae as a bloom at Gulf of Mannar (GoM) and Palk Bay (PB) which are protected and unprotected reefs, respectively. The two reefs varying in their protection level enabled to conduct an assessment on the response of coral communities and their recovery potential during and after the macroalgal bloom. Surveys in 2012 revealed a live coral cover of 36.8 and 14.6% in GoM and PB, respectively. Live coral cover was lost at an annual rate of 4% in PB due to the Caulerpa racemosa blooms that occurred in 2013 and 2014. In GoM, the loss of live coral cover was estimated to be 16.5% due to C. taxifolia bloom in 2013. Tissue regeneration by the foliose and branching coral morphotypes aided the recovery of live coral cover in GoM, whereas the chances for the recovery of live coral cover in PB reef were low, primarily due to frequent algal blooms, and the existing live coral cover was mainly due to the abundance of slow-growing massive corals. In combination, results of this study suggested that the recovery of a coral reef after a macroalgal bloom largely depends on coral species composition and the frequency of stress events. A further study linking macroalgal bloom to its specific cause is essential for the successful intervention and management.

Carry over effects of nutrient addition on the recovery of an invasive seaweed from the winter die-back.

Nutrient enrichment of coastal waters can enhance the invasibility and regrowth of non-native species. The invasive alga Caulerpa cylindracea has two distinct phases: a well-studied fast-growing summer phase, and a winter latent phase. To investigate the effects of nutrient enrichment on the regrowth of the seaweed after the winter resting-phase, a manipulative experiment was carried out in intertidal rockpools in the North-western Mediterranean. Nutrients were supplied under different temporal regimes: press (constant release from January to May), winter pulse (January to March) and spring pulse (March to May). Independently from the temporal characteristics of their addition, nutrients accelerated the re-growth of C. cylindracea after the winter die-back, resulting in increased percentage covers at the peak of the growing season. Nutrient addition did not influence the number and length of fronds and the biomass. Native components of the algal community did not respond to nutrient additions. Our results show that nutrient supply can favour the spread of C. cylindracea even when occurring at a time of the year at which the seaweed is not actively growing.