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Diseases of the commercially cultivated brown seaweed Saccharina japonica have been noticed since the 1960s in China. However, the usually opportunistic nature of the pathogenic bacteria of seaweeds is still a main obstacle for isolating and identifying pathogenic bacteria responsible for causing diseases of the cultivated S. japonica. In this study, a Gram-positive bacterium XP-2 was isolated from the abnormal diseased mature sporophytes of S. japonica. Pathogenicity test indicated that XP-2 could cause green-rotten disease in healthy juvenile sporophytes and significant algal ultrastructural changes were observed after S. japonica was infected by XP-2. The cells shrank, and the thylakoids of the chloroplasts became blurred after 12 h of re-infection. At 24 h slight deformation was observed in the structure of the mitochondria, chloroplasts, and nucleus. There also were many vacuoles in the infected cells. After 31 h of re-infection, mitochondrial structure and thylakoids became more indistinct. SEM and TEM observations indicated that XP-2 was rod-shaped with no flagella. Analysis of the full length 16S rRNA gene sequence and physiological and chemical characteristics indicated that XP-2 was closely related to Exiguobacterium oxidotolerans 12280 T. Based on these results, XP-2 was designated Exiguobacterium oxidotolerans XP-2. The pathogenicity of E. oxidotolerans XP-2 was identified by Koch’s Postulate under laboratory conditions. These results enrich the growing list of pathogenic bacteria in commercially cultivated seaweeds and also provide a foundation for investigating the virulence mechanisms of pathogenic bacteria of commercially cultivated seaweeds in the near future.

Seaweed bioinvasions increasingly affect coastal environments around the world, which increases the need for predictive models and mitigation strategies. The biotic interactions between seaweed invaders and invaded communities are often considered a key determinant of invasion success and failure and we here revise the current evidence that the capacity of seaweed invaders to deter enemies in newly reached environments correlates with their invasion success. Particularly efficient chemical defences have been described for several of the more problematic seaweed invaders during the last decades. However, confirmed cases in which seaweed invaders confronted un-adapted enemies in newly gained environments with deterrents that were absent from these environments prior to the invasion (so-called “novel weapons”) are scarce, although an increasing number of invasive and non-invasive seaweeds are screened for defence compounds. More evidence exists that seaweeds may adapt defence intensities to changing pressure by biological enemies in newly invaded habitats. However, most of this evidence of shifting defence was gathered with only one particular model seaweed, the Asia-endemic red alga Agarophyton vermiculophyllum, which is particularly accessible for direct comparisons of native and non-native populations in common garden experiments. A. vermiculophyllum interacts with consumers, epibionts and bacterial pathogens and in most of these interactions, non-native populations have rather gained than lost defensive capacity relative to native conspecifics. The increases in the few examined cases were due to an increased production of broad-spectrum deterrents and the relative scarcity of specialized deterrents perhaps reflects the circumstance that seaweed consumers and epibionts are overwhelmingly generalists.

Lipids are key constituents of all cells, which express thousands of different lipid species. In most cases, it is not known why cells synthesize such diverse lipidomes, nor what regulates their metabolism. Although it is known that dividing cells specifically regulate their lipid content and that the correct lipid complement is required for successful division, it is unclear how lipids connect with the cell division machinery. Here, we report that the membrane protein stomatin is involved in the cytokinesis step of cell division. Although it is not a lipid biosynthetic enzyme, depletion of stomatin causes cells to change their lipidomes. These changes include specific lipid species, like ether lipids, and lipid families like phosphatidylcholines. Addition of exogenous phosphatidylcholines rescues stomatin-induced defects. These data suggest that stomatin interfaces with lipid metabolism. Stomatin has multiple contacts with the plasma membrane and we identify which sites are required for its role in cell division, as well as associated lipid shifts. We also show that stomatin's mobility on the plasma membrane changes during division, further supporting the requirement for a highly regulated physical interaction between membrane lipids and this newly identified cell division protein.

The supply of healthy sporelings plays a crucial role in the successful aquaculture of commercially cultivated brown kelp Saccharina japonica. Epibacteria are of critical importance for maintaining seaweed health and development. Thus, this study aimed to investigate the epibacterial community associated with mature sporophytes and 1.5- and 2.0-month-old sporelings, as well as juvenile sporophytes of S. japonica by using 16S rRNA gene amplicon sequencing and predicted the bacterial functional profiles. We found that Proteobacteria, Verrucomicrobia, Bacteroidetes, and Planctomycetes were the main phyla, while Loktanella and Rubritalea and Persicirhabdus were the dominant genera for all four developmental stages. Highly abundant core microbiota included Persicirhabdus, Loktanella, Litorimonas, and Rubritalea. Further analysis of the predicted functions showed that different metabolic functions were involved at different developmental stages. More metabolic functions were found in 2-month-old sporelings and mature sporophytes. Both the diversity, composition of the epiphytic bacterial communities, and metabolic functions changed according to different developmental stages at least in 1.5- and 2-month-old sporelings. Our results not only provide fundamental knowledge for the epibacteria associated with S. japonica but will also help nursery farms in preventing and mitigating the disease outbreaks by monitoring the variations of the epibacterial communities at the nursery stage.

Disease outbreaks cause severe economic losses in commercially cultivated Saccharina japonica. Epiphytic bacteria are increasingly recognized to play an important role in S. japonica health and may also become opportunistic pathogens when the environment deteriorates. However, there is little knowledge about the epiphytic bacterial communities associated with commercially cultivated S. japonica and how these communities change over the harvest season. We analyzed the diversity, composition, and temporal dynamics of epiphytic bacterial communities associated with both healthy and diseased tissue of S. japonica over a harvest season (from April to June, 2015) at Ailian Bay, Rongcheng, China. Our results showed that Proteobacteria and Bacteroidetes were the main phyla and Halomonas spp. were the most dominant bacterial species. Halomonadaceae, Hyphomonadaceae, Rhodobacteraceae, and Phyllobacteriaceae families were abundant in both healthy and diseased S. japonica, but the relative abundance of these bacteria differed between healthy and diseased groups. Spatial (site 1, site 2, and site 3 in April) and temporal comparisons (site 3 from April to June) revealed that the bacterial community associated with cultivated S. japonica was influenced by sampling time and sampling site independent of the health status. Several OTUs from the Roseobacter group, including members of the genera Sulfitobacter and Loktanella, were enriched in diseased tissues suggesting these taxa could be candidate opportunistic pathogens. In contrast, OTUs belonging to the genera Halomonas were enriched in healthy tissues, indicating a potential beneficial role for group during the cultivation of S. japonica. Our results provide important baseline knowledge of the epiphytic bacterial communities and possibly pathogenic bacteria associated with commercially cultivated S. japonica.

Saccharina japonica is a commercially farmed seaweed of global importance. However, disease occurrence during different stages of cultivation can result in substantial economic losses. Identification of the causative agents of disease remains a significant bottleneck to the large scale cultivation of S. japonica. In this study, an aerobic heterotrophic, flagellated, rod-shaped Gram-negative bacterial strain X-8 was isolated from the bleaching diseased S. japonica sporelings. Pathogenecity of strain X-8 was tested by re-infection assay. The ultrastructural changes of infected S. japonica cells by strain X-8 indicated that chloroplasts were the first organelle responding to X-8 infection with deformed structure and later followed by fragmented nucleus. However, the ultra-structure of mitochondria and cell wall remained intact during the re-infection. Based on 16S rRNA gene sequence, morphological and biochemical characteristics, strain X-8 was designated as Pseudoalteromonas piscicida X-8. The pathogenicity of P. piscicida X-8 was identified by Koch's Postulate under laboratory conditions. Our results will not only help to establish a stable experimental model between the pathogenic bacteria and the host S. japonica to further elucidate the virulence mechanisms, but will also provide information for disease management to effectively prevent and mitigate the occurrence of bleaching disease of S. japonica at nursery stage.

Invasive species are co-introduced with microbiota from their native range and also interact with microbiota found in the novel environment to which they are introduced. Host flexibility toward microbiota, or host promiscuity, is an important trait underlying terrestrial plant invasions. To test whether host promiscuity may be important in macroalgal invasions, we experimentally simulated an invasion in a common garden setting, using the widespread invasive macroalga Agarophyton vermiculophyllum as a model invasive seaweed holobiont. After disturbing the microbiota of individuals from native and non-native populations with antibiotics, we monitored the microbial succession trajectories in the presence of a new source of microbes. Microbial communities were strongly impacted by the treatment and changed compositionally and in terms of diversity but recovered functionally by the end of the experiment in most respects. Beta-diversity in disturbed holobionts strongly decreased, indicating that different populations configure more similar –or more common– microbial communities when exposed to the same conditions. This decline in beta-diversity occurred not only more rapidly, but was also more pronounced in non-native populations, while individuals from native populations retained communities more similar to those observed in the field. This study demonstrates that microbial communities of non-native A. vermiculophyllum are more flexibly adjusted to the environment and suggests that an intraspecific increase in host promiscuity has promoted the invasion process of A. vermiculophyllum. This phenomenon may be important among invasive macroalgal holobionts in general.

ABSRTRACTIncreased activity and excitability (sensitisation) of a series of molecules including the transient receptor potential ion channel, vanilloid subfamily, member 1 (TRPV1) in pain-sensing (nociceptive) primary sensory neurons are pivotal for developing pathological pain experiences in tissue injuries. TRPV1 sensitisation is induced and maintained by two major mechanisms; post-translational and transcriptional changes in TRPV1 induced by inflammatory mediators produced and accumulated in injured tissues, and TRPV1 activation-induced feed-forward signalling. The latter mechanism includes synthesis of TRPV1 agonists within minutes, and upregulation of various receptors functionally linked to TRPV1 within a few hours, in nociceptive primary sensory neurons. Here, we report that a novel mechanism, which contributes to TRPV1 activation-induced TRPV1-sensitisation within ~30 minutes in at least ~30% of TRPV1-expressing cultured murine primary sensory neurons, is mediated through upregulation in cyclooxygenase 2 (COX2) expression and increased synthesis of a series of COX2 products. These findings highlight the importance of feed-forward signalling in sensitisation, and the value of inhibiting COX2 activity to control pain, in nociceptive primary sensory neurons in tissue injuries.

Sargassum thunbergii, a common brown macroalga that occurs in the Asian Northwest Pacific, has important ecological services and economic values. In recent years, the distribution range and biomass of wild S. thunbergii resources have declined rapidly due to severe anthropogenic impacts and habitat loss. To slow the decline of this natural resource by taking appropriate measures, it is very important to evaluate the population genetic structure and diversity of S. thunbergii. In this study, we used amplified fragment length polymorphism (AFLP) markers to investigate population genetic differentiation and diversity of S. thunbergii along the coast of China at a genome-scanning scale. Five primer combinations that yielded high polymorphisms were identified in ten populations of S. thunbergii. Genetic analyses revealed low genetic diversity within populations and high levels of genetic differentiation among populations. Moreover, apparent gene flow occurred between S. thunbergii populations in the Yellow-Bohai Sea, whereas no such exchange was detected in the populations of the East China Sea. Genetic clustering, principal coordinates analysis (PCoA), and pairwise FST estimates consistently indicated that S. thunbergii populations divided into two genetic groups, coinciding with their geographical distribution (Yellow-Bohai Sea vs. East China Sea). The detected genetic diversity of S. thunbergii along the coast of China provides important insights not only for selecting high-yielding varieties for commercial cultivation, but also for proposing strategic management plans to protect and restore declining natural resources under anthropogenic climate change.

Communities are shaped by scale dependent processes. To study the diversity and variation of microbial communities across scales, the invasive and widespread seaweed Agarophyton vermiculophyllum presents a unique opportunity. We characterized pro- and eukaryotic communities associated with this holobiont across its known distribution range, which stretches over the northern hemisphere. Our data reveal that community composition and diversity in the holobiont vary at local but also larger geographic scales. While processes acting at the local scale (i.e., within population) are the main structuring drivers of associated microbial communities, changes in community composition also depend on processes acting at larger geographic scales. Interestingly, the largest analysed scale (i.e., native and non-native ranges) explained variation in the prevalence of predicted functional groups, which could suggest a functional shift in microbiota occurred over the course of the invasion process. While high variability in microbiota at the local scale supports A. vermiculophyllum to be a generalist host, we also identified a number of core taxa. These geographically independent holobiont members imply that cointroduction of specific microbiota may have additionally promoted the invasion process.

Summary 1.Epibiosis in the marine environment is a stressor that may determine invasion success in introduced species. Previous comparisons showed 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. To elucidate this question, a comparison between native and non-native populations of the same species is needed. 2.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 G. vermiculophylla from four populations in each range and used a fully-crossed design with the factors a) ‘Origin of Gracilaria’, b) ‘Origin of epiphytes’, c) ‘Season’ and d) ‘Solvent used for extraction’. 3.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. 4.Synthesis. Our results show for the first time that non-native individuals of a marine organism are better defended against epiphytes than native conspecifics. Furthermore, we found evidence that at least a part of the defense is based on extractable secondary metabolites. We discuss several mechanisms that could explain the increased resistance to epiphytes in non-native individuals, including the release from enemies in the non-native range, which could lead to an increase in algal performance during the invasion process. We suggest that an enhanced defense against epiphytes after introduction is one reason for G. vermiculophylla's invasion success. Our observation may also apply to other basibiont-epibiont and host-enemy systems, including plant-plant, plant-animal and animal-animal interactions, in aquatic environments and could be a key feature of bioinvasions. This article is protected by copyright. All rights reserved.

Highlights: • Wound activated prostaglandin production was quantified in Gracilaria vermiculophylla. • Specimens from non-native populations contained higher prostaglandin concentrations. • Prostaglandins and related compounds deter animal consumers. • Activated defense through Prostaglandins was selected during Gracilaria's invasion history. • As a consequence specimens from invasive populations are not safe for human consumption. Abstract: The capacity of the East Asian seaweed Gracilaria vermiculophylla (“Ogonori”) for production of prostaglandin E2 from arachidonic acid occasionally causes food poisoning after ingestion. During the last two decades the alga has been introduced to Europe and North America. Non-native populations have been shown to be generally less palatable to marine herbivores than native populations. We hypothesized that the difference in palatability among populations could be due to differences in the algal content of prostaglandins. We therefore compared the capacity for wound-activated production of prostaglandins and other eicosatetraenoid oxylipins among five native populations in East Asia and seven non-native populations in Europe and NW Mexico, using a targeted metabolomics approach. In two independent experiments non-native populations exhibited a significant tendency to produce more eicosatetraenoids than native populations after acclimation to identical conditions and subsequent artificial wounding. Fourteen out of 15 eicosatetraenoids that were detected in experiment I and all 19 eicosatetraenoids that were detected in experiment II reached higher mean concentrations in non-native than in native specimens. Wounding of non-native specimens resulted on average in 390% more 15-keto-PGE2, in 90% more PGE2, in 37% more PGA2 and in 96% more 7,8-di-hydroxy-eicosatetraenoic acid than wounding of native specimens. Not only PGE2, but also PGA2 and dihydroxylated eicosatetraenoic acid are known to deter various biological enemies of G. vermiculophylla that cause tissue or cell wounding, and in the present study the latter two compounds also repelled the mesograzer Littorina brevicula. Non-native populations of G. vermiculophylla are thus more defended against herbivory than native populations. This increased capacity for activated chemical defense may have contributed to their invasion success and at the same time it poses an elevated risk for human food safety.

Plants and algae can detect the presence of bacteria via sensing of proteins or peptides of bacterial origin. Flg22, a fragment of bacterial flagellin, is one of these peptides and has been shown to be an effective elicitor in both plants and algae. Here, we investigated the elicitor activity of flg22-derived peptides in the brown alga, Saccharina japonica. By monitoring luminol-dependent fluorescence, we could observe that the release of H2O2 induced by flg22-derived peptides is maximal at 2 h after induction. The elicitor activity was depending on the length of the peptides in the order of flg22 > flg15 > flg14. Cytological observations regarding the presence of reactive oxygen species (ROS) after induction were consistent with quantitative measurements of H2O2 generation using a 2′,7′-dichlorofluorescein diacetate (DCFH-DA) fluorescent probe. Addition of 1 μM each of flg22 and flg15 was sufficient to inhibit growth of female gametophytes. Furthermore, the elicitor activity of C-terminally shortened flg15-derived peptides suggests that flg15 apparently is the smallest peptide with elicitor activity. Amino acid position D43 at the N-terminus of a flagellin was demonstrated to be involved in the elicitor activity. Finally, H2O2 was localized in the plasma membranes of female gametophytes by an NADPH oxidase inhibitor and electron-dense deposits of cerium perhydroxide by transmission electron microscope. published

The susceptibility of native and non-native populations of the red alga Gracilaria vermiculophylla to fouling was compared in common garden experiments. Native and non-native algae were enclosed into dialysis membrane tubes, and the tubes were exposed to natural fouling. Fouling on the outside of the tubes was mediated by chemical compounds excreted by G. vermiculophylla that diffused through the membranes. Fouling pressure was significantly higher in the Kiel Fjord (non-native range) than in Akkeshi Bay (native range), but, at both sites, tubes containing non-native G. vermiculophylla were less fouled than those with native conspecifics. This is the first in situ evidence that susceptibility to fouling differs between native and non-native populations of an aquatic organism. The technique of enclosing organisms into dialysis tubes represents a simple, efficient and accurate way to test chemical antifouling defenses and could possibly be applied to other organisms.

Saccharina japonica is an economically important alga worldwide as a source of of alginate, mannitol and iodine etc. China contributes 60 % of S. japonica production volume and 90 % of its alginate globally. Like the land crops, S. japonica is also plagued by diseases initi- ated by causative bacteria, fungi or viruses. In this article, we review investigations over the last 50 years on microbial diseases in nursery and fi eld-cultivated S. japonica in China, including the etilology of two different sources of diseases occurring in S. japonica cultivation, the prev- alence of causative alginic acid-decomposing bacteria and the recent progress in the under- standing of innate immunity mechanisms of S. japonica. Meanwhile, we highlight the questions for future research and advocate that researchers from different disciplines apply traditional technologies and newly emerging ones to elucidate the S. japonica microbial diseases. The full understanding of interactions between S. japonica, causative bacteria and environmental fac- tors will help to develop management strategies for preventing and mitigating the disease out- breaks in S. japonica aquaculture in the long term.

Silvetia babingtonii is a potentially economic brown alga for sources of food and high-value added utilization. So far, sporeling nursery and field cultivation has not been successful. The lack of knowledge on development and life cycle of this alga hinder the development of techniques for the sporeings and cultivation. In this study, internal structure of oogonium and antherium of S. babingtonii was observed with hematoxylin and eosin staining and through microscope. Meanwhile, early development from zygotes to juvenile sporelings was studied at 20 degrees C under 60-100 mu mol photons m(-2) s(-1). Zygotes germinated and divided into thallus and rhizoid cells. The larger thallus cells further divided and developed into juvenile sporelings; while the smaller rhizoid cells divided and elongated into rhizoid hairs. These findings documented the life cycle of S. babingtonii and provided fundamental knowledge for sporeling nursery in the near future.

In this study, variations in the chemical composition of Costaria costata collected during 3 months of the harvest period were analyzed. Moisture (4.94–10.50 %), ash (29.25–38.19 %), protein (9.77–18.15 %), lipid (0.60–2.21 %), crude fiber (4.45–5.68 %), alginate (22.49–29.13 %), fucoxanthin (0.07–0.32 mg g−1), polyphenol (1.579–4.796 mg g−1) were analyzed from dried alga. Six mineral elements were analyzed and the most abundant were calcium (6.64–11.56 mg g−1) and magnesium (7.02–7.92 mg g−1). Analysis of fatty acid composition indicated that the polyunsaturated fatty acids palmitoleic acid and linoleic acid were abundant in May and June, whereas the saturated fatty acid palmitic acid was abundant in July. Amino acid composition was also analyzed and the most abundant amino acids were aspartic acid, glutamic acid, glycine, and alanine. The ratio of mannuronic acid to guluronic acid of alginate was 2.57, 2.17, and 1.66 in May, June, and July, respectively. The gel strength of alginate was 1,449.0, 1,935.0, and 980.5 g cm−1 in May, June, and July, respectively. The results of this study indicate that C. costata is an excellent resource that provides extensively applications in the industrial areas of chemicals, food, cosmetics, and pharmacy.

Pigment variation within and among algal species may have important ecological consequences because small changes in the concentration and composition of pigments can influence the photosynthetic efficiency and rate as well as the spectra of light utilized. Toward the goal of developing a rapid method for comparing pigment composition among algal thalli, we characterized the relationship between visual color information taken from photographs (e.g., red, green, and blue color values) and photopigment composition in the non-native red alga Agarophyton vermiculophyllum (Ohmi) Gurgel, J.N.Norris & Fredericq. We used a set of 19 thalli, collected from across the known native and non-native range in the Northern Hemisphere, which exhibited substantial color variation at the time of field collection, and sustained this variation after being maintained in a common garden. We identified a set of ecologically interesting pigment traits that are readily predicted by color information, including chlorophyll a and phycobilin concentration. Finally, we demonstrated the repeatability of estimating color phenotypes from photographs of thalli taken under a range of light conditions in order to evaluate the utility of this approach for field studies. We suggest this method could be useful for the rapid, high-throughput phenotyping of photopigments in other red algae as well.

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.

Grateloupia turuturu Yamada is an economically valuable red alga with great potential in nutraceuticals and pharmaceuticals. Filaments of G. turuturu are of primary importance in germplasm preservation and sporeling culture, although filaments were not present in its life cycle. In this study, effects of temperature (10, 15, 20, 25, and 30 °C) and irradiance (10, 30, 60, and 90 μmol photons m−2 s−1) with photoperiod 10:14 h (light/dark) on filament development were investigated. Our results indicated that 25 °C was the optimal temperature for the formation of discoid crusts regardless of the irradiance. Conditions of 20 °C and 60 μmol photons m−2 s−1 promoted the development of discoid crusts and formation of upright thalli.

Most macroalgae and other aquatic organisms feature life cycles without a stage of dormancy and elevated stress resistance that is comparable to plant seeds. During anthropogenic transportation, they are therefore probably more exposed to stress than plant invaders, which could result in a more rigorous selection of stress-resistant phenotypes. However, selection of stress resistance during invasion processes of aquatic organisms has rarely been studied. We therefore compared the responses of native and non-native populations of the seaweed Gracilaria vermiculophylla to heat shock in common garden-type experiments. Specimens from six native populations in East Asia and from eight non-native populations in Europe and on the Mexican Pacific coast were acclimated to two sets of identical conditions before their resistance to heat shock was examined. The experiments were carried out twice—one time in the native range in Qingdao, China, and one time in the invaded range in Kiel, Germany—to rule out effects of specific local conditions. In both testing sites the non-native populations survived heat shock significantly better than the native populations. After three hours of heat shock G. vermiculophylla exhibited increased levels of heat-shock protein 70 (HSP70) and of a specific isoform of haloperoxidase, suggesting that both enzymes could be required for heat-shock stress management. However, the elevated resistance toward heat shock of non-native populations only correlated with an increased constitutive expression of heat-shock protein 70 (HSP70). The haloperoxidase isoform was more prominent in native populations, suggesting that not only increased HSP70 expression, but also reduced allocation into haloperoxidase expression after heat shock was selected during the invasion history. This selection probably happened early because the same combination of traits was detected in all realms within the non-native range.

Flg22, which is the most conserved 22-amino acid peptide in the N-terminal part of flagellin, functions as an effective elicitor in higher plants. Marine algae and higher plants share some conserved characteristics on defense response pathways. Flg22-induced defense responses were investigated in female gametophytes of Saccharina japonica. Condensation of chloroplasts and thickened cell walls, as well as relatively stable structures of mitochondria and nucleus indicated that there were hypersensitive programmed cell death occurred after induction by flg22. By using luminol-dependent luminescence detection method, rapid release of H2O2 was detected in the induced female gametophytes and reached a peak concentration of about 46 μM at 2 h. Reactive oxygen species production was also observed histologically using the fluorescent dye 2′,7′-dichlorofluorescein diacetate, showing a consistent result with quantitative analysis of H2O2. Furthermore, results of antioxidant enzyme activities indicated that there was a trend in the order of catalase > superoxide dismutase > glutathione peroxidase. Finally, high level phenol content of cell-free extracts was found after flg22 induction. According to our results, flg22 could be an effective elicitor which could induce defense responses in female gametophytes of S. japonica.

Grateloupia turuturu is a commercial red alga with potential value in nutraceuticals and pharmaceuticals. To supplement information on its life history and verify whether carpospores can be used for seedling culture, early development of G. turuturu was investigated under culture conditions (27°C, 10–13 μol/(m2·s) in irradiance, photoperiod 10:14 h L:D). Three physiological stages were recognized by continuous microscopic observation: division stage, discoid crust stage, and juvenile seedling stage. At the beginning of the division stage, the carpospores developed germ tubes into which the carpospore protoplasm was evacuated, and then the carpospore protoplasm in the germ tubes began to divide continuously until discoid crusts formed. Finally, upright thalli appeared on the discoid crusts and developed into juvenile seedlings. It took about 60 days for carpospores to develop into juvenile seedlings. The growth parameters, including germination rate for carpospores and discoid crust diameter, were recorded. These results contribute more information on the life cycle, and at the same time are of great significance in the scaling-up of artificial seedling cultures of G. turuturu.

Costaria costata (C. Agardh) Saunders is one of common kelps distributed in many coastal areas worldwide; however, in China, no reports have been made on cultivation of the genus. To investigate potential cultivation of the species in the northern part of China, trials on isolation and preservation of the gametophytes were conducted using C. costata from Korea; growth and development of the gametophytes were observed. We showed that at 10±1°C, 60 μmol m−2s−1 and 12:12 h (L:D), freshly released zoospores settled down within 1 hour, and then developed into the primary cell during the following 2 days. After a vegetative growth phase lasting 6–8 days, female gametophytes became 3–4 times larger in diameter than that of the primary cell, but still remained at a unicellular stage, while male gametophytes divided into 4–10 cells with only a slight change in size. Fertilization occurred within 10 days after the zoospores were released from the sporangia, and the apical and basal tissues of the juvenile sporophyte divided and differentiated into the blade and stipe. Temperature and irradiance influenced gametophytic vegetative growth and developmental patterns. Generally, low irradiance (15 μmol m−2s−1 and 30 μmol m−2s−1) was unfavorable to the induction of fertility, but it enhanced female gametophyte division. The optimal conditions for vegetative growth were 15°C and 30 μmol m−2s−1. After transplantation of the juvenile seedlings and after eight months cultivation, the harvested mature blade reached 194 cm in length and 32.7 cm in width. Our study proves that it is feasible to implement propagation and large scale cultivation of C. costata in northern China.

Chondrus is a type of commercially produced red seaweed that widely used for food and carrageen extraction. Although the natural life history of the alga had been well understood, the factors influencing development of the tetraspore and carpospore remain poorly understood. In the perspective of seedling resources, the regulation of early development is crucial for the seedling nursing; therefore, it is necessary to understand the physiological influences during its early development. In this study, we studied the effects of temperature and irradiance on the early development of Chondrus ocellatus Holm under laboratory conditions. The released tetraspores and carpospores were cultivated at different temperatures (10-28 degrees C) and irradiances ( 10, 60 mu mol photons m(-2)s(-1)) with a photoperiod of 12L:12D. The results indicate that both tetraspores and carpospores are tolerant to temperatures of 10-25 degrees C, and have the highest relative growth rate at 20 degrees C. Irradiance variances influenced the growth of the discoid crusts, and the influence was more significant with increasing temperature; 60 mu mol photons m(-2)s(-1) was more suitable than 10 mu mol photons m(-2)s(-1). The optimum temperature and irradiance for the development of seedlings was 20 degrees C and 60 mu mol photons m(-2)s(-1), respectively.

The capacity of the East Asian seaweed Gracilaria vermiculophylla ("Ogonori") for production of prostaglandin E

TdT-mediated dUTP-biotin nick end labeling (TUNEL) is a sensitive and valid method for detecting DNA cleavage in programmed cell death (PCD). Using this method, DNA cleavage was observed in Laminaria japonica sporophytic tissues, which were infected with alginic acid decomposing bacterium. It was found that DNA cleavage occurred 5 min after the infection, the fragments with 3'-OH groups of cleaved nuclear DNA increased with time of infection and spread from the infection site. Although no typical DNA ladder (200 bp/ 180 bp) was detected by routine agarose gel electrophoresis, the cleavage of nuclear DNA fragments of 97 similar to 48.5 kb could be detected by pulsed field gel electrophoresis (PFGE). By using CaspGLOW(TM) fluorescein active caspase-3 staining method, caspase-3 activity has been detected in response to the infection of alginic acid decomposing bacterium. Our results are similar to the observations in hypersensitive response (HR) of higher plant, suggesting that the rapid cell death of L. japonica infected by alginic acid decomposing bacterium might be involved in PCD, and indicating that the occurrence of PCD is an active defense process against the pathogen's infection.

Fertility and flower development are both controlled in part by jasmonates, fatty acid-derived mediators produced via the activity of 13-lipoxygenases (13-LOXs). The Arabidopsis thaliana Columbia-0 reference genome is predicted to encode four of these enzymes and it is already known that one of these, LOX2, is dispensable for fertility. In this study, the roles of the other three 13-LOXs (LOX3, LOX4 and LOX6) were investigated in single and double mutants. Four independent lox3 lox4 double mutants assembled with different mutated lox3 and lox4 alleles had fully penetrant floral phenotypes, displaying abnormal anther maturation and defective dehiscence. The plants were no longer self-fertile and pollen was not viable. Fertility in the double mutant was restored genetically by complementation with either the LOX3 or the LOX4 cDNAs and biochemically with exogenous jasmonic acid. Furthermore, deficiency in LOX3 and LOX4 causes developmental dysfunctions, compared to wild type; lox3 lox4 double mutants are taller and develop more inflorescence shoots and flowers. Further analysis revealed that developmental arrest in the lox3 lox4 inflorescence occurs with the production of an abnormal carpelloid flower. This distinguishes lox3 lox4 mutants from the wild type where developmentally typical flower buds are the terminal inflorescence structures observed in both the laboratory and in nature. Our studies of lox3 lox4 as well as other jasmonic acid biosynthesis and perception mutants show that this plant hormone is not only required for male fertility but also involved in global proliferative arrest.